Sample records for optimal ph temperature

  1. Effects of temperature, pH and NaCl on protease activity in digestive tract of young turbot, Scophthalmus maximus

    NASA Astrophysics Data System (ADS)

    Chen, Muyan; Zhang, Xiumei; Gao, Tianxiang; Chen, Chao

    2006-09-01

    The protease activity in digestive tract of young turbot Scophthalmus maximum was studied, and the optimal pH, temperature and NaCl concentration were determined for different portions of the fish's internal organs. The optimal activity in the fish's stomach was at pH of 2.2, while that in the intestinal extracts was within the alkaline range from 9.5 to 10.0. In hepatopancreas, the optimal pH was in low alkalinity at 8.5. The optimal reaction temperature was above 40°C in stomach, intestine and hepatopancreas. With increasing temperature, the pH value increased in stomach, while in the intestine, an opposite tendency was observed due to combined effect of pH and temperature. NaCl concentration showed inhibitory impact on protein digestion in hepatopancreas. The main protease for protein digestion in turbot seemed to be pepsin. Moreover, the maximum protease activity in different segments of intestine existed in the hindgut.

  2. [Chemotaxis response of Erwinia carotovora on sugars and amino acids of root exudates of Panax ginseng].

    PubMed

    Zhang, Ai-Hua; An, Ning-Bo; Lei, Feng-Jie; Ma, Wen-Li; Chi, Kun; Zhang, Lian-Xue

    2016-11-01

    The chemotaxis response of Erwinia carotovora to different sugars and amino acids in four kinds of chemotactic parameters (concentration, time, temperature and pH ) was determined by capillary method. The results showed that when pH was 8, concentration was 0.025 mg•L ⁻¹, culture temperature was 25 ℃ and the duration was 60 minutes, the optimal chemotaxis rate of lysine was 2.509,when pH was 6, concentration was 0.25 mg•L ⁻¹, culture temperature was 25 ℃ and the duration was 60 minutes, the optimal chemotaxis rate of arginine was 2.218 8,when pH was 7, concentration was 0.25 mg•L ⁻¹, culture temperature was 30 ℃ and the duration was 60 minutes, the optimal chemotaxis rate of L-rhamnose was 3.091 2, when pH was 6, concentration was 0.25 mg•L ⁻¹, culture temperature was 30 ℃ and the duration was 45 minutes, the optimal chemotaxis rate of D-arabinose was 3.026 3. Sugars and amino acids had obvious chemotaxis with E. carotovora,the high concentration of carbohydrate and amino acid exited an inhibitory effect on chemotaxis response of E. carotovora, and the chemotaxis response decreased with the increase of concentration of carbohydrates and amino acids. Copyright© by the Chinese Pharmaceutical Association.

  3. Effect of temperature, water activity, and pH on growth and production of ochratoxin A by Aspergillus niger and Aspergillus carbonarius from Brazilian grapes.

    PubMed

    Passamani, Fabiana Reinis Franca; Hernandes, Thais; Lopes, Noelly Alves; Bastos, Sabrina Carvalho; Santiago, Wilder Douglas; Cardoso, Maria das Graças; Batista, Luís Roberto

    2014-11-01

    The growth of ochratoxigenic fungus and the presence of ochratoxin A (OTA) in grapes and their derivatives can be caused by a wide range of physical, chemical, and biological factors. The determination of interactions between these factors and fungal species from different climatic regions is important in designing models for minimizing the risk of OTA in wine and grape juice. This study evaluated the influence of temperature, water activity (aw), and pH on the development and production of OTA in a semisynthetic grape culture medium by Aspergillus carbonarius and Aspergillus niger strains. To analyze the growth conditions and production of OTA, an experimental design was conducted using response surface methodology as a tool to assess the effects of these abiotic variables on fungal behavior. A. carbonarius showed the highest growth at temperatures from 20 to 33°C, aw between 0.95 and 0.98, and pH levels between 5 and 6.5. Similarly, for A. niger, temperatures between 24 and 37°C, aw greater than 0.95, and pH levels between 4 and 6.5 were optimal. The greatest toxin concentrations for A. carbonarius and A. niger (10 μg/g and 7.0 μg/g, respectively) were found at 15°C, aw 0.99, and pH 5.35. The lowest pH was found to contribute to greater OTA production. These results show that the evaluated fungi are able to grow and produce OTA in a wide range of temperature, aw, and pH. However, the optimal conditions for toxin production are generally different from those optimal for fungal growth. The knowledge of optimal conditions for fungal growth and production of OTA, and of the stages of cultivation in which these conditions are optimal, allows a more precise assessment of the potential risk to health from consumption of products derived from grapes.

  4. Optimizing isothiocyanate formation during enzymatic glucosinolate breakdown by adjusting pH value, temperature and dilution in Brassica vegetables and Arabidopsis thaliana

    NASA Astrophysics Data System (ADS)

    Hanschen, Franziska S.; Klopsch, Rebecca; Oliviero, Teresa; Schreiner, Monika; Verkerk, Ruud; Dekker, Matthijs

    2017-01-01

    Consumption of glucosinolate-rich Brassicales vegetables is associated with a decreased risk of cancer with enzymatic hydrolysis of glucosinolates playing a key role. However, formation of health-promoting isothiocyanates is inhibited by the epithiospecifier protein in favour of nitriles and epithionitriles. Domestic processing conditions, such as changes in pH value, temperature or dilution, might also affect isothiocyanate formation. Therefore, the influences of these three factors were evaluated in accessions of Brassica rapa, Brassica oleracea, and Arabidopsis thaliana. Mathematical modelling was performed to determine optimal isothiocyanate formation conditions and to obtain knowledge on the kinetics of the reactions. At 22 °C and endogenous plant pH, nearly all investigated plants formed nitriles and epithionitriles instead of health-promoting isothiocyanates. Response surface models, however, clearly demonstrated that upon change in pH to domestic acidic (pH 4) or basic pH values (pH 8), isothiocyanate formation considerably increases. While temperature also affects this process, the pH value has the greatest impact. Further, a kinetic model showed that isothiocyanate formation strongly increases due to dilution. Finally, the results show that isothiocyanate intake can be strongly increased by optimizing the conditions of preparation of Brassicales vegetables.

  5. Effect of the culture media optimization, pH and temperature on the biohydrogen production and the hydrogenase activities by Klebsiella pneumoniae ECU-15.

    PubMed

    Xiao, Yan; Zhang, Xu; Zhu, Minglong; Tan, Wensong

    2013-06-01

    The low yield of the biohydrogen production is the main constraint for its industrialization process. In order to improve its production, medium compositions of the hydrogen fermentation by Klebsiella pneumoniae ECU-15 were optimized through the response surface methodology (RSM). Experimental results showed that the optimum hydrogen production of 5363.8 ml/L was obtained when the concentration of glucose, the ammonium sulfate and the trace elements were 35.62 g/L, 2.78 g/L and 23.15 ml/L at temperature 37.0°C, pH 6.0. H2 evolving hydrogenase was greatly enhanced by the optimization of the medium compositions. The activity of H2 evolving hydrogenase increased with the temperature, and decreased with the pH, while the activity of the uptake hydrogenase increased with the temperature and the pH. So the biohydrogen production process of the K. pneumoniae ECU-15 was the comprehensive results of the evolution hydrogen process and the uptake hydrogen process. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Optimization study of ethanolic fermentation from oil palm trunk, rubberwood and mixed hardwood hydrolysates using Saccharomyces cerevisiae.

    PubMed

    Chin, K L; H'ng, P S; Wong, L J; Tey, B T; Paridah, M T

    2010-05-01

    Ethanolic fermentation using Saccharomyces cerevisiae was carried out on three types of hydrolysates produced from lignocelulosic biomass which are commonly found in Malaysia such as oil palm trunk, rubberwood and mixed hardwood. The effect of fermentation temperature and pH of hydrolysate was evaluated to optimize the fermentation efficiency which defined as maximum ethanol yield in minimum fermentation time. The fermentation process using different temperature of 25 degrees Celsius, 30 degrees Celsius and 40 degrees Celsius were performed on the prepared fermentation medium adjusted to pH 4, pH 6 and pH 7, respectively. Results showed that the fermentation time was significantly reduced with the increase of temperature but an adverse reduction in ethanol yield was observed using temperature of 40 degrees Celsius. As the pH of hydrolysate became more acidic, the ethanol yield increased. Optimum fermentation efficiency for ethanolic fermentation of lignocellulosic hydrolysates using S. cerevisiae can be obtained using 33.2 degrees Celsius and pH 5.3. Copyright 2009 Elsevier Ltd. All rights reserved.

  7. Optimized postweld heat treatment procedures for 17-4 PH stainless steels

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bhaduri, A.K.; Sujith, S.; Srinivasan, G.

    1995-05-01

    The postweld heat treatment (PWHT) procedures for 17-4 PH stainless steel weldments of matching chemistry was optimized vis-a-vis its microstructure prior to welding based on microstructural studies and room-temperature mechanical properties. The 17-4 PH stainless steel was welded in two different prior microstructural conditions (condition A and condition H 1150) and then postweld heat treated to condition H900 or condition H1150, using different heat treatment procedures. Microstructural investigations and room-temperature tensile properties were determined to study the combined effects of prior microstructural and PWHT procedures.

  8. [Induce of laccase from Trametes gallica and its degradation on neutral dyes and organophosphorus pesticides].

    PubMed

    Jing, De-Jun; Huang, Jian-Bo; Yang, Zhou-Ping; Hu, Rong; Cheng, Zi-Zhang; Huang, Qian-Ming

    2011-12-01

    The characteristics of the induction of laccase in Trametes gallica under different initial cultural pH, incubation time by different inducers were discussed, as well as the effects of temperature, pH and time on laccase degradation of six dyes and four organophosphors. The results showed that RB-bright blue, ABTS and o-toluidine affected the production of laccase at different levels, and ABTS was the best inductive agent in our test conditions, whose optimal initial pH and incubation time were 4.0 and 13 days, respectively. The appropriate reaction temperature of the laccase produced was 38 degrees C, and it got a good stability, for it could retain 78.6% of the enzyme activity after 20 min holding at 40 degrees C. Mediated by ABTS, the optimal temperature for laccase to degrade the six types of neutral dyes could be divided into two cases, that was 30 degrees C (neutral black, neutral bordeaux, neutral pink, methyl orange) and 60 degrees C (neutral dark yellow, cresol red), the optimal pH were 6.0 (neutral black), 2.0 (neutral bordeaux, neutral pink) and 4.0 (methyl orange, neutral dark yellow, cresol red), respectively, while the optimal times separately were 6 h (methyl orange, neutral dark yellow, cresol red), 12 h (neutral pink) and 24 h (neutral bordeaux). And using the same inductive agent, the best temperature for laccase to degrade dimethoate, chlorpyrifos, trichlorfon and parathion-pyridazine was 25 degrees C, the suitable time was 9 h, and the optimal pH was 10.0 for dimethoate, chlorpyrifos and parathion-pyridazine, and 8.0 for trichlorfon.

  9. Temperature and pH Conditions That Prevail during Fermentation of Sausages Are Optimal for Production of the Antilisterial Bacteriocin Sakacin K

    PubMed Central

    Leroy, Frédéric; de Vuyst, Luc

    1999-01-01

    Sakacin K is an antilisterial bacteriocin produced by Lactobacillus sake CTC 494, a strain isolated from Spanish dry fermented sausages. The biokinetics of cell growth and bacteriocin production of L. sake CTC 494 in vitro during laboratory fermentations were investigated by making use of MRS broth. The data obtained from the fermentations was used to set up a predictive model to describe the influence of the physical factors temperature and pH on microbial behavior. The model was validated successfully for all components. However, the specific bacteriocin production rate seemed to have an upper limit. Both cell growth and bacteriocin activity were very much influenced by changes in temperature and pH. The production of biomass was closely related to bacteriocin activity, indicating primary metabolite kinetics, but was not the only factor of importance. Acidity dramatically influenced both the production and the inactivation of sakacin K; the optimal pH for cell growth did not correspond to the pH for maximal sakacin K activity. Furthermore, cells grew well at 35°C but no bacteriocin production could be detected at this temperature. L. sake CTC 494 shows special promise for implementation as a novel bacteriocin-producing sausage starter culture with antilisterial properties, considering the fact that the temperature and acidity conditions that prevail during the fermentation process of dry fermented sausages are optimal for the production of sakacin K. PMID:10049850

  10. Multi-Response Optimization of Granaticinic Acid Production by Endophytic Streptomyces thermoviolaceus NT1, Using Response Surface Methodology

    PubMed Central

    Roy, Sudipta; Halder, Suman Kumar; Banerjee, Debdulal

    2016-01-01

    Streptomyces thermoviolaceus NT1, an endophytic isolate, was studied for optimization of granaticinic acid production. It is an antimicrobial metabolite active against even drug resistant bacteria. Different media, optimum glucose concentration, initial media pH, incubation temperature, incubation period, and inoculum size were among the selected parameters optimized in the one-variable-at-a-time (OVAT) approach, where glucose concentration, pH, and temperature were found to play a critical role in antibiotic production by this strain. Finally, the Box–Behnken experimental design (BBD) was employed with three key factors (selected after OVAT studies) for response surface methodological (RSM) analysis of this optimization study.RSM analysis revealed a multifactorial combination; glucose 0.38%, pH 7.02, and temperature 36.53 °C as the optimum conditions for maximum antimicrobial yield. Experimental verification of model analysis led to 3.30-fold (61.35 mg/L as compared to 18.64 mg/L produced in un-optimized condition) enhanced granaticinic acid production in ISP2 medium with 5% inoculum and a suitable incubation period of 10 days. So, the conjugated optimization study for maximum antibiotic production from Streptomyces thermoviolaceus NT1 was found to result in significantly higher yield, which might be exploited in industrial applications. PMID:28952581

  11. Normalization of doxorubicin release from graphene oxide: New approach for optimization of effective parameters on drug loading.

    PubMed

    Hashemi, Mohadeseh; Yadegari, Amir; Yazdanpanah, Ghasem; Omidi, Meisam; Jabbehdari, Sayena; Haghiralsadat, Fatemeh; Yazdian, Fatemeh; Tayebi, Lobat

    2017-05-01

    Graphene oxide (GO) has been recently introduced as a suitable anticancer drug carrier, which could be loaded with doxorubicin (DOX) as a general chemotherapy agent. Herein, the attempts were made to optimize the effective parameters on both loading and release of DOX on GO. GO and GO-DOX were characterized using transition electron microscopy , zeta potential, Raman spectroscopy, UV-visible spectroscopy, and Fourier transform infrared spectroscopy. In addition, loading and releasing behaviors of DOX on GO were studied in terms of different temperature and pH values. The primary optimized values of pH and temperature for best-loaded amount of DOX were 8.9 and 309 K, respectively. Moreover, we found that the smallest amount of released DOX, in pH of cancer microenvironment (5.4), occurs when DOX had been previously loaded in pH 7.8 and 310 K. Although the highest amount of loaded DOX was in basic pH, the results of efficient release of DOX from the GO-DOX complex and also cellular toxicity assay revealed that the best pH for loading of DOX on GO was 7.8. Therefore, in addition to optimization of parameters for efficient loading of DOX on GO, this study suggested that normalization of a released drug compared with the amount of a loaded drug could be a new approach for optimization of drug loading on nanocarriers. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

  12. Modeling the Effect of pH and Temperature for Cellulases Immobilized on Enzymogel Nanoparticles.

    PubMed

    Samaratunga, Ashani; Kudina, Olena; Nahar, Nurun; Zakharchenko, Andrey; Minko, Sergiy; Voronov, Andriy; Pryor, Scott W

    2015-06-01

    Production costs of cellulosic biofuels can be lowered if cellulases are recovered and reused using particulate carriers that can be extracted after biomass hydrolysis. Such enzyme recovery was recently demonstrated using enzymogel nanoparticles with grafted polymer brushes loaded with cellulases. In this work, cellulase (NS50013) and β-glucosidase (Novozyme 188) were immobilized on enzymogels made of poly(acrylic acid) polymer brushes grafted to the surface of silica nanoparticles. Response surface methodology was used to model effects of pH and temperature on hydrolysis and recovery of free and attached enzymes. Hydrolysis yields using both enzymogels and free cellulase and β-glucosidase were highest at the maximum temperature tested, 50 °C. The optimal pH for cellulase enzymogels and free enzyme was 5.0 and 4.4, respectively, while both free β-glucosidase and enzymogels had an optimal pH near 4.4. Highest hydrolysis sugar concentrations with cellulase and β-glucosidase enzymogels were 69 and 53 % of those with free enzymes, respectively. Enzyme recovery using enzymogels decreased with increasing pH, but cellulase recovery remained greater than 88 % throughout the operating range of pH values less than 5.0 and was greater than 95 % at pH values below 4.3. Recovery of β-glucosidase enzymogels was not affected by temperature and had little impact on cellulase recovery.

  13. Modelling lamb carcase pH and temperature decline parameters: relationship to shear force and abattoir variation.

    PubMed

    Hopkins, David L; Holman, Benjamin W B; van de Ven, Remy J

    2015-02-01

    Carcase pH and temperature decline rates influence lamb tenderness; therefore pH decline parameters are beneficial when modelling tenderness. These include pH at temperature 18 °C (pH@Temp18), temperature when pH is 6 (Temp@pH6), and pH at 24 h post-mortem (pH24). This study aimed to establish a relationship between shear force (SF) as a proxy for tenderness and carcase pH decline parameters estimated using both linear and spline estimation models for the m. longissimus lumborum (LL). The study also compared abattoirs regarding their achievement of ideal pH decline, indicative of optimal tenderness. Based on SF measurements of LL and m. semimembranosus collected as part of the Information Nucleus slaughter programme (CRC for Sheep Industry Innovation) this study found significant relationships between tenderness and pH24LL, consistent across the meat cuts and ageing periods examined. Achievement of ideal pH decline was shown not to have significantly differed across abattoirs, although rates of pH decline varied significantly across years within abattoirs.

  14. Production of fibrinolytic protease from Streptomyces lusitanus isolated from marine sediments

    NASA Astrophysics Data System (ADS)

    SudeshWarma, S.; Merlyn keziah, S.; Subathra Devi, C.

    2017-11-01

    This study aim was to isolate, screen, characterize and optimize marine Streptomyces for fibrinolytic enzyme production. The potent actinomycete isolate was subjected to optimization. The parameters for optimization included pH, temperature, carbon, nitrogen sources. The crude supernatant produced was purified using size exclusion gel filtration chromatography. The optimized parameters for maximum productivity were found to be pH 7, 37°C, maltose and peptone respectively. The molecular weight of the purified enzyme was found to be 21kDa.

  15. Applying Central Composite Design and Response Surface Methodology to Optimize Growth and Biomass Production of Haemophilus influenzae Type b.

    PubMed

    Momen, Seyed Bahman; Siadat, Seyed Davar; Akbari, Neda; Ranjbar, Bijan; Khajeh, Khosro

    2016-06-01

    Haemophilus influenzae type b (Hib) is the leading cause of bacterial meningitis, otitis media, pneumonia, cellulitis, bacteremia, and septic arthritis in infants and young children. The Hib capsule contains the major virulence factor, and is composed of polyribosyl ribitol phosphate (PRP) that can induce immune system response. Vaccines consisting of Hib capsular polysaccharide (PRP) conjugated to a carrier protein are effective in the prevention of the infections. However, due to costly processes in PRP production, these vaccines are too expensive. To enhance biomass, in this research we focused on optimizing Hib growth with respect to physical factors such as pH, temperature, and agitation by using a response surface methodology (RSM). We employed a central composite design (CCD) and a response surface methodology to determine the optimum cultivation conditions for growth and biomass production of H. influenzae type b. The treatment factors investigated were initial pH, agitation, and temperature, using shaking flasks. After Hib cultivation and determination of dry biomass, analysis of experimental data was performed by the RSM-CCD. The model showed that temperature and pH had an interactive effect on Hib biomass production. The dry biomass produced in shaking flasks was about 5470 mg/L, which was under an initial pH of 8.5, at 250 rpm and 35° C. We found CCD and RSM very effective in optimizing Hib culture conditions, and Hib biomass production was greatly influenced by pH and incubation temperature. Therefore, optimization of the growth factors to maximize Hib production can lead to 1) an increase in bacterial biomass and PRP productions, 2) lower vaccine prices, 3) vaccination of more susceptible populations, and 4) lower risk of Hib infections.

  16. Rational design of Bacillus stearothermophilus US100 L-arabinose isomerase: potential applications for D-tagatose production.

    PubMed

    Rhimi, Moez; Aghajari, Nushin; Juy, Michel; Chouayekh, Hichem; Maguin, Emmanuelle; Haser, Richard; Bejar, Samir

    2009-05-01

    L-arabinose isomerases catalyze the bioconversion of D-galactose into D-tagatose. With the aim of producing an enzyme optimized for D-tagatose production, three Bacillus stearothermophilus US100 L-arabinose isomerase mutants were constructed, purified and characterized. Our results indicate that mutant Q268K was significantly more acidotolerant and more stable at acidic pH than the wild-type enzyme. The N175H mutant has a broad optimal temperature range from 50 to 65 degrees C. With the aim of constructing an acidotolerant mutant working at relatively low temperatures we generated the Q268K/N175H construct. This double mutant displays an optimal pH in the range 6.0-7.0 and an optimal activity around 50-65 degrees C, temperatures at which the enzyme was stable without addition of metal ions.

  17. Response surface methodology as an approach to determine optimal activities of lipase entrapped in sol-gel matrix using different vegetable oils.

    PubMed

    Pinheiro, Rubiane C; Soares, Cleide M F; de Castro, Heizir F; Moraes, Flavio F; Zanin, Gisella M

    2008-03-01

    The conditions for maximization of the enzymatic activity of lipase entrapped in sol-gel matrix were determined for different vegetable oils using an experimental design. The effects of pH, temperature, and biocatalyst loading on lipase activity were verified using a central composite experimental design leading to a set of 13 assays and the surface response analysis. For canola oil and entrapped lipase, statistical analyses showed significant effects for pH and temperature and also the interactions between pH and temperature and temperature and biocatalyst loading. For the olive oil and entrapped lipase, it was verified that the pH was the only variable statistically significant. This study demonstrated that response surface analysis is a methodology appropriate for the maximization of the percentage of hydrolysis, as a function of pH, temperature, and lipase loading.

  18. How pH, Temperature, and Time of Incubation Affect False-Positive Responses and Uncertainty of the LAL Gel-Clot Test.

    PubMed

    Lourenço, Felipe Rebello; Botelho, Túlia De Souza; Pinto, Terezinha De Jesus Andreoli

    2012-01-01

    The limulus amebocyte lysate (LAL) test is the simplest and most widely used procedure for detection of endotoxin in parenteral drugs. The LAL test demands optimal pH, ionic strength, temperature, and time of incubation. Slight changes in these parameters may increase the frequency of false-positive responses and the estimated uncertainty of the LAL test. The aim of this paper is to evaluate how changes in the pH, temperature, and time of incubation affect the occurrence of false-positive responses in the LAL test. LAL tests were performed in nominal conditions (37 °C, 60 min, and pH 7) and in different conditions of temperature (36 °C and 38 °C), time of incubation (58 and 62 min), and pH (6 and 8). Slight differences in pH increase the frequency of false-positive responses 5-fold (relative risk 5.0), resulting in an estimated of uncertainty 7.6%. Temperature and time of incubation affect the LAL test less, showing relative risks of 1.5 and 1.0, respectively. Estimated uncertainties in 36 °C or 38 °C temperatures and 58 or 62 min of incubation were found to be 2.0% and 1.0%, respectively. Simultaneous differences in these parameters significantly increase the frequency of false-positive responses. The limulus amebocyte lysate (LAL) gel-clot test is a simple test for detection of endotoxin from Gram-negative bacteria. The test is based on a gel formation when a certain amount of endotoxin is present; it is a pass/fail test. The LAL test requires optimal pH, ionic strength, temperature, and time of incubation. Slight difference in these parameters may increase the frequency of false-positive responses. The aim of this paper is to evaluate how changes in the pH, temperature, and time of incubation affect the occurrence of false-positive responses in the LAL test. We find that slight differences in pH increase the frequency of false-positive responses 5-fold. Temperature and time of incubation affect the LAL test less. Simultaneous differences in these parameters significantly increase the frequency of false-positive responses.

  19. Superlattice photonic crystal as broadband solar absorber for high temperature operation.

    PubMed

    Rinnerbauer, Veronika; Shen, Yichen; Joannopoulos, John D; Soljačić, Marin; Schäffler, Friedrich; Celanovic, Ivan

    2014-12-15

    A high performance solar absorber using a 2D tantalum superlattice photonic crystal (PhC) is proposed and its design is optimized for high-temperature energy conversion. In contrast to the simple lattice PhC, which is limited by diffraction in the short wavelength range, the superlattice PhC achieves solar absorption over broadband spectral range due to the contribution from two superposed lattices with different cavity radii. The superlattice PhC geometry is tailored to achieve maximum thermal transfer efficiency for a low concentration system of 250 suns at 1500 K reaching 85.0% solar absorptivity. In the high concentration case of 1000 suns, the superlattice PhC absorber achieves a solar absorptivity of 96.2% and a thermal transfer efficiency of 82.9% at 1500 K, amounting to an improvement of 10% and 5%, respectively, versus the simple square lattice PhC absorber. In addition, the performance of the superlattice PhC absorber is studied in a solar thermophotovoltaic system which is optimized to minimize absorber re-emission by reducing the absorber-to-emitter area ratio and using a highly reflective silver aperture.

  20. Optimization of CMCase production from sorghum straw by Aspergillus terreus SUK-1 under solid substrate fermentation using response surface methodology

    NASA Astrophysics Data System (ADS)

    Tibin, El Mubarak Musa; Al-Shorgani, Najeeb Kaid Naseer; Abuelhassan, Nawal Noureldaim; Hamid, Aidil Abdul; Kalil, Mohd Sahaid; Yusoff, Wan Mohtar Wan

    2013-11-01

    The cellulase production using sorghum straw as substrate by fungal culture of Aspergillus terreus SUK-1 was investigated in solid substrate fermentation (SSF). The optimum CMCase was achieved by testing most effective fermentation parameters which were: incubation temperature, pH and moisture content using Response Surface Methodology (RSM) based on Central Composite Design (CCD). The carboxymethyl cellulase activity (CMCase) was measured as the defining factor. The results were analysed by analysis of variance (ANOVA) and the regression quadratic model was obtained. The model was found to be significant (p<0.05) and the effect of temperature (25-40°C) and pH (4-7) was found to be not significant on CMCase activity whereas the moisture content was significant in the SSF conditions employed. The high yield of predicted CMCase activity (0.2 U/ml) was obtained under the optimized conditions (temperature 40 □C, pH 5.4 and moisture content of 80%). The model was validated by applying the optimized conditions and it was found that the model was valid.

  1. Effect of pH and temperature on the uptake of cadmium by Lemna minor L

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chawla, G.; Singh, J.; Viswanathan, P.N.

    1991-07-01

    Many aquatic macrophytes have the capacity to take up toxic heavy metals from polluted water and accumulate them. Cut leaves and intact plants have been suggested for clearing polluted water bodies of heavy metals. However, uptake of metal ion from water is dependent on concentration, pH, temperature, presence of other substances and functional and morphological status of the biotic species. In an attempt to understand any correlation between metal bioconcentration, pH and temperature, the optimal conditions for the removal of cadmium ions by duckweed, Lemna minor (L.) were studied.

  2. Preparation of starch-poly-glutamic acid graft copolymers by microwave irradiation and the characterization of their properties

    USDA-ARS?s Scientific Manuscript database

    Graft copolymers of waxy maize starch and poly-y-glutamic acid (PGA) were produced in an aqueous solution using microwave irradiation. The microwave reaction conditions were optimized with regard to temperature and pH. The temperature of 180 deg C and pH 7.0 were the best reaction conditions resulti...

  3. Optimization of pH, temperature and CaCl2 concentrations for Ricotta cheese production from Buffalo cheese whey using Response Surface Methodology.

    PubMed

    Rashid, Abdul Ahid; Huma, Nuzhat; Zahoor, Tahir; Asgher, Muhammad

    2017-02-01

    The recovery of milk constituents from cheese whey is affected by various processing conditions followed during production of Ricotta cheese. The objective of the present investigation was to optimize the temperature (60-90 °C), pH (3-7) and CaCl2 concentration (2·0-6·0 mm) for maximum yield/recovery of milk constituents. The research work was carried out in two phases. In 1st phase, the influence of these processing conditions was evaluated through 20 experiments formulated by central composite design (CCD) keeping the yield as response factor. The results obtained from these experiments were used to optimize processing conditions for maximum yield using response surface methodology (RSM). The three best combinations of processing conditions (90 °C, pH 7, CaCl2 6 mm), (100 °C, pH 5, CaCl2 4 mm) and (75 °C, pH 8·4, CaCl2 4 mm) were exploited in the next phase for Ricotta cheese production from a mixture of Buffalo cheese whey and skim milk (9 : 1) to determine the influence of optimized conditions on the cheese composition. Ricotta cheeses were analyzed for various physicochemical (moisture, fat, protein, lactose, total solids, pH and acidity indicated) parameters during storage of 60 d at 4 ± 2 °C after every 15 d interval. Ricotta cheese prepared at 90 °C, pH 7 and CaCl2 6 mm exhibited the highest cheese yield, proteins and total solids, while high fat content was recorded for cheese processed at 100 °C, pH 5 and 4 mm CaCl2 concentration. A significant storage-related increase in acidity and NPN was recorded for all cheese samples.

  4. Optimization of ultrasound-assisted extraction of pectinase enzyme from guava (Psidium guajava) peel: Enzyme recovery, specific activity, temperature, and storage stability.

    PubMed

    Amid, Mehrnoush; Murshid, Fara Syazana; Manap, Mohd Yazid; Islam Sarker, Zaidul

    2016-01-01

    This study aimed to investigate the effects of the ultrasound-assisted extraction conditions on the yield, specific activity, temperature, and storage stability of the pectinase enzyme from guava peel. The ultrasound variables studied were sonication time (10-30 min), ultrasound temperature (30-50 °C), pH (2.0-8.0), and solvent-to-sample ratio (2:1 mL/g to 6:1 mL/g). The main goal was to optimize the ultrasound-assisted extraction conditions to maximize the recovery of pectinase from guava peel with the most desirable enzyme-specific activity and stability. Under the optimum conditions, a high yield (96.2%), good specific activity (18.2 U/mg), temperature stability (88.3%), and storage stability (90.3%) of the extracted enzyme were achieved. The optimal conditions were 20 min sonication time, 40 °C temperature, at pH 5.0, using a 4:1 mL/g solvent-to-sample ratio. The study demonstrated that optimization of ultrasound-assisted process conditions for the enzyme extraction could improve the enzymatic characteristics and yield of the enzyme.

  5. Optimized production of vanillin from green vanilla pods by enzyme-assisted extraction combined with pre-freezing and thawing.

    PubMed

    Zhang, Yanjun; Mo, Limei; Chen, Feng; Lu, Minquan; Dong, Wenjiang; Wang, Qinghuang; Xu, Fei; Gu, Fenglin

    2014-02-19

    Production of vanillin from natural green vanilla pods was carried out by enzyme-assisted extraction combined with pre-freezing and thawing. In the first step the green vanilla pods were pre-frozen and then thawed to destroy cellular compartmentation. In the second step pectinase from Aspergillus niger was used to hydrolyze the pectin between the glucovanillin substrate and β-glucosidase. Four main variables, including enzyme amount, reaction temperature, time and pH, which were of significance for the vanillin content were studied and a central composite design (CCD) based on the results of a single-factor tests was used. Response surface methodology based on CCD was employed to optimize the combination of enzyme amount, reaction temperature, time, and pH for maximum vanillin production. This resulted in the optimal condition in regards of the enzyme amount, reaction temperature, time, and pH at 84.2 mg, 49.5 °C, 7.1 h, and 4.2, respectively. Under the optimal condition, the experimental yield of vanillin was 4.63% ± 0.11% (dwb), which was in good agreement with the value predicted by the model. Compared to the traditional curing process (1.98%) and viscozyme extract (2.36%), the optimized method for the vanillin production significantly increased the yield by 133.85% and 96%, respectively.

  6. Ultrasound assisted extraction of pectin from waste Artocarpus heterophyllus fruit peel.

    PubMed

    Moorthy, I Ganesh; Maran, J Prakash; Ilakya, S; Anitha, S L; Sabarima, S Pooja; Priya, B

    2017-01-01

    Four factors three level face centered central composite response surface design was employed in this study to investigate and optimize the effect of process variables (liquid-solid (LS) ratio (10:1-20:1ml/g), pH (1-2), sonication time (15-30min) and extraction temperature (50-70°C)) on the maximum extraction yield of pectin from waste Artocarpus heterophyllus (Jackfruit) peel by ultrasound assisted extraction method. Numerical optimization method was adapted in this study and the following optimal condition was obtained as follows: Liquid-solid ratio of 15:1ml/g, pH of 1.6, sonication time of 24min and temperature of 60°C. The optimal condition was validated through experiments and the observed value was interrelated with predicted value. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Biphasic cultivation strategy to avoid Epo-Fc aggregation and optimize protein expression.

    PubMed

    Kaisermayer, Christian; Reinhart, David; Gili, Andreas; Chang, Martina; Aberg, Per-Mikael; Castan, Andreas; Kunert, Renate

    2016-06-10

    In biphasic cultivations, the culture conditions are initially kept at an optimum for rapid cell growth and biomass accumulation. In the second phase, the culture is shifted to conditions ensuring maximum specific protein production and the protein quality required. The influence of specific culture parameters is cell line dependent and their impact on product quality needs to be investigated. In this study, a biphasic cultivation strategy for a Chinese hamster ovary (CHO) cell line expressing an erythropoietin fusion protein (Epo-Fc) was developed. Cultures were run in batch mode and after an initial growth phase, cultivation temperature and pH were shifted. Applying a DoE (Design of Experiments) approach, a fractional factorial design was used to systematically evaluate the influence of cultivation temperature and pH as well as their synergistic effect on cell growth as well as on recombinant protein production and aggregation. All three responses were influenced by the cultivation temperature. Additionally, an interaction between pH and temperature was found to be related to protein aggregation. Compared with the initial standard conditions of 37°C and pH 7.05, a parameter shift to low temperature and acidic pH resulted in a decrease in the aggregate fraction from 75% to less than 1%. Furthermore, the synergistic effect of temperature and pH substantially lowered the cell-specific rates of glucose and glutamine consumption as well as lactate and ammonium production. The optimized culture conditions also led to an increase of the cell-specific rates of recombinant Epo-Fc production, thus resulting in a more economic bioprocess. Copyright © 2016. Published by Elsevier B.V.

  8. Screening and optimization of pectin lyase and polygalacturonase activity from ginseng pathogen Cylindrocarpon Destructans

    PubMed Central

    Sathiyaraj, Gayathri; Srinivasan, Sathiyaraj; Kim, Ho-Bin; Subramaniyam, Sathiyamoorthy; Lee, Ok Ran; Kim, Yeon-Ju; Yang, Deok Chun

    2011-01-01

    Cylindrocarpon destructans isolated from ginseng field was found to produce pectinolytic enzymes. A Taguchi’s orthogonal array experimental design was applied to optimize the preliminary production of polygalacturonase (PG) and pectin lyase (PL) using submerged culture condition. This method was applied to evaluate the significant parameters for the production of enzymes. The process variables were pH, pectin concentration, incubation time and temperature. Optimization of process parameters resulted in high levels of enzyme (PG and PL) production after ten days of incubation at a pH of 5.0 at 25°C in the presence of 1.5% pectin. Among different nitrogen sources, urea and peptone showed high production of PG and PL, respectively. The enzyme production and mycelial growth seems to have direct influence on the culture conditions; therefore, at stationary state high enzyme production and mycelial growth were obtained than agitation state. Along with this, optimization of enzyme activity was also determined using various physiological parameters like, temperature, incubation time and pH. Taguchi’s data was also analyzed using one step ANOVA statistical method. PMID:24031695

  9. Effect of pH on simultaneous saccharification and isomerization by glucoamylase and glucose isomerase.

    PubMed

    Mishra, Abha; Debnath Das, Meera

    2002-01-01

    pH and temperature play critical roles in multistep enzymatic conversions. In such conversions, the optimal pH for individual steps differs greatly. In this article, we describe the production of glucoamylase (from Aspergillus oryzae MTCC152 in solid-state fermentation) and glucose isomerase (from Streptomyces griseus NCIM2020 in submerged fermentation), used in industries for producing high-fructose syrup. Optimum pH for glucoamylase was found to be 5.0. For glucose isomerase, the optimum pH ranged between 7.0 and 8.5, depending on the type of buffer used. Optimum temperature for glucoamylase and glucose isomerase was 50 and 60 degrees C, respectively. When both the enzymatic conversions were performed simultaneously at a compromised pH of 6.5, both the enzymes showed lowered activity. We also studied the kinetics at different pHs, which allows the two-step reaction to take place simultaneously. This was done by separating two steps by a thin layer of urease. Ammonia generated by the hydrolysis of urea consumed the hydrogen ions, thereby allowing optimal activity of glucose isomerase at an acidic pH of 5.0.

  10. Optimization of hydrolysis conditions for bovine plasma protein using response surface methodology.

    PubMed

    Seo, Hyun-Woo; Jung, Eun-Young; Go, Gwang-Woong; Kim, Gap-Don; Joo, Seon-Tea; Yang, Han-Sul

    2015-10-15

    The purpose of this study was to establish optimal conditions for the hydrolysis of bovine plasma protein. Response surface methodology was used to model and optimize responses [degree of hydrolysis (DH), 2,2-diphenyl-1-picrydrazyl (DPPH) radical-scavenging activity and Fe(2+)-chelating activity]. Hydrolysis conditions, such as hydrolysis temperature (46.6-63.4 °C), hydrolysis time (98-502 min), and hydrolysis pH (6.32-9.68) were selected as the main processing conditions in the hydrolysis of bovine plasma protein. Optimal conditions for maximum DH (%), DPPH radical-scavenging activity (%) and Fe(2+)-chelating activity (%) of the hydrolyzed bovine plasma protein, were respectively established. We discovered the following three conditions for optimal hydrolysis of bovine plasma: pH of 7.82-8.32, temperature of 54.1 °C, and time of 338.4-398.4 min. We consequently succeeded in hydrolyzing bovine plasma protein under these conditions and confirmed the various desirable properties of optimal hydrolysis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Optimization of trehalose production by a novel strain Brevibacterium sp. SY361.

    PubMed

    Wang, Lei; Huang, Rui; Gu, Guanbin; Fang, Hongying

    2008-10-01

    Trehalose production by a novel strain of Brevibacterium sp. SY361 was optimized in submerged fermentation. Different chemical and physical parameters such as carbon and nitrogen sources, inoculum level, initial pH, incubation temperature, aeration and time-course of fermentation, were studied in order to increase trehalose productivity. An optimal production medium containing 3% (w/v) glucose, 0.9% (v/v) corn steep liquor, 0.5% (w/v) KH(2)PO(4) and 0.4% (w/v) MgSO(4).7 H(2)O was found suitable for trehalose production. An optimal volume of medium in a 500 ml flask was 80 ml. The optimal levels of other parameters were 4.0% (v/v) of inoculum, initial pH of 6.0, incubation temperature of 28-32 degrees C and time-course of 60 h. Optimized parameters gave a maximum trehalose of 12.2 mg/ml with a conversion rate of 58.4%. (c) 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Cloning, Codon Optimization, and Expression of Yersinia intermedia Phytase Gene in E. coli.

    PubMed

    Mirzaei, Maryam; Saffar, Behnaz; Shareghi, Behzad

    2016-06-01

    Phytate is an anti-nutritional factor in plants, which catches the most phosphorus contents and some vital minerals. Therefore, Phytase is added mainly as an additive to the monogastric animals' foods to hydrolyze phytate and increase absorption of phosphorus. Y. intermedia phytase is a new phytase with special characteristics such as high specific activity, pH stability, and thermostability. Our aim was to clone, express, and characterizea codon optimized Y. intermedia phytase gene in E. coli . The Y. intermedia phytase gene was optimized according to the codon usage in E. coli . The sequence was synthesized and sub-cloned in pET-22b (+) vector and transformed into E. coli Bl21 (DE3). The protein was expressed in the presence of IPTG at a final concentration of 1 mM at 30°C. The purification of recombinant protein was performed by Ni 2+ affinity chromatography. Phytase activity and stability were determined in various pH and temperatures. The codon optimized Y. intermedia phytase gene was sub-cloned successfully.The expression was confirmed by SDS-PAGE and Western blot analysis. The recombinant enzyme (approximately 45 kDa) was purified. Specific activity of enzyme was 3849 (U.mg -1 ) with optimal pH 5 and optimal temperature of 55°C. Thermostability (80°C for 15 min) and pH stability (3-6) of the enzyme were 56 and more than 80%, respectively. The results of the expression and enzyme characterization revealed that the optimized Y. intermedia phytase gene has a good potential to be produced commercially andto be applied in animals' foodsindustry.

  13. A Statistical Approach for Optimization of Simultaneous Production of β-Glucosidase and Endoglucanase by Rhizopus oryzae from Solid-State Fermentation of Water Hyacinth Using Central Composite Design

    PubMed Central

    Karmakar, Moumita; Ray, Rina Rani

    2011-01-01

    The production cost of β-glucosidase and endoglucanase could be reduced by using water hyacinth, an aquatic weed, as the sole carbon source and using cost-efficient fermentation strategies like solid-state fermentation (SSF). In the present study, the effect of different production conditions on the yield of β-glucosidase and endoglucanase by Rhizopus oryzae MTCC 9642 from water hyacinth was investigated systematically using response surface methodology. A Central composite experimental design was applied to optimize the impact of three variables, namely, substrate concentration, pH, and temperature, on enzyme production. The optimal level of each parameter for maximum enzyme production by the fungus was determined. Highest activity of endoglucanase of 495 U/mL was achieved at a substrate concentration of 1.23%, pH 7.29, and temperature 29.93°C whereas maximum β-glucosidase activity of 137.32 U/ml was achieved at a substrate concentration of 1.25%, pH 6.66, and temperature 32.09°C. There was a direct correlation between the levels of enzymatic activities and the substrate concentration of water hyacinth as carbon source. PMID:21687577

  14. Statistical medium optimization of an alkaline protease from Pseudomonas aeruginosa MTCC 10501, its characterization and application in leather processing.

    PubMed

    Boopathy, Naidu Ramachandra; Indhuja, Devadas; Srinivasan, Krishnan; Uthirappan, Mani; Gupta, Rishikesh; Ramudu, Kamini Numbi; Chellan, Rose

    2013-04-01

    Proteases are shown to have greener mode of application in leather processing for dehairing of goat skins and cow hides. Production of protease by submerged fermentation with potent activity is reported using a new isolate P. aeruginosa MTCC 10501. The production parameters were optimized by statistical methods such as Plackett-Burman and response surface methodology. The optimized production medium contained (g/L); tryptone, 2.5; yeast extract, 3.0; skim milk 30.0; dextrose 1.0; inoculum concentration 4%: initial pH 6.0; incubation temperature 30 degrees C and optimum production at 48 h with protease activity of 7.6 U/mL. The protease had the following characteristics: pH optima, 9.0; temperature optima 50 degrees C; pH stability between 5.0-10.0 and temperature stability between 10-40 degrees C. The protease was observed to have high potential for dehairing of goat skins in the pre- tanning process comparable to that of the chemical process as evidenced by histology. The method offers cleaner processing using enzyme only instead of toxic chemicals in the pre-tanning process of leather manufacture.

  15. Rapid development of xylanase assay conditions using Taguchi methodology.

    PubMed

    Prasad Uday, Uma Shankar; Bandyopadhyay, Tarun Kanti; Bhunia, Biswanath

    2016-11-01

    The present investigation is mainly concerned with the rapid development of extracellular xylanase assay conditions by using Taguchi methodology. The extracellular xylanase was produced from Aspergillus niger (KP874102.1), a new strain isolated from a soil sample of the Baramura forest, Tripura West, India. Four physical parameters including temperature, pH, buffer concentration and incubation time were considered as key factors for xylanase activity and were optimized using Taguchi robust design methodology for enhanced xylanase activity. The main effect, interaction effects and optimal levels of the process factors were determined using signal-to-noise (S/N) ratio. The Taguchi method recommends the use of S/N ratio to measure quality characteristics. Based on analysis of the S/N ratio, optimal levels of the process factors were determined. Analysis of variance (ANOVA) was performed to evaluate statistically significant process factors. ANOVA results showed that temperature contributed the maximum impact (62.58%) on xylanase activity, followed by pH (22.69%), buffer concentration (9.55%) and incubation time (5.16%). Predicted results showed that enhanced xylanase activity (81.47%) can be achieved with pH 2, temperature 50°C, buffer concentration 50 Mm and incubation time 10 min.

  16. Development of new UV-vis spectroscopic microwave-assisted method for determination of glucose in pharmaceutical samples

    NASA Astrophysics Data System (ADS)

    Mabood, Fazal; Hussain, Z.; Haq, H.; Arian, M. B.; Boqué, R.; Khan, K. M.; Hussain, K.; Jabeen, F.; Hussain, J.; Ahmed, M.; Alharasi, A.; Naureen, Z.; Hussain, H.; Khan, A.; Perveen, S.

    2016-01-01

    A new UV-Visible spectroscopic method assisted with microwave for the determination of glucose in pharmaceutical formulations was developed. In this study glucose solutions were oxidized by ammonium molybdate in the presence of microwave energy and reacted with aniline to produce a colored solution. Optimum conditions of the reaction including wavelength, temperature, and pH of the medium and relative concentration ratio of the reactants were investigated. It was found that the optimal wavelength for the reaction is 610 nm, the optimal reaction time is 80 s, the optimal reaction temperature is 160 °C, the optimal reaction pH is 4, and the optimal concentration ratio aniline/ammonium molybdate solution was found to be 1:1. The limits of detection and quantification of the method are 0.82 and 2.75 ppm for glucose solution, respectively. The use of microwaves improved the speed of the method while the use of aniline improved the sensitivity of the method by shifting the wavelength.

  17. Optimal experimental condition for hemicellulosic hydrolyzate treatment with activated charcoal for xylitol production.

    PubMed

    Mussatto, Solange I; Roberto, Inês C

    2004-01-01

    Rice straw was hydrolyzed into a mixture of sugars using diluted H(2)SO(4). During hydrolysis, a variety of inhibitors was also produced, including acetic acid, furfural, hydroxymethylfurfural, and lignin degradation products (several aromatic and phenolic compounds). To reduce the toxic compounds concentration in the hydrolyzate and to improve the xylitol yield and volumetric productivity, rice straw hemicellulosic hydrolyzate was treated with activated charcoal under different pH values, stirring rates, contact times, and temperatures, employing a 2(4) full-factorial design. Fermentative assays were conducted with treated hydrolyzates containing 90 g/L xylose. The results indicated that temperature, pH, and stirring rate strongly influenced the hydrolyzate treatment, temperature and pH interfering with all of the responses analyzed (removal of color and lignin degradation products, xylitol yield factor, and volumetric productivity). The combination of pH 2.0, 150 rpm, 45 degrees C, and 60 min was considered an optimal condition, providing significant removal rates of color (48.9%) and lignin degradation products (25.8%), as well as a xylitol production of 66 g/L, a volumetric productivity of 0.57 g/L.h, and a yield factor of 0.72 g/g.

  18. Low-temperature-active and salt-tolerant β-mannanase from a newly isolated Enterobacter sp. strain N18.

    PubMed

    You, Jia; Liu, Jin-Feng; Yang, Shi-Zhong; Mu, Bo-Zhong

    2016-02-01

    A low-temperature-active and salt-tolerant β-mannanase produced by a novel mannanase-producer, Enterobacter sp. strain N18, was isolated, purified and then evaluated for its potential application as a gel-breaker in relation to viscosity reduction of guar-based hydraulic fracturing fluids used in oil field. The enzyme could lower the viscosity of guar gum solution by more than 95% within 10 min. The purified β-mannanase with molecular mass of 90 kDa displayed high activity in a broad range of pH and temperature: more than 70% of activity was retained in the pH range of 3.0-8.0 with the optimal pH 7.5, about 50% activity at 20°C with the optimal temperature 50°C. Furthermore, the enzyme retained >70% activity in the presence of 0.5-4.0 M NaCl. These properties implied that the enzyme from strain N18 had potential for serving as a gel-breaker for low temperature oil wells and other industrial fields, where chemical gel breakers were inactive due to low temperature. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  19. Optimization of Extraction Conditions for the 6-Shogaol-rich Extract from Ginger (Zingiber officinale Roscoe).

    PubMed

    Ok, Seon; Jeong, Woo-Sik

    2012-06-01

    6-Shogaol, a dehydrated form of 6-gingerol, is a minor component in ginger (Zingiber officinale Roscoe) and has recently been reported to have more potent bioactivity than 6-gingerol. Based on the thermal instability of gingerols (their dehydration to corresponding shogaols at high temperature), we aimed to develop an optimal process to maximize the 6-shogaol content during ginger extraction by modulating temperature and pH. Fresh gingers were dried under various conditions: freeze-, room temperature (RT)- or convection oven-drying at 60 or 80°C, and extracted by 95% ethanol at RT, 60 or 80°C. The content of 6-shogaol was augmented by increasing both drying and extraction temperatures. The highest production of 6-shogaol was achieved at 80°C extraction after drying at the same temperature and the content of 6-shogaol was about 7-fold compared to the lowest producing process by freezing and extraction at RT. Adjustment of pH (pH 1, 4, 7 and 10) for the 6-shogaol-richest extract (dried and extracted both at 80°C) also affected the chemical composition of ginger and the yield of 6-shogaol was maximized at the most acidic condition of pH 1. Taken together, the current study shows for the first time that a maximized production of 6-shogaol can be achieved during practical drying and extraction process of ginger by increasing both drying and extracting temperatures. Adjustment of pH to extraction solvent with strong acid also helps increase the production of 6-shogaol. Our data could be usefully employed in the fields of food processing as well as nutraceutical industry.

  20. Laccase production by Monotospora sp., an endophytic fungus in Cynodon dactylon.

    PubMed

    Wang, J W; Wu, J H; Huang, W Y; Tan, R X

    2006-03-01

    The effects of the carbon and nitrogen sources, initial pH and incubation temperature on laccase production by the endophytic fungus Monotospora sp. were evaluated. The optimal temperature and initial pH for laccase production by Monotospora sp. in submerged culture were found to be 30 degrees C and 8.5, respectively. Maltose (2 g l(-1)) and ammonium tartrate (10 g l(-1)) were the most suitable carbon and nitrogen source for laccase production. Under optimal culture medium, the maximum laccase activity was determined to be 13.55 U ml(-1), which was approximately four times higher than that in basal medium. This is the first report on laccase production by an endophytic fungus.

  1. Larvae of the coral eating crown-of-thorns starfish, Acanthaster planci in a warmer-high CO2 ocean.

    PubMed

    Kamya, Pamela Z; Dworjanyn, Symon A; Hardy, Natasha; Mos, Benjamin; Uthicke, Sven; Byrne, Maria

    2014-11-01

    Outbreaks of crown-of-thorns starfish (COTS), Acanthaster planci, contribute to major declines of coral reef ecosystems throughout the Indo-Pacific. As the oceans warm and decrease in pH due to increased anthropogenic CO2 production, coral reefs are also susceptible to bleaching, disease and reduced calcification. The impacts of ocean acidification and warming may be exacerbated by COTS predation, but it is not known how this major predator will fare in a changing ocean. Because larval success is a key driver of population outbreaks, we investigated the sensitivities of larval A. planci to increased temperature (2-4 °C above ambient) and acidification (0.3-0.5 pH units below ambient) in flow-through cross-factorial experiments (3 temperature × 3 pH/pCO2 levels). There was no effect of increased temperature or acidification on fertilization or very early development. Larvae reared in the optimal temperature (28 °C) were the largest across all pH treatments. Development to advanced larva was negatively affected by the high temperature treatment (30 °C) and by both experimental pH levels (pH 7.6, 7.8). Thus, planktonic life stages of A. planci may be negatively impacted by near-future global change. Increased temperature and reduced pH had an additive negative effect on reducing larval size. The 30 °C treatment exceeded larval tolerance regardless of pH. As 30 °C sea surface temperatures may become the norm in low latitude tropical regions, poleward migration of A. planci may be expected as they follow optimal isotherms. In the absence of acclimation or adaptation, declines in low latitude populations may occur. Poleward migration will be facilitated by strong western boundary currents, with possible negative flow-on effects on high latitude coral reefs. The contrasting responses of the larvae of A. planci and those of its coral prey to ocean acidification and warming are considered in context with potential future change in tropical reef ecosystems. © 2014 John Wiley & Sons Ltd.

  2. Immobilization of cross-linked tannase enzyme on multiwalled carbon nanotubes and its catalytic behavior.

    PubMed

    Ong, Chong-Boon; Annuar, Mohamad S M

    2018-02-07

    Immobilization of cross-linked tannase on pristine multiwalled carbon nanotubes (MWCNT) was successfully performed. Cross-linking of tannase molecules was made through glutaraldehyde. The immobilized tannase exhibited significantly improved pH, thermal, and recycling stability. The optimal pH for both free and immobilized tannase was observed at pH 5.0 with optimal operating temperature at 30°C. Moreover, immobilized enzyme retained greater biocatalytic activities upon 10 repeated uses compared to free enzyme in solution. Immobilization of tannase was accomplished by strong hydrophobic interaction most likely between hydrophobic amino acid moieties of the glutaraldehyde-cross-linked tannase to the MWCNT.

  3. Influence of culture media, pH and temperature on growth and bacteriocin production of bacteriocinogenic lactic acid bacteria.

    PubMed

    Yang, En; Fan, Lihua; Yan, Jinping; Jiang, Yueming; Doucette, Craig; Fillmore, Sherry; Walker, Bradley

    2018-01-24

    There has been continued interest in bacteriocins research from an applied perspective as bacteriocins have potential to be used as natural preservative. Four bacteriocinogenic lactic acid bacteria (LAB) strains of Lactobacillus curvatus (Arla-10), Enterococcus faecium (JFR-1), Lactobacillus paracasei subsp. paracasei (JFR-5) and Streptococcus thermophilus (TSB-8) were previously isolated and identified in our lab. The objective of this study was to determine the optimal growth conditions for both LAB growth and bacteriocins production. In this study, various growth conditions including culture media (MRS and BHI), initial pH of culture media (4.5, 5.5, 6.2, 7.4 and 8.5), and incubation temperatures (20, 37 and 44 °C) were investigated for LAB growth measured as optical density (OD), bacteriocin activity determined as arbitrary unit and viability of LAB expressed as log CFU ml -1 . Growth curves of the bacteriocinogenic LAB were generated using a Bioscreen C. Our results indicated that Arla-10, JFR-1, and JFR-5 strains grew well on both MRS and BHI media at growth temperature tested whereas TSB-8 strain, unable to grow at 20 °C. LAB growth was significantly affected by the initial pH of culture media (p < 0.001) and the optimal pH was found ranging from 6.2 to 8.5. Bacteriocin activity was significantly different in MRS versus BHI (p < 0.001), and the optimal condition for LAB to produce bacteriocins was determined in MRS broth, pH 6.2 at 37 °C. This study provides useful information on potential application of bacteriocinogenic LAB in food fermentation processes.

  4. Optimization of hydrolysis conditions for the production of glucomanno-oligosaccharides from konjac using β-mannanase by response surface methodology.

    PubMed

    Chen, Junfan; Liu, Desheng; Shi, Bo; Wang, Hai; Cheng, Yongqiang; Zhang, Wenjing

    2013-03-01

    Glucomanno-oligosaccharides (GMO), usually produced from hydrolysis of konjac tubers with a high content of glucomannan, have a positive effect on Bifidobacterium as well as a variety of other physiological activities. Response surface methodology (RSM) was employed to optimize the hydrolysis time, hydrolysis temperature, pH and enzyme to substrate ratio (E/S) to obtain a high GMO yield from konjac tubers. From the signal-factor experiments, it was concluded that the change in the direct reducing sugar (DRS) is consistent with total reducing sugar (TRS) but contrary to the degree of polymerization (DP). DRS was used as an indicator of the content of GMO in the RSM study. The optimum RSM operating conditions were: reaction time of 3.4 h, reaction temperature of 41.0°C, pH of 7.1 and E/S of 0.49. The results suggested that the enzymatic hydrolysis was enhanced by temperature, pH and incubation time. Model validation showed good agreement between experimental results and the predicted responses. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Impacts of Ocean Acidification and Temperature Change on Zooxanthellae Density in Coral Stylophora pistillata

    NASA Astrophysics Data System (ADS)

    Pantaleo, G. E.; Martínez Fernández, A.; Paytan, A.

    2016-12-01

    As ocean conditions continue to change, marine ecosystems are significantly impacted. Many calcifying organisms are being affected by the gradual changes in ocean pH and temperature that continue to occur over time. Corals are organisms that engage in a symbiotic relationship with Symbiodinium dinoflagellates (zooxanthellae). Symbiodinium are responsible for photosynthetic activity within oligotrophic waters. Corals depend on high levels of aragonite saturation state of seawater in order to build their skeletal structure. Most corals have a relatively narrow optimal range of temperature and pH in which they thrive. However, it is thought that corals residing in the Gulf of Aqaba (Red Sea) are resilient to the effects of increasing temperature. Stylophora pistillata's response to environmental impacts was tested via a simulation of ocean conditions at a high temperature and high CO2 emission scenario (pH 7.65) and lower CO2 emission scenario (pH 7.85) that are predicted for the end of this century. We present the difference in zooxanthellae density following a short term experiment where corals were placed in seawater tanks at pH 7.65, 7.85 and 8.1 and temperature was increased by 4 degrees C above seawater temperature in order to measure the response of Stylophora pistillata to potential future ocean conditions.

  6. Research of Isolation and Degradation Conditions of Petroleum Degrading Marine

    NASA Astrophysics Data System (ADS)

    Fangrui, Guo

    2017-01-01

    A novel petroleum-degrading microbial strain was isolated from sediment samples in estuary of Bohai Sea estuary beaches. The strain was primarily identified as Alcanivorax sp. and named Alcanivorax sp. H34. Effect of PH values, temperature, nitrogen and phosphorus concentrations on degradation of H34 were investigated. The paraffinic components average degradation rate of H34 ungrowth cells under optimized conditions was studied. The results showed that the optimal growth conditions of H34 are were temperature of 30°C, initial PH of 7.0, nitrogen concentration of 3g/L, phosphorus concentration of 3g/L, and paraffinic components average degradation rates of H34 ungrowth cells was 41.6%, while total degradation rate was 45.5%.

  7. Optimization of monomethoxy polyethyleneglycol-modified oxalate decarboxylase by response surface methodology.

    PubMed

    Long, Han; Cai, XingHua; Yang, Hui; He, JunBin; Wu, Jia; Lin, RiHui

    2017-09-01

    In order to improve the stability of oxalate decarboxylase (Oxdc), response surface methodology (RSM), based on a four-factor three-level Box-Behnken central composite design was used to optimize the reaction conditions of oxalate decarboxylase (Oxdc) modified with monomethoxy polyethyleneglycol (mPEG5000). Four independent variables such as the ratio of mPEG-aldehyde to Oxdc, reaction time, temperature, and reaction pH were investigated in this work. The structure of modified Oxdc was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared (FTIR) spectroscopy, the stability of the modified Oxdc was also investigated. The optimal conditions were as follows: the mole ratio of mPEG-aldehyde to Oxdc of 1:47.6, time of 13.1 h, temperature at 29.9 °C, and the reaction pH of 5.3. Under optimal conditions, experimental modified rate (MR = 73.69%) and recovery rate (RR = 67.58%) were matched well with the predicted value (MR = 75.11%) and (RR = 69.17%). SDS-PAGE and FTIR analysis showed that mPEG was covalently bound to the Oxdc. Compared with native Oxdc, the modified Oxdc (mPEG-Oxdc) showed higher thermal stability and better tolerance to trypsin or different pH treatment. This work will provide a further theoretical reference for enzyme modification and conditional optimization.

  8. Optimization of probiotic and lactic acid production by Lactobacillus plantarum in submerged bioreactor systems.

    PubMed

    Brinques, Graziela Brusch; do Carmo Peralba, Maria; Ayub, Marco Antônio Záchia

    2010-02-01

    Biomass and lactic acid production by a Lactobacillus plantarum strain isolated from Serrano cheese, a microorganism traditionally used in foods and recognized as a potent probiotic, was optimized. Optimization procedures were carried out in submerged batch bioreactors using cheese whey as the main carbon source. Sequential experimental Plackett-Burman designs followed by central composite design (CCD) were used to assess the influence of temperature, pH, stirring, aeration rate, and concentrations of lactose, peptone, and yeast extract on biomass and lactic acid production. Results showed that temperature, pH, aeration rate, lactose, and peptone were the most influential variables for biomass formation. Under optimized conditions, the CCD for temperature and aeration rate showed that the model predicted maximal biomass production of 14.30 g l(-1) (dw) of L. plantarum. At the central point of the CCD, a biomass of 10.2 g l(-1) (dw), with conversion rates of 0.10 g of cell g(-1) lactose and 1.08 g lactic acid g(-1) lactose (w/w), was obtained. These results provide useful information about the optimal cultivation conditions for growing L. plantarum in batch bioreactors in order to boost biomass to be used as industrial probiotic and to obtain high yields of conversion of lactose to lactic acid.

  9. Influence of prebiotics on Lactobacillus reuteri death kinetics under sub-optimal temperatures and pH.

    PubMed

    Altieri, Clelia; Iorio, Maria Clara; Bevilacqua, Antonio; Sinigaglia, Milena

    2016-01-01

    Eaten foodstuffs are usually fortified with prebiotic ingredients, such as inulin and oligofructose (FOS). The main goal of this study was to evaluate the combined effects of inulin and FOS with either suboptimal pH or storage temperature on the viability of Lactobacillus reuteri DSM 20016. Data were modeled through Weibull equation for the evaluation of the microbiological shelf life and the survival time. Prebiotics enhanced the microbiological shelf life and enhanced the survival time of the target bacterium. The use of the factorial ANOVA highlighted that inulin and FOS exerted a different effect as a function of pH and temperature. Inulin prolonged survival time under acidic conditions, while the effect of glucose + FOS was significant at pH 8. Finally, temperature could act by increasing or decreasing the effect of prebiotics, as they could exert a protective effect at 30 °C but not at 44 °C. As the main output of this research, we could suggest that the effect of prebiotics on L. reuteri could be significantly affected by pH and temperature, thus pinpointing that the design of a symbiotic food should also rely on these factors.

  10. Optimum production and characterization of an acid protease from marine yeast Metschnikowia reukaufii W6b

    NASA Astrophysics Data System (ADS)

    Li, Jing; Peng, Ying; Wang, Xianghong; Chi, Zhenming

    2010-12-01

    The marine yeast strain W6b isolated from sediment of the South China Sea was found to produce a cell-bound acid protease. The crude acid protease produced by this marine yeast showed the highest activity at pH 3.5 and 40 °C. The optimal pH and temperature for the crude acid protease were in agreement with those for acid protease produced by the terrestrial yeasts. The optimal medium of the acid protease production was seawater containing 1.0% glucose, 1.5% casein, and 0.5% yeast extract, and the optimal cultivation conditions of the acid protease production were pH 4.0, a temperature of 25 °C and a shaking speed of 140 rmin-1. Under the optimal conditions, 72.5 UmL-1 of acid protease activity could be obtained in cell suspension within 48 h of fermentation at shake flask level. The acid protease production was induced by high-molecular-weight nitrogen sources and repressed by low-molecular-weight nitrogen sources. Skimmed-milk-clotting test showed that the crude acid protease from the cell suspension of the yeast W6b had high skimmed milk coagulability. The acid protease produced by M. reukaufii W6b may have highly potential applications in cheese, food and fermentation industries.

  11. Sequentially Integrated Optimization of the Conditions to Obtain a High-Protein and Low-Antinutritional Factors Protein Isolate from Edible Jatropha curcas Seed Cake.

    PubMed

    León-López, Liliana; Dávila-Ortiz, Gloria; Jiménez-Martínez, Cristian; Hernández-Sánchez, Humberto

    2013-01-01

    Jatropha curcas seed cake is a protein-rich byproduct of oil extraction which could be used to produce protein isolates. The purpose of this study was the optimization of the protein isolation process from the seed cake of an edible provenance of J. curcas by an alkaline extraction followed by isoelectric precipitation method via a sequentially integrated optimization approach. The influence of four different factors (solubilization pH, extraction temperature, NaCl addition, and precipitation pH) on the protein and antinutritional compounds content of the isolate was evaluated. The estimated optimal conditions were an extraction temperature of 20°C, a precipitation pH of 4, and an amount of NaCl in the extraction solution of 0.6 M for a predicted protein content of 93.3%. Under these conditions, it was possible to obtain experimentally a protein isolate with 93.21% of proteins, 316.5 mg 100 g(-1) of total phenolics, 2891.84 mg 100 g(-1) of phytates and 168 mg 100 g(-1) of saponins. The protein content of the this isolate was higher than the content reported by other authors.

  12. Sequentially Integrated Optimization of the Conditions to Obtain a High-Protein and Low-Antinutritional Factors Protein Isolate from Edible Jatropha curcas Seed Cake

    PubMed Central

    León-López, Liliana; Dávila-Ortiz, Gloria; Jiménez-Martínez, Cristian; Hernández-Sánchez, Humberto

    2013-01-01

    Jatropha curcas seed cake is a protein-rich byproduct of oil extraction which could be used to produce protein isolates. The purpose of this study was the optimization of the protein isolation process from the seed cake of an edible provenance of J. curcas by an alkaline extraction followed by isoelectric precipitation method via a sequentially integrated optimization approach. The influence of four different factors (solubilization pH, extraction temperature, NaCl addition, and precipitation pH) on the protein and antinutritional compounds content of the isolate was evaluated. The estimated optimal conditions were an extraction temperature of 20°C, a precipitation pH of 4, and an amount of NaCl in the extraction solution of 0.6 M for a predicted protein content of 93.3%. Under these conditions, it was possible to obtain experimentally a protein isolate with 93.21% of proteins, 316.5 mg 100 g−1 of total phenolics, 2891.84 mg 100 g−1 of phytates and 168 mg 100 g−1 of saponins. The protein content of the this isolate was higher than the content reported by other authors. PMID:25937971

  13. Optimization of Pumpkin Oil Recovery by Using Aqueous Enzymatic Extraction and Comparison of the Quality of the Obtained Oil with the Quality of Cold-Pressed Oil

    PubMed Central

    Roszkowska, Beata; Czaplicki, Sylwester; Tańska, Małgorzata

    2016-01-01

    Summary The study was carried out to optimize pumpkin oil recovery in the process of aqueous extraction preceded by enzymatic maceration of seeds, as well as to compare the quality of the obtained oil to the quality of cold-pressed pumpkin seed oil. Hydrated pulp of hulless pumpkin seeds was macerated using a 2% (by mass) cocktail of commercial pectinolytic, cellulolytic and proteolytic preparations (Rohapect® UF, Rohament® CL and Colorase® 7089). The optimization procedure utilized response surface methodology based on Box- -Behnken plan of experiment. The optimized variables of enzymatic pretreatment were pH, temperature and maceration time. The results showed that the pH value, temperature and maceration time of 4.7, 54 °C and 15.4 h, respectively, were conducive to maximize the oil yield up to 72.64%. Among these variables, the impact of pH was crucial (above 73% of determined variation) for oil recovery results. The oil obtained by aqueous enzymatic extraction was richer in sterols, squalene and tocopherols, and only slightly less abundant in carotenoids than the cold-pressed one. However, it had a lower oxidative stability, with induction period shortened by approx. 30% in relation to the cold-pressed oil. PMID:28115898

  14. Optimization of Cellulase Production from Bacteria Isolated from Soil

    PubMed Central

    Sethi, Sonia; Datta, Aparna; Gupta, B. Lal; Gupta, Saksham

    2013-01-01

    Cellulase-producing bacteria were isolated from soil and identified as Pseudomonas fluorescens, Bacillus subtilIs, E. coli, and Serratia marcescens. Optimization of the fermentation medium for maximum cellulase production was carried out. The culture conditions like pH, temperature, carbon sources, and nitrogen sources were optimized. The optimum conditions found for cellulase production were 40°C at pH 10 with glucose as carbon source and ammonium sulphate as nitrogen source, and coconut cake stimulates the production of cellulase. Among bacteria, Pseudomonas fluorescens is the best cellulase producer among the four followed by Bacillus subtilis, E. coli, and Serratia marscens. PMID:25937986

  15. Optimal Conditions for the Mycelial Growth of Coprinus comatus Strains

    PubMed Central

    Lee, Yun-Hae; Liu, Jun-Jie; Ju, Young-Cheol

    2009-01-01

    The principal objective of this study was to acquire basic data regarding the mycelial growth characteristics for the artificial cultivation of Coprinus comatus. 12 URP primers were employed to evaluate the genetic relationships of C. comatus, and the results were divided into three groups. Among six kinds of mushroom media, MYP medium was selected as the most favorable culture medium for C. comatus. The optimal temperature and pH ranges for the mycelial growth of C. comatus were 23~26℃ and pH 6~8, respectively. The carbon and nitrogen sources for optimal mycelial growth were sucrose and tryptone, respectively. PMID:23983517

  16. Comprehensive model of microalgae photosynthesis rate as a function of culture conditions in photobioreactors.

    PubMed

    Costache, T A; Acién Fernández, F Gabriel; Morales, M M; Fernández-Sevilla, J M; Stamatin, I; Molina, E

    2013-09-01

    In this paper, the influence of culture conditions (irradiance, temperature, pH, and dissolved oxygen) on the photosynthesis rate of Scenedesmus almeriensis cultures is analyzed. Short-run experiments were performed to study cell response to variations in culture conditions, which take place in changing environments such as outdoor photobioreactors. Experiments were performed by subjecting diluted samples of cells to different levels of irradiance, temperature, pH, and dissolved oxygen concentration. Results demonstrate the existence of photoinhibition phenomena at irradiances higher than 1,000 μE/m(2) s; in addition to reduced photosynthesis rates at inadequate temperatures or pH-the optimal values being 35 °C and 8, respectively. Moreover, photosynthesis rate reduction at dissolved oxygen concentrations above 20 mg/l is demonstrated. Data have been used to develop an integrated model based on considering the simultaneous influence of irradiance, temperature, pH, and dissolved oxygen. The model fits the experimental results in the range of culture conditions tested, and it was validated using data obtained by the simultaneous variation of two of the modified variables. Furthermore, the model fits experimental results obtained from an outdoor culture of S. almeriensis performed in an open raceway reactor. Results demonstrate that photosynthetic efficiency is modified as a function of culture conditions, and can be used to determine the proximity of culture conditions to optimal values. Optimal conditions found (T = 35 °C, pH = 8, dissolved oxygen concentration <20 mg/l) allows to maximize the use of light by the cells. The developed model is a powerful tool for the optimal design and management of microalgae-based processes, especially outdoors, where the cultures are subject to daily culture condition variations.

  17. Parameterization of hyperpolarized (13)C-bicarbonate-dissolution dynamic nuclear polarization.

    PubMed

    Scholz, David Johannes; Otto, Angela M; Hintermair, Josef; Schilling, Franz; Frank, Annette; Köllisch, Ulrich; Janich, Martin A; Schulte, Rolf F; Schwaiger, Markus; Haase, Axel; Menzel, Marion I

    2015-12-01

    (13)C metabolic MRI using hyperpolarized (13)C-bicarbonate enables preclinical detection of pH. To improve signal-to-noise ratio, experimental procedures were refined, and the influence of pH, buffer capacity, temperature, and field strength were investigated. Bicarbonate preparation was investigated. Bicarbonate was prepared and applied in spectroscopy at 1, 3, 14 T using pure dissolution, culture medium, and MCF-7 cell spheroids. Healthy rats were imaged by spectral-spatial spiral acquisition for spatial and temporal bicarbonate distribution, pH mapping, and signal decay analysis. An optimized preparation technique for maximum solubility of 6 mol/L and polarization levels of 19-21% is presented; T1 and SNR dependency on field strength, buffer capacity, and pH was investigated. pH mapping in vivo is demonstrated. An optimized bicarbonate preparation and experimental procedure provided improved T1 and SNR values, allowing in vitro and in vivo applications.

  18. Optimization of γ-aminobutyric acid production by Lactobacillus plantarum Taj-Apis362 from honeybees.

    PubMed

    Tajabadi, Naser; Ebrahimpour, Afshin; Baradaran, Ali; Rahim, Raha Abdul; Mahyudin, Nor Ainy; Manap, Mohd Yazid Abdul; Bakar, Fatimah Abu; Saari, Nazamid

    2015-04-15

    Dominant strains of lactic acid bacteria (LAB) isolated from honey bees were evaluated for their γ-aminobutyric acid (GABA)-producing ability. Out of 24 strains, strain Taj-Apis362 showed the highest GABA-producing ability (1.76 mM) in MRS broth containing 50 mM initial glutamic acid cultured for 60 h. Effects of fermentation parameters, including initial glutamic acid level, culture temperature, initial pH and incubation time on GABA production were investigated via a single parameter optimization strategy. The optimal fermentation condition for GABA production was modeled using response surface methodology (RSM). The results showed that the culture temperature was the most significant factor for GABA production. The optimum conditions for maximum GABA production by Lactobacillus plantarum Taj-Apis362 were an initial glutamic acid concentration of 497.97 mM, culture temperature of 36 °C, initial pH of 5.31 and incubation time of 60 h, which produced 7.15 mM of GABA. The value is comparable with the predicted value of 7.21 mM.

  19. Citrobacter amalonaticus Phytase on the Cell Surface of Pichia pastoris Exhibits High pH Stability as a Promising Potential Feed Supplement

    PubMed Central

    Li, Cheng; Lin, Ying; Huang, Yuanyuan; Liu, Xiaoxiao; Liang, Shuli

    2014-01-01

    Phytase expressed and anchored on the cell surface of Pichia pastoris avoids the expensive and time-consuming steps of protein purification and separation. Furthermore, yeast cells with anchored phytase can be used as a whole-cell biocatalyst. In this study, the phytase gene of Citrobacter amalonaticus was fused with the Pichia pastoris glycosylphosphatidylinositol (GPI)-anchored glycoprotein homologue GCW61. Phytase exposed on the cell surface exhibits a high activity of 6413.5 U/g, with an optimal temperature of 60°C. In contrast to secreted phytase, which has an optimal pH of 5.0, phytase presented on the cell surface is characterized by an optimal pH of 3.0. Moreover, our data demonstrate that phytase anchored on the cell surface exhibits higher pH stability than its secreted counterpart. Interestingly, our in vitro digestion experiments demonstrate that phytase attached to the cell surface is a more efficient enzyme than secreted phytase. PMID:25490768

  20. Citrobacter amalonaticus phytase on the cell surface of Pichia pastoris exhibits high pH stability as a promising potential feed supplement.

    PubMed

    Li, Cheng; Lin, Ying; Huang, Yuanyuan; Liu, Xiaoxiao; Liang, Shuli

    2014-01-01

    Phytase expressed and anchored on the cell surface of Pichia pastoris avoids the expensive and time-consuming steps of protein purification and separation. Furthermore, yeast cells with anchored phytase can be used as a whole-cell biocatalyst. In this study, the phytase gene of Citrobacter amalonaticus was fused with the Pichia pastoris glycosylphosphatidylinositol (GPI)-anchored glycoprotein homologue GCW61. Phytase exposed on the cell surface exhibits a high activity of 6413.5 U/g, with an optimal temperature of 60°C. In contrast to secreted phytase, which has an optimal pH of 5.0, phytase presented on the cell surface is characterized by an optimal pH of 3.0. Moreover, our data demonstrate that phytase anchored on the cell surface exhibits higher pH stability than its secreted counterpart. Interestingly, our in vitro digestion experiments demonstrate that phytase attached to the cell surface is a more efficient enzyme than secreted phytase.

  1. Cloning and high-level expression of β-xylosidase from Selenomonas ruminantium in Pichia pastoris by optimizing of pH, methanol concentration and temperature conditions.

    PubMed

    Dehnavi, Ehsan; Ranaei Siadat, Seyed Omid; Fathi Roudsari, Mehrnoosh; Khajeh, Khosro

    2016-08-01

    β-xylosidase and several other glycoside hydrolase family members, including xylanase, cooperate together to degrade hemicelluloses, a commonly found xylan polymer of plant-cell wall. β-d-xylosidase/α-l-arabinofuranosidase from the ruminal anaerobic bacterium Selenomonas ruminantium (SXA) has potential utility in industrial processes such as production of fuel ethanol and other bioproducts. The optimized synthetic SXA gene was overexpressed in methylotrophic Pichia pastoris under the control of alcohol oxidase I (AOX1) promoter and secreted into the medium. Recombinant protein showed an optimum pH 4.8 and optimum temperature 50 °C. Furthermore, optimization of growth and induction conditions in shake flask was carried out. Using the optimum expression condition (pH 6, temperature 20 °C and 1% methanol induction), protein production was increased by about three times in comparison to the control. The recombinant SXA we have expressed here showed higher turnover frequency using ρ-nitrophenyl β-xylopyranoside (PNPX) substrate, in contrast to most xylosidase experiments reported previously. This is the first report on the cloning and expression of a β-xylosidase gene from glycoside hydrolase (GH) family 43 in Pichia pastoris. Our results confirm that P. pastoris is an appropriate host for high level expression and production of SXA for industrial applications. Copyright © 2016. Published by Elsevier Inc.

  2. Culture Conditions for Production of Biomass, Adenosine, and Cordycepin from Cordyceps sinensis CS1197: Optimization by Desirability Function Method

    PubMed Central

    Ghatnur, Shashidhar M.; Parvatam, Giridhar; Balaraman, Manohar

    2015-01-01

    Background: Cordyceps sinensis (CS) is a traditional Chinese medicine contains potent active metabolites such as nucleosides and polysaccharides. The submerged cultivation technique is studied for the large scale production of CS for biomass and metabolites production. Objective: To optimize culture conditions for large-scale production of CS1197 biomass and metabolites production. Materials and Methods: The CS1197 strain of CS was isolated from dead larvae of natural CS and the authenticity was assured by the presence of two major markers adenosine and cordycepin by high performance liquid chromatography and mass spectrometry. A three-level Box-Behnken design was employed to optimize process parameters culturing temperature, pH, and inoculum volume for the biomass yield, adenosine and cordycepin. The experimental results were regressed to a second-order polynomial equation by a multiple regression analysis for the prediction of biomass yield, adenosine and cordycepin production. Multiple responses were optimized based on desirability function method. Results: The desirability function suggested the process conditions temperature 28°C, pH 7 and inoculum volume 10% for optimal production of nutraceuticals in the biomass. The water extracts from dried CS1197 mycelia showed good inhibition for 2 diphenyl-1-picrylhydrazyl and 2,2-azinobis-(3-ethyl-benzo-thiazoline-6-sulfonic acid-free radicals. Conclusion: The result suggests that response surface methodology-desirability function coupled approach can successfully optimize the culture conditions for CS1197. SUMMARY Authentication of CS1197 strain by the presence of adenosine and cordycepin and culturing period was determined to be for 14 daysContent of nucleosides in natural CS was found higher than in cultured CS1197 myceliumBox-Behnken design to optimize critical cultural conditions: temperature, pH and inoculum volumeWater extract showed better antioxidant activity proving credible source of natural antioxidants. PMID:26929580

  3. Enhanced activity and stability of L-arabinose isomerase by immobilization on aminopropyl glass.

    PubMed

    Zhang, Ye-Wang; Jeya, Marimuthu; Lee, Jung-Kul

    2011-03-01

    Immobilization of Bacillus licheniformis L: -arabinose isomerase (BLAI) on aminopropyl glass modified with glutaraldehyde (4 mg protein g support⁻¹) was found to enhance the enzyme activity. The immobilization yield of BLAI was proportional to the quantity of amino groups on the surface of support. Reducing particle size increased the adsorption capacity (q(m)) and affinity (k(a)). The pH and temperature for immobilization were optimized to be pH 7.1 and 33 °C using response surface methodology (RSM). The immobilized enzyme was characterized and compared to the free enzyme. There is no change in optimal pH and temperature before and after immobilization. However, the immobilized BLAI enzyme achieved 145% of the activity of the free enzyme. Correspondingly, the catalytic efficiency (k(cat)/K(m)) was improved 1.47-fold after immobilization compared to the free enzyme. The thermal stability was improved 138-fold (t₁/₂) increased from 2 to 275 h) at 50 °C following immobilization.

  4. Formation of glycosidases in batch and continuous culture of Bacteroides fragilis.

    PubMed Central

    Berg, J O; Nord, C E; Wadström, T

    1978-01-01

    Nine strains of bacteroides fragilis were cultivated in stirred fermentors and tested for their ability to produce glycosidases. B. fragilis subsp. vulgatus B70 was used for optimizing the production of glycosidases. The highest bacterial yield was obtained in proteose peptone-yeast extract medium. The optimum pH for maximal bacterial yield was 7.0, and the optimum temperature for growth was 37 degrees C. The formation of glycosidases was optimal between pH 6.5 and 7.5, and the optimum temperature for synthesis of glycosidases was between 33 and 37 degrees C. Culture under controlled conditions in fermentors gave more reproducible production of glycosidases than static cultures in bottles. The strain was also grown in continuous culture at a dilution rate of 0.1 liter/h at pH 7.0 and 37 degrees C with a yield of 2.0 mg of dry weight per ml in the complex medium. The formation of glycosidases remained constant during the entire continuous process. PMID:25044

  5. Monoterpene alcohol metabolism: identification, purification, and characterization of two geraniol dehydrogenase isoenzymes from Polygonum minus leaves.

    PubMed

    Hassan, Maizom; Maarof, Nur Diyana; Ali, Zainon Mohd; Noor, Normah Mohd; Othman, Roohaida; Mori, Nobuhiro

    2012-01-01

    NADP(+)-dependent geraniol dehydrogenase (EC 1.1.1.183) is an enzyme that catalyzes the oxidation of geraniol to geranial. Stable, highly active cell-free extract was obtained from Polygonum minus leaves using polyvinylpolypyrrolidone, Amberlite XAD-4, glycerol, 2-mercaptoethanol, thiourea, and phenylmethylsulfonylfluoride in tricine-NaOH buffer (pH 7.5). The enzyme preparation was separated into two activity peaks, geraniol-DH I and II, by DEAE-Toyopearl 650M column chromatography at pH 7.5. Both isoenzymes were purified to homogeneity in three chromatographic steps. The geraniol-DH isoenzymes were similar in molecular mass, optimal temperature, and pH, but the isoelectric point, substrate specificity, and kinetic parameters were different. The K(m) values for geraniol of geraniol-DH I and II appeared to be 0.4 mM and 0.185 mM respectively. P. minus geraniol-DHs are unusual among geraniol-DHs in view of their thermal stability and optimal temperatures, and also their high specificity for allylic alcohols and NADP(+).

  6. Fuel alcohol biosynthesis by Zymomonas anaerobia: optimization studies

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kosaric, N.; Ong, S.L.; Davnjak, Z.

    1982-03-01

    The optimum operating conditions for growth and ethanol production of Zymomonas anaerobia ATCC 29501 were established. The optimum pH range and temperature were found to be 5.0-6.0 and 35/sup 0/C, respectively. Based on the results obtained from the temperature optimization study, an Arrhenius-type temperature relationship for the specific growth rate was developed. The growth and ethanol production of this microbe also have been optimized in terms of concentrations of glucose, essential nutrients, and minerals. With optimum medium and operating conditions, an ethanol concentration of 96 g/L was obtained in 23h. Both growth and ethanol yield coefficients in dependence on initialmore » glucose concentrations were determined.« less

  7. Fuel alcohol biosynthesis by Zymomonas anaerobia: optimization studies

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kosaric, N.; Ong, S.L.; Duvnjak, Z.

    1982-03-01

    The optimum operating conditions for growth and ethanol production of Zymomonas anaerobia ATCC 29501 were established. The optimum pH range and temperature were found to be 5.0-6.0 and 35 degrees C, respectively. Based on the results obtained from the temperature optimization study, an Arrhenius-type temperature relationship for the specific growth rate was developed. The growth and ethanol production of this microbe also have been optimized in terms of concentrations of glucose, essential nutrients, and minerals. With optimum medium and operating conditions, an ethanol concentration of 96 g/L was obtained in 23 hours. Both growth and ethanol yield coefficients in dependencemore » on initial glucose concentrations were determined. (Refs. 16).« less

  8. The Effect of Temperature and Solution pH on the Nucleation of Tetragonal Lysozyme Crystals

    NASA Technical Reports Server (NTRS)

    Judge, Russell A.; Jacobs, Randolph S.; Frazier, Tyralynn; Snell, Edward H.; Pusey, Marc L.

    1999-01-01

    Part of the challenge of macromolecular crystal growth for structure determination is obtaining crystals with a volume suitable for x-ray analysis. In this respect an understanding of the effect of solution conditions on macromolecule nucleation rates is advantageous. This study investigated the effects of supersaturation, temperature, and pH on the nucleation rate of tetragonal lysozyme crystals. Batch crystallization plates were prepared at given solution concentrations and incubated at set temperatures over 1 week. The number of crystals per well with their size and axial ratios were recorded and correlated with solution conditions. Crystal numbers were found to increase with increasing supersaturation and temperature. The most significant variable, however, was pH; crystal numbers changed by two orders of magnitude over the pH range 4.0-5.2. Crystal size also varied with solution conditions, with the largest crystals obtained at pH 5.2. Having optimized the crystallization conditions, we prepared a batch of crystals under the same initial conditions, and 50 of these crystals were analyzed by x-ray diffraction techniques. The results indicate that even under the same crystallization conditions, a marked variation in crystal properties exists.

  9. Realm of Thermoalkaline Lipases in Bioprocess Commodities.

    PubMed

    Lajis, Ahmad Firdaus B

    2018-01-01

    For decades, microbial lipases are notably used as biocatalysts and efficiently catalyze various processes in many important industries. Biocatalysts are less corrosive to industrial equipment and due to their substrate specificity and regioselectivity they produced less harmful waste which promotes environmental sustainability. At present, thermostable and alkaline tolerant lipases have gained enormous interest as biocatalyst due to their stability and robustness under high temperature and alkaline environment operation. Several characteristics of the thermostable and alkaline tolerant lipases are discussed. Their molecular weight and resistance towards a range of temperature, pH, metal, and surfactants are compared. Their industrial applications in biodiesel, biodetergents, biodegreasing, and other types of bioconversions are also described. This review also discusses the advance of fermentation process for thermostable and alkaline tolerant lipases production focusing on the process development in microorganism selection and strain improvement, culture medium optimization via several optimization techniques (i.e., one-factor-at-a-time, surface response methodology, and artificial neural network), and other fermentation parameters (i.e., inoculums size, temperature, pH, agitation rate, dissolved oxygen tension (DOT), and aeration rate). Two common fermentation techniques for thermostable and alkaline tolerant lipases production which are solid-state and submerged fermentation methods are compared and discussed. Recent optimization approaches using evolutionary algorithms (i.e., Genetic Algorithm, Differential Evolution, and Particle Swarm Optimization) are also highlighted in this article.

  10. Optimization of pectin extraction from banana peels with citric acid by using response surface methodology.

    PubMed

    Oliveira, Túlio Ítalo S; Rosa, Morsyleide F; Cavalcante, Fabio Lima; Pereira, Paulo Henrique F; Moates, Graham K; Wellner, Nikolaus; Mazzetto, Selma E; Waldron, Keith W; Azeredo, Henriette M C

    2016-05-01

    A central composite design was used to determine effects of pH (2.0-4.5), extraction temperature (70-90 °C) and time (120-240 min) on the yield, degree of methoxylation (DM) and galacturonic acid content (GA) of pectins extracted from banana peels with citric acid. Changes in composition during the main steps of pectin extraction were followed by Fourier transform infrared (FTIR) spectroscopy. FTIR was also used to determine DM and GA of pectins. Harsh temperature and pH conditions enhanced the extraction yield, but decreased DM. GA presented a maximum value at 83 °C, 190 min, and pH 2.7. The yield of galacturonic acid (YGA), which took into account both the extraction yield and the pectin purity, was improved by higher temperature and lower pH values. The optimum extraction conditions, defined as those resulting in a maximum YGA while keeping DM at a minimum of 51%, were: 87 °C, 160 min, pH 2.0. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. The effects of pH and temperature on the in vitro bindings of delta-9-tetrahydrocannabinol and other cannabinoids to bovine serum albumin.

    PubMed

    Papa, V M; Shen, M L; Ou, D W

    1990-01-01

    Albumin is a major carrier of drugs and fatty acids in biological fluids. These protein-drug complexes serve to solubilize, transport these compounds to sites of action, and have been associated with increased half-life for these compounds. The authors are interested in the pH and temperature effects of the binding of delta-9-tetrahydrocannabinol to albumin. Ultrafiltration techniques were used in the separation of free to bound compounds. Cannabinoids bind to bovine serum albumin rapidly. The cannabinoid binding sites are more sensitive to temperature changes (37-47 degrees C) than changes in pH with 37 degrees C and pH 7.4 resulting in optimal binding. These conditions would result in the greatest viability in the cells, while allowing for the use of a variety of compounds in in vitro studies for the administration of compounds to isolated cells and cell lines.

  12. [Optimization of prokaryotic expression conditions of Leptospira interrogans trigeminy genus-specific protein antigen based on surface response analysis].

    PubMed

    Wang, Jiang; Luo, Dongjiao; Sun, Aihua; Yan, Jie

    2008-07-01

    Lipoproteins LipL32 and LipL21 and transmembrane protein OMPL1 have been confirmed as the superficial genus-specific antigens of Leptospira interrogans, which can be used as antigens for developing a universal genetic engineering vaccine. In order to obtain high expression of an artificial fusion gene lipL32/1-lipL21-ompL1/2, we optimized prokaryotic expression conditions. We used surface response analysis based on the central composite design to optimize culture conditions of a new antigen protein by recombinant Escherichia coli DE3.The culture conditions included initial pH, induction start time, post-induction time, Isopropyl beta-D-thiogalactopyranoside (IPTG) concentration, and temperature. The maximal production of antigen protein was 37.78 mg/l. The optimal culture conditions for high recombinant fusion protein was determined: initial pH 7.9, induction start time 2.5 h, a post-induction time of 5.38 h, 0.20 mM IPTG, and a post-induction temperature of 31 degrees C. Surface response analysis based on CCD increased the target production. This statistical method reduced the number of experiments required for optimization and enabled rapid identification and integration of the key culture condition parameters for optimizing recombinant protein expression.

  13. Decolorization of Anthraquinonic Dyes from Textile Effluent Using Horseradish Peroxidase: Optimization and Kinetic Study

    PubMed Central

    Šekuljica, Nataša Ž.; Prlainović, Nevena Ž.; Stefanović, Andrea B.; Žuža, Milena G.; Čičkarić, Dragana Z.; Mijin, Dušan Ž.; Knežević-Jugović, Zorica D.

    2015-01-01

    Two anthraquinonic dyes, C.I. Acid Blue 225 and C.I. Acid Violet 109, were used as models to explore the feasibility of using the horseradish peroxidase enzyme (HRP) in the practical decolorization of anthraquinonic dyes in wastewater. The influence of process parameters such as enzyme concentration, hydrogen peroxide concentration, temperature, dye concentration, and pH was examined. The pH and temperature activity profiles were similar for decolorization of both dyes. Under the optimal conditions, 94.7% of C.I. Acid Violet 109 from aqueous solution was decolorized (treatment time 15 min, enzyme concentration 0.15 IU/mL, hydrogen peroxide concentration 0.4 mM, dye concentration 30 mg/L, pH 4, and temperature 24°C) and 89.36% of C.I. Acid Blue 225 (32 min, enzyme concentration 0.15 IU/mL, hydrogen peroxide concentration 0.04 mM, dye concentration 30 mg/L, pH 5, and temperature 24°C). The mechanism of both reactions has been proven to follow the two substrate ping-pong mechanism with substrate inhibition, revealing the formation of a nonproductive or dead-end complex between dye and HRP or between H2O2 and the oxidized form of the enzyme. Both chemical oxygen demand and total organic carbon values showed that there was a reduction in toxicity after the enzymatic treatment. This study verifies the viability of use of horseradish peroxidase for the wastewaters treatment of similar anthraquinonic dyes. PMID:25685837

  14. Enhancement of 2,3-Butanediol Production by Klebsiella oxytoca PTCC 1402

    PubMed Central

    Anvari, Maesomeh; Safari Motlagh, Mohammad Reza

    2011-01-01

    Optimal operating parameters of 2,3-Butanediol production using Klebsiella oxytoca under submerged culture conditions are determined by using Taguchi method. The effect of different factors including medium composition, pH, temperature, mixing intensity, and inoculum size on 2,3-butanediol production was analyzed using the Taguchi method in three levels. Based on these analyses the optimum concentrations of glucose, acetic acid, and succinic acid were found to be 6, 0.5, and 1.0 (% w/v), respectively. Furthermore, optimum values for temperature, inoculum size, pH, and the shaking speed were determined as 37°C, 8 (g/L), 6.1, and 150 rpm, respectively. The optimal combinations of factors obtained from the proposed DOE methodology was further validated by conducting fermentation experiments and the obtained results revealed an enhanced 2,3-Butanediol yield of 44%. PMID:21318172

  15. Optimization of the mineralization of a mixture of phenolic pollutants under a ferrioxalate-induced solar photo-Fenton process.

    PubMed

    Monteagudo, J M; Durán, A; Aguirre, M; San Martín, I

    2011-01-15

    The mineralization of solutions containing a mixture of three phenolic compounds, gallic, p-coumaric and protocatechuic acids, in a ferrioxalate-induced solar photo-Fenton process was investigated. The reactions were carried out in a pilot plant consisting of a compound parabolic collector (CPC) solar reactor. An optimization study was performed combining a multivariate experimental design and neuronal networks that included the following variables: pH, temperature, solar power, air flow and initial concentrations of H(2)O(2), Fe(II) and oxalic acid. Under optimal conditions, total elimination of the original compounds and 94% TOC removal of the mixture were achieved in 5 and 194 min, respectively. pH and initial concentrations of H(2)O(2) and Fe(II) were the most significant factors affecting the mixture mineralization. The molar correlation between consumed hydrogen peroxide and removed TOC was always between 1 and 3. A detailed analysis of the reaction was presented. The values of the pseudo-first-order mineralization kinetic rate constant, k(TOC), increased as initial Fe(II) and H(2)O(2) concentrations and temperature increased. The optimum pH value also slightly increased with greater Fe(II) and hydrogen peroxide concentrations but decreased when temperature increased. OH and O(2)(-) radicals were the main oxidative intermediate species in the process, although singlet oxygen ((1)O(2)) also played a role in the mineralization reaction. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Optimization of pectinase immobilization on grafted alginate-agar gel beads by 24 full factorial CCD and thermodynamic profiling for evaluating of operational covalent immobilization.

    PubMed

    Abdel Wahab, Walaa A; Karam, Eman A; Hassan, Mohamed E; Kansoh, Amany L; Esawy, Mona A; Awad, Ghada E A

    2018-07-01

    Pectinase produced by a honey derived from the fungus Aspergillus awamori KX943614 was covalently immobilized onto gel beads made of alginate and agar. Polyethyleneimine, glutaraldehyde, loading time and enzyme's units were optimized by 2 4 full factorial central composite design (CCD). The immobilization process increased the optimal working pH for the free pectinase from 5 to a broader range of pH4.5-5.5 and the optimum operational temperature from 55°C to a higher temperature, of 60°C, which is favored to reduce the enzyme's microbial contamination. The thermodynamics studies showed a thermal stability enhancement against high temperature for the immobilized formula. Moreover, an increase in half-lives and D-values was achieved. The thermodynamic studies proved that immobilization of pectinase made a remarkable increase in enthalpy and free energy because of enzyme stability enhancement. The reusability test revealed that 60% of pectinase's original activity was retained after 8 successive cycles. This gel formula may be convenient for immobilization of other industrial enzymes. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. Effect of fermentation parameters on bio-alcohols production from glycerol using immobilized Clostridium pasteurianum: an optimization study.

    PubMed

    Khanna, Swati; Goyal, Arun; Moholkar, Vijayanand S

    2013-01-01

    This article addresses the issue of effect of fermentation parameters for conversion of glycerol (in both pure and crude form) into three value-added products, namely, ethanol, butanol, and 1,3-propanediol (1,3-PDO), by immobilized Clostridium pasteurianum and thereby addresses the statistical optimization of this process. The analysis of effect of different process parameters such as agitation rate, fermentation temperature, medium pH, and initial glycerol concentration indicated that medium pH was the most critical factor for total alcohols production in case of pure glycerol as fermentation substrate. On the other hand, initial glycerol concentration was the most significant factor for fermentation with crude glycerol. An interesting observation was that the optimized set of fermentation parameters was found to be independent of the type of glycerol (either pure or crude) used. At optimum conditions of agitation rate (200 rpm), initial glycerol concentration (25 g/L), fermentation temperature (30°C), and medium pH (7.0), the total alcohols production was almost equal in anaerobic shake flasks and 2-L bioreactor. This essentially means that at optimum process parameters, the scale of operation does not affect the output of the process. The immobilized cells could be reused for multiple cycles for both pure and crude glycerol fermentation.

  18. Conformal self-assembled thin films for optical pH sensors

    NASA Astrophysics Data System (ADS)

    Topasna, Daniela M.; Topasna, Gregory A.; Liu, Minghanbo; Tseng, Ching-Hung

    2016-04-01

    Simple, reliable, lightweight, and inexpensive thin films based sensors are still in intense development and high demand in many applications such as biomedical, industrial, environmental, military, and consumer products. One important class of sensors is the optical pH sensor. In addition, conformal thin film based sensors extend the range of application for pH optical sensors. We present the results on the fabrication and characterization of optical pH sensing coatings made through ionic self-assembled technique. These thin films are based on the combination of a polyelectrolyte and water-soluble organic dye molecule Direct Yellow 4. A series of films was fabricated and characterized in order to determine the optimized parameters of the polymer and of the organic dye solutions. The optical pH responses of these films were also studied. The transparent films were immersed in solutions at various temperature and pH values. The films are stable when immersed in solutions with pH below 9.0 and temperatures below 90 °C and they maintain their performance after longer immersion times. We also demonstrate the functionality of these coatings as conformal films.

  19. Stability of urea in solution and pharmaceutical preparations.

    PubMed

    Panyachariwat, Nattakan; Steckel, Hartwig

    2014-01-01

    The stability of urea in solution and pharmaceutical preparations was analyzed as a function of temperature (25°-60°C), pH (3.11-9.67), and initial urea concentration (2.5%-20%). This study was undertaken to (i) obtain more extensive, quantitative information relative to the degradation of urea in both aqueous and non-aqueous solutions and in pharmaceutical preparations, and (ii) test the effects of initial urea concentration, pH, buffer, and temperature values on urea degradation. The stability analysis shows that urea is more stable at the pH range of 4-8 and the stability of urea decreases by increase in temperature for all pH values. Within the experimental range of temperature and initial urea concentration values, the lowest urea degradation was found with lactate buffer pH 6.0. The urea decomposition rate in solution and pharmaceutical preparations shows the dependence of the initial urea concentrations. At higher initial urea concentrations, the rate of degradation is a decreasing function with time. This suggests that the reverse reaction is a factor in the degradation of concentrated urea solution. For non-aqueous solvents, isopropanol showed the best effort in retarding the decomposition of urea. Since the losses in urea is directly influenced by its stability at a given temperature and pH, the stability analysis of urea by the proposed model can be used to prevent the loss and optimize the operating condition for urea-containing pharmaceutical preparations.

  20. Optimization of furfural and 5-hydroxymethylfurfural production from wheat straw by a microwave-assisted process.

    PubMed

    Yemiş, Oktay; Mazza, Giuseppe

    2012-04-01

    Optimization of acid-catalyzed conversion conditions of wheat straw into furfural, 5-hydroxymethylfurfural (HMF), glucose, and xylose was studied by response surface methodology (RSM). A central composite design (CCD) was used to determine the effects of independent variables, including reaction temperature (140-200 °C), residence time (1-41 min), pH (0.1-2.1), and liquid:solid ratio (15-195 mL/g) on furan and sugar production. The surface response analysis revealed that temperature, time and pH had a strong influence on the furfural, HMF, xylose and glucose yield, whereas liquid to solid ratio was found not to be significant. The initial pH of solution was the most important variable in acid-catalyzed conversion of wheat straw to furans. The maximum predicted furfural, HMF, xylose and glucose yields were 66%, 3.4%, 100%, and 65%, respectively. This study demonstrated that the microwave-assisted process was a very effective method for the xylose production from wheat straw by diluted acid catalysis. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

  1. Optimization of the SPME parameters and its online coupling with HPLC for the analysis of tricyclic antidepressants in plasma samples.

    PubMed

    Alves, Claudete; Fernandes, Christian; Dos Santos Neto, Alvaro José; Rodrigues, José Carlos; Costa Queiroz, Maria Eugênia; Lanças, Fernando Mauro

    2006-07-01

    Solid-phase microextraction (SPME)-liquid chromatography (LC) is used to analyze tricyclic antidepressant drugs desipramine, imipramine, nortriptyline, amitriptyline, and clomipramine (internal standard) in plasma samples. Extraction conditions are optimized using a 2(3) factorial design plus a central point to evaluate the influence of the time, temperature, and matrix pH. A Polydimethylsiloxane-divinylbenzene (60-mum film thickness) fiber is selected after the assessment of different types of coating. The chromatographic separation is realized using a C(18) column (150 x 4.6 mm, 5-microm particles), ammonium acetate buffer (0.05 mol/L, pH 5.50)-acetonitrile (55:45 v/v) with 0.1% of triethylamine as mobile phase and UV-vis detection at 214 nm. Among the factorial design conditions evaluated, the best results are obtained at a pH 11.0, temperature of 30 degrees C, and extraction time of 45 min. The proposed method, using a lab-made SPME-LC interface, allowed the determination of tricyclic antidepressants in in plasma at therapeutic concentration levels.

  2. High-yield production of a low-temperature-active polygalacturonase for papaya juice clarification.

    PubMed

    Tu, Tao; Meng, Kun; Bai, Yingguo; Shi, Pengjun; Luo, Huiying; Wang, Yaru; Yang, Peilong; Zhang, Yuhong; Zhang, Wei; Yao, Bin

    2013-12-01

    A novel endo-polygalacturonase (endo-PG I) from Achaetomium sp. Xz8 was identified, overexpressed in Pichia pastoris, and characterized in this report. Recombinant endo-PG I is distinguished from other enzyme counterparts by its high activity towards polygalacturonic acid (49,934 U/ml) and high yield in the 15-l fermentor (2.13 g/l). It exhibits optimal activity at 45 °C and remained active over a broad temperature range of 0-80 °C. Distinct from most fungal polygalacturonases that have acidic pH optima, endo-PG I is optimally active at pH 6, similar to the pH of fresh papaya juice (5.7). Endo-PG I alone reduced the viscosity of papaya juice by 17.6%, and increased its transmittance by 59.1%. When combined with a commercial pectin methylesterase, it showed much higher efficiency with a synergy degree of more than 1.25. All these favourable enzymatic properties make endo-PG I attractive for potential applications in the juice industry. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. Low molecular weight thermostable {beta}-D-glucosidase from Acidothermus cellulolyticus

    DOEpatents

    Himmel, M.E.; Tucker, M.P.; Adney, W.S.; Nieves, R.A.

    1995-07-11

    A purified low molecular weight {beta}-D-glucosidase is produced from Acidothermus cellulolyticus ATCC 43068. The enzyme is water soluble, possesses activity against pNP-{beta}-D-glucopyranoside, has a high of degree of stability toward heat, exhibits optimal temperature activity at about 65 C at a pH range of from about 2 to about 7, has an inactivation temperature of about 80 C at a pH range of from about 2 to about 7 and has a molecular weight of about 50.5--54.5 kD as determined by SDS-PAGE. 6 figs.

  4. Low molecular weight thermostable .beta.-D-glucosidase from acidothermus cellulolyticus

    DOEpatents

    Himmel, Michael E.; Tucker, Melvin P.; Adney, William S.; Nieves, Rafael A.

    1995-01-01

    A purified low molecular weight .beta.-D-glucosidase is produced from Acidothermus cellulolyticus ATCC 43068. The enzyme is water soluble, possesses activity against pNP-.beta.-D-glucopyranoside, has a high of degree of stability toward heat, exhibits optimal temperature activity at about 65.degree. C. at a pH range of from about 2 to about 7, has an inactivation temperature of about 80.degree. C. at a pH range of from about 2 to about 7 and has a molecular weight of about 50.5-54.5 kD as determineded by SDS-PAGE.

  5. The Effect of Temperature and Solution pH on Tetragonal Lysozyme Nucleation Kinetics

    NASA Technical Reports Server (NTRS)

    Judge, Russell A.; Jacobs, Randolph S.; Frazier, Tyralynn; Snell, Edward H.; Pusey, Marc L.

    1998-01-01

    Part of the challenge of macromolecular crystal growth for structure determination is obtaining an appropriate number of crystals with a crystal volume suitable for x-ray analysis. In this respect an understanding of the effect of solution conditions on macromolecule nucleation rates is advantageous. This study investigated the effects of supersaturation, temperature and pH on the nucleation rate of tetragonal lysozyme crystals. Batch crystallization plates were prepared at given solution concentrations and incubated at set temperatures over one week. The number of crystals per well with their size and axial ratios were recorded and correlated with solution conditions, Duplicate experiments indicate the reproducibility of the technique, Crystal numbers were found to increase with increasing supersaturation and temperature. The most significant variable however, was pH, where crystal numbers changed by two orders of magnitude over the pH range 4.0 to 5.2. Crystal size varied also with solution conditions, with the largest crystals being obtained at pH 5.2. Having optimized the crystallization conditions, a batch of crystals were prepared under exactly the same conditions and fifty of these crystals were analyzed by x-ray techniques. The results indicate that even under the same crystallization conditions, a marked variation in crystal properties exists.

  6. Biomimetic synthesis of hierarchical crystalline hydroxyapatite fibers in large-scale

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Xing, Chaogang; Ge, Suxiang; Huang, Baojun

    Highlights: ► Crystalline hierarchical hydroxyapatite (HAp) fibers are synthesized. ► We employ a biomimetic route by using cotton cloth as a natural bio-template. ► We study the effects of pH, ultrasonic cleaning time, and calcination temperature. ► We obtain an optimized reaction condition. ► This is a low cost method for production of hierarchical HAp fibers. -- Abstract: Crystalline hierarchical hydroxyapatite [Ca{sub 10}(PO{sub 4}){sub 6}(OH){sub 2}, HAp)] fibers were successfully synthesized via a biomimetic route by using cotton cloth as a natural bio-template. The effects of pH value, aging time, ultrasonic cleaning time, and calcination temperature on the purity andmore » morphology of the resulting hydroxyapatite (HAp) were monitored by scanning election microscope (SEM), X-ray diffraction (XRD), and infrared spectrophotometer (IR) to obtain an optimized reaction condition, namely, pH 9, ultrasonic cleaning for 1 min, aging for 24 h, and calcination at 600 °C for 4 h. We found that the natural cellulose could not only control the morphology of HAp but also lower its phase transformation temperature. The impact of this method lies in its low cost and successful production of large-scale patterning of three-dimensional hierarchical HAp fibers.« less

  7. Identification and phylogenetic analysis of new sulfate-reducing bacteria isolated from oilfield samples.

    PubMed

    Chen, Wu; Xiang, Fu; Fu, Jie; Wang, Qiang; Wang, Wenjun; Zeng, Qingfu; Yu, Longjiang

    2009-01-01

    Microbiologically influenced corrosion (MIC) caused by sulfate-reducing bacteria (SRB) has been investigated in an oilfield injection water system. Strain CW-01 was isolated from an oilfield and strain CW-04 was isolated from biofilm dirt of pipeline walls. The strains were facultative anaerobes, non-motile, Gram-positive, pole flagellum, and spore-forming curved rods. The growth was observed over the temperature range 20-70 degrees C. Strain CW-01 grew optimally at 37 degrees C. The pH range for growth was 3.0-11, optimal at pH 6.0. Strain CW-04 grew optimally at 48 degrees C. The pH range for growth was 3.0-10, optimal at pH 7.2. The strains grew at a very broad range of salt concentrations. Optimal growth was observed with 1.5 g/L NaCl for strain CW-01 and 0.7 g/L NaCl for strain CW-04. The strains showed most similarity in physiological characteristics, except for acetone and saccharose. Analysis of the 16S rDNA sequences allowed strains CW-01 and CW-04 to be classified into the genus Desulfotomaculum. The corrosion speciality of the strains had been comparatively investigated. Especially SRB's growth curve, bearable oxygen capability, drug fastness and corrosion rate had been analyzed. The results showed that it is difficult to prevent bacterial corrosion caused by these two strains.

  8. Simultaneously saccharification and fermentation approach as a tool for enhanced fossil fuels biodesulfurization.

    PubMed

    Paixão, Susana M; Arez, Bruno F; Roseiro, José C; Alves, Luís

    2016-11-01

    Biodesulfurization can be a complementary technology to the hydrodesulfurization, the commonly physical-chemical process used for sulfur removal from crude oil. The desulfurizing bacterium Gordonia alkanivorans strain 1B as a fructophilic microorganism requires fructose as C-source. In this context, the main goal of this work was the optimization of a simultaneous saccharification and fermentation (SSF) approach using the Zygosaccharomyces bailii strain Talf1 crude enzymes with invertase activity and sucrose as a cheaper fructose-rich commercial C-source (50% fructose) towards dibenzothiophene (DBT) desulfurization by strain 1B. The determination of optimal conditions, for both sucrose hydrolysis and DBT desulfurization was carried out through two sequential experimental uniform designs according to the Doehlert distribution for two factors: pH (5.5-7.5) and temperature (28-38 °C), with the enzyme load of 1.16 U/g/L; and enzyme load (0-4 U/g/L) and temperature (28-38 °C), with pH at 7.5. Based on 2-hydroxybiphenyl production, the analysis of the response surfaces obtained pointed out for pH 7.5, 32 °C and 1.8 U/g/L as optimal conditions. Further optimized SSF of sucrose during the DBT desulfurization process permitted to attain a 4-fold enhanced biodesulfurization. This study opens a new focus of research through the exploitation of sustainable low cost sucrose-rich feedstocks towards a more economical viable bioprocess scale-up. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Realm of Thermoalkaline Lipases in Bioprocess Commodities

    PubMed Central

    2018-01-01

    For decades, microbial lipases are notably used as biocatalysts and efficiently catalyze various processes in many important industries. Biocatalysts are less corrosive to industrial equipment and due to their substrate specificity and regioselectivity they produced less harmful waste which promotes environmental sustainability. At present, thermostable and alkaline tolerant lipases have gained enormous interest as biocatalyst due to their stability and robustness under high temperature and alkaline environment operation. Several characteristics of the thermostable and alkaline tolerant lipases are discussed. Their molecular weight and resistance towards a range of temperature, pH, metal, and surfactants are compared. Their industrial applications in biodiesel, biodetergents, biodegreasing, and other types of bioconversions are also described. This review also discusses the advance of fermentation process for thermostable and alkaline tolerant lipases production focusing on the process development in microorganism selection and strain improvement, culture medium optimization via several optimization techniques (i.e., one-factor-at-a-time, surface response methodology, and artificial neural network), and other fermentation parameters (i.e., inoculums size, temperature, pH, agitation rate, dissolved oxygen tension (DOT), and aeration rate). Two common fermentation techniques for thermostable and alkaline tolerant lipases production which are solid-state and submerged fermentation methods are compared and discussed. Recent optimization approaches using evolutionary algorithms (i.e., Genetic Algorithm, Differential Evolution, and Particle Swarm Optimization) are also highlighted in this article. PMID:29666707

  10. Cellulase production in a new mutant strain of Penicillium decumbens ML-017 by solid state fermentation with rice bran.

    PubMed

    Liu, Yun-Tao; Luo, Ze-Yu; Long, Chuan-Nan; Wang, Hai-Dong; Long, Min-Nan; Hu, Zhong

    2011-10-01

    To produce cellulolytic enzyme efficiently, Penicillium decumbens strain L-06 was used to prepare mutants with ethyl methane sulfonate (EMS) and UV-irradiation. A mutant strain ML-017 is shown to have a higher cellulase activity than others. Box-Behnken's design (BBD) and response surface methodology (RSM) were adopted to optimize the conditions of cellulase (filter paper activity, FPA) production in strain ML-017 by solid-state fermentation (SSF) with rice bran as the substrate. And the result shows that the initial pH, moisture content and culture temperature all have significant effect on the production of cellulase. The optimized condition shall be initial pH 5.7, moisture content 72% and culture temperature 30°C. The maximum cellulase (FPA) production was obtained under the optimized condition, which is 5.76 IU g(-1), increased by 44.12% to its original strain. It corresponded well with the calculated results (5.15 IU g(-1)) by model prediction. The result shows that both BBD and RSM are the cellulase optimization methods with good prospects. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. Effect of cultivating conditions on alpha-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation.

    PubMed

    Liu, Cai-qin; Chen, Qi-he; Cheng, Qian-jun; Wang, Jin-ling; He, Guo-qing

    2007-05-01

    The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of alpha-galactosidase production in SSF were 60% initial moisture of medium, 28 degrees C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0 approximately 6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum alpha-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation.

  12. Effect of cultivating conditions on α-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation

    PubMed Central

    Liu, Cai-qin; Chen, Qi-he; Cheng, Qian-jun; Wang, Jin-ling; He, Guo-qing

    2007-01-01

    The work is intended to achieve optimum culture conditions of α-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of α-galactosidase production in SSF were 60% initial moisture of medium, 28 °C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0~6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum α-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation. PMID:17542067

  13. Biophysical characterization and conformational stability of Ebola and Marburg virus-like particles.

    PubMed

    Hu, Lei; Trefethen, Jared M; Zeng, Yuhong; Yee, Luisa; Ohtake, Satoshi; Lechuga-Ballesteros, David; Warfield, Kelly L; Aman, M Javad; Shulenin, Sergey; Unfer, Robert; Enterlein, Sven G; Truong-Le, Vu; Volkin, David B; Joshi, Sangeeta B; Middaugh, C Russell

    2011-12-01

    The filoviruses, Ebola virus and Marburg virus, cause severe hemorrhagic fever with up to 90% human mortality. Virus-like particles of EBOV (eVLPs) and MARV (mVLPs) are attractive vaccine candidates. For the development of stable vaccines, the conformational stability of these two enveloped VLPs produced in insect cells was characterized by various spectroscopic techniques over a wide pH and temperature range. Temperature-induced aggregation of the VLPs at various pH values was monitored by light scattering. Temperature/pH empirical phase diagrams (EPDs) of the two VLPs were constructed to summarize the large volume of data generated. The EPDs show that both VLPs lose their conformational integrity above about 50°C-60°C, depending on solution pH. The VLPs were maximally thermal stable in solution at pH 7-8, with a significant reduction in stability at pH 5 and 6. They were much less stable in solution at pH 3-4 due to increased susceptibility of the VLPs to aggregation. The characterization data and conformational stability profiles from these studies provide a basis for selection of optimized solution conditions for further vaccine formulation and long-term stability studies of eVLPs and mVLPs. Copyright © 2011 Wiley-Liss, Inc.

  14. Evaluation of Low-Temperature Phosphine Fumigation for Control of Oriental Fruit Fly in Loquat Fruit.

    PubMed

    Liu, Tao; Li, Li; Li, Baishu; Zhan, Guoping; Wang, Yuejin

    2018-05-28

    Oriental fruit fly, Bactrocera dorsalis (Hendel; Diptera: Tephritidae), is recognized as a quarantine pest and a threat to Chinese loquat (Eriobotrya japonica Lindl.) fruit exports. Since loquat fruit is very sensitive to methyl bromide (MB) fumigation and cold treatment, in this study, low-temperature phosphine (PH3) fumigation was investigated to develop an alternative phytosanitary treatment method. Tolerance tests showed that the third instar was the most tolerant of all life stages of B dorsalis to PH3 gas at 8°C. Toxicity assay with 500-3000 ppm PH3 and subsequent probit analysis showed that 2000 ppm PH3 was optimal for fumigation and 152.75 h of treatment duration were required to achieve 99.9968% mortality. In the verification test, 144 and 168 h of treatment duration with 2000 ppm PH3 completely killed 35,277 and 35,134 B. dorsalis third instars, respectively. However, 13 live larvae were found after 120 h of treatment. Furthermore, these treatments reduced fruit respiration rates while causing no adverse effects on other fruit quality parameters, including firmness, soluble solid content, and titratable acidity over 192 h storage at 8°C. The results strongly suggest that low-temperature PH3 fumigation could be used for the postharvest control of B. dorsalis in loquat fruit.

  15. A Facile pH Controlled Citrate-Based Reduction Method for Gold Nanoparticle Synthesis at Room Temperature.

    PubMed

    Tyagi, Himanshu; Kushwaha, Ajay; Kumar, Anshuman; Aslam, Mohammed

    2016-12-01

    The synthesis of gold nanoparticles using citrate reduction process has been revisited. A simplified room temperature approach to standard Turkevich synthesis is employed to obtain fairly monodisperse gold nanoparticles. The role of initial pH alongside the concentration ratio of reactants is explored for the size control of Au nanoparticles. The particle size distribution has been investigated using UV-vis spectroscopy and transmission electron microscope (TEM). At optimal pH of 5, gold nanoparticles obtained are highly monodisperse and spherical in shape and have narrower size distribution (sharp surface plasmon at 520 nm). For other pH conditions, particles are non-uniform and polydisperse, showing a red-shift in plasmon peak due to aggregation and large particle size distribution. The room temperature approach results in highly stable "colloidal" suspension of gold nanoparticles. The stability test through absorption spectroscopy indicates no sign of aggregation for a month. The rate of reduction of auric ionic species by citrate ions is determined via UV absorbance studies. The size of nanoparticles under various conditions is thus predicted using a theoretical model that incorporates nucleation, growth, and aggregation processes. The faster rate of reduction yields better size distribution for optimized pH and reactant concentrations. The model involves solving population balance equation for continuously evolving particle size distribution by discretization techniques. The particle sizes estimated from the simulations (13 to 25 nm) are close to the experimental ones (10 to 32 nm) and corroborate the similarity of reaction processes at 300 and 373 K (classical Turkevich reaction). Thus, substitution of experimentally measured rate of disappearance of auric ionic species into theoretical model enables us to capture the unusual experimental observations.

  16. Kinetic characterization of glucose aerodehydrogenase from Aspergillus niger EMS-150-F after optimizing the dose of mutagen for enhanced production of enzyme

    PubMed Central

    Umbreen, Huma; Zia, Muhammad Anjum; Rasul, Samreen

    2013-01-01

    In the present study enhanced production of glucose aerodehydrogenase from Aspergillus niger has been achieved after optimizing the dose of chemical mutagen ethyl methane sulfonate (EMS) that has not been reported earlier. Different doses of mutagen were applied and a strain was developed basing upon the best production. The selected strain Aspergillus niger EMS-150-F was optimized for nutrient requirements in order to produce enzyme through fermentation and the results showed the best yield at 2% corn steep liquor (CSL), 36 hours fermentation time, pH 5, 30°C temperature, 0.3% KH2PO4, 0.3% urea and 0.06% CaCO3. The enzyme was then purified and resulted in 57.88 fold purification with 52.12% recovery. On kinetic characterization, the enzyme showed optimum activity at pH 6 and temperature 30°C. The Michaelis-Menton constants (Km, Vmax, Kcat and Kcat/Km) were 20 mM, 45.87 U mL−1, 1118.81 s−1 and 55.94 s−1 mM−1, respectively. The enzyme was found to be thermaly stable and the enthalpy and free energy showed an increase with increase in temperature and ΔS* was highly negative proving the enzyme from A. niger EMS-150-F resistant to temperature and showing a very little disorderliness. PMID:24688499

  17. Isolation of fish skin and bone gelatin from tilapia (Oreochromis niloticus): Response surface approach

    NASA Astrophysics Data System (ADS)

    Arpi, N.; Fahrizal; Novita, M.

    2018-03-01

    In this study, gelatin from fish collagen, as one of halal sources, was extracted from tilapia (Oreochromis niloticus) skin and bone, by using Response Surface Methodology to optimize gelatin extraction conditions. Concentrations of alkaline NaOH and acid HCl, in the pretreatment process, and temperatures in extraction process were chosen as independent variables, while dependent variables were yield, gel strength, and emulsion activity index (EAI). The result of investigation showed that lower NaOH pretreatment concentrations provided proper pH extraction conditions which combine with higher extraction temperatures resulted in high gelatin yield. However, gelatin emulsion activity index increased proportionally to the decreased in NaOH concentrations and extraction temperatures. No significant effect of the three independent variables on the gelatin gel strength. RSM optimization process resulted in optimum gelatin extraction process conditions using alkaline NaOH concentration of 0.77 N, acid HCl of 0.59 N, and extraction temperature of 66.80 °C. The optimal solution formula had optimization targets of 94.38%.

  18. Optimization of growth and bacteriocin production by Lactobacillus sakei subsp. sakei2a.

    PubMed

    Malheiros, Patrícia S; Sant'Anna, Voltaire; Todorov, Svetoslav D; Franco, Bernadette D G M

    2015-01-01

    Lactobacillus sakei subsp. sakei 2a is a bacteriocinogenic lactic acid bacterium isolated from Brazilian pork sausage, capable of inhibiting the growth of microbial pathogens, mainly Listeria monocytogenes. In order to optimize bacteriocin production for industrial applications, this study evaluated the effect of supplementation of MRS broth with glucose, Tween 20, Tween 80, sodium citrate, potassium chloride and cysteine, and effect of the initial pH and temperature of incubation of the medium on production of bacteriocins by L. sakei 2a. Adding glucose and Tween 20 to the medium, an initial pH of 5.0 or 5.5, and incubation temperatures of 25 °C or 30 °C resulted to the highest bacteriocin yields. Thus, a 2(4) factorial design with the four variables was performed, and statistical analysis showed that it was an adequate model (R (2) = 0.8296). In the studied range, the four parameters significantly influenced bacteriocin production, with the maximum yield produced at an initial pH between 5.5 and 7.0, a temperature between 25 and 30 °C and supplementation of the MRS broth with glucose from 3.25 to 6.0 g L(-1) and Tween 20 from 0.575 to 1.15% (v/v). Response Surface Methodology analysis indicated that the highest bacteriocin production (12800 AU mL(-1)) occurred in the MRS broth supplemented with 5.5 g L(-1) glucose and 1.05% Tween 20 at an initial pH of 6.28 and an incubation temperature of 25 °C. The amount of bacteriocin produced in commercial MRS broths under the same conditions was only 5600AU mL(-1).

  19. Optimization of growth and bacteriocin production by Lactobacillus sakei subsp. sakei2a

    PubMed Central

    Malheiros, Patrícia S.; Sant’Anna, Voltaire; Todorov, Svetoslav D.; Franco, Bernadette D.G.M.

    2015-01-01

    Lactobacillus sakei subsp. sakei 2a is a bacteriocinogenic lactic acid bacterium isolated from Brazilian pork sausage, capable of inhibiting the growth of microbial pathogens, mainly Listeria monocytogenes. In order to optimize bacteriocin production for industrial applications, this study evaluated the effect of supplementation of MRS broth with glucose, Tween 20, Tween 80, sodium citrate, potassium chloride and cysteine, and effect of the initial pH and temperature of incubation of the medium on production of bacteriocins by L. sakei 2a. Adding glucose and Tween 20 to the medium, an initial pH of 5.0 or 5.5, and incubation temperatures of 25 °C or 30 °C resulted to the highest bacteriocin yields. Thus, a 24 factorial design with the four variables was performed, and statistical analysis showed that it was an adequate model (R 2 = 0.8296). In the studied range, the four parameters significantly influenced bacteriocin production, with the maximum yield produced at an initial pH between 5.5 and 7.0, a temperature between 25 and 30 °C and supplementation of the MRS broth with glucose from 3.25 to 6.0 g L−1 and Tween 20 from 0.575 to 1.15% (v/v). Response Surface Methodology analysis indicated that the highest bacteriocin production (12800 AU mL−1) occurred in the MRS broth supplemented with 5.5 g L−1 glucose and 1.05% Tween 20 at an initial pH of 6.28 and an incubation temperature of 25 °C. The amount of bacteriocin produced in commercial MRS broths under the same conditions was only 5600AU mL−1. PMID:26413066

  20. Occurrence of killer Candida glabrata clinical isolates

    PubMed Central

    Arroyo-Helguera, O; Penas Alejandro, De Las; Irene, Castaño

    2012-01-01

    In this work we characterized the occurrence of killer activity in 64 Candida glabrata clinical isolates under different conditions. We found that only 6.25 % of the clinical isolates tested were positive for killer activity against a Saccharomyces cerevisiae W303 sensitive strain. Sensitivity of killer activity to different values of pH and temperatures was analyzed. We found that the killer activity presented by all isolates was resistant to every pH and temperature tested, although optimal activity was found at a range of pH values from 4 to 7 and at 37°C. We did not observe extrachromosomal genetic elements associated with killer activity in any of the positive C. glabrata isolates. The killer effect was due to a decrease in viability and DNA fragmentation in sensitive yeast. PMID:24031902

  1. Batch technique to evaluate the efficiency of different natural adsorbents for defluoridation from groundwater

    NASA Astrophysics Data System (ADS)

    Kumar, Pankaj; Saraswat, Chitresh; Mishra, Binaya Kumar; Avtar, Ram; Patel, Hiral; Patel, Asha; Sharma, Tejal; Patel, Roshni

    2017-09-01

    Fluoride pollution (with concentration >1.0 mg/L) in groundwater has become a global threat in the recent past due to the lesser availability of potable groundwater resource. In between several defluoridation techniques discovered so far, the adsorption process proved to be most economic and efficient. This study is an effort to evaluate defluoridation efficiency of powdered rice husk, fine chopped rice husk and sawdust by the batch adsorption process. Optimum defluoridation capacity is achieved by optimizing various parameters, viz. dose of adsorbent, pH, contact time and initial concentration. It was found that all three materials can be employed for the defluoridation technique, but powdered rice husk is the best adsorbent in the midst of all three. Powdered rice husk showed fluoride removal efficiency ranging between 85 and 90 % in the contact period of 7 h only in conditions of all optimized parameter. Following this parameter optimization, adsorption efficiency was also evaluated at natural pH of groundwater to minimize the cost of defluoridation. No significant difference was found between fluoride adsorption at optimized pH (pH = 4) and natural one (pH = 7), which concludes that powdered rice husk can be efficiently used for the defluoridation technique at field scale. The adsorption isotherm using this adsorbent perfectly followed Langmuir isotherms. The value of calculated separation factor also suggests the favourable adsorption of fluoride onto this adsorbent under the conditions used for the experiments. The field application for defluoridation of groundwater using this adsorbent (based on pH of natural groundwater there and seasonal variation of temperature) showed the high success rate.

  2. Optimization of microwave-assisted extraction of analgesic and anti-inflammatory drugs from human plasma and urine using response surface experimental designs.

    PubMed

    Fernández, Purificación; Fernández, Ana M; Bermejo, Ana M; Lorenzo, Rosa A; Carro, Antonia M

    2013-04-01

    The performance of microwave-assisted extraction and HPLC with photodiode array detection method for determination of six analgesic and anti-inflammatory drugs from plasma and urine, is described, optimized, and validated. Several parameters affecting the extraction technique were optimized using experimental designs. A four-factor (temperature, phosphate buffer pH 4.0 volume, extraction solvent volume, and time) hybrid experimental design was used for extraction optimization in plasma, and three-factor (temperature, extraction solvent volume, and time) Doehlert design was chosen to extraction optimization in urine. The use of desirability functions revealed the optimal extraction conditions as follows: 67°C, 4 mL phosphate buffer pH 4.0, 12 mL of ethyl acetate and 9 min, for plasma and the same volume of buffer and ethyl acetate, 115°C and 4 min for urine. Limits of detection ranged from 4 to 45 ng/mL in plasma and from 8 to 85 ng/mL in urine. The reproducibility evaluated at two concentration levels was less than 6.5% for both specimens. The recoveries were from 89 to 99% for plasma and from 83 to 99% for urine. The proposed method was successfully applied in plasma and urine samples obtained from analgesic users. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. A rapid and low energy consumption method to decolorize the high concentration triphenylmethane dye wastewater: operational parameters optimization for the ultrasonic-assisted ozone oxidation process.

    PubMed

    Zhou, Xian-Jiao; Guo, Wan-Qian; Yang, Shan-Shan; Ren, Nan-Qi

    2012-02-01

    This research set up an ultrasonic-assisted ozone oxidation process (UAOOP) to decolorize the triphenylmethane dyes wastewater. Five factors - temperature, initial pH, reaction time, ultrasonic power (low frequency 20 kHz), and ozone concentration - were investigated. Response surface methodology was used to find out the major factors influencing color removal rate and the interactions between these factors, and optimized the operating parameters as well. Under the experimental conditions: reaction temperature 39.81 °C, initial pH 5.29, ultrasonic power 60 W and ozone concentration 0.17 g/L, the highest color removals were achieved with 10 min reaction time and the initial concentration of the MG solution was 1000 mg/L. The optimal results indicated that the UAOOP was a rapid, efficient and low energy consumption technique to decolorize the high concentration MG wastewater. The predicted model was approximately in accordance with the experimental cases with correlation coefficients R(2) and R(adj)(2) of 0.9103 and 0.8386. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

  4. Optimization and Immobilization of Purified Labeo rohita Visceral Protease by Entrapment Method

    PubMed Central

    Geethanjali, S.; Subash, Anitha

    2013-01-01

    The purified fish visceral protease enzyme was immobilized by using various concentrations of sodium alginate and calcium chloride to optimize the best concentration for the formation of the beads. Then it was characterized by assaying the optimal pH, temperature, storage stability and reusability. The results on immobilization with sodium alginate and calcium chloride showed that a combination of 2% sodium alginate and 0.3 M calcium chloride weas found to be the optimum concentration for the formation of spherical and stable beads, this gave a maximal entrapped activity of 48.31%, and there was no change in the optimum pH 8.0 and temperature 40°C of protease before and after entrapment. The results on stability and reusability indicated that it was stable at 4°C retaining 100% residual activity after 5 days of storage and 67% loss of activity after ten days of storage and it retained 100% residual activity on the first reuse, 75% residual activity on the second reuse, 25% residual activity on the third use and complete loss in the activity on the fourth reuse. PMID:23533718

  5. Isolation and identification of chitinolytic bacteria of pohara river of South East Sulawesi and the optimization production of chitinase enzyme

    NASA Astrophysics Data System (ADS)

    Halimahtussadiyah, R.; Natsir, Muh.; Kurniawati, Desy; Utamy, Sukma Puspita

    2017-03-01

    Isolation and identification of chitinolytic bacteria from pohara river and optimation of chitinase enzyme production has been conducted. The aims of the study were isolation, characterize and optimaze of chitinase enzyme production. This study was carried out in three stages; isolation and selection of chitinolytic bacteria, characterization and identification of selected bacteria; optimization of the production of the enzyme (substrate concentration, temperature, and pH), and the determination of growth curve of T3 isolate. The chitinase activity assay was carried out using Schales method. The results of the screening obtained 6 isolates of potential bacteria of chitinolytic. The T3 isolate then was selected for the enzyme production, because it had the highest chitinolytic index of 22.31 mm. The morphological and biochemical observation showed that T3 isolate as a group of bacteria Aerobacter with Gram-negative nature, and shaped bacillus. The optimum condition for chitinase enzyme production was in chitin substrat concentration 0.06%, temperature of 30°C, and pH of 6.

  6. Color removal from distillery spent wash through coagulation using Moringa oleifera seeds: use of optimum response surface methodology.

    PubMed

    Prasad, R Krishna

    2009-06-15

    The effects of dosage, pH and concentration of salts were investigated for an optimized condition of color removal from the distillery spent wash. The optimization process was analyzed using custom response surface methodology (RSM). The design was employed to derive a statistical model for the effect of parameters studied on removal of color using Moringa oleifera coagulant (MOC). The dosage (20 and 60 ml), pH (7 and 8.5) and concentration of 0.25 M had been found to be the optimum conditions for maximum 56% and 67% color removal using sodium chloride (NaCl) and potassium chloride (KCl) salts respectively. The actual color removal at optimal conditions was found to be 53% and 64% respectively for NaCl and KCl salts which confirms close to RSM results. The effects of storage duration and temperature on MOC studied reveal that coagulation efficiency of MOC kept at room temperature was effective for 3 days and at 4 degrees C it performed coagulation up to 5 days.

  7. Optimization of tannase production by a novel Klebsiella pneumoniae KP715242 using central composite design.

    PubMed

    Kumar, Mukesh; Rana, Shiny; Beniwal, Vikas; Salar, Raj Kumar

    2015-09-01

    A novel tannase producing bacterial strain was isolated from rhizospheric soil of Acacia species and identified as Klebsiella pneumoniae KP715242. A 3.25-fold increase in tannase production was achieved upon optimization with central composite design using response surface methodology. Four variables namely pH, temperature, incubation period, and agitation speed were used to optimize significant correlation between the effects of these variables on tannase production. A second-order polynomial was fitted to data and validated by ANOVA. The results showed a complex relationship between variables and response given that all factors were significant and could explain 99.6% of the total variation. The maximum production was obtained at 5.2 pH, 34.97 °C temperature, 103.34 rpm agitation speed and 91.34 h of incubation time. The experimental values were in good agreement with the predicted ones and the models were highly significant with a correlation coefficient ( R 2 ) of 0.99 and a highly significant F-value of 319.37.

  8. Removal of mercury (II), elemental mercury and arsenic from simulated flue gas by ammonium sulphide.

    PubMed

    Ning, Ping; Guo, Xiaolong; Wang, Xueqian; Wang, Ping; Ma, Yixing; Lan, Yi

    2015-01-01

    A tubular resistance furnace was used as a reactor to simulate mercury and arsenic in smelter flue gases by heating mercury and arsenic compounds. The flue gas containing Hg(2+), Hg(0) and As was treated with ammonium sulphide. The experiment was conducted to investigate the effects of varying the concentration of ammonium sulphide, the pH value of ammonium sulphide, the temperature of ammonium sulphide, the presence of SO2 and the presence of sulphite ion on removal efficiency. The prepared adsorption products were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, X-ray photoelectron spectroscopy and scanning electron microscopy. The results showed that the optimal concentration of ammonium sulphide was 0.8 mol/L. The optimal pH value of ammonium sulphide was 10, and the optimal temperature of ammonium sulphide was 20°C.Under the optimum conditions, the removal efficiency of Hg(2+), Hg(0) and As could reach 99%, 88.8%, 98%, respectively. In addition, SO2 and sulphite ion could reduce the removal efficiency of mercury and arsenic from simulated flue gas.

  9. Bioleaching of nickel from spent petroleum catalyst using Acidithiobacillus thiooxidans DSM- 11478.

    PubMed

    Sharma, Mohita; Bisht, Varsha; Singh, Bina; Jain, Pratiksha; Mandal, Ajoy K; Lal, Banwari; Sarma, Priyangshu M

    2015-06-01

    The present work deals with optimization of culture conditions and process parameters for bioleaching of spent petroleum catalyst collected from a petroleum refinery. The efficacy of Ni bioleaching from spent petroleum catalyst was determined using pure culture of Acidithiobacillus thiooxidans DSM- 11478. The culture conditions of pH, temperature and headspace volume to media volume ratio were optimized. EDX analysis was done to confirm the presence of Ni in the spent catalyst after roasting it to decoke its surface. The optimum temperature for A. thiooxidans DSM-11478 growth was found to be 32 degrees C. The enhanced recovery of nickel at very low pH was attributed to the higher acidic strength of sulfuric acid produced in the culture medium by the bacterium. During the bioleaching process, 89% of the Ni present in the catalyst waste could be successfully recovered in optimized conditions. This environment friendly bioleaching process proved efficient than the chemical method. Taking leads from the lab scale results, bioleaching in larger volumes (1, 5 and 10 L) was also performed to provide guidelines for taking up this technology for in situ industrial waste management.

  10. Low-temperature solution processing of palladium/palladium oxide films and their pH sensing performance.

    PubMed

    Qin, Yiheng; Alam, Arif U; Pan, Si; Howlader, Matiar M R; Ghosh, Raja; Selvaganapathy, P Ravi; Wu, Yiliang; Deen, M Jamal

    2016-01-01

    Highly sensitive, easy-to-fabricate, and low-cost pH sensors with small dimensions are required to monitor human bodily fluids, drinking water quality and chemical/biological processes. In this study, a low-temperature, solution-based process is developed to prepare palladium/palladium oxide (Pd/PdO) thin films for pH sensing. A precursor solution for Pd is spin coated onto pre-cleaned glass substrates and annealed at low temperature to generate Pd and PdO. The percentages of PdO at the surface and in the bulk of the electrodes are correlated to their sensing performance, which was studied by using the X-ray photoelectron spectroscope. Large amounts of PdO introduced by prolonged annealing improve the electrode's sensitivity and long-term stability. Atomic force microscopy study showed that the low-temperature annealing results in a smooth electrode surface, which contributes to a fast response. Nano-voids at the electrode surfaces were observed by scanning electron microscope, indicating a reason for the long-term degradation of the pH sensitivity. Using the optimized annealing parameters of 200°C for 48 h, a linear pH response with sensitivity of 64.71±0.56 mV/pH is obtained for pH between 2 and 12. These electrodes show a response time shorter than 18 s, hysteresis less than 8 mV and stability over 60 days. High reproducibility in the sensing performance is achieved. This low-temperature solution-processed sensing electrode shows the potential for the development of pH sensing systems on flexible substrates over a large area at low cost without using vacuum equipment. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Precision Sensing by Two Opposing Gradient Sensors: How Does Escherichia coli Find its Preferred pH Level?

    PubMed Central

    Hu, Bo; Tu, Yuhai

    2013-01-01

    It is essential for bacteria to find optimal conditions for their growth and survival. The optimal levels of certain environmental factors (such as pH and temperature) often correspond to some intermediate points of the respective gradients. This requires the ability of bacteria to navigate from both directions toward the optimum location and is distinct from the conventional unidirectional chemotactic strategy. Remarkably, Escherichia coli cells can perform such a precision sensing task in pH taxis by using the same chemotaxis machinery, but with opposite pH responses from two different chemoreceptors (Tar and Tsr). To understand bacterial pH sensing, we developed an Ising-type model for a mixed cluster of opposing receptors based on the push-pull mechanism. Our model can quantitatively explain experimental observations in pH taxis for various mutants and wild-type cells. We show how the preferred pH level depends on the relative abundance of the competing sensors and how the sensory activity regulates the behavioral response. Our model allows us to make quantitative predictions on signal integration of pH and chemoattractant stimuli. Our study reveals two general conditions and a robust push-pull scheme for precision sensing, which should be applicable in other adaptive sensory systems with opposing gradient sensors. PMID:23823247

  12. Optimization of polyhydroxybutyrate production utilizing waste water as nutrient source by Botryococcus braunii Kütz using response surface methodology.

    PubMed

    Kavitha, Ganapathy; Kurinjimalar, Chidambaram; Sivakumar, Krishnan; Kaarthik, Muthukumar; Aravind, Rajamani; Palani, Perumal; Rengasamy, Ramasamy

    2016-12-01

    Investigations have been made to optimize various factors including pH, temperature, and substrate for enhanced polyhydroxybutyrate (PHB) production in Botryococcus braunii which serves as a pioneer for production of bioplastic (PHB). Polyhydroxybutyrate is a natural, decomposable polymers accumulated by the microorganism under different nutritional condition. Strain selection was done by staining method using Sudan black and Nile red dye. Using response surface methodology (RSM), three level- three variables Box Behnken design (BBD), the best potential combination of pH (4-11), temperature (30-50°C) and sewage waste water as substrate fed at different concentrations at 20%-100% for maximum PHB production was investigated. Maximum yield (247±0.42mg/L) of PHB dry weight was achieved from the 60% concentration of sewage waste water as a growth medium at pH 7.5 at 40°C. It was well in close agreement with the value predicted by RSM model yield (246± 0.32mg/L). Thus the study shows the production of PHB by B. braunii along with the basic characterization of PHB by using FTIR and TEM analysis. These preliminary studies indicated that PHB can also be produced by B. braunii utilizing waste water. There is no report on the optimization of PHB production in this microalgae have been documented. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Optimization of covalent immobilization of pectinase on sodium alginate support.

    PubMed

    Li, Tuoping; Wang, Na; Li, Suhong; Zhao, Qiancheng; Guo, Mei; Zhang, Cheyun

    2007-09-01

    Pectinase was immobilized on a sodium alginate support using glutaraldehyde and retained 66% activity. The optimal pH for activity shifted from 3.0 to 3.5 after immobilization; however, the optimum temperature remained unchanged at 40 degrees C. The immobilized enzyme also had a higher thermal stability and reusability than the free enzyme, and retained 80% of initial activity after 11 batch reactions.

  14. Identification of lipolytic enzymes isolated from bacteria indigenous to Eucalyptus wood species for application in the pulping industry.

    PubMed

    Ramnath, L; Sithole, B; Govinden, R

    2017-09-01

    This study highlights the importance of determining substrate specificity at variable experimental conditions. Lipases and esterases were isolated from microorganisms cultivated from Eucalyptus wood species and then concentrated (cellulases removed) and characterized. Phenol red agar plates supplemented with 1% olive oil or tributyrin was ascertained to be the most favourable method of screening for lipolytic activity. Lipolytic activity of the various enzymes were highest at 45-61 U/ml at the optimum temperature and pH of between at 30-35 °C and pH 4-5, respectively. Change in pH influenced the substrate specificity of the enzymes tested. The majority of enzymes tested displayed a propensity for longer aliphatic acyl chains such as dodecanoate (C 12 ), myristate (C 14 ), palmitate (C 16 ) and stearate (C 18 ) indicating that they could be characterised as potential lipases. Prospective esterases were also detected with specificity towards acetate (C 2 ), butyrate (C 4 ) and valerate (C 5 ). Enzymes maintained up to 95% activity at the optimal pH and temperature for 2-3 h. It is essential to test substrates at various pH and temperature when determining optimum activity of lipolytic enzymes, a method rarely employed. The stability of the enzymes at acidic pH and moderate temperatures makes them excellent candidates for application in the treatment of pitch during acid bi-sulphite pulping, which would greatly benefit the pulp and paper industry.

  15. Study on the technology of compound enzymatic hydrolysis of whole passion fruit

    NASA Astrophysics Data System (ADS)

    Yang, Yu-xia; Duan, Zhen-hua; Kang, Chao; Zhu, Xiang-hao; Li, Ding-jin

    2017-12-01

    Fresh Whole Passion Fruit was used as raw material, The enzymatic hydrolysis technology of Passion Fruit by Complex enzyme were studied, The effects of enzyme dosage, Enzyme ratio(cellulose: pectinase), pH, temperature and time on the hydrolysis were investigated by single-tests and orthogonal tests, the hydrolysis indicators of single-factor tests and orthogonal tests were juice yield. The optimal hydrolysis conditions of Passion Fruit by Complex enzyme were enzyme dosage 0.12%, Enzyme ratio 5:1, hydrolysis temperature 50°C, pH4.0 and time 3.5 h. Under such conditions, juice yield of Passion Fruit was 92.91%.

  16. Isolation and Characterization of Biosurfactant Producing Bacteria for the Application in Enhanced Oil Recovery

    NASA Astrophysics Data System (ADS)

    Prasad, Niraj; Dasgupta, Sumita; Chakraborty, Mousumi; Gupta, Smita

    2017-07-01

    In the present study, a biosurfactant producing bacterial strain was isolated, screened and identified. Further, various fermentation conditions (such as pH (5-10), incubation period (24-96h) and incubation temperature (20-60 °C) were optimized for maximum production of biosurfactant. The produced biosurfactant was characterized by measuring emulsification index, foaming characteristics, rhamnolipid detection, interfacial tension between water and oil and stability against pH and temperature for its potential application in oil recovery process. The additional oil recovery for two different sand, sand1 and sand2, was found to be 49% and 38%, respectively.

  17. Screening for Cellulase Encoding Clones in Metagenomic Libraries.

    PubMed

    Ilmberger, Nele; Streit, Wolfgang R

    2017-01-01

    For modern biotechnology there is a steady need to identify novel enzymes. In biotechnological applications, however, enzymes often must function under extreme and nonnatural conditions (i.e., in the presence of solvents, high temperature and/or at extreme pH values). Cellulases have many industrial applications from the generation of bioethanol, a realistic long-term energy source, to the finishing of textiles. These industrial processes require cellulolytic activity under a wide range of pH, temperature, and ionic conditions, and they are usually carried out by mixtures of cellulases. Investigation of the broad diversity of cellulolytic enzymes involved in the natural degradation of cellulose is necessary for optimizing these processes.

  18. Production of cellulase from Pestalotiopsis versicolor

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rao, M.N.A.; Mithal, B.M.; Thakur, R.N.

    1983-01-01

    Production of cellulase from Pestalotiopsis versicolor was studied in a shake flask culture. The cellulase system was found to be rich in beta-glucosidase. Kinetic parameters such as pH and temperature have been optimized for the various enzyme components. 9 references.

  19. Optimization of enzymatic hydrolysis of guar gum using response surface methodology.

    PubMed

    Mudgil, Deepak; Barak, Sheweta; Khatkar, B S

    2014-08-01

    Guar gum is a polysaccharide obtained from guar seed endosperm portion. Enzymatically hydrolyzed guar gum is low in viscosity and has several health benefits as dietary fiber. In this study, response surface methodology was used to determine the optimum conditions for hydrolysis that give minimum viscosity of guar gum. Central composite was employed to investigate the effects of pH (3-7), temperature (20-60 °C), reaction time (1-5 h) and cellulase concentration (0.25-1.25 mg/g) on viscosity during enzymatic hydrolysis of guar (Cyamopsis tetragonolobus) gum. A second order polynomial model was developed for viscosity using regression analysis. Results revealed statistical significance of model as evidenced from high value of coefficient of determination (R(2) = 0.9472) and P < 0.05. Viscosity was primarily affected by cellulase concentration, pH and hydrolysis time. Maximum viscosity reduction was obtained when pH, temperature, hydrolysis time and cellulase concentration were 6, 50 °C, 4 h and 1.00 mg/g, respectively. The study is important in optimizing the enzymatic process for hydrolysis of guar gum as potential source of soluble dietary fiber for human health benefits.

  20. Simultaneous production of detergent stable keratinolytic protease, amylase and biosurfactant by Bacillus subtilis PF1 using agro industrial waste.

    PubMed

    Bhange, Khushboo; Chaturvedi, Venkatesh; Bhatt, Renu

    2016-06-01

    The present study is an attempt to optimize simultaneous production of keratinolytic protease, amylase and biosurfactant from feather meal, potato peel and rape seed cake in a single media by response surface methodology to evaluate their biochemical properties for detergent additive. The optimization was carried out using 20 run, 3 factor and 5-level of central composite design on design expert software which resulted in a 1.2, 0.84 and 2.28 fold increase in protease, amylase and biosurfactant production. The proteolytic activity was found to be optimum at pH 9.0 and 60 °C while optimum amylolytic activity was recorded at pH 6.0 and 70 °C respectively. Both enzymes were found to be stable in the presence of organic solvents, ionic and commercial detergent and oxidizing agents. The biosurfactant was extracted with chloroform and was found to be stable at varying pH and temperature; however a reduction in the activity was observed at temperature higher than 70 °C. The isolated enzymes and biosurfactants may find applications in the effective removal of stains.

  1. Analysis of problems with dry fermentation process for biogas production

    NASA Astrophysics Data System (ADS)

    Pilát, Peter; Patsch, Marek; Jandačka, Jozef

    2012-04-01

    The technology of dry anaerobic fermentation is still meeting with some scepticism, and therefore in most biogas plants are used wet fermentation technology. Fermentation process would be not complete without an optimal controlled condition: dry matter content, density, pH, and in particular the reaction temperature. If is distrust of dry fermentation eligible it was on the workplace of the Department of Power Engineering at University of Zilina built an experimental small-scale biogas station that allows analysis of optimal parameters of the dry anaerobic fermentation, in particular, however, affect the reaction temperature on yield and quality of biogas.

  2. Bio-conversion of apple pomace into ethanol and acetic acid: Enzymatic hydrolysis and fermentation.

    PubMed

    Parmar, Indu; Rupasinghe, H P Vasantha

    2013-02-01

    Enzymatic hydrolysis of cellulose present in apple pomace was investigated using process variables such as enzyme activity of commercial cellulase, pectinase and β-glucosidase, temperature, pH, time, pre-treatments and end product separation. The interaction of enzyme activity, temperature, pH and time had a significant effect (P<0.05) on release of glucose. Optimal conditions of enzymatic saccharification were: enzyme activity of cellulase, 43units; pectinase, 183units; β-glucosidase, 41units/g dry matter (DM); temperature, 40°C; pH 4.0 and time, 24h. The sugars were fermented using Saccharomyces cerevisae yielding 19.0g ethanol/100g DM. Further bio-conversion using Acetobacter aceti resulted in the production of acetic acid at a concentration of 61.4g/100g DM. The present study demonstrates an improved process of enzymatic hydrolysis of apple pomace to yield sugars and concomitant bioconversion to produce ethanol and acetic acid. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. Draft Genome Sequence of thermoalkaliphilic Caldalkalibacillus thermarum strain TA2.A1 Reveals Molecular Adaptations to Extreme pH and Temperature

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kalamorz, Falk; Keis, Stefanie; Stanton, Jo-Ann

    The genes and molecular machines that allow for a thermoalkaliphilic lifestyle have not been defined. To address this goal, we report on the improved high-quality draft genome sequence of Caldalkalibacillus thermarum strain TA2.A1, an obligately aerobic bacterium that grows optimally at pH 9.5 and 65 to 70 C on a wide variety of carbon and energy sources.

  4. Using an artificial neural network to predict the optimal conditions for enzymatic hydrolysis of apple pomace.

    PubMed

    Gama, Repson; Van Dyk, J Susan; Burton, Mike H; Pletschke, Brett I

    2017-06-01

    The enzymatic degradation of lignocellulosic biomass such as apple pomace is a complex process influenced by a number of hydrolysis conditions. Predicting optimal conditions, including enzyme and substrate concentration, temperature and pH can improve conversion efficiency. In this study, the production of sugar monomers from apple pomace using commercial enzyme preparations, Celluclast 1.5L, Viscozyme L and Novozyme 188 was investigated. A limited number of experiments were carried out and then analysed using an artificial neural network (ANN) to model the enzymatic hydrolysis process. The ANN was used to simulate the enzymatic hydrolysis process for a range of input variables and the optimal conditions were successfully selected as was indicated by the R 2 value of 0.99 and a small MSE value. The inputs for the ANN were substrate loading, enzyme loading, temperature, initial pH and a combination of these parameters, while release profiles of glucose and reducing sugars were the outputs. Enzyme loadings of 0.5 and 0.2 mg/g substrate and a substrate loading of 30% were optimal for glucose and reducing sugar release from apple pomace, respectively, resulting in concentrations of 6.5 g/L glucose and 28.9 g/L reducing sugars. Apple pomace hydrolysis can be successfully carried out based on the predicted optimal conditions from the ANN.

  5. Validation of a method for the quantitation of ghrelin and unacylated ghrelin by HPLC.

    PubMed

    Staes, Edith; Rozet, Eric; Ucakar, Bernard; Hubert, Philippe; Préat, Véronique

    2010-02-05

    An HPLC/UV method was first optimized for the separation and quantitation of human acylated and unacylated (or des-acyl) ghrelin from aqueous solutions. This method was validated by an original approach using accuracy profiles based on tolerance intervals for the total error measurement. The concentration range that achieved adequate accuracy extended from 1.85 to 59.30microM and 1.93 to 61.60microM for acylated and unacylated ghrelin, respectively. Then, optimal temperature, pH and buffer for sample storage were determined. Unacylated ghrelin was found to be stable in all conditions tested. At 37 degrees C acylated ghrelin was stable at pH 4 but unstable at pH 7.4, the main degradation product was unacylated ghrelin. Finally, this validated HPLC/UV method was used to evaluate the binding of acylated and unacylated ghrelin to liposomes.

  6. Optimization of Crude Oil and PAHs Degradation by Stenotrophomonas rhizophila KX082814 Strain through Response Surface Methodology Using Box-Behnken Design

    PubMed Central

    Virupakshappa, Praveen Kumar Siddalingappa; Mishra, Gaurav; Mehkri, Mohammed Ameenuddin

    2016-01-01

    The present paper describes the process optimization study for crude oil degradation which is a continuation of our earlier work on hydrocarbon degradation study of the isolate Stenotrophomonas rhizophila (PM-1) with GenBank accession number KX082814. Response Surface Methodology with Box-Behnken Design was used to optimize the process wherein temperature, pH, salinity, and inoculum size (at three levels) were used as independent variables and Total Petroleum Hydrocarbon, Biological Oxygen Demand, and Chemical Oxygen Demand of crude oil and PAHs as dependent variables (response). The statistical analysis, via ANOVA, showed coefficient of determination R 2 as 0.7678 with statistically significant P value 0.0163 fitting in second-order quadratic regression model for crude oil removal. The predicted optimum parameters, namely, temperature, pH, salinity, and inoculum size, were found to be 32.5°C, 9, 12.5, and 12.5 mL, respectively. At this optimum condition, the observed and predicted PAHs and crude oil removal were found to be 71.82% and 79.53% in validation experiments, respectively. The % TPH results correlate with GC/MS studies, BOD, COD, and TPC. The validation of numerical optimization was done through GC/MS studies and % removal of crude oil. PMID:28116165

  7. Extending the applicability of pressurized hot water extraction to compounds exhibiting limited water solubility by pH control: curcumin from the turmeric rhizome.

    PubMed

    Euterpio, Maria Anna; Cavaliere, Chiara; Capriotti, Anna Laura; Crescenzi, Carlo

    2011-11-01

    Pressurized hot water extraction (PHWE, also known as subcritical water extraction) is commonly considered to be an environmentally friendly extraction technique that could potentially replace traditional methods that use organic solvents. Unfortunately, the applicability of this technique is often limited by the very low water solubility of the target compounds, even at high temperatures. In this paper, the scope for broadening the applicability of PHWE by adjusting the pH of the water used in the extraction is demonstrated in the extraction of curcumin (which exhibits very limited water solubility) from untreated turmeric (Curcuma longa L.) rhizomes. Although poor extraction yields were obtained, even at high temperatures when using degassed water or neutral phosphate buffer as the extraction medium, yields exceeding those obtained by Soxhlet extraction were achieved using highly acidic pH buffers due to curcumin protonation. The influence of the temperature, pH, and buffer concentration on the extraction yield were investigated in detail by means of a series of designed experiments. Optimized conditions for the extraction of curcumin from turmeric by PHWE were estimated at 197 °C using 62 g/L buffer concentration at pH 1.6. The relationships between these variables were subjected to statistical analysis using response surface methodology.

  8. Microbial β-Galactosidase of Pediococcus pentosaceus ID-7: Isolation, Cloning, and Molecular Characterization.

    PubMed

    Lee, Ji-Yeong; Kwak, Mi-Sun; Roh, Jong-Bok; Kim, Kwang; Sung, Moon-Hee

    2017-03-28

    Pediococcus pentosaceus ID-7 was isolated from kimchi, a Korean fermented food, and it showed high activity for lactose hydrolysis. The β-galactosidase of P. pentosaceus ID-7 belongs to the GH2 group, which is composed of two distinct proteins. The heterodimeric LacLM type of β-galactosidase found in P. pentosaceus ID-7 consists of two genes partially overlapped, lacL and lacM encoding LacL (72.2 kDa) and LacM (35.4 kDa). In this study, Escherichia coli MM294 was used for the production of LacL, LacM, and LacLM. These three types of recombinant proteins were expressed, purified, and characterized. The specific activities of LacLM and LacL were 339 and 31 U/mg, respectively. However, activity was not detected with LacM alone. The optimal pH of LacLM and LacL was pH 7.5 and pH 7.0, and the optimal temperature of LacLM and LacL was 40°C and 50°C, respectively. The optimal temperature changes indicate that LacLM is able to achieve higher activity at a relatively lower temperature. LacLM was strongly activated by Mg 2+ , Mn 2+ , and Zn 2+ , which was not true for LacL. Consistent with this, EDTA strongly inactivated LacLM and LacL, but the presence of reducing agents did not dramatically alter the activity. Taken together, multiple alignment of amino acid sequences and phylogenetic analysis results of LacL and LacM of P. pentosaceus ID-7 suggest the evolution of LacL into LacLM and that the use of divalent metal ions results in higher activity.

  9. Optimization of process variables by central composite design for the immobilization of urease enzyme on functionalized gold nanoparticles for various applications.

    PubMed

    Talat, Mahe; Singh, Ashwani Kumar; Srivastava, O N

    2011-08-01

    In the present study, enzyme urease has been immobilized on amine-functionalized gold nanoparticles (AuNPs). AuNPs were synthesized using natural precursor, i.e., clove extract and amine functionalized through 0.004 M L: -cysteine. Enzyme (urease) was extracted and purified from the vegetable waste, i.e., seeds of pumpkin to apparent homogeneity (sp. activity 353 U/mg protein). FTIR spectroscopy and transmission electron microscopy was used to characterize the immobilized enzyme. The immobilized enzyme exhibited enhanced activity as compared with the enzyme in the solution, especially, at lower enzyme concentration. Based on the evaluation of activity assay of the immobilized enzyme, it was found that the immobilized enzyme was quite stable for about a month and could successfully be used even after eight cycles having enzyme activity of about 47%. In addition to this central composite design (CCD) with the help of MINITAB version 15 Software was utilized to optimize the process variables viz., pH and temperature affecting the enzyme activity upon immobilization on AuNPs. The results predicted by the design were found in good agreement (R2 = 96.38%) with the experimental results indicating the applicability of proposed model. The multiple regression analysis and ANOVA showed the individual and cumulative effect of pH and temperature on enzyme activity indicating that the activity increased with the increase of pH up to 7.5 and temperature 75 °C. The effects of each variables represented by main effect plot, 3D surface plot, isoresponse contour plot and optimized plot were helpful in predicting results by performing a limited set of experiments.

  10. Optimized conditions for primary culture of pituitary cells from the Atlantic cod (Gadus morhua). The importance of osmolality, pCO₂, and pH.

    PubMed

    Hodne, Kjetil; von Krogh, Kristine; Weltzien, Finn-Arne; Sand, Olav; Haug, Trude M

    2012-09-01

    Protocols for primary cultures of teleost cells are commonly only moderately adjusted from similar protocols for mammalian cells, the main adjustment often being of temperature. Because aquatic habitats are in general colder than mammalian body temperatures and teleosts have gills in direct contact with water, pH and buffer capacity of blood and extracellular fluid are different in fish and mammals. Plasma osmolality is generally higher in marine teleosts than in mammals. Using Atlantic cod (Gadus morhua) as a model, we have optimized these physiological parameters to maintain primary pituitary cells in culture for an extended period without loosing key properties. L-15 medium with adjusted osmolality, adapted to low pCO(2) (3.8mm Hg) and temperature (12°C), and with pH 7.85, maintained the cells in a physiologically sounder state than traditional culture medium, significantly improving cell viability compared to the initial protocol. In the optimized culture medium, resting membrane potential and response to releasing hormone were stable for at least two weeks, and the proportion of cells firing action potentials during spawning season was about seven times higher than in the original culture medium. The cells were moderately more viable when the modified medium was supplemented with newborn calf serum or artificial serum substitute. Compared to serum-free L-15 medium, expression of key genes (lhb, fshb, and gnrhr2a) was better maintained in medium containing SSR, whereas NCS tended to decrease the expression level. Although serum-free medium is adequate for many applications, serum supplement may be preferable for experiments dependent on membrane integrity. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. [Optimization of fermentation conditions for cold-adapted amylase production by Micrococcus antarcticus and its enzymatic properties].

    PubMed

    Fan, Hong-xi; Liu, Ying; Liu, Zhi-pei

    2009-08-15

    By single factor experiments, the fermentation conditions for cold-adapted amylase production from Micrococcus antarcticus were determined as follows(medium g/L): Na2 HPO4 2.0, KH2PO4 1.0, MgSO4 x 7H2O 0.1, NaCl 5.0, (NH4)2SO4 2.5, maltose 5.0, trace element solution 5.0 mL, pH 8.0, 100 mL/Erlenmeyer flask (500 mL); cultivation was in a rotating shaker at 12 degrees C and 160 r/min for 64 h.Under those conditions,the highest total enzyme activity (2.6 U/mL) was obtained and increased by 10.8 fold compared with the original value of 0.24 U/mL before optimization. This amylase was purified by concentration with ultrafiltration membrane module, Hitrap Q anion exchange chromatography and Superdex 200 gel filtration chromatography. The optimal temperature and pH for the purified amylase were 30 degrees C and 6.0, respectively.It still showed high activity at low temperature 10-15 degrees C. It was sensitive to high temperature but was stable at pH 6.0-10.0 with at least 70% activity remained. These results indicated that it was a typical cold-adapted enzyme. The enzyme activity was stimulated by Ca2+, Mn2+, Co2+ and Mg2+; but inhibited by Zn2+, Ba2+, Ag+, Cu2+, Al3+, Fe2, Fe3+, Hg2+, EDTA and citrate. This cold-adapted amylase showed resistance to inactivation of 0.1% nonionic surfactants such as Tween 80, TrintonX-100, etc. Its Km was 0.90 mg/mL.

  12. Comparative study of energy-transducing properties of cytoplasmic membranes from mesophilic and thermophilic Bacillus species.

    PubMed Central

    De Vrij, W; Bulthuis, R A; Konings, W N

    1988-01-01

    The properties of enzymes involved in energy transduction from a mesophilic (Bacillus subtilis) and a thermophilic (B. stearothermophilus) bacterium were compared. Membrane preparations of the two organisms contained dehydrogenases for NADH, succinate, L-alpha-glycerophosphate, and L-lactate. Maximum NADH and cytochrome c oxidation rates were obtained at the respective growth temperatures of the two bacteria. The enzymes involved in the oxidation reactions in membranes of the thermophilic species were more thermostable than those of the mesophilic species. The apparent microviscosities of the two membrane preparations were studied at different temperatures. At the respective optimal growth temperatures, the apparent microviscosities of the membranes of the two organisms were remarkably similar. The transition from the gel to the liquid-crystalline state occurred at different temperatures in the two species. In the two species, the oxidation of physiological (NADH) and nonphysiological (N,N,N',N'-tetramethyl-p-phenylenediamine or phenazine methosulfate) electron donors led to generation of a proton motive force which varied strongly with temperature. At increasing temperatures, the efficiency of energy transduction declined because of increasing H+ permeability. At the growth temperature, the efficiency of energy transduction was lower in B. stearothermophilus than in the mesophilic species. Extremely high respiratory activities enabled B. stearothermophilus to maintain a high proton motive force at elevated temperatures. The pH dependence of proton motive force generation appeared to be similar in the two membrane preparations. The highest proton motive forces were generated at low external pH, mainly because of a high pH gradient. At increasing external pH, the proton motive force declined. PMID:2834342

  13. Statistical Methodologies for the Optimization of Lipase and Biosurfactant by Ochrobactrum intermedium Strain MZV101 in an Identical Medium for Detergent Applications.

    PubMed

    Ebrahimipour, Gholamhossein; Sadeghi, Hossein; Zarinviarsagh, Mina

    2017-09-11

    The Plackett-Burman design and the Box-Behnken design, statistical methodologies, were employed for the optimization lipase and biosurfactant production by Ochrobactrum intermedium strain MZV101 in an identical broth medium for detergent applications. Environmental factor pH determined to be most mutual significant variables on production. A high concentration of molasses at high temperature and pH has a negative effect on lipase and biosurfactant production by O. intermedium strain MZV101. The chosen mathematical method of medium optimization was sufficient for improving the industrial production of lipase and biosurfactant by bacteria, which were respectively increased 3.46- and 1.89-fold. The duration of maximum production became 24 h shorter, so it was fast and cost-saving. In conclusion, lipase and biosurfactant production by O. intermedium strain MZV101 in an identical culture medium at pH 10.5-11 and 50-60 °C, with 1 g/L of molasses, seemed to be economical, fast, and effective for the enhancement of yield percentage for use in detergent applications.

  14. Understanding short-chain fatty acids accumulation enhanced in waste activated sludge alkaline fermentation: kinetics and microbiology.

    PubMed

    Zhang, Peng; Chen, Yinguang; Zhou, Qi; Zheng, Xiong; Zhu, Xiaoyu; Zhao, Yuxiao

    2010-12-15

    Most of the studies on sewage sludge treatment in literature were conducted for methane generation under acidic or near neutral pH conditions. It was reported in our previous studies that the accumulation of short-chain fatty acids (SCFAs), the preferred carbon source of biological wastewater nutrient removal, was significantly enhanced when sludge was fermented under alkaline conditions, but the optimal pH was temperature-dependent (pH 10 at ambient temperature, pH 9 at mesophilic, and pH 8 at thermophilic), and the maximal SCFAs yields were in the following order: thermophilic pH 8 > mesophilic pH 9 > ambient pH 10 > ambient uncontrolled pH. In this study the kinetic and microbiological features of waste activated sludge fermented in the range of pH 7-10 were investigated to understand the mechanism of remarkably high SCFAs accumulation under alkaline conditions. The developed sludge alkaline fermentation model could be applied to predicate the experimental data in either batch or semicontinuous sludge alkaline fermentation tests, and the relationships among alkaline pH, kinetic parameters, and SCFAs were discussed. Further analyses with fluorescence in situ hybridization (FISH) and PCR-based 16S rRNA gene clone library indicated that both the ratio of bacteria to archaea and the fraction of SCFAs producer accounting for bacteria were in the sequence of thermophilic pH 8 > mesophilic pH 9 > ambient pH 10 > ambient uncontrolled pH, which was in correspondence with the observed order of maximal SCFAs yields.

  15. Low-temperature thermal pre-treatment of municipal wastewater sludge: Process optimization and effects on solubilization and anaerobic degradation.

    PubMed

    Nazari, Laleh; Yuan, Zhongshun; Santoro, Domenico; Sarathy, Siva; Ho, Dang; Batstone, Damien; Xu, Chunbao Charles; Ray, Madhumita B

    2017-04-15

    The present study examines the relationship between the degree of solubilization and biodegradability of wastewater sludge in anaerobic digestion as a result of low-temperature thermal pre-treatment. The main effect of thermal pre-treatment is the disintegration of cell membranes and thus solubilization of organic compounds. There is an established correlation between chemical oxygen demand (COD) solubilization and temperature of thermal pre-treatment, but results of thermal pre-treatment in terms of biodegradability are not well understood. Aiming to determine the impact of low temperature treatments on biogas production, the thermal pre-treatment process was first optimized based on an experimental design study on waste activated sludge in batch mode. The optimum temperature, reaction time and pH of the process were determined to be 80 °C, 5 h and pH 10, respectively. All three factors had a strong individual effect (p < 0.001), with a significant interaction effect for temp. pH 2 (p = 0.002). Thermal pre-treatments, carried out on seven different municipal wastewater sludges at the above optimum operating conditions, produced increased COD solubilization of 18.3 ± 7.5% and VSS reduction of 27.7 ± 12.3% compared to the untreated sludges. The solubilization of proteins was significantly higher than carbohydrates. Methane produced in biochemical methane potential (BMP) tests, indicated initial higher rates (p = 0.0013) for the thermally treated samples (k hyd up to 5 times higher), although the ultimate methane yields were not significantly affected by the treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Optimized preparation and characterization of CLEA-lipase from cocoa pod husk.

    PubMed

    Khanahmadi, Soofia; Yusof, Faridah; Amid, Azura; Mahmod, Safa Senan; Mahat, Mohd Khairizal

    2015-05-20

    Cross-linked enzyme aggregate (CLEA) is easily prepared from crude enzyme and has many advantages to the environment and it is considered as an economic method in the context of industrial biocatalysis compared to free enzyme. In this work, a highly active and stable CLEA-lipase from cocoa pod husk (CPH) which is a by-product after removal of cocoa beans, were assayed for their hydrolytic activity and characterized under the optimum condition successfully. Face centered central composite design (FCCCD) under response surface methodology (RSM) was used to get the optimal conditions of the three significant factors (concentration of ammonium sulfate, concentration of glutaraldehyde and concentration of additive) to achieve higher enzyme activity of CLEA. From 20 runs, the highest activity recorded was around 9.407U (83% recovered activity) under the condition of using 20% saturated ammonium sulfate, 60mM glutaraldehyde as cross-linker and 0.17mM bovine serum albumin as feeder. Moreover, the optimal reaction temperature and pH value in enzymatic reaction for both crude enzyme and immobilized were found to be 45°C at pH 8 and 60°C at pH 8.2, respectively. A systematic study of the stability of CLEA and crude enzyme was taken with regards to temperature (25-60°C) and pH (5-10) value and in both factors, CLEA-lipase showed more stability than free lipase. The Km value of CLEA was higher compared to free enzyme (0.55mM vs. 0.08mM). The CLEA retained more than 60% of the initial activity after six cycles of reuse compared to free enzyme. The high stability and recyclability of CLEA-lipase from CPH make it efficient for different industrial applications. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Precision sensing by two opposing gradient sensors: how does Escherichia coli find its preferred pH level?

    PubMed

    Hu, Bo; Tu, Yuhai

    2013-07-02

    It is essential for bacteria to find optimal conditions for their growth and survival. The optimal levels of certain environmental factors (such as pH and temperature) often correspond to some intermediate points of the respective gradients. This requires the ability of bacteria to navigate from both directions toward the optimum location and is distinct from the conventional unidirectional chemotactic strategy. Remarkably, Escherichia coli cells can perform such a precision sensing task in pH taxis by using the same chemotaxis machinery, but with opposite pH responses from two different chemoreceptors (Tar and Tsr). To understand bacterial pH sensing, we developed an Ising-type model for a mixed cluster of opposing receptors based on the push-pull mechanism. Our model can quantitatively explain experimental observations in pH taxis for various mutants and wild-type cells. We show how the preferred pH level depends on the relative abundance of the competing sensors and how the sensory activity regulates the behavioral response. Our model allows us to make quantitative predictions on signal integration of pH and chemoattractant stimuli. Our study reveals two general conditions and a robust push-pull scheme for precision sensing, which should be applicable in other adaptive sensory systems with opposing gradient sensors. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  18. Effects of pH and calcination temperature on structural and optical properties of alumina nanoparticles

    NASA Astrophysics Data System (ADS)

    Amirsalari, A.; Farjami Shayesteh, S.

    2015-06-01

    In this study, we describe the synthesis of alumina nanoparticles using a chemical wet method in at varying pH. The optimized prepared particles with pH equals to 9 were calcined at various temperatures. For characterization of structural and optical properties of nanoparticles had been used X-ray diffraction, Infrared Fourier transform spectroscopy, field effect-scanning electron microscopy, photoluminescence and ultraviolet-visible spectroscopy. The results revealed that the nanoparticles calcined at 500 °C consist of an Al2O3 tetragonal structure and tetragonal distortion decreases with increasing calcination temperature up to 750 °C then increased with increasing temperature. Another phase similar to γ-Al2O3 was formed instead of δ-Al2O3 in the transition sequence from the γ to θ phase. FT-IR analysis; suggests that there are a few different types of functional groups on the surface of the alumina nanoparticles such as hydroxy groups and oxy groups. The transmittance spectra showed that the absorption bands in the UV region strongly depend on the calcination temperature. Moreover, the results showed that alumina has an optical direct band gap and that the energy gap decreases with increasing the calcination temperature and pH of the reaction. Luminescence spectra showed that some luminescent centers such as OH-related radiative centers and oxygen vacancies (F, F22+ and F2 centers) centers exist in the nanoparticles.

  19. Surface treatment process of Al-Mg alloy powder by BTSPS

    NASA Astrophysics Data System (ADS)

    Zhao, Ran; Gao, Xinbao; Lu, Yanling; Du, Fengzhen; Zhang, Li; Liu, Dazhi; Chen, Xuefang

    2018-04-01

    The surface of Al-Mg alloy powder was treated by BTSPS(bis(triethoxysilylpropyl)tetrasulfide) in order to avoid easy oxidation in air. The pH value, reaction temperature, reaction time, and reaction concentration were used as test conditions. The results show that the BTSPS can form a protected film on the surface of Al-Mg alloy powder. Select the best test solution by orthogonal test. The study found that the reaction time and reaction temperature have the biggest influence on the two indexes of the orthogonal test (melting enthalpy of heat and enthalpy of oxidation). The optimal conditions were as follows: pH value is 8, reaction concentration is 2%, reaction temperature is 25 °C, reaction time is 2 h. The oxidation weight gain of the alloy reached 74.45% and the decomposition temperature of silane film is 181.8 °C.

  20. Effect of ferrate on green algae removal.

    PubMed

    Kubiňáková, Emília; Híveš, Ján; Gál, Miroslav; Fašková, Andrea

    2017-09-01

    Green algae Cladophora aegagropila, present in cooling water of thermal power plants, causes many problems and complications, especially during summer. However, algae and its metabolites are rarely eliminated by common removal methods. In this work, the elimination efficiency of electrochemically prepared potassium ferrate(VI) on algae from cooling water was investigated. The influence of experimental parameters, such as Fe(VI) dosage, application time, pH of the system, temperature and hydrodynamics of the solution on removal efficiency, was optimized. This study demonstrates that algae C. aegagropila can be effectively removed from cooling water by ferrate. Application of ferrate(VI) at the optimized dosage and under the suitable conditions (temperature, pH) leads to 100% removal of green algae Cladophora from the system. Environmentally friendly reduction products (Fe(III)) and coagulation properties favour the application of ferrate for the treatment of water contaminated with studied microorganisms compared to other methods such as chlorination and use of permanganate, where harmful products are produced.

  1. [Purification and physicochemical properties of Bacillus thuringiensis IMB B-7324 peptidase with elastolytic and fibrinolytic activity].

    PubMed

    Matseliukh, O V; Nidialkova, N A; Varbanets', L D

    2012-01-01

    The scheme of isolation and purification of Bacillus thuringiensis IMV B-7324 peptidase has been developed. This scheme includes ammonium sulfate precipitation and chromatography on neutral and charged TSK-gels. It was found that the enzyme hydrolyzes elastin and fibrin. The molecular weight is 26 kDa. It was shown that the enzyme is an alkaline serine peptidase. The optimal pH of hydrolysis of elastin and fibrin were 9.0 and 10.0, respectively. The optimal temperature of elastin and fibrin hydrolysis are 40 and 50 degrees C, respectively. The high stability of the purified preparation in the studied range of pH and temperature was shown. The stabilizing effect of zinc at a concentration of 1 mM on the elastase activity, and the inhibitory effect of other divalent cations under study have been established. The investigated chloride and acetate anions reduced activity by 20%, while phosphate anions increased activity by 15-30%.

  2. Improving industrial full-scale production of baker's yeast by optimizing aeration control.

    PubMed

    Blanco, Carlos A; Rayo, Julia; Giralda, José M

    2008-01-01

    This work analyzes the control of optimum dissolved oxygen of an industrial fed-batch procedure in which baker's yeast (Saccharomyces cerevisiae) is grown under aerobic conditions. Sugar oxidative metabolism was controlled by monitoring aeration, molasses flows, and yeast concentration in the propagator along the later stage of the propagation, and keeping pH and temperature under controlled conditions. A large number of fed-batch growth experiments were performed in the tank for a period of 16 h, for each of the 3 manufactured commercial products. For optimization and control of cultivations, the growth and metabolite formation were quantified through measurement of specific growth and ethanol concentration. Data were adjusted to a model of multiple lineal regression, and correlations representing dissolved oxygen as a function of aeration, molasses, yeast concentration in the broth, temperature, and pH were obtained. The actual influence of each variable was consistent with the mathematical model, further justified by significant levels of each variable, and optimum aeration profile during the yeast propagation.

  3. Improved in situ saccharification of cellulose pretreated by dimethyl sulfoxide/ionic liquid using cellulase from a newly isolated Paenibacillus sp. LLZ1.

    PubMed

    Hu, Dongxue; Ju, Xin; Li, Liangzhi; Hu, Cuiying; Yan, Lishi; Wu, Tianyun; Fu, Jiaolong; Qin, Ming

    2016-02-01

    A cellulase producing strain was newly isolated from soil samples and identified as Paenibacillus sp. LLZ1. A novel aqueous-dimethyl sulfoxide (DMSO)/1-ethyl-3-methylimidazolium diethyl phosphate ([Emin]DEP)-cellulase system was designed and optimized. In the pretreatment, DMSO was found to be a low-cost substitute of up to 70% ionic liquid to enhance the cellulose dissolution. In the enzymatic saccharification, the optimum pH and temperature of the Paenibacillus sp. LLZ1 cellulase were identified as 6.0 and 40°C, respectively. Under the optimized reaction condition, the conversion of microcrystalline cellulose and bagasse cellulose increased by 39.3% and 37.6%, compared with unpretreated cellulose. Compared to current methods of saccharification, this new approach has several advantages including lower operating temperature, milder pH, and less usage of ionic liquid, indicating a marked progress in environmental friendly hydrolysis of biomass-based materials. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Optimization and Performance parameters for adsorption of Cr6+ by microwave assisted carbon from Sterculia foetida shells

    NASA Astrophysics Data System (ADS)

    Gnanasundaram, N.; Loganathan, M.; Singh, A.

    2017-06-01

    Modeling of adsorption of Cr6+ on to activated carbon prepared from Sterculia foetida dried seed shells under different drying techniques namely sun, oven, and microwave drying (450W, 600W, 900W power). Optimization of process parameters such as pH, adsorbent dosage (g/ml), temperature (°C), contact time (min) were evaluated using Central Composite Rotatable Design (CCRD) of Response Surface Methodology (RSM). For batch adsorption studies at pH 3, adsorbent dosage of 1.5 g/ml, temperature 35°C and contact time 90 min were found to be optimum for the system under consideration and Microwave Activated Carbonized Sterculia foetida (MACSF) at 450W was found to be best suited for the adsorption of Cr+6 ions. The system was found to follow Langmuir type monolayer adsorption for the given operational parameters. SEM analysis was used to study the surface morphology of the carbon samples and the effect of pretreatment on carbonization.

  5. Optimization of Fe2+ Removal from Coal Mine Wastewater using Activated Biochar of Colocasia esculenta.

    PubMed

    Banerjee, Soumya; LaminKa-Ot, Augustine; Joshi, S R; Mandal, Tamal; Halder, Gopinath

    2017-09-01

      The present study investigates the sorptive removal of Fe2+ from simulated coal mine waste water using steam activated biochar (SABC) developed from the roots of Colocasia esculenta. The process was optimized by response surface methodology (RSM) under the influence of pH, temperature, adsorbent dosage and contact time at a constant shaking speed of 180 rpm with an initial concentration of 3 mg/L. The uptake performance of the biosorbent was assessed following a 24 full factorial experimental matrix developed by central composite design approach. Adsorbent was characterised by SEM, EDAX, XRD and B.E.T surface area analyzer. Maximum removal of 72.96% of Fe2+ was observed at pH 7.75, temperature 37.5 °C, adsorbent dosage 1.5 g/L for a time period of 180 mins. The study suggested that SABC prepared from roots of Colocasia esculenta could be used as an efficient and cost effective sorbent for removal of Fe2+ from coal mine wastewater.

  6. Struvite Crystallization of Anaerobic Digestive Fluid of Swine Manure Containing Highly Concentrated Nitrogen

    PubMed Central

    Lee, Eun Young; Oh, Min Hwan; Yang, Seung-Hak; Yoon, Tae Han

    2015-01-01

    In this study, the optimal operation factors for struvite crystallization for removing and recovering nitrogen and phosphorus from anaerobic digestive fluid of swine manure containing highly concentrated nitrogen was determined. Every experiment for the struvite crystallization reaction was conducted by placing 1,000 mL of digestion fluid in a 2,000 mL Erlenmeyer flask at various temperatures, pH, and mixing speed. Except for special circumstances, the digestion fluid was centrifuged (10,000 rpm, 10 min) and then the supernatant was used for the experiment at room temperature and 100 rpm. The optimal mole ratio of PO43−:Mg2+ was 1:1.5, and the pH effect ranging from 9 to 11 was similar, when mixed for 1 hour. Under this condition, the removal efficiency of NH4+-N and PO43−-P was 40% and 88.6%, respectively. X-shaped crystal was observed by light and scanning electron microscopy. In addition, struvite crystal structure was confirmed through X-ray diffraction analysis. PMID:26104412

  7. Aspects of the "Design Space" in high pressure liquid chromatography method development.

    PubMed

    Molnár, I; Rieger, H-J; Monks, K E

    2010-05-07

    The present paper describes a multifactorial optimization of 4 critical HPLC method parameters, i.e. gradient time (t(G)), temperature (T), pH and ternary composition (B(1):B(2)) based on 36 experiments. The effect of these experimental variables on critical resolution and selectivity was carried out in such a way as to systematically vary all four factors simultaneously. The basic element is a gradient time-temperature (t(G)-T) plane, which is repeated at three different pH's of the eluent A and at three different ternary compositions of eluent B between methanol and acetonitrile. The so-defined volume enables the investigation of the critical resolution for a part of the Design Space of a given sample. Further improvement of the analysis time, with conservation of the previously optimized selectivity, was possible by reducing the gradient time and increasing the flow rate. Multidimensional robust regions were successfully defined and graphically depicted. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  8. Ultrasound-assisted enzymatic extraction and antioxidant activity of polysaccharides from pumpkin (Cucurbita moschata).

    PubMed

    Wu, Hao; Zhu, Junxiang; Diao, Wenchao; Wang, Chengrong

    2014-11-26

    An efficient ultrasound-assisted enzymatic extraction (UAEE) of Cucurbita moschata polysaccharides (CMCP) was established and the CMCP antioxidant activities were studied. The UAEE operating parameters (extraction temperature, ultrasonic power, pH, and liquid-to-material ratio) were optimized using the central composite design (CCD) and the mass transfer kinetic study in UAEE procedure was used to select the optimal extraction time. Enzymolysis and ultrasonication that were simultaneously conducted was selected as the UAEE synergistic model and the optimum extraction conditions with a maximum polysaccharide yield of 4.33 ± 0.15% were as follows: extraction temperature, 51.5 °C; ultrasonic power, 440 W; pH, 5.0; liquid-to-material ratio, 5.70:1 mL/g; and extraction time, 20 min. Evaluation of the antioxidant activity in vitro suggested that CMCP has good potential as a natural antioxidant used in the food or medicine industry because of their high reducing power and positive radical scavenging activity for DPPH radical. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Effects of culture conditions on growth and docosahexaenoic acid production from Schizochytrium limacinum

    NASA Astrophysics Data System (ADS)

    Zhu, Luying; Zhang, Xuecheng; Ren, Xueying; Zhu, Qinghua

    2008-02-01

    The effects of temperature, initial pH, salinity of culture medium, and carbon and nitrogen sources on growth and docosahexaenoic acid (C22: 6 n-3, DHA) production from Schizochytrium limacinum OUC88 were investigated in the present study. The results revealed that the optimal temperature, initial pH and salinity level of the medium for DHA production were 23°C, 7.0 and 18, respectively. Glucose was proved the best carbon source for the growth and DHA production from S. limacinum. Among the nitrogen sources tested, soybean cake hydrolysate, a cheap by-product, was found to be effective for the accumulation of DHA in S. limacinum cells. In addition, increasing the concentration of carbon sources in the medium caused a significant increase in cell biomass; however, accumulation of DHA in cells was mainly stimulated by the ratio of C/N in the medium. Under the optimal culture conditions, the maximum DHA yield achieved in flasks was 4.08 g L-1 after 5 d of cultivation.

  10. Optimization of Processing Parameters for Extraction of Amylase Enzyme from Dragon (Hylocereus polyrhizus) Peel Using Response Surface Methodology

    PubMed Central

    Abdul Manap, Mohd Yazid; Zohdi, Norkhanani

    2014-01-01

    The main goal of this study was to investigate the effect of extraction conditions on the enzymatic properties of thermoacidic amylase enzyme derived from dragon peel. The studied extraction variables were the buffer-to-sample (B/S) ratio (1 : 2 to 1 : 6, w/w), temperature (−18°C to 25°), mixing time (60 to 180 seconds), and the pH of the buffer (2.0 to 8.0). The results indicate that the enzyme extraction conditions exhibited the least significant (P < 0.05) effect on temperature stability. Conversely, the extraction conditions had the most significant (P < 0.05) effect on the specific activity and pH stability. The results also reveal that the main effect of the B/S ratio, followed by its interaction with the pH of the buffer, was significant (P < 0.05) among most of the response variables studied. The optimum extraction condition caused the amylase to achieve high enzyme activity (648.4 U), specific activity (14.2 U/mg), temperature stability (88.4%), pH stability (85.2%), surfactant agent stability (87.2%), and storage stability (90.3%). PMID:25050403

  11. Facile synthesis of concentrated gold nanoparticles with low size-distribution in water: temperature and pH controls

    NASA Astrophysics Data System (ADS)

    Li, Chunfang; Li, Dongxiang; Wan, Gangqiang; Xu, Jie; Hou, Wanguo

    2011-07-01

    The citrate reduction method for the synthesis of gold nanoparticles (GNPs) has known advantages but usually provides the products with low nanoparticle concentration and limits its application. Herein, we report a facile method to synthesize GNPs from concentrated chloroauric acid (2.5 mM) via adding sodium hydroxide and controlling the temperature. It was found that adding a proper amount of sodium hydroxide can produce uniform concentrated GNPs with low size distribution; otherwise, the largely distributed nanoparticles or instable colloids were obtained. The low reaction temperature is helpful to control the nanoparticle formation rate, and uniform GNPs can be obtained in presence of optimized NaOH concentrations. The pH values of the obtained uniform GNPs were found to be very near to neutral, and the pH influence on the particle size distribution may reveal the different formation mechanism of GNPs at high or low pH condition. Moreover, this modified synthesis method can save more than 90% energy in the heating step. Such environmental-friendly synthesis method for gold nanoparticles may have a great potential in large-scale manufacturing for commercial and industrial demand.

  12. Combined Homogeneous Surface Diffusion Model - Design of experiments approach to optimize dye adsorption considering both equilibrium and kinetic aspects.

    PubMed

    Muthukkumaran, A; Aravamudan, K

    2017-12-15

    Adsorption, a popular technique for removing azo dyes from aqueous streams, is influenced by several factors such as pH, initial dye concentration, temperature and adsorbent dosage. Any strategy that seeks to identify optimal conditions involving these factors, should take into account both kinetic and equilibrium aspects since they influence rate and extent of removal by adsorption. Hence rigorous kinetics and accurate equilibrium models are required. In this work, the experimental investigations pertaining to adsorption of acid orange 10 dye (AO10) on activated carbon were carried out using Central Composite Design (CCD) strategy. The significant factors that affected adsorption were identified to be solution temperature, solution pH, adsorbent dosage and initial solution concentration. Thermodynamic analysis showed the endothermic nature of the dye adsorption process. The kinetics of adsorption has been rigorously modeled using the Homogeneous Surface Diffusion Model (HSDM) after incorporating the non-linear Freundlich adsorption isotherm. Optimization was performed for kinetic parameters (color removal time and surface diffusion coefficient) as well as the equilibrium affected response viz. percentage removal. Finally, the optimum conditions predicted were experimentally validated. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Simultaneous microemulsion liquid chromatographic analysis of fat-soluble vitamins in pharmaceutical formulations: optimization using genetic algorithm.

    PubMed

    Momenbeik, Fariborz; Roosta, Mostafa; Nikoukar, Ali Akbar

    2010-06-11

    An environmentally benign and simple method has been proposed for separation and determination of fat-soluble vitamins using isocratic microemulsion liquid chromatography. Optimization of parameters affecting the separation selectivity and efficiency including surfactant concentration, percent of cosurfactant (1-butanol), and percent of organic oily solvent (diethyl ether), temperature and pH were performed simultaneously using genetic algorithm method. A new software package, MLR-GA, was developed for this purpose. The results indicated that 73.6mM sodium dodecyl sulfate, 13.64% (v/v) 1-butanol, 0.48% (v/v) diethyl ether, column temperature of 32.5 degrees C and 0.02M phosphate buffer of pH 6.99 are the best conditions for separation of fat-soluble vitamins. At the optimized conditions, the calibration plots for the vitamins were obtained and detection limits (1.06-3.69microgmL(-1)), accuracy (recoveries>94.3), precision (RSD<3.96) and linearity (0.01-10mgmL(-1)) were estimated. Finally, the amount of vitamins in multivitamin syrup and a sample of fish oil capsule were determined. The results showed a good agreement with those reported by manufactures. Copyright 2010 Elsevier B.V. All rights reserved.

  14. Optimization of process variables for decolorization of Disperse Yellow 211 by Bacillus subtilis using Box-Behnken design.

    PubMed

    Sharma, Praveen; Singh, Lakhvinder; Dilbaghi, Neeraj

    2009-05-30

    Decolorization of textile azo dye Disperse Yellow 211 (DY 211) was carried out from simulated aqueous solution by bacterial strain Bacillus subtilis. Response surface methodology (RSM), involving Box-Behnken design matrix in three most important operating variables; temperature, pH and initial dye concentration was successfully employed for the study and optimization of decolorization process. The total 17 experiments were conducted in the study towards the construction of a quadratic model. According to analysis of variance (ANOVA) results, the proposed model can be used to navigate the design space. Under optimized conditions the bacterial strain was able to decolorize DY 211 up to 80%. Model indicated that initial dye concentration of 100 mgl(-1), pH 7 and a temperature of 32.5 degrees C were found optimum for maximum % decolorization. Very high regression coefficient between the variables and the response (R(2)=0.9930) indicated excellent evaluation of experimental data by polynomial regression model. The combination of the three variables predicted through RSM was confirmed through confirmatory experiments, hence the bacterial strain holds a great potential for the treatment of colored textile effluents.

  15. Production of Laccase by a New Myrothecium verrucaria MD-R-16 Isolated from Pigeon Pea [Cajanus cajan (L.) Millsp.] and its Application on Dye Decolorization.

    PubMed

    Sun, Jiao; Guo, Na; Niu, Li-Li; Wang, Qing-Fang; Zang, Yu-Ping; Zu, Yuan-Gang; Fu, Yu-Jie

    2017-04-23

    The present study was conducted to screen a laccase-producing fungal endophyte, optimize fermentation conditions, and evaluate the decolorization ability of the laccase. A new fungal endophyte capable of laccase-producing was firstly isolated from pigeon pea and identified as Myrothecium verrucaria based on a ITS-rRNA sequences analysis. Meanwhile, various fermentation parameters on the laccase production were optimized via response surface methodology (RSM). The optimal fermentation conditions were a fermentation time of five days, temperature 30 °C and pH 6.22. Laccase activity reached 16.52 ± 0.18 U/mL under the above conditions. Furthermore, the laccase showed effective decolorization capability toward synthetic dyes (Congo red, Methyl orange, Methyl red, and Crystal violet) in the presence of the redox mediator ABTS, with more than 70% of dyes decolorizing after 24 h of incubation. Additionally, the activity of laccase was relatively stable with pH (4.5-6.5) and a temperature range of 35-55 °C. Therefore, the high laccase production of the strain and the new fungal laccase could provide a promising alterative approach for industrial and environmental applications.

  16. Crystal Structure and Superconductivity of PH 3 at High Pressures

    DOE PAGES

    Liu, Hanyu; Li, Yinwei; Gao, Guoying; ...

    2016-01-20

    Here, we performed systematic structure search on solid PH 3 at high pressures using particle swarm optimization method. Furthermore, at 100-200 GPa, the search led to two structures consisting of P-P bonds that different from these predicted for H 2S. Phonon and electron-phonon calculations indicate both structures are dynamically stable and superconductive. Particularly, the estimated critical temperature for the monoclinic (C2/m) phase of 83 K at 200 GPa is in excellent agreement with a recent experimental report.

  17. Process optimization for green synthesis of silver nanoparticles by Sclerotinia sclerotiorum MTCC 8785 and evaluation of its antibacterial properties.

    PubMed

    Saxena, Juhi; Sharma, Prashant Kumar; Sharma, Madan Mohan; Singh, Abhijeet

    2016-01-01

    Eco-friendly synthesis of nanoparticles is viewed as an alternative to the chemical method and initiated the use of microorganisms for synthesis. The present study has been designed to utilize plant pathogenic fungi Sclerotinia sclerotiorum MTCC 8785 strain for synthesis and optimization of silver nanoparticles (AgNPs) production as well as evaluation of antibacterial properties. The AgNPs were synthesized by reduction of aqueous silver nitrate (AgNO3) solution after incubation of 3-5 days at room temperature. The AgNPs were further characterized using UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM). Reaction parameters including media, fungal biomass, AgNO3 concentration, pH and temperature were further optimized for rapid AgNPs production. The antibacterial efficacy of AgNPs was evaluated against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 by disc diffusion and growth kinetics assay at the concentration determined by the minimum inhibitory concentration (MIC). AgNPs synthesis was initially marked by the change in colour from pale white to brown and was confirmed by UV-Vis spectroscopy. Optimization studies showed that potato dextrose broth (PDB) media, 10 g of biomass, addition of 2 mM AgNO3, pH 11 and 80 °C temperature resulted in enhanced AgNPs synthesis through extracellular route. TEM data revealed spherical shape AgNPs with size in the range of 10 nm. Presence of proteins capped to AgNPs was confirmed by FTIR. AgNPs showed antibacterial activity against E. coli and S. aureus at 100 ppm concentration, corresponding MIC value. S. sclerotiorum MTCC 8785 mediated AgNPs was synthesized rapidly under optimized conditions, which showed antibacterial activity.

  18. Different efficiency of ozonated water washing to inactivate Salmonella enterica typhimurium on green onions, grape tomatoes, and green leaf lettuces.

    PubMed

    Xu, Wenqing; Wu, Changqing

    2014-03-01

    Ozonated water washing is one of the emerging techniques to inactivate foodborne pathogens on produce, and limited information is available to optimize processing parameters (treatment time, temperature, and pH) to improve ozone efficacy on Salmonella inactivation for different produce. The efficacy of ozonated water washing for inactivation of Salmonella enterica Typhimurium on green onions, grape tomatoes and green leaf lettuces were studied in our research. Surface inoculated fresh produce were washed by ozonated water for 1, 5, or 10 min at room temperature and pH 5.60 ± 0.03. Then efficacy of ozonated water washing at mild heated (50 °C) and refrigerated (4 °C) temperature for 5 min with pH 5.60 ± 0.03 was investigated. Salmonella inactivation efficacy under pH 5.60 ± 0.03 and 2.64 ± 0.02 with 5 min washing at room temperature were also compared. Our results showed that Salmonella inactivation by ozonated water was time-dependent for 3 fresh produce. Mild heated temperature (50 °C) and pH 2.64 ± 0.02 improved efficacy of ozonated water to inactivate Salmonella on tomatoes and lettuces, but not on green onions. It is suggested that different surface structures of fresh produce significantly impact the antimicrobial efficacy of ozonated water washing operated under various parameters (time, temperature, and pH). Washing is the essential step for green onions and lettuces in the packinghouse and grape tomatoes in the restaurants and grocery stores having salad bars. Ozonated water can be used as disinfectant to reduce microbial contamination (FDA). The effectiveness of this disinfectant depends on the type of product and treatment conditions, such as water temperature, acidity, contact time. Our study showed that Salmonella inactivation by ozonated water washing was time-dependent. Mild heat and low pH improved inactivation efficacy on tomatoes and lettuces, but not on green onions. Processors should consider adjustments that are most appropriate for their produce. © 2014 Institute of Food Technologists®

  19. Factors affecting Archaeal Lipid Compositions of the Sulfolobus Species

    NASA Astrophysics Data System (ADS)

    He, L.; Han, J.; Wei, Y.; Lin, L.; Wei, Y.; Zhang, C.

    2010-12-01

    Temperature is the best known variable affecting the distribution of the archaeal glycerol dibiphytanyl glycerol tetraethers (GDGTs) in marine and freshwater systems. Other variables such as pH, ionic strength, or bicarbonate concentration may also affect archaeal GDGTs in terrestrial systems. Studies of pure cultures can help us pinpoint the specific effects these variables may have on archaeal lipid distribution in natural environments. In this study, three Sulfolobus species (HG4, HB5-2, HB9-6) isolated from Tengchong hot springs (pH 2-3, temperature 73-90°C) in China were used to investigate the effects of temperature, pH, substrate, and type of strain on the composition of GDGTs. Results showed that increase in temperature had negative effects on the relative contents of GDGT-0 (no cyclopentyl rings), GDGT-1 (one cyclopentyl ring), GDGT-2 and GDGT-3 but positive effects on GDGT-4, GDGT-4', GDGT-5 and GDGT-5'. Increase in pH, on the other hand, had negative effects on GDGT-0, GDGT-1, GDGT-4', GDGT-5 and GDGT-5', and positive effects on GDGT-3 and GDGT-4. GDGT-2 remained relatively constant with changing pH. When the HG4 was grown on different substrates, GDGT-5 was five time more abundant in sucrose-grown cultures than in yeast extract- or sulfur- grown cultures, suggesting that carbohydrates may stimulate the production of GDGT-5. For all three species, the ring index (average number of rings) of GDGTs correlated positively with incubation temperature. In HG4, ring index was much lower at optimal pH (3.5) than at other pH values. Ring index of HB5-2 or HB9-6 is higher than that of HG4, suggesting that speciation may affect the degree of cyclization of GDGT of the Sulfolobus. These results indicate that individual archaeal lipids respond differently to changes in environmental variables, which may be also species specific.

  20. Optimization of extraction of novel pectinase enzyme discovered in red pitaya (Hylocereus polyrhizus) peel.

    PubMed

    Zohdi, Nor Khanani; Amid, Mehrnoush

    2013-11-20

    Plant peels could be a potential source of novel pectinases for use in various industrial applications due to their broad substrate specificity with high stability under extreme conditions. Therefore, the extraction conditions of a novel pectinase enzyme from pitaya peel was optimized in this study. The effect of extraction variables, namely buffer to sample ratio (2:1 to 8:1, X₁), extraction temperature (-15 to +25 °C, X₂) and buffer pH (4.0 to 12.0, X₃) on specific activity, temperature stability, storage stability and surfactant agent stability of pectinase from pitaya peel was investigated. The study demonstrated that the optimum conditions for the extraction of pectinase from pitaya sources could improve the enzymatic characteristics of the enzyme and protect its activity and stability during the extraction procedure. The optimum extraction conditions cause the pectinase to achieve high specific activity (15.31 U/mg), temperature stability (78%), storage stability (88%) and surfactant agent stability (83%). The most desirable conditions to achieve the highest activity and stability of pectinase enzyme from pitaya peel were the use of 5:1 buffer to sample ratio at 5 °C and pH 8.0.

  1. Removal of Emulsified Oil from Water by Fruiting Bodies of Macro-Fungus (Auricularia polytricha)

    PubMed Central

    Yang, Xunan; Guo, Mengting; Wu, Yinghai; Wu, Qunhe; Zhang, Renduo

    2014-01-01

    The aim of this study was to investigate the feasibility of utilizing the fruiting bodies of a jelly macro-fungus Auricularia polytricha as adsorbents to remove emulsified oil from water. The effects of several factors, including temperature, initial pH, agitation speed, and adsorbent dosage, were taken into account. Results showed that the optimized conditions for adsorption of A. polytricha were a temperature of 35°C, pH of 7.5, and agitation speed of 100 rpm. The adsorption kinetics were characterized by the pseudo-first order model, which showed the adsorption to be a fast physical process. The Langmuir-Freundlich isotherm described the adsorption very well and predicted the maximum adsorption capacity of 398 mg g−1, under optimized conditions. As illustrated by scanning electron micrographs, the oil particles were adsorbed onto the hairs covering the bottom surface and could be desorbed by normal temperature volatilization. The material could be used as an emulsified oil adsorbent at least three times, retaining more than 95% of the maximum adsorption capacity. The results demonstrated that the fruiting bodies of A. polytricha can be a useful adsorbent to remove emulsified oil from water. PMID:24743498

  2. Solid-substrate bioprocessing of cow dung for the production of carboxymethyl cellulase by Bacillus halodurans IND18.

    PubMed

    Vijayaraghavan, P; Prakash Vincent, S G; Dhillon, G S

    2016-02-01

    The production of carboxymethyl cellulase (CMCase) by Bacillus halodurans IND18 under solid substrate fermentation (SSF) using cow dung was optimized through two level full factorial design and second order response surface methodology (RSM). The central composite design (CCD) was employed to optimize the vital fermentation parameters, such as pH of the substrate, concentration of nitrogen source (peptone) and ion (sodium dihydrogen phosphate) sources in medium for achieving higher enzyme production. The optimum medium composition was found to be 1.46% (w/w) peptone, 0.095% (w/w) sodium dihydrogen phosphate and pH 8.0. The model prediction of 4210IU/g enzyme activity at optimum conditions was verified experimentally as 4140IU/g. The enzyme was active over a broad temperature range (40-60±1°C) and pH (7.0-9.0) with maximal activity at 60±1°C and pH 8.0. This study demonstrated the potential of cow dung as novel substrate for CMCase production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Overexpression and characterization of laccase from Trametes versicolor in Pichia pastoris.

    PubMed

    Li, Q; Pei, J; Zhao, L; Xie, J; Cao, F; Wang, G

    2014-01-01

    A laccase-encoding gene of Trametes versicolor, lccA, was cloned and expressed in Pichia pastoris X33. The lccA gene consists ofa 1560 bp open reading frame encoding 519 amino acids, which was classified into family copper blue oxidase. To improve the expression level of recombinant laccase in P. pastoris, conditions of the fermentation were optimized by the single factor experiments. The optimal fermentation conditions for the laccase production in shake flask cultivation using BMGY medium were obtained: the optimal initial pH 7.0, the presence of 0.5 mM Cu2+, 0.6% methanol added into the culture every 24 h. The laccase activity was up to 11.972 U/L under optimal conditions after 16 days of induction in a medium with 4% peptone. After 100 h of large scale production in 5 L fermenter the enzyme activity reached 18.123 U/L. The recombinant laccase was purified by ultrafiltration and (NH4)2SO4 precipitation showing a single band on SDS-PAGE, which had a molecular mass of 58 kDa. The optimum pH and temperature for the laccase were pH 2.0 and 50 degrees C with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as a substrate. The recombinant laccase was stable over a pH range of 2.0-7.0. The K(m) and the V(max) value of LccA were 0.43 mM and 82.3 U/mg for ABTS, respectively.

  4. Adaptive Neuro-Fuzzy Inference system analysis on adsorption studies of Reactive Red 198 from aqueous solution by SBA-15/CTAB composite

    NASA Astrophysics Data System (ADS)

    Aghajani, Khadijeh; Tayebi, Habib-Allah

    2017-01-01

    In this study, the Mesoporous material SBA-15 were synthesized and then, the surface was modified by the surfactant Cetyltrimethylammoniumbromide (CTAB). Finally, the obtained adsorbent was used in order to remove Reactive Red 198 (RR 198) from aqueous solution. Transmission electron microscope (TEM), Fourier transform infra-red spectroscopy (FTIR), Thermogravimetric analysis (TGA), X-ray diffraction (XRD), and BET were utilized for the purpose of examining the structural characteristics of obtained adsorbent. Parameters affecting the removal of RR 198 such as pH, the amount of adsorbent, and contact time were investigated at various temperatures and were also optimized. The obtained optimized condition is as follows: pH = 2, time = 60 min and adsorbent dose = 1 g/l. Moreover, a predictive model based on ANFIS for predicting the adsorption amount according to the input variables is presented. The presented model can be used for predicting the adsorption rate based on the input variables include temperature, pH, time, dosage, concentration. The error between actual and approximated output confirm the high accuracy of the proposed model in the prediction process. This fact results in cost reduction because prediction can be done without resorting to costly experimental efforts. SBA-15, CTAB, Reactive Red 198, adsorption study, Adaptive Neuro-Fuzzy Inference systems (ANFIS).

  5. Performance Study of Chromium (VI) Removal in Presence of Phenol in a Continuous Packed Bed Reactor by Escherichia coli Isolated from East Calcutta Wetlands

    PubMed Central

    Chakraborty, Bhaswati; Indra, Suvendu; Hazra, Ditipriya; Betai, Rupal; Ray, Lalitagauri; Basu, Srabanti

    2013-01-01

    Organic pollutants, like phenol, along with heavy metals, like chromium, are present in various industrial effluents that pose serious health hazard to humans. The present study looked at removal of chromium (VI) in presence of phenol in a counter-current continuous packed bed reactor packed with E. coli cells immobilized on clay chips. The cells removed 85% of 500 mg/L of chromium (VI) from MS media containing glucose. Glucose was then replaced by 500 mg/L phenol. Temperature and pH of the MS media prior to addition of phenol were 30°C and 7, respectively. Hydraulic retention times of phenol- and chromium (VI)-containing synthetic media and air flow rates were varied to study the removal efficiency of the reactor system. Then temperature conditions of the reactor system were varied from 10°C to 50°C, the optimum being 30°C. The pH of the media was varied from pH 1 to pH 12, and the optimum pH was found to be 7. The maximum removal efficiency of 77.7% was achieved for synthetic media containing phenol and chromium (VI) in the continuous reactor system at optimized conditions, namely, hydraulic retention time at 4.44 hr, air flow rate at 2.5 lpm, temperature at 30°C, and pH at 7. PMID:24073400

  6. Preparation and optimization of self-assembled chondroitin sulfate-nisin nanogel based on quality by design concept.

    PubMed

    Mohtashamian, Shahab; Boddohi, Soheil; Hosseinkhani, Saman

    2018-02-01

    Self-assembled nanogel was prepared by electrostatic complexation of two oppositely charged biological macromolecules, which were cationic nisin and anionic chondroitin sulfate (ChS). The critical factors affected the physical properties of ChS-nisin nanogel was screened and optimized by Plackett-Burman design (PB) and central composite design (CCD). The independent factors selected were: concentration ratio of nisin to ChS, injection rate of nisin solution, buffer solvent type, magnetic stirring rate, pH of initial buffer solution, centrifuge-cooling temperature, and centrifuge rotation speed. Among these factors, concentration ratio changed the entrapment efficiency and loading capacity significantly. In addition, the hydrodynamic diameter and loading capacity were significantly influenced by injection rate and pH of initial buffer solution. The optimized nanogel structure was obtained by concentration ratio of 6.4mg/mL nisin to 1mg/mL ChS, pH of buffer solution at 4.6, and nisin solution injection rate of 0.2mL/min. The observed values of dependent responses were close to predicted values confirmed by model from response surface methodology. The results obviously showed that quality by design concept (QbD) could be effectively applied to optimize the developed ChS-nisin nanogel. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Delineating Spatial Patterns in the Yellowstone Hydrothermal System using Geothermometry

    NASA Astrophysics Data System (ADS)

    King, J.; Hurwitz, S.; Lowenstern, J. B.

    2015-12-01

    Yellowstone National Park is unmatched with regard to its quantity of active hydrothermal features. Origins of thermal waters in its geyser basins have been traced to mixing of a deep parent water with meteoric waters in shallow local reservoirs (Fournier, 1989). A mineral-solution equilibrium model was developed to calculate water-rock chemical re-equilibration temperatures in these shallow reservoirs. We use the GeoT program, which uses water composition data as input to calculate saturation indices of selected minerals; the "best-clustering" minerals are then statistically determined to infer reservoir temperatures (Spycher et al., 2013). We develop the method using water composition data from Heart Lake Geyser Basin (HLGB), for which both chemical and isotopic geothermometers predict a reservoir water temperature of 205°C ± 10°C (Lowenstern et al., 2012), and minerals found in drill cores in Yellowstone's geyser basins. We test the model for sensitivity to major element composition, pH, Total Inorganic Carbon (TIC) and selected minerals to optimize model parameters. Calculated temperatures are most accurate at pH values below 9.0, and closely match the equilibrium saturation indices of quartz, stilbite, microcline, and albite. The model is optimized with a TIC concentration that is consistent with the mass of diffuse CO2 flux in HLGB (Lowenstern et al., 2012). We then use water compositions from other thermal basins in Yellowstone in search of spatial variations in reservoir temperatures. We then compare the calculated temperatures with various SiO2 and cation geothermometers.

  8. Impact of reaction conditions on the laccase-catalyzed conversion of bisphenol A.

    PubMed

    Kim, Young-Jin; Nicell, James A

    2006-08-01

    The oxidative conversion of aqueous BPA catalyzed by laccase from Trametes versicolor was conducted in a closed, temperature-controlled system containing buffer for pH control. The effects of medium pH, buffer concentration, temperature and mediators and the impacts of dissolved wastewater constituents on BPA conversion were investigated. The optimal pH for BPA conversion was approximately 5, with greater than half maximal conversion and good enzyme stability in the range of 4-7. The stability of the enzyme was not impacted by buffer concentration, nor was BPA conversion. Despite the observation that the enzyme tended to be inactivated at elevated temperatures, enhanced conversion of BPA was observed up until a reaction temperature of 45 degrees C. Of the mediators studied, ABTS was most successful at enhancing the conversion of BPA. Dissolved wastewater constituents that were studied included various inorganic salts, organic compounds and heavy metal ions. BPA conversion was inhibited in the presence of anions such as sulfite, thiosulfate, sulfide, nitrite and cyanide. The metal ions Fe(III) and Cu(II) and the halogens chloride and fluoride substantially suppressed BPA conversion, but the presence of selected organic compounds did not significantly reduce the conversion of BPA.

  9. Optimization of cultivation conditions of fermented shaggy ink cap culinary-medicinal mushroom, Coprinus comatus (O.Mull.:Fr.) Pers. (higher Basidiomycetes) rich in Vanadium.

    PubMed

    Zhang, Chunjing; Qi, Xiaodan; Shi, Yan; Sun, Yan; Li, Shuyan; Gao, Xiulan; Yu, Haitao

    2012-01-01

    The present paper is mainly aimed at optimization of cultivation conditions of fermented mushrooms of Coprinus comatus rich in vanadium (CCRV). Initial screening of effects of carbon source, temperature, pH, and inoculum size were done by using a one-factor-at-a-time method. The results obtained in that study showed that the optimal medium composition was 30 g glucose/Lin YEPG medium, initial pH 6.0, inoculum volume 10%, and incubation time 120 h. Then the medium was subjected to screening of the most significant parameters using the L9 orthogonal array to solve multivariable equations simultaneously. The results obtained in this study showed that the optimal medium composition was 0.4% V and 30 g glucose/Lin YEPG medium, initial pH 5.0, inoculum volume 15%, and incubation time 120 h. At this medium composition, the mycelial biomass and V content were 7.18 ± 0.24 g/L and 3786.0 ± 17 μg/g, respectively. The anti-diabetic potential of CCRV produced with the optimal level was tested in alloxan-induced diabetes. After the mice were administered (i.g.) with CCRV, the level of blood sugar in the CCRV group was very close to that of the control group. These findings suggested that CCRV produced with the optimal level is useful in the control of diabetes mellitus.

  10. Modeling and verification of process parameters for the production of tannase by Aspergillus oryzae under submerged fermentation using agro-wastes.

    PubMed

    Varadharajan, Venkatramanan; Vadivel, Sudhan Shanmuga; Ramaswamy, Arulvel; Sundharamurthy, Venkatesaprabhu; Chandrasekar, Priyadharshini

    2017-01-01

    Tannase production by Aspergillus oryzae using various agro-wastes as substrates by submerged fermentation was studied in this research. The microbe was isolated from degrading corn kernel obtained from the corn fields at Tiruchengode, India. The microbial identification was done using 18S rRNA gene analysis. The agro-wastes chosen for the study were pomegranate rind, Cassia auriculata flower, black gram husk, and tea dust. The process parameters chosen for optimization study were substrate concentration, pH, temperature, and incubation period. During one variable at a time optimization, the pomegranate rind extract produced maximum tannase activity of 138.12 IU/mL and it was chosen as the best substrate for further experiments. The quadratic model was found to be the effective model for prediction of tannase production by A. oryzae. The optimized conditions predicted by response surface methodology (RSM) with genetic algorithm (GA) were 1.996% substrate concentration, pH of 4.89, temperature of 34.91 °C, and an incubation time of 70.65 H with maximum tannase activity of 138.363 IU/mL. The confirmatory experiment under optimized conditions showed tannase activity of 139.22 IU/mL. Hence, RSM-GA pair was successfully used in this study to optimize the process parameters required for the production of tannase using pomegranate rind. © 2015 International Union of Biochemistry and Molecular Biology, Inc.

  11. Oleosins (24 and 18 kDa) are hydrolyzed not only in extracted soybean oil bodies but also in soybean germination.

    PubMed

    Chen, Yeming; Zhao, Luping; Cao, Yanyun; Kong, Xiangzhen; Hua, Yufei

    2014-01-29

    After oil bodies (OBs) were extracted from ungerminated soybean by pH 6.8 extraction, it was found that 24 and 18 kDa oleosins were hydrolyzed in the extracted OBs, which contained many OB extrinsic proteins (i.e., lipoxygenase, β-conglycinin, γ-conglycinin, β-amylase, glycinin, Gly m Bd 30K (Bd 30K), and P34 probable thiol protease (P34)) as well as OB intrinsic proteins. In this study, some properties (specificity, optimal pH and temperature) of the proteases of 24 and 18 kDa oleosins and the oleosin hydrolysis in soybean germination were examined, and the high relationship between Bd 30K/P34 and the proteases was also discussed. The results showed (1) the proteases were OB extrinsic proteins, which had high specificity to hydrolyze 24 and 18 kDa oleosins, and cleaved the specific peptide bonds to form limited hydrolyzed products; (2) 24 and 18 kDa oleosins were not hydrolyzed in the absence of Bd 30K and P34 (or some Tricine-SDS-PAGE undetectable proteins); (3) the protease of 24 kDa oleosin had strong resistance to alkaline pH while that of 18 kDa oleosin had weak resistance to alkaline pH, and Bd 30K and P34, resolved into two spots on two-dimensional electrophoresis gel, also showed the same trend; (4) 16 kDa oleosin as well as 24 and 18 kDa oleosins were hydrolyzed in soybean germination, and Bd 30K and P34 were always contained in the extracted OBs from germinated soybean even when all oleosins were hydrolyzed; (5) the optimal temperature and pH of the proteases were respectively determined as in the ranges of 35-50 °C and pH 6.0-6.5, while 60 °C or pH 11.0 could denature them.

  12. Laccase Production from a Temperature and pH Tolerant Fungal Strain of Trametes hirsuta (MTCC 11397).

    PubMed

    Dhakar, Kusum; Pandey, Anita

    2013-01-01

    Laccase production by a temperature and pH tolerant fungal strain (GBPI-CDF-03) isolated from a glacial site in Indian Himalayan Region (IHR) has been investigated. The fungus developed white cottony mass on potato dextrose agar and revealed thread-like mycelium under microscope. ITS region analysis of fungus showed its 100% similarity with Trametes hirsuta. The fungus tolerated temperature from 4 to 48°C ± 2 (25°C opt.) and pH 3-13 (5-7 opt.). Molecular weight of laccase was determined approximately 45 kDa by native PAGE. Amplification of laccase gene fragment (corresponding to the copper-binding conserved domain) contained 200 bp. The optimum pH for laccase production, at optimum growth temperature, was determined between 5.5 and 7.5. In optimization experiments, fructose and ammonium sulfate were found to be the best carbon and nitrogen sources, respectively, for enhancing the laccase production. Production of laccase was favored by high carbon/nitrogen ratio. Addition of CuSO4 (up to 1.0 mM) induced laccase production up to 2-fold, in case of 0.4 mM concentration. Addition of organic solvents also induced the production of laccase; acetone showed the highest (2-fold) induction. The study has implications in bioprospecting of ecologically resilient microbial strains.

  13. Optimizing culture conditions for production of intra and extracellular inulinase and invertase from Aspergillus niger ATCC 20611 by response surface methodology (RSM).

    PubMed

    Dinarvand, Mojdeh; Rezaee, Malahat; Foroughi, Majid

    The aim of this study was obtain a model that maximizes growth and production of inulinase and invertase by Aspergillus niger ATCC 20611, employing response surface methodology (RSM). The RSM with a five-variable and three-level central composite design (CCD) was employed to optimize the medium composition. Results showed that the experimental data could be appropriately fitted into a second-order polynomial model with a coefficient of determination (R 2 ) more than 0.90 for all responses. This model adequately explained the data variation and represented the actual relationships between the parameters and responses. The pH and temperature value of the cultivation medium were the most significant variables and the effects of inoculum size and agitation speed were slightly lower. The intra-extracellular inulinase, invertase production and biomass content increased 10-32 fold in the optimized medium condition (pH 6.5, temperature 30°C, 6% (v/v), inoculum size and 150rpm agitation speed) by RSM compared with medium optimized through the one-factor-at-a-time method. The process development and intensification for simultaneous production of intra-extracellular inulinase (exo and endo inulinase) and invertase from A. niger could be used for industrial applications. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  14. Modeling and optimization of trihalomethanes formation potential of surface water (a drinking water source) using Box-Behnken design.

    PubMed

    Singh, Kunwar P; Rai, Premanjali; Pandey, Priyanka; Sinha, Sarita

    2012-01-01

    The present research aims to investigate the individual and interactive effects of chlorine dose/dissolved organic carbon ratio, pH, temperature, bromide concentration, and reaction time on trihalomethanes (THMs) formation in surface water (a drinking water source) during disinfection by chlorination in a prototype laboratory-scale simulation and to develop a model for the prediction and optimization of THMs levels in chlorinated water for their effective control. A five-factor Box-Behnken experimental design combined with response surface and optimization modeling was used for predicting the THMs levels in chlorinated water. The adequacy of the selected model and statistical significance of the regression coefficients, independent variables, and their interactions were tested by the analysis of variance and t test statistics. The THMs levels predicted by the model were very close to the experimental values (R(2) = 0.95). Optimization modeling predicted maximum (192 μg/l) TMHs formation (highest risk) level in water during chlorination was very close to the experimental value (186.8 ± 1.72 μg/l) determined in laboratory experiments. The pH of water followed by reaction time and temperature were the most significant factors that affect the THMs formation during chlorination. The developed model can be used to determine the optimum characteristics of raw water and chlorination conditions for maintaining the THMs levels within the safe limit.

  15. Optimization of antifungal production by an alkaliphilic and halotolerant actinomycete, Streptomyces sp. SY-BS5, using response surface methodology.

    PubMed

    Souagui, Y; Tritsch, D; Grosdemange-Billiard, C; Kecha, M

    2015-06-01

    Optimization of medium components and physicochemical parameters for antifungal production by an alkaliphilic and salt-tolerant actinomycete designated Streptomyces sp. SY-BS5; isolated from an arid region in south of Algeria. The strain showed broad-spectrum activity against pathogenic and toxinogenic fungi. Identification of the actinomycete strain was realized on the basis of 16S rRNA gene sequencing. Antifungal production was optimized following one-factor-at-a-time (OFAT) and response surface methodology (RSM) approaches. The most suitable medium for growth and antifungal production was found using one-factor-at-a-time methodology. The individual and interaction effects of three nutritional variables, carbon source (glucose), nitrogen source (yeast extract) and sodium chloride (NaCl) were optimized by Box-Behnken design. Finally, culture conditions for the antifungal production, pH and temperature were studied and determined. Analysis of the 16S rRNA gene sequence (1454 nucleotides) assigned this strain to Streptomyces genus with 99% similarity with Streptomyces cyaneofuscatus JCM4364(T), the most closely related. The results of the optimization study show that concentrations 3.476g/L of glucose, 3.876g/L of yeast extract and 41.140g/L of NaCl are responsible for the enhancement of antifungal production by Streptomyces sp. SY-BS5. The preferable culture conditions for antifungal production were pH 10, temperature 30°C for 09 days. This study proved that RSM is usual and powerful tool for the optimization of antifungal production from actinomycetes. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  16. Anodic oxidation of coke oven wastewater: Multiparameter optimization for simultaneous removal of cyanide, COD and phenol.

    PubMed

    Sasidharan Pillai, Indu M; Gupta, Ashok K

    2016-07-01

    Anodic oxidation of industrial wastewater from a coke oven plant having cyanide including thiocyanate (280 mg L(-1)), chemical oxygen demand (COD - 1520 mg L(-1)) and phenol (900 mg L(-1)) was carried out using a novel PbO2 anode. From univariate optimization study, low NaCl concentration, acidic pH, high current density and temperature were found beneficial for the oxidation. Multivariate optimization was performed with cyanide including thiocyanate, COD and phenol removal efficiencies as a function of changes in initial pH, NaCl concentration and current density using Box-Behnken experimental design. Optimization was performed for maximizing the removal efficiencies of these three parameters simultaneously. The optimum condition was obtained as initial pH 3.95, NaCl as 1 g L(-1) and current density of 6.7 mA cm(-2), for which the predicted removal efficiencies were 99.6%, 86.7% and 99.7% for cyanide including thiocyanate, COD and phenol respectively. It was in agreement with the values obtained experimentally as 99.1%, 85.2% and 99.7% respectively for these parameters. The optimum conditions with initial pH constrained to a range of 6-8 was initial pH 6, NaCl as 1.31 g L(-1) and current density as 6.7 mA cm(-2). The predicted removal efficiencies were 99%, 86.7% and 99.6% for the three parameters. The efficiencies obtained experimentally were in agreement at 99%, 87.8% and 99.6% respectively. The cost of operation for degradation at optimum conditions was calculated as 21.4 USD m(-3). Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Response surface methodological approach for the decolorization of simulated dye effluent using Aspergillus fumigatus fresenius.

    PubMed

    Sharma, Praveen; Singh, Lakhvinder; Dilbaghi, Neeraj

    2009-01-30

    The aim of our research was to study, effect of temperature, pH and initial dye concentration on decolorization of diazo dye Acid Red 151 (AR 151) from simulated dye solution using a fungal isolate Aspergillus fumigatus fresenius have been investigated. The central composite design matrix and response surface methodology (RSM) have been applied to design the experiments to evaluate the interactive effects of three most important operating variables: temperature (25-35 degrees C), pH (4.0-7.0), and initial dye concentration (100-200 mg/L) on the biodegradation of AR 151. The total 20 experiments were conducted in the present study towards the construction of a quadratic model. Very high regression coefficient between the variables and the response (R(2)=0.9934) indicated excellent evaluation of experimental data by second-order polynomial regression model. The RSM indicated that initial dye concentration of 150 mg/L, pH 5.5 and a temperature of 30 degrees C were optimal for maximum % decolorization of AR 151 in simulated dye solution, and 84.8% decolorization of AR 151 was observed at optimum growth conditions.

  18. Environmental Factors Affecting Ammonium Oxidation Under Iron Reducing Conditions

    NASA Astrophysics Data System (ADS)

    Jaffe, P. R.; Huang, S.; Ruiz-Urigüen, M.

    2014-12-01

    Ammonium (NH4+) oxidation coupled to iron (Fe) reduction in the absence of oxygen and nitrate/nitrite (NO3-/NO2-) has been reported by several investigators and referred to as Feammox. Feammox is a biological reaction, where Fe(III) is the electron acceptor, which is reduced to Fe(II), and NH4+ is the electron donor, which is oxidized to NO2-. Through a 180-day anaerobic incubation experiment, and using PCR-DGGE, 454-pyosequecing and qPCR analysis, we have shown that an Acidimicrobiaceae bacterium A6, a previously unreported species in the Acidimicrobiaceae family, might be either responsible or plays a key role in the Feammox process, We have enriched these Feammox bacteria (65.8% in terms of cell numbers) in a membrane reactor, and isolated the pure Acidimicrobiaceae bacterium A6 strain in an autotrophic medium. In samples collected and then incubated from a series of local wetland-, upland-, as well as storm-water detention pond-sediments, Feammox activity was only detected in acidic soil environments that contain Fe oxides. Using primers we developed for this purpose, Acidimicrobiaceae bacterium A6 was detected in all incubations where Feammox was observed. Anaerobic incubations of Feammox enrichment cultures adjusted to different pH, revealed that the optimal pH for Feammox is 4 ~ 5, and the reaction does not proceed when pH > 7. Feammox was still proceeding at pH as low as 2. In Feammox culture amended with different Fe(III) sources, Feammox reaction proceeded only when Fe oxides (ferrihydrite or goethite ) were supplied, whereas samples incubated with ferric chloride or ferric citrate showed no measurable NH4+ oxidation. Furthermore, we have also determined from incubation experiments conducted with a temperature gradient (10 ~ 35℃), that the Feammox process was active when the temperature is above 15℃, and the optimal temperature is 20℃. Incubations of enrichment culture with 79% Feammox bacteria appeared to remove circa 8% more NH4+ at 20ºC than at 35ºC. This is in contrast to anammox, another anaerobic ammonium oxidation pathway, for which optimal NH4+ oxidation is at temperatures ~ 30ºC. Hence, a Feammox-based process is an attractive candidate for wastewater treatment that could result in further energy savings, by requiring no aeration or heating of the wastewater in temperate climates.

  19. Effect of temperature and pH on the properties of skim milk gels made from a tamarillo (Cyphomandra betacea) coagulant and rennet.

    PubMed

    Li, Z; Scott, K; Otter, D; Zhou, P; Hemar, Y

    2018-06-01

    Reconstituted skim milk was gelled with a crude protease extract from tamarillo [Cyphomandra betacea or Solanum betacea (syn.)] fruit and compared with gels prepared with calf rennet. The effects of temperature and pH on the gelation of skim milk were investigated by small deformation oscillatory rheology. The tamarillo extract-induced gels had a faster rate of increase in the elastic modulus (G') at the early stage of gelation than rennet-induced milk gels. This was probably due to the broader proteolytic activity of tamarillo protease extracts as shown by sodium dodecyl sulfate-PAGE analysis. Confocal microscopy also showed that the milk gels resulting from the addition of tamarillo extracts had larger voids than rennet-induced milk gels. The proteolytic activity of tamarillo extracts was found to be optimal at pH 11. For both rennet and tamarillo extracts, the aggregation time was similar between pH 6.7 and 6.5, but the aggregation time of rennet-induced milk gels was lower than that of milk gels obtained by the addition of tamarillo extracts at pH lower than 6.5. An increase in temperature was found to have a significant effect on aggregation time, particularly at 20°C, where rennet did not coagulate milk in 3 h but the tamarillo extracts coagulated milk within 2 h. The results of this study suggest that extracts from tamarillo fruit could be used for milk gelation, particularly under lower temperature or high pH conditions. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  20. [Immobilized ammonia-oxidizing bacteria Comamonas aquatic LNL3 and its partial nitrification characterization].

    PubMed

    Li, Zheng-kui; Shi, Lu-na; Yang, Zhu-you; Zhang, Xiao-jiao; Wang, Yue-ming; Chen, Qi-chun; Wu, Kai

    2009-10-15

    A new kind of ammonia-oxidizing bacteria (AOB)-Comamonas aquatic LNL3 was screened out and immobilized by Poly (HEA)-Poly (HEMA) copolymer carrier using irradiation techniques. Four kinds of impact factors on short-cut nitrification, including temperature, pH, DO and free ammonia (FA) concentration had been investigated. The result showed that AOB-Comamonas aquatic LNL3 had short-cut nitrification capability and the optimal temperature, pH, DO and FA concentration were 30 degrees C, 8.5, 4.03 mg/L and 9 mg/L respectively. Corresponding to above results, ammonia nitrogen removal rate and short-cut nitrification efficiency were 93.52%, 94.73%; 79.74%, 94.67%; 91.17%, 94.66% and 90%, 94.4% respectively.

  1. Trained sensory perception of pork eating quality as affected by fresh and cooked pork quality attributes and end-point cooked temperature.

    PubMed

    Moeller, S J; Miller, R K; Aldredge, T L; Logan, K E; Edwards, K K; Zerby, H N; Boggess, M; Box-Steffensmeier, J M; Stahl, C A

    2010-05-01

    The present study evaluated individual and interactive influences of pork loin (n=679) ultimate ph (pH), intramuscular fat (IMF), Minolta L* color (L*), Warner-Bratzler shear force (WBSF), and internal cooked temperatures (62.8 degrees C, 68.3 degrees C, 73.9 degrees C, and 79.4 degrees C) on trained sensory perception of palatability. Logistical regression analyses were used, fitting sensory responses as dependent variables and quality and cooked temperature as independent variables, testing quadratic and interactive effects. Incremental increases in cooked temperature reduced sensory juiciness and tenderness scores by 3.8% and 0.9%, respectively, but did not influence sensory flavor or saltiness scores. An increase of 4.9N in WBSF, from a base of 14.7N (lowest) to 58.8N (greatest) was associated with a 3.7% and 1.8% reduction in sensory tenderness and juiciness scores, respectively, with predicted sensory tenderness scores reduced by 3.55 units when comparing ends of the WBSF range. Modeled sensory responses for loins with pH of 5.40 and 5.60 had reduced tenderness, chewiness, and fat flavor ratings when compared with responses for loins with pH of 5.80 to 6.40, the range indicative of optimal sensory response. Loin IMF and L* were significant model effects; however, their influence on sensory attributes was small, with predicted mean sensory responses measurably improved only when comparing 6% and 1% IMF and L* values of 46.9 (dark) when compared with 65.0 (pale). Tenderness and juiciness scores, were related to a greater extent to loin WBSF and pH, and to a lesser extent to cooked temperature, IMF and L*. Copyright (c) 2009 Elsevier Ltd. All rights reserved.

  2. Potential phosphorus eutrophication mitigation strategy: Biochar carbon composition, thermal stability and pH influence phosphorus sorption.

    PubMed

    Ngatia, L W; Hsieh, Y P; Nemours, D; Fu, R; Taylor, R W

    2017-08-01

    Phosphorus (P) eutrophication is a major pollution problem globally, with unprecedented amount of P emanating from agricultural sources. But little is known about the optimization of soil-biochar P sorption capacity. The study objective was to determine how biochar feedstocks and pyrolysis conditions influences carbon (C) thermal stability, C composition and pH and in turn influence the phosphorus sorption optimization. Biochar was produced from switchgrass, kudzu and Chinese tallow at 200, 300, 400, 500, 550, 650,750 °C. Carbon thermal stability was determined by multi-element scanning thermal analysis (MESTA), C composition was determined using solid state 13 C NMR. Phosphorus sorption was determined using a mixture of 10% biochar and 90% sandy soil after incubation. Results indicate increased P sorption (P < 0.0001) and decreased P availability (P < 0.0001) with increasing biochar pyrolysis temperature. However, optimum P sorption was feedstock specific with switchgrass indicating P desorption between 200 and 550 °C. Phosphorus sorption was in the order of kudzu > switchgrass > Chinese tallow. Total C, C thermal stability, aromatic C and alkalinity increased with elevated pyrolysis temperature. Biochar alkalinity favored P sorption. There was a positive relationship between high thermal stable C and P sorption for Kudzu (r = 0.62; P = 0.0346) and Chinese tallow (r = 0.73; P = 0.0138). In conclusion, biochar has potential for P eutrophication mitigation, however, optimum biochar pyrolysis temperature for P sorption is feedstock specific and in some cases might be out of 300-500 °C temperature range commonly used for agronomic application. High thermal stable C dominated by aromatic C and alkaline pH seem to favor P sorption. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Optimization of protein solution by a novel experimental design method using thermodynamic properties.

    PubMed

    Kim, Nam Ah; An, In Bok; Lee, Sang Yeol; Park, Eun-Seok; Jeong, Seong Hoon

    2012-09-01

    In this study, the structural stability of hen egg white lysozyme in solution at various pH levels and in different types of buffers, including acetate, phosphate, histidine, and Tris, was investigated by means of differential scanning calorimetry (DSC). Reasonable pH values were selected from the buffer ranges and were analyzed statistically through design of experiment (DoE). Four factors were used to characterize the thermograms: calorimetric enthalpy (ΔH), temperature at maximum heat flux (T( m )), van't Hoff enthalpy (ΔH( V )), and apparent activation energy of protein solution (E(app)). It was possible to calculate E(app) through mathematical elaboration from the Lumry-Eyring model by changing the scan rate. The transition temperature of protein solution, T( m ), increased when the scan rate was faster. When comparing the T( m ), ΔH( V ), ΔH, and E(app) of lysozyme in various pH ranges and buffers with different priorities, lysozyme in acetate buffer at pH 4.767 (scenario 9) to pH 4.969 (scenario 11) exhibited the highest thermodynamic stability. Through this experiment, we found a significant difference in the thermal stability of lysozyme in various pH ranges and buffers and also a new approach to investigate the physical stability of protein by DoE.

  4. Influence of pH and temperature of dip-coating solution on the properties of cellulose acetate-ceramic composite membrane for ultrafiltration.

    PubMed

    Kaur, Harjot; Bulasara, Vijaya Kumar; Gupta, Raj Kumar

    2018-09-01

    Polymer-ceramic composite membranes were prepared by dip coating technique using 5 wt.% cellulose acetate (CA) solution at different temperatures (15 °C, 25 °C and 40 °C). The effect of pH (2-12) of the polymeric solution on the properties of the membranes was studied using SEM, EDAX, FTIR, gas and liquid permeation. The thickness of the polymeric layer depended on the interaction of CA solution with the surface of ceramic support. Membrane permeability decreased with increase in pH because of decrease in pore size and porosity resulting from strong interaction of the polymer layer with the ceramic support. The porosity and mean pore size of the prepared membranes were found to be 28-60% and 30-47 nm (ultrafiltration range), respectively. The optimized membrane (pH 7) was used for ultrafiltration of oil in water emulsions (100 and 200 mg/L). Oil rejection of 99.61% was obtained for 100 mg/L of oil concentration in water. Copyright © 2018 Elsevier Ltd. All rights reserved.

  5. Purification and characterisation of a novel chitinase from persimmon (Diospyros kaki) with antifungal activity.

    PubMed

    Zhang, Jianzhi; Kopparapu, Narasimha Kumar; Yan, Qiaojuan; Yang, Shaoqing; Jiang, Zhengqiang

    2013-06-01

    A novel chitinase from the persimmon fruit was isolated, purified and characterised in this report. The Diospyros kaki chitinase (DKC) was found to be a monomer with a molecular mass of 29 kDa. It exhibited optimal activity at pH 4.5 with broad pH stability from pH 4.0-9.0. It has an optimal temperature of 60°C and thermostable up to 60°C when incubated for 30 min. The internal peptide sequences of DKC showed similarity with other reported plant chitinases. It has the ability to hydrolyse colloidal chitin into chito-oligomers such as chitotriose, chitobiose and into its monomer N-acetylglucosamine. It can be used to degrade chitin waste into useful products such as chito-oligosacchaarides. DKC exhibited antifungal activity towards pathogenic fungus Trichoderma viride. Chitinases with antifungal property can be used as biocontrol agents replacing chemical fungicides. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Statistical optimization for lipase production from solid waste of vegetable oil industry.

    PubMed

    Sahoo, Rajesh Kumar; Kumar, Mohit; Mohanty, Swati; Sawyer, Matthew; Rahman, Pattanathu K S M; Sukla, Lala Behari; Subudhi, Enketeswara

    2018-04-21

    The production of biofuel using thermostable bacterial lipase from hot spring bacteria out of low-cost agricultural residue olive oil cake is reported in the present paper. Using a lipase enzyme from Bacillus licheniformis, a 66.5% yield of methyl esters was obtained. Optimum parameters were determined, with maximum production of lipase at a pH of 8.2, temperature 50.8°C, moisture content of 55.7%, and biosurfactant content of 1.693 mg. The contour plots and 3D surface responses depict the significant interaction of pH and moisture content with biosurfactant during lipase production. Chromatographic analysis of the lipase transesterification product was methyl esters, from kitchen waste oil under optimized conditions, generated methyl palmitate, methyl stearate, methyl oleate, and methyl linoleate.

  7. Immobilization and stabilization of pectinase by multipoint attachment onto an activated agar-gel support.

    PubMed

    Li, Tuoping; Li, Suhong; Wang, Na; Tain, Lirui

    2008-08-15

    Pectinase was immobilized on an activated agar-gel support by multipoint attachment. The maximal activity of immobilized pectinase was obtained at 5°C, pH 3.6, with a 24h reaction time at an enzyme dose of 0.52mg protein/g gel, and the gel was activated with 1.0M glycidol. These conditions increased the thermal stability of the immobilized pectinase 19-fold compared with the free enzyme at 65°C. The optimal temperature for pectinase activity changed from 40 to 50°C after immobilization; however, the optimal pH remained unchanged. The immobilized enzyme also exhibited great operational stability, and an 81% residual activity was observed in the immobilized enzyme after 10 batch reactions. Copyright © 2008 Elsevier Ltd. All rights reserved.

  8. Organic solvent tolerance of an α-amylase from haloalkaliphilic bacteria as a function of pH, temperature, and salt concentrations.

    PubMed

    Pandey, Sandeep; Singh, S P

    2012-04-01

    A haloalkaliphilic bacterium was isolated from salt-enriched soil of Mithapur, Gujarat (India) and identified as Bacillus agaradhaerens Mi-10-6₂ based on 16S rRNA sequence analysis (NCBI gene bank accession, GQ121032). The bacterium was studied for its α-amylase characteristic in the presence of organic solvents. The enzyme was quite active and it retained considerable activity in 30% (v/v) organic solvents, dodecane, decane, heptane, n-hexane, methanol, and propanol. At lower concentrations of solvents, the catalysis was quite comparable to control. Enzyme catalysis at wide range of alkanes and alcohol was an interesting finding of the study. Mi-10-6₂ amylase retained activity over a broader alkaline pH range, with the optimal pH at 10-11. Two molars of salt was optimum for catalysis in the presence of most of the tested solvents, though the enzyme retained significant activity even at 4 M salt. With dodecane, the optimum temperature shifted from 50 °C to 60 °C, while the enzyme was active up to 80 °C. Over all, the present study focused on the effect of organic solvents on an extracellular α-amylase from haloalkaliphilic bacteria under varying conditions of pH, temperature, and salt.

  9. Production and Properties of a Thermostable, pH-Stable Exo-Polygalacturonase Using Aureobasidium pullulans Isolated from Saharan Soil of Algeria Grown on Tomato Pomace.

    PubMed

    Bennamoun, Leila; Hiligsmann, Serge; Dakhmouche, Scheherazad; Ait-Kaki, Amel; Labbani, Fatima-Zohra Kenza; Nouadri, Tahar; Meraihi, Zahia; Turchetti, Benedetta; Buzzini, Pietro; Thonart, Philippe

    2016-10-29

    Polygalacturonase is a valuable biocatalyst for several industrial applications. Production of polygalacturonase using the Aureobasidium pullulans stain isolated from Saharan soil of Algeria was investigated. Its capacity to produce polygalacturonase was assessed under submerged culture using tomato pomace as an abundant agro-industrial substrate. Optimization of the medium components, which enhance polygalacturonase activity of the strain Aureobasidium pullulans , was achieved with the aid of response surface methodology. The composition of the optimized medium was as follows: tomato pomace 40 g/L, lactose 1.84 g/L, CaCl₂0.09 g/L and pH 5.16. Practical validation of the optimum medium provided polygalacturonase activity of 22.05 U/mL, which was 5-fold higher than in unoptimized conditions. Batch cultivation in a 20 L bioreactor performed with the optimal nutrients and conditions resulted in a high polygalacturonase content (25.75 U/mL). The enzyme showed stability over a range of temperature (5-90 °C) with an optimum temperature of 60 °C with pH 5.0, exhibiting 100% residual activity after 1h at 60 °C. This enzyme was stable at a broad pH range (5.0-10). The enzyme proved to be an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of polygalacturonic acid. Moreover, the exo-polygalacturonase was able to enhance the clarification of both apple and citrus juice. As a result, an economical polygalacturonase production process was defined and proposed using an industrial food by-product.

  10. Production and Properties of a Thermostable, pH—Stable Exo-Polygalacturonase Using Aureobasidium pullulans Isolated from Saharan Soil of Algeria Grown on Tomato Pomace

    PubMed Central

    Bennamoun, Leila; Hiligsmann, Serge; Dakhmouche, Scheherazad; Ait-Kaki, Amel; Labbani, Fatima-Zohra Kenza; Nouadri, Tahar; Meraihi, Zahia; Turchetti, Benedetta; Buzzini, Pietro; Thonart, Philippe

    2016-01-01

    Polygalacturonase is a valuable biocatalyst for several industrial applications. Production of polygalacturonase using the Aureobasidium pullulans stain isolated from Saharan soil of Algeria was investigated. Its capacity to produce polygalacturonase was assessed under submerged culture using tomato pomace as an abundant agro-industrial substrate. Optimization of the medium components, which enhance polygalacturonase activity of the strain Aureobasidium pullulans, was achieved with the aid of response surface methodology. The composition of the optimized medium was as follows: tomato pomace 40 g/L, lactose 1.84 g/L, CaCl20.09 g/L and pH 5.16. Practical validation of the optimum medium provided polygalacturonase activity of 22.05 U/mL, which was 5-fold higher than in unoptimized conditions. Batch cultivation in a 20 L bioreactor performed with the optimal nutrients and conditions resulted in a high polygalacturonase content (25.75 U/mL). The enzyme showed stability over a range of temperature (5–90 °C) with an optimum temperature of 60 °C with pH 5.0, exhibiting 100% residual activity after 1h at 60 °C. This enzyme was stable at a broad pH range (5.0–10). The enzyme proved to be an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of polygalacturonic acid. Moreover, the exo-polygalacturonase was able to enhance the clarification of both apple and citrus juice. As a result, an economical polygalacturonase production process was defined and proposed using an industrial food by-product. PMID:28231166

  11. Anaerobic treatment of complex chemical wastewater in a sequencing batch biofilm reactor: process optimization and evaluation of factor interactions using the Taguchi dynamic DOE methodology.

    PubMed

    Venkata Mohan, S; Chandrasekhara Rao, N; Krishna Prasad, K; Murali Krishna, P; Sreenivas Rao, R; Sarma, P N

    2005-06-20

    The Taguchi robust experimental design (DOE) methodology has been applied on a dynamic anaerobic process treating complex wastewater by an anaerobic sequencing batch biofilm reactor (AnSBBR). For optimizing the process as well as to evaluate the influence of different factors on the process, the uncontrollable (noise) factors have been considered. The Taguchi methodology adopting dynamic approach is the first of its kind for studying anaerobic process evaluation and process optimization. The designed experimental methodology consisted of four phases--planning, conducting, analysis, and validation connected sequence-wise to achieve the overall optimization. In the experimental design, five controllable factors, i.e., organic loading rate (OLR), inlet pH, biodegradability (BOD/COD ratio), temperature, and sulfate concentration, along with the two uncontrollable (noise) factors, volatile fatty acids (VFA) and alkalinity at two levels were considered for optimization of the anae robic system. Thirty-two anaerobic experiments were conducted with a different combination of factors and the results obtained in terms of substrate degradation rates were processed in Qualitek-4 software to study the main effect of individual factors, interaction between the individual factors, and signal-to-noise (S/N) ratio analysis. Attempts were also made to achieve optimum conditions. Studies on the influence of individual factors on process performance revealed the intensive effect of OLR. In multiple factor interaction studies, biodegradability with other factors, such as temperature, pH, and sulfate have shown maximum influence over the process performance. The optimum conditions for the efficient performance of the anaerobic system in treating complex wastewater by considering dynamic (noise) factors obtained are higher organic loading rate of 3.5 Kg COD/m3 day, neutral pH with high biodegradability (BOD/COD ratio of 0.5), along with mesophilic temperature range (40 degrees C), and low sulfate concentration (700 mg/L). The optimization resulted in enhanced anaerobic performance (56.7%) from a substrate degradation rate (SDR) of 1.99 to 3.13 Kg COD/m3 day. Considering the obtained optimum factors, further validation experiments were carried out, which showed enhanced process performance (3.04 Kg COD/m3-day from 1.99 Kg COD/m3 day) accounting for 52.13% improvement with the optimized process conditions. The proposed method facilitated a systematic mathematical approach to understand the complex multi-species manifested anaerobic process treating complex chemical wastewater by considering the uncontrollable factors. Copyright (c) 2005 Wiley Periodicals, Inc.

  12. Injectable Chitosan/β-Glycerophosphate System for Sustained Release: Gelation Study, Structural Investigation, and Erosion Tests.

    PubMed

    Dalmoro, Annalisa; Abrami, Michela; Galzerano, Barbara; Bochicchio, Sabrina; Barba, Anna Angela; Grassi, Mario; Larobina, Domenico

    2017-01-01

    Hydrogels can constitute reliable delivery systems of drugs, including those based on nucleic acids (NABDs) such as small interfering ribonucleic acid (siRNA). Their nature, structure, and response to physiological or external stimuli strongly influence the delivery mechanisms of entrapped active molecules, and, in turn, their possible uses in pharmacological and biomedical applications. In this study, a thermo-gelling chitosan/β-glycero-phosphate system has been optimized in order to assess its use as injectable system able to: i) gelling at physiological pH and temperature, and ii) modulate the release of included active ingredients. To this aim, we first analyzed the effect of acetic acid concentration on the gelation temperature. We then found the "optimized composition", namely, the one in which the Tgel is equal to the physiological temperature. The resulting gel was tested, by low field nuclear magnetic resonance (LF-NMR), to evaluate its average mesh-size, which can affect release kinetics of loaded drug. Finally, films of gelled chitosan, loaded with a model drug, have been tested in vitro to monitor their characteristic times, i.e. diffusion and erosion time, when they are exposed to a medium mimicking a physiological environment (buffer solution at pH 7.4). Results display that the optimized system is deemed to be an ideal candidate as injectable gelling material for a sustained release. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  13. Optimization of Fermentation Conditions and Rheological Properties of Exopolysaccharide Produced by Deep-Sea Bacterium Zunongwangia profunda SM-A87

    PubMed Central

    Liu, Sheng-Bo; Qiao, Li-Ping; He, Hai-Lun; Zhang, Qian; Chen, Xiu-Lan; Zhou, Wei-Zhi; Zhou, Bai-Cheng; Zhang, Yu-Zhong

    2011-01-01

    Zunongwangia profunda SM-A87 isolated from deep-sea sediment can secrete large quantity of exopolysaccharide (EPS). Response surface methodology was applied to optimize the culture conditions for EPS production. Single-factor experiment showed that lactose was the best carbon source. Based on the Plackett–Burman design, lactose, peptone and temperature were selected as significant variables, which were further optimized by the steepest ascent (descent) method and central composite design. The optimal culture conditions for EPS production and broth viscosity were determined as 32.21 g/L lactose, 8.87 g/L peptone and an incubation temperature of 9.8°C. Under these conditions, the maximum EPS yield and broth viscosity were 8.90 g/L and 6551 mPa•s, respectively, which is the first report of such high yield of EPS from a marine bacterium. The aqueous solution of the EPS displayed high viscosity, interesting shearing thinning property and great tolerance to high temperature, a wide range of pH, and high salinity. PMID:22096500

  14. Addendum: Development of a preprototype times wastewater recovery subsystem

    NASA Technical Reports Server (NTRS)

    Dehner, G. F.

    1984-01-01

    The results of the second generation operational improvements and the TIMES (Thermoelectric Integrated Membrane Evaporation Subsystem) 2 study are covered. Areas covered in the second generation operational improvements are improved temperature control, water quality improvements, subsytem operational improvements, solid handling improvements, wastewater pretreatment optimization, and membrane rejuvenation concepts. The task for the TIMES 2 study are thermoelectric regenerator improvement, recycle loop pH operational criteria, recycle loop component optimization, and hollow fiber membrane evaporator improvement. Results are presented and conclusions are drawn from both studies.

  15. Remediation and recycling of WBP-treated lumber for use as flakeboard

    Treesearch

    Ronald Sabo; Jerrold E. Winandy; Carol A. Clausen; Altaf Basta

    2008-01-01

    Laboratory-scale experiments were conducted in which preservative metals (As, Cr, & Cu) were thermochemically extracted from CCA-treated spruce (Picea engelmannii) using oxalic acid and sodium hydroxide. The effects of extraction time, temperature, and pH were examined and laboratory scale optimization was achieved. Two series of experiments were carried out. In...

  16. Optimizing the synthesis conditions of silver nanoparticles using corn starch and their catalytic reduction of 4-nitrophenol

    NASA Astrophysics Data System (ADS)

    Salaheldin, Hosam I.

    2018-06-01

    In this study, silver nanoparticles (SNPs) were synthesised in an aqueous solution of corn starch. To fabricate the SNPs, reaction conditions, such as varying silver nitrate () concentration, time, temperature and solution pH of the reaction, were optimized. Since, the optimum reaction conditions were found 1 mmo l‑1, 15 min and , respectively. Then, to study the role of pH on SNP synthesis, varying pH values of the solution (3, 5, 7, 9 and 11) were investigated. Subsequently, the obtained silver/starch nanocomposites were characterised using different techniques. The x-ray diffraction (XRD) results revealed that the particles were face-centred cubic (FCC), and had an average particle size of 7.5 nm. This was confirmed by high-resolution transmission electron microscopy (HR-TEM) images. Moreover, the synthesised SNPs, at different pH values, were used as nanocatalyst for the reduction of 4-nitrophenol to 4-aminophenol in the presence of sodium borohydride. Under optimum reaction conditions, the higher catalytic activity was obtained with SNPs synthesised at pH 11 compared to lower pH of 7 or 9. Therefore, the rapid, reproducible, cost-effective silver/starch nanocomposite can be widely used for various applications such as drug manufacturing (e.g. analgesics and antipyretics) and the removal of pollutants from wastewater.

  17. Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests.

    PubMed

    Karam, Amanda L; McMillan, Catherine C; Lai, Yi-Chun; de Los Reyes, Francis L; Sederoff, Heike W; Grunden, Amy M; Ranjithan, Ranji S; Levis, James W; Ducoste, Joel J

    2017-06-14

    The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic performance of microalgae-based biofuel production. Models that estimate microalgal growth under different conditions can help to optimize PBR design and operation. To be effective, the growth parameters used in these models must be accurately determined. Algal growth experiments are often constrained by the dynamic nature of the culture environment, and control systems are needed to accurately determine the kinetic parameters. The first step in setting up a controlled batch experiment is live data acquisition and monitoring. This protocol outlines a process for the assembly and operation of a bench-scale photosynthetic bioreactor that can be used to conduct microalgal growth experiments. This protocol describes how to size and assemble a flat-plate, bench-scale PBR from acrylic. It also details how to configure a PBR with continuous pH, light, and temperature monitoring using a data acquisition and control unit, analog sensors, and open-source data acquisition software.

  18. Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests

    PubMed Central

    Karam, Amanda L.; McMillan, Catherine C.; Lai, Yi-Chun; de los Reyes, Francis L.; Sederoff, Heike W.; Grunden, Amy M.; Ranjithan, Ranji S.; Levis, James W.; Ducoste, Joel J.

    2017-01-01

    The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic performance of microalgae-based biofuel production. Models that estimate microalgal growth under different conditions can help to optimize PBR design and operation. To be effective, the growth parameters used in these models must be accurately determined. Algal growth experiments are often constrained by the dynamic nature of the culture environment, and control systems are needed to accurately determine the kinetic parameters. The first step in setting up a controlled batch experiment is live data acquisition and monitoring. This protocol outlines a process for the assembly and operation of a bench-scale photosynthetic bioreactor that can be used to conduct microalgal growth experiments. This protocol describes how to size and assemble a flat-plate, bench-scale PBR from acrylic. It also details how to configure a PBR with continuous pH, light, and temperature monitoring using a data acquisition and control unit, analog sensors, and open-source data acquisition software. PMID:28654054

  19. Production of cellulose by Aspergillus niger under submerged and solid state fermentation using coir waste as a substrate

    PubMed Central

    Mrudula, Soma; Murugammal, Rangasamy

    2011-01-01

    Aspergillus niger was used for cellulase production in submerged (SmF) and solid state fermentation (SSF). The maximum production of cellulase was obtained after 72 h of incubation in SSF and 96 h in Smf. The CMCase and FPase activities recorded in SSF were 8.89 and 3.56 U per g of dry mycelial bran (DBM), respectively. Where as in Smf the CMase & FPase activities were found to be 3.29 and 2.3 U per ml culture broth, respectively. The productivity of extracellular cellulase in SSF was 14.6 fold higher than in SmF. The physical and nutritional parameters of fermentation like pH, temperature, substrate, carbon and nitrogen sources were optimized. The optimal conditions for maximum biosynthesis of cellulase by A. niger were shown to be at pH 6, temperature 30 °C. The additives like lactose, peptone and coir waste as substrate increased the productivity both in SmF and SSF. The moisture ratio of 1:2 (w/v) was observed for optimum production of cellulase in SSF. PMID:24031730

  20. High-level expression of recombinant thermostable β-glucosidase in Escherichia coli by regulating acetic acid.

    PubMed

    Shi, Xuejia; Xie, Jingcong; Liao, Shiyong; Wu, Tao; Zhao, Lin-Guo; Ding, Gang; Wang, Zhenzhong; Xiao, Wei

    2017-10-01

    In the fermentation progress, fermentation parameters including the feed rate, induction temperature, and induction pH evidently regulate the accumulation of acetic acid generated by recombinant E. coli in the medium. The production of thermostable β-glucosidase (Tpebgl3) was increased by optimizing the parameters mentioned step by step. The optimal conditions were obtained with the highest enzyme expression (560.4U/mL) and the maximum DCW (65g/L) at the pre-induction specific growth rate of 0.2h -1 followed by a post-induction specific growth rate (0.18h -1 ); induction temperature is 39°C; the pH is 7.2; the concentration of acetic acid was maintained all along below 0.9g/L. Results show it is necessary for the synthesis of Tpebgl3 to regulate the accumulation of acetic acid at the premise of feeding to meet the normal growth of E. coli. The production of Tpebgl3 by recombinant E. coli is the highest reported to date. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Optimization of Bromelain-Aided Production of Angiotensin I-Converting Enzyme Inhibitory Hydrolysates from Stone Fish Using Response Surface Methodology.

    PubMed

    Muhammad Auwal, Shehu; Zarei, Mohammad; Abdul-Hamid, Azizah; Saari, Nazamid

    2017-03-31

    The stone fish is an under-utilized sea cucumber with many nutritional and ethno-medicinal values. This study aimed to establish the conditions for its optimum hydrolysis with bromelain to generate angiotensin I-converting enzyme (ACE)-inhibitory hydrolysates. Response surface methodology (RSM) based on a central composite design was used to model and optimize the degree of hydrolysis (DH) and ACE-inhibitory activity. Process conditions including pH (4-7), temperature (40-70 °C), enzyme/substrate (E/S) ratio (0.5%-2%) and time (30-360 min) were used. A pH of 7.0, temperature of 40 °C, E/S ratio of 2% and time of 240 min were determined using a response surface model as the optimum levels to obtain the maximum ACE-inhibitory activity of 84.26% at 44.59% degree of hydrolysis. Hence, RSM can serve as an effective approach in the design of experiments to improve the antihypertensive effect of stone fish hydrolysates, which can thus be used as a value-added ingredient for various applications in the functional foods industries.

  2. Extraction of Oil from Flaxseed (Linum usitatissimum L.) Using Enzyme-Assisted Three-Phase Partitioning.

    PubMed

    Tan, Zhi-Jian; Yang, Zi-Zhen; Yi, Yong-Jian; Wang, Hong-Ying; Zhou, Wan-Lai; Li, Fen-Fang; Wang, Chao-Yun

    2016-08-01

    In this study, enzyme-assisted three-phase partitioning (EATPP) was used to extract oil from flaxseed. The whole procedure is composed of two parts: the enzymolysis procedure in which the flaxseed was hydrolyzed using an enzyme solution (the influencing parameters such as the type and concentration of enzyme, temperature, and pH were optimized) and three-phase partitioning (TPP), which was conducted by adding salt and t-butanol to the crude flaxseed slurry, resulting in the extraction of flaxseed oil into alcohol-rich upper phase. The concentration of t-butanol, concentration of salt, and the temperature were optimized to maximize the extraction yield. Under optimized conditions of a 49.29 % t-butanol concentration, 30.43 % ammonium sulfate concentration, and 35 °C extraction temperature, a maximum extraction yield of 71.68 % was obtained. This simple and effective EATPP can be used to achieve high extraction yields and oil quality, and thus, it is potential for large-scale oil production.

  3. Pretreatment of forest residues of Douglas fir by wet explosion for enhanced enzymatic saccharification.

    PubMed

    Biswas, Rajib; Teller, Philip J; Ahring, Birgitte K

    2015-09-01

    The logging and lumbering industry in the Pacific Northwest region generates huge amount of forest residues, offering an inexpensive raw material for biorefineries. Wet explosion (WEx) pretreatment was applied to the recalcitrant biomass to optimize process conditions including temperature (170-190 °C), time (10-30 min), and oxygen loading (0.5-7.5% of DM) through an experimental design. Optimal pH for enzymatic hydrolysis of the optimized samples and a complete mass balance have been evaluated. Results indicated that cellulose digestibility improved in all conditions tested with maximum digestibility achieved at 190 °C, time 30 min, and oxygen loading of 7.5%. Glucose yield at optimal pH of 5.5 was 63.3% with an excellent recovery of cellulose and lignin of 99.9% and 96.3%, respectively. Hemicellulose sugars recovery for xylose and mannose was found to be 69.2% and 76.0%, respectively, indicating that WEx is capable of producing relative high sugar yield even from the recalcitrant forest residues. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Photonic crystal enhanced silicon cell based thermophotovoltaic systems

    DOE PAGES

    Yeng, Yi Xiang; Chan, Walker R.; Rinnerbauer, Veronika; ...

    2015-01-30

    We report the design, optimization, and experimental results of large area commercial silicon solar cell based thermophotovoltaic (TPV) energy conversion systems. Using global non-linear optimization tools, we demonstrate theoretically a maximum radiative heat-to-electricity efficiency of 6.4% and a corresponding output electrical power density of 0.39 W cm⁻² at temperature T = 1660 K when implementing both the optimized two-dimensional (2D) tantalum photonic crystal (PhC) selective emitter, and the optimized 1D tantalum pentoxide – silicon dioxide PhC cold-side selective filter. In addition, we have developed an experimental large area TPV test setup that enables accurate measurement of radiative heat-to-electricity efficiency formore » any emitter-filter-TPV cell combination of interest. In fact, the experimental results match extremely well with predictions of our numerical models. Our experimental setup achieved a maximum output electrical power density of 0.10W cm⁻² and radiative heat-to-electricity efficiency of 1.18% at T = 1380 K using commercial wafer size back-contacted silicon solar cells.« less

  5. Pretreatment of corn stover with diluted acetic acid for enhancement of acidogenic fermentation.

    PubMed

    Zhao, Xu; Wang, Lijuan; Lu, Xuebin; Zhang, Shuting

    2014-04-01

    A Box-Behnken design of response surface method was used to optimize acetic acid-catalyzed hydrothermal pretreatment of corn stover, in respect to acid concentration (0.05-0.25%), treatment time (5-15 min) and reaction temperature (180-210°C). Acidogenic fermentations with different initial pH and hydrolyzates were also measured to evaluate the optimal pretreatment conditions for maximizing acid production. The results showed that pretreatment with 0.25% acetic acid at 191°C for 7.74 min was found to be the most optimal condition for pretreatment of corn stover under which the production of acids can reach the highest level. Acidogenic fermentation with the hydrolyzate of pretreatment at the optimal condition at the initial pH=5 was shown to be butyric acid type fermentation, producing 21.84 g acetic acid, 7.246 g propionic acid, 9.170 butyric acid and 1.035 g isovaleric acid from 100g of corn stover in 900 g of water containing 2.25 g acetic acid. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Coagulation efficiency and flocs characteristics of recycling sludge during treatment of low temperature and micro-polluted water.

    PubMed

    Zhou, Zhiwei; Yang, Yanling; Li, Xing; Gao, Wei; Liang, Heng; Li, Guibai

    2012-01-01

    Drinking water treatment sludge, characterized as accumulated suspended solids and organic and inorganic matter, is produced in large quantities during the coagulation process. The proper disposal, regeneration or reuse of sludge is, therefore, a significant environmental issue. Reused sludge at low temperatures is an alternative method to enhance traditional coagulation efficiency. In the present study, the recycling mass of mixed sludge and properties of raw water (such as pH and turbidity) were systematically investigated to optimize coagulation efficiency. We determined that the appropriate dosage of mixed sludge was 60 mL/L, effective initial turbidity ranges were below 45.0 NTU, and optimal pH for DOMs and turbidity removal was 6.5-7.0 and 8.0, respectively. Furthermore, by comparing the flocs characteristics with and without recycling sludge, we found that floc structures with sludge were more irregular with average size growth to 64.7 microm from 48.1 microm. Recycling sludge was a feasible and successful method for enhancing pollutants removal, and the more irregular flocs structure after recycling might be caused by breakage of reused flocs and incorporation of powdered activated carbon into larger flocs structure. Applied during the coagulation process, recycling sludge could be significant for the treatment of low temperature and micro-polluted source water.

  7. Improvement of Functional Properties of Wheat Gluten Using Acid Protease from Aspergillus usamii

    PubMed Central

    Deng, Lingli; Wang, Zhaoxia; Yang, Sheng; Song, Junmei; Que, Fei; Zhang, Hui; Feng, Fengqin

    2016-01-01

    Hydrolysis parameters (temperature, E/S ratio, pH, and time) for acid protease (from Aspergillus usamii) hydrolysis of wheat gluten were optimized by response surface methodology (RSM) using emulsifying activity index (EAI) as the response factor. A temperature of 48.9°C, E/S ratio of 1.60%, pH 3.0, hydrolysis time of 2.5 h was found to be the optimum condition to obtain wheat gluten hydrolysate with higher EAI. The solubility of wheat gluten was greatly improved by hydrolysis and became independent of pH over the studied range. Enzymatic hydrolysis resulted in dramatically increase in EAI, water and oil holding capacity. Molecular weight distribution results showed that most of the peptides above 10 kDa have been hydrolyzed into smaller peptides. The results of FTIR spectra and disulfide bond (SS) and sulfhydryl (SH) content suggested that a more extensional conformation was formed after hydrolysis, which could account for the improved functional properties. PMID:27467884

  8. Improvement of Functional Properties of Wheat Gluten Using Acid Protease from Aspergillus usamii.

    PubMed

    Deng, Lingli; Wang, Zhaoxia; Yang, Sheng; Song, Junmei; Que, Fei; Zhang, Hui; Feng, Fengqin

    2016-01-01

    Hydrolysis parameters (temperature, E/S ratio, pH, and time) for acid protease (from Aspergillus usamii) hydrolysis of wheat gluten were optimized by response surface methodology (RSM) using emulsifying activity index (EAI) as the response factor. A temperature of 48.9°C, E/S ratio of 1.60%, pH 3.0, hydrolysis time of 2.5 h was found to be the optimum condition to obtain wheat gluten hydrolysate with higher EAI. The solubility of wheat gluten was greatly improved by hydrolysis and became independent of pH over the studied range. Enzymatic hydrolysis resulted in dramatically increase in EAI, water and oil holding capacity. Molecular weight distribution results showed that most of the peptides above 10 kDa have been hydrolyzed into smaller peptides. The results of FTIR spectra and disulfide bond (SS) and sulfhydryl (SH) content suggested that a more extensional conformation was formed after hydrolysis, which could account for the improved functional properties.

  9. Optimized conditions for chelation of yttrium-90-DOTA immunoconjugates.

    PubMed

    Kukis, D L; DeNardo, S J; DeNardo, G L; O'Donnell, R T; Meares, C F

    1998-12-01

    Radioimmunotherapy (RIT) with 90Y-labeled immunoconjugates has shown promise in clinical trials. The macrocyclic chelating agent 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA) binds 90Y with extraordinary stability, minimizing the toxicity of 90Y-DOTA immunoconjugates arising from loss of 90Y to bone. However, reported 90Y-DOTA immunoconjugate product yields have been typically only < or =50%. Improved yields are needed for RIT with 90Y-DOTA immunoconjugates to be practical. (S) 2-[p-(bromoacetamido)benzyl]-DOTA (BAD) was conjugated to the monoclonal antibody Lym-1 via 2-iminothiolane (2IT). The immunoconjugate product, 2IT-BAD-Lym-1, was labeled in excess yttrium in various buffers over a range of concentrations and pH. Kinetic studies were performed in selected buffers to estimate radiolabeling reaction times under prospective radiopharmacy labeling conditions. The effect of temperature on reaction kinetics was examined. Optimal radiolabeling conditions were identified and used in eight radiolabeling experiments with 2IT-BAD-Lym-1 and a second immunoconjugate, DOTA-peptide-chimeric L6, with 248-492 MBq (6.7-13.3 mCi) of 90Y. Ammonium acetate buffer (0.5 M) was associated with the highest uptake of yttrium. On the basis of kinetic data, the time required to chelate 94% of 90Y (four half-times) under prospective radiopharmacy labeling conditions in 0.5 M ammonium acetate was 17-148 min at pH 6.5, but it was only 1-10 min at pH 7.5. Raising the reaction temperature from 25 degrees C to 37 degrees C markedly increased the chelation rate. Optimal radiolabeling conditions were identified as: 30-min reaction time, 0.5 M ammonium acetate buffer, pH 7-7.5 and 37 degrees C. In eight labeling experiments under optimal conditions, a mean product yield (+/- s.d.) of 91%+/-8% was achieved, comparable to iodination yields. The specific activity of final products was 74-130 MBq (2.0-3.5 mCi) of 90Y per mg of monoclonal antibody. The immunoreactivity of 90Y-labeled immunoconjugates was 100%+/-11%. The optimization of 90Y-DOTA chelation conditions represents an important advance in 90Y RIT because it facilitates the dependable and cost-effective preparation of 90Y-DOTA pharmaceuticals.

  10. Polyphenol oxidase activity from three sicilian artichoke [ Cynara cardunculus L. Var. scolymus L. (Fiori)] cultivars: studies and technological application on minimally processed production.

    PubMed

    Todaro, Aldo; Peluso, Orazio; Catalano, Anna Eghle; Mauromicale, Giovanni; Spagna, Giovanni

    2010-02-10

    Several papers helped with the development of more methods to control browning, or study thermal polyphenol oxidase (PPO) inactivation, but did not provide any solutions to technological process problems and food process improvement. Artichokes [ Cynara cardunculus L. var. scolymus L. (Fiori)] are susceptible to browning; this alteration could affect and reduce the suitability for its use, fresh or processed. Within this study, the catecholase and cresolase activities of PPO from three different Sicilian artichokes cultivar were characterized with regard to substrate specificity and enzyme kinetics, optimum pH and temperature, temperature and pH stability, and inhibitor test; all of the results were used for technological purposes, particularly to optimize minimally processed productions (ready-to-eat and cook-chilled artichokes).

  11. Biosynthesis of inulin from sucrose using inulosucrase from Lactobacillus gasseri DSM 20604.

    PubMed

    Ni, Dawei; Zhu, Yingying; Xu, Wei; Bai, Yuxiang; Zhang, Tao; Mu, Wanmeng

    2018-04-01

    Inulin is composed of fructose residues connected by β-(2, 1) glycosidic linkages with many promising physiochemical and physiological properties. In this study, an inulin-producing inulosucrase gene from Lactobacillus gasseri DSM 20604 was cloned, expressed and purified. SDS-PAGE and gel filtration found that the recombinant inulosucrase is a monomeric protein with a molecular weight of 63KDa. The optimal pH for its sucrose hydrolysis and transfructosylation activities was pH 5.5. The optimal temperatures were measured to be 45, 25, and 35°C for sucrose hydrolysis, transfructosylation, and total activity, respectively. Biosynthesis studies showed that the optimal enzyme dosage was 4.5U/g sucrose. Higher sucrose concentrations immensely contributed to inulin biosynthesis; the inulin yield reached its maximum after 1.5h of reaction. Structural analyses of the polysaccharide produced by the recombinant enzyme from sucrose revealed that it is an inulin-type fructan with a molecular weight of 5.858×10 6 Da. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Rapid Synthesis of Silver Nanoparticles from Fusarium oxysporum by Optimizing Physicocultural Conditions

    PubMed Central

    Birla, Sonal S.; Gaikwad, Swapnil C.; Gade, Aniket K.; Rai, Mahendra K.

    2013-01-01

    Synthesis of silver nanoparticles (SNPs) by fungi is emerging as an important branch of nanotechnology due to its ecofriendly, safe, and cost-effective nature. In order to increase the yield of biosynthesized SNPs of desired shape and size, it is necessary to control the cultural and physical parameters during the synthesis. We report optimum synthesis of SNPs on malt extract glucose yeast extract peptone (MGYP) medium at pH 9–11, 40–60°C, and 190.7 Lux and in sun light. The salt concentrations, volume of filtrate and biomass quantity were found to be directly proportional to the yield. The optimized conditions for the stable and rapid synthesis will help in large scale synthesis of monodispersed SNPs. The main aim of the present study was to optimize different media, temperature, pH, light intensity, salt concentration, volume of filtrate, and biomass quantity for the synthesis of SNPs by Fusarium oxysporum. PMID:24222751

  13. [Optimization of expression conditions and activity identification of hepatocyte-targeting peptide-human endostatin].

    PubMed

    Ma, Yan; Li, Wei; Li, Xiaobo; Bao, Dongmei; Lu, Jianpei

    2016-12-25

    To obtain sufficient purified and active fusion protein-hepatocyte-targeting peptide-human endostatin (HTP-rES), we studied the growth curve and the optimal induction timing of BL21/pET21b-HTP-rES. Different conditions of pH value, induction time, induction concentration and induction temperature were optimized by univariate analysis. After washing, refolding and purifying, the activity of fusion protein was identified by flow cytometry and 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT). Results show that the logarithmic growth phase of BL21/pET21b-HTP-rES was from 1.5 h to 3.5 h, the optimum expression conditions were pH 8.0, 0.06 mmol/L IPTG, at 42 ℃ for 5 h. The purity of inclusion bodies was up to 60% after washing. The purity of target protein was more than 95% after refolding and purification. Our findings provide the foundation for further biological activity and drug development.

  14. Culture conditions and medium components for the production of mycelial biomass and exo-polysaccharides with Paecilomyces japonica in liquid culture.

    PubMed

    Lee, Jong Seok; Jung, Woo Chul; Park, Seok Jae; Lee, Keun Eok; Shin, Won Cheol; Hong, Eock Kee

    2013-04-01

    In this study, the liquid culture conditions were optimized for maximal production of mycelial biomass and exo-polysaccharide by Paecilomyces japonica. The effects of medium composition, C/N ratio and physical parameters were investigated. From these experiments, 30 g glucose, 20 g yeast extract, 0.5 g KH2PO4, and 0.1 g CuCl2 2H2O in 1-l distilled water were found to be the most suitable carbon, nitrogen, and mineral sources, respectively. The optimal temperature, initial pH, agitation, and aeration were determined to be 27°C, uncontrolled pH, 400 rpm, and 1.0 vvm, respectively. Under these optimal conditions, the maximum mycelial growth and polysaccharides production were 23.1 g/l and 2.5 g/l, respectively. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  15. Isolation, identification, and culture optimization of a novel glycinonitrile-hydrolyzing fungus-Fusarium oxysporum H3.

    PubMed

    Gong, Jin-Song; Lu, Zhen-Ming; Shi, Jing-Song; Dou, Wen-Fang; Xu, Hong-Yu; Zhou, Zhe-Min; Xu, Zheng-Hong

    2011-10-01

    Microbial transformation of glycinonitrile into glycine by nitrile hydrolase is of considerable interest to green chemistry. A novel fungus with high nitrile hydrolase was newly isolated from soil samples and identified as Fusarium oxysporum H3 through 18S ribosomal DNA, 28S ribosomal DNA, and the internal transcribed spacer sequence analysis, together with morphology characteristics. After primary optimization of culture conditions including pH, temperature, carbon/nitrogen sources, inducers, and metal ions, the enzyme activity was greatly increased from 326 to 4,313 U/L. The preferred carbon/nitrogen sources, inducer, and metal ions were glucose and yeast extract, caprolactam, and Cu(2+), Mn(2+), and Fe(2+), respectively. The maximum enzyme formation was obtained when F. oxysporum H3 was cultivated at 30 °C for 54 h with the initial pH of 7.2. There is scanty report about the optimization of nitrile hydrolase production from nitrile-converting fungus.

  16. Molecular Identification of a Newly Isolated Bacillus subtilis BI19 and Optimization of Production Conditions for Enhanced Production of Extracellular Amylase

    PubMed Central

    Dash, Biplab Kumar; Rahman, M. Mizanur; Sarker, Palash Kumar

    2015-01-01

    A study was carried out with a newly isolated bacterial strain yielding extracellular amylase. The phylogenetic tree constructed on the basis of 16S rDNA gene sequences revealed this strain as clustered with the closest members of Bacillus sp. and identified as Bacillus subtilis BI19. The effect of various fermentation conditions on amylase production through shake-flask culture was investigated. Rice flour (1.25%) as a cheap natural carbon source was found to induce amylase production mostly. A combination of peptone and tryptone as organic and ammonium sulfate as inorganic nitrogen sources gave highest yield. Maximum production was obtained after 24 h of incubation at 37°C with an initial medium pH 8.0. Addition of surfactants like Tween 80 (0.25 g/L) and sodium lauryl sulfate (0.2 g/L) resulted in 28% and 15% increase in enzyme production, respectively. Amylase production was 3.06 times higher when optimized production conditions were used. Optimum reaction temperature and pH for crude amylase activity were 50°C and 6.0, respectively. The crude enzyme showed activity and stability over a fair range of temperature and pH. These results suggest that B. subtilis BI19 could be exploited for production of amylase at relatively low cost and time. PMID:26180814

  17. Purification and characterization of polyphenol oxidase from rape flower.

    PubMed

    Sun, Han-Ju; Wang, Jing; Tao, Xue-Ming; Shi, Juan; Huang, Mei-Ying; Chen, Zhe

    2012-01-25

    The purification and partial enzymology characteristics of polyphenol oxidase (PPO) from rape flower were studied. After preliminary treatments, the crude enzyme solution was in turn purified with ammonium sulfate, dialysis, and Sephadex G-75 gel chromatography. The optimal conditions and stability of PPO were examined at different pH values and temperatures. Subsequently, PPO was also characterized by substrate (catechol) concentrations, inhibitors, kinetic parameters, and molecular weight. Results showed that the optimal pH for PPO activity was 5.5 in the presence of catechol and that PPO was relatively stable at pH 3.5-5.5. PPO was moderately stable at temperatures from 60 to 70 °C, whereas it was easily denatured at 80-90 °C. Ethylenediaminetetraacetic acid, sodium chloride, and calcium chloride had little inhibitive effects on PPO, whereas citric acid, sodium sulfite, and ascorbic acid had strongly inhibitive effects. The Michaelis-Menten constant (K(m)) and maximal reaction velocity (V(max)) of PPO were 0.767 mol/L and 0.519 Ab/min/mL of the crude PPO solution, respectively. PPO was finally purified to homogeneity with a purification factor of 4.41-fold and a recovery of 12.41%. Its molecular weight was 60.4 kDa, indicating that the PPO is a dimer. The data obtained in this research may help to prevent the enzymatic browning of rape flower during its storage and processing.

  18. Enhanced stability and permeation potential of nanoemulsion containing sefsol-218 oil for topical delivery of amphotericin B.

    PubMed

    Hussain, Afzal; Samad, Abdus; Singh, Sandeep Kumar; Ahsan, Mohd Neyaz; Faruk, Abdul; Ahmed, Farhan Jalees

    2015-05-01

    To characterize the enhanced stability and permeation potential of amphotericin B nanoemulsion comprising sefsol-218 oil at varying pH and temperature of aqueous continuous phase. Several batches of amphotericin B loaded nanoemulsion were prepared and evaluated for their physical and chemical stability at different pH and temperature. Also, a comparative study of ex vivo drug permeation across the albino rat skin was investigated with commercial Fungisome® and drug solution at 37 °C for 24 h. The extent of drug penetrated through the rat skin was thereby evaluated using the confocal laser scanning microscopy (CLSM). The optimized nanoemulsion demonstrated the highest flux rate 17.85 ± 0.5 µg/cm(2)/h than drug solution (5.37 ± 0.01 µg/cm(2)/h) and Fungisome® (7.97 ± 0.01 µg/cm(2)/h). Ex vivo drug penetration mechanism from the developed formulations at pH 6.8 and pH 7.4 of aqueous phase pH using the CLSM revealed enhanced penetration. Ex vivo drug penetration studies of developed formulation comprising of CLSM revealed enhanced penetration in aqueous phase at pH 6.8 and 7.4. The aggregation behavior of nanoemulsion at both the pH was found to be minimum and non-nephrotoxic. The stability of amphotericin B was obtained in terms of pH, optical density, globular size, polydispersity index and zeta potential value at different temperature for 90 days. The slowest drug degradation was observed in aqueous phase at pH 7.4 with shelf life 20.03-folds higher when stored at 4 °C (3.8 years) and 5-fold higher at 25 °C (0.951 years) than at 40 °C. The combined results suggested that nanoemulsion may hold an alternative for enhanced and sustained topical delivery system for amphotericin B.

  19. Multivariate optimization for the simultaneous bioremoval of BTEX and chlorinated aliphatic hydrocarbons by Pseudomonas plecoglossicida.

    PubMed

    Li, Junhui; de Toledo, Renata Alves; Shim, Hojae

    2017-01-05

    This study aimed to evaluate the effects of some major parameters on the cometabolic removal of cis-1,2-dichloroethylene (cis-DCE) and trichloroethylene (TCE), mixed with benzene, toluene, ethylbenzene, and xylenes, by an indigenous bacterial isolate Pseudomonas plecoglossicida. Such statistical methodologies as hierarchical cluster analysis heat map and principal component analysis were applied to better evaluate the effects of major parameters (soil pH, temperature, moisture, and cis-DCE/TCE concentrations) on the biological process. The bioremoval experiments were carried out in microcosms containing soil slurry, and the headspace concentrations of contaminants were analyzed by gas chromatography. The optimal bioremoval conditions for the mixture were soil water content >110%, pH 8-9, and temperature 15-20°C, while the cis-DCE/TCE concentration did not significantly affect the mixture bioremoval within the tested range (∼10mg per kg soil). Under the optimal conditions, benzene (97.7%), toluene (96.3%), and ethylbenzene (89.8%) were almost completely removed, while cis-DCE (24.5%), TCE (29.0%), m,p-xylene (36.3%), and o-xylene (29.6%) showed lower removal efficiencies. The obtained results would help to better design a remediation technology to be applied to the sites contaminated with mixed wastes, and the statistical methodologies used in this study appear to be very efficient and could serve as a template for optimization. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Cloning, Overexpression, and Characterization of a Metagenome-Derived Phytase with Optimal Activity at Low pH.

    PubMed

    Tan, Hao; Wu, Xiang; Xie, Liyuan; Huang, Zhongqian; Gan, Bingcheng; Peng, Weihong

    2015-06-01

    A phytase gene was identified in a publicly available metagenome derived from subsurface groundwater, which was deduced to encode for a protein of the histidine acid phosphatase (HAP) family. The nucleotide sequence of the phytase gene was chemically synthesized and cloned, in order to further overexpress the phytase in Escherichia coli. Purified protein of the recombinant phytase demonstrated an activity for phytic acid of 298 ± 17 μmol P/min/mg, at the pH optimum of 2.0 with the temperature of 37 °C. Interestingly, the pH optimum of this phytase is much lower in comparison with most HAP phytases known to date. It suggests that the phytase could possess improved adaptability to the low pH condition caused by the gastric acid in livestock and poultry stomachs.

  1. Optimized Production of Xylitol from Xylose Using a Hyper-Acidophilic Candida tropicalis.

    PubMed

    Tamburini, Elena; Costa, Stefania; Marchetti, Maria Gabriella; Pedrini, Paola

    2015-08-19

    The yeast Candida tropicalis DSM 7524 produces xylitol, a natural, low-calorie sweetener, by fermentation of xylose. In order to increase xylitol production rate during the submerged fermentation process, some parameters-substrate (xylose) concentration, pH, aeration rate, temperature and fermentation strategy-have been optimized. The maximum xylitol yield reached at 60-80 g/L initial xylose concentration, pH 5.5 at 37 °C was 83.66% (w/w) on consumed xylose in microaerophilic conditions (kLa = 2·h(-1)). Scaling up on 3 L fermenter, with a fed-batch strategy, the best xylitol yield was 86.84% (w/w), against a 90% of theoretical yield. The hyper-acidophilic behaviour of C. tropicalis makes this strain particularly promising for industrial application, due to the possibility to work in non-sterile conditions.

  2. Optimized Production of Xylitol from Xylose Using a Hyper-Acidophilic Candida tropicalis

    PubMed Central

    Tamburini, Elena; Costa, Stefania; Marchetti, Maria Gabriella; Pedrini, Paola

    2015-01-01

    The yeast Candida tropicalis DSM 7524 produces xylitol, a natural, low-calorie sweetener, by fermentation of xylose. In order to increase xylitol production rate during the submerged fermentation process, some parameters-substrate (xylose) concentration, pH, aeration rate, temperature and fermentation strategy-have been optimized. The maximum xylitol yield reached at 60–80 g/L initial xylose concentration, pH 5.5 at 37 °C was 83.66% (w/w) on consumed xylose in microaerophilic conditions (kLa = 2·h−1). Scaling up on 3 L fermenter, with a fed-batch strategy, the best xylitol yield was 86.84% (w/w), against a 90% of theoretical yield. The hyper-acidophilic behaviour of C. tropicalis makes this strain particularly promising for industrial application, due to the possibility to work in non-sterile conditions. PMID:26295411

  3. Evaluation of the effects of various culture condition on Cr (VI)reduction by Shewanella oneidensis MR-1 in a novel high-throughputmini-bioreactor

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tang, Yinjie J.; Laidlaw, David; Gani, Kishen

    2006-03-16

    The growth and Cr(VI) reduction by Shewanella oneidensisMR-1 was examined using a mini-bioreactor system that independentlymonitors and controls pH, dissolved oxygen, and temperature for each ofits 24, 10-mL reactors. Independent monitoring and control of eachreactor in the cassette allows the exploration of a matrix ofenvironmental conditions known to influence S. oneidensis chromiumreduction. S. oneidensis MR-1 grew in minimal medium without amino acidor vitamin supplementation under aerobic conditions but required serineand glycine supplementation under anaerobic conditions. Growth wasinhibited by dissolved oxygen concentrations>80 percent. Lactatetransformation to acetate was enhanced by low concentration of dissolvedoxygen during the logarithmic growth phase. Between 11 andmore » 35oC, thegrowth rate obeyed the Arrhenius reaction rate-temperature relationship,with a maximum growth rate occurring at 35oC. S. oneidensis MR-1 was ableto grow over a wide range of pH (6-9). At neutral pH and temperaturesranging from 30-35oC, S. oneidensis MR-1 reduced 100 mu M Cr(VI) toCr(III) within 20 minutes in the exponential growth phase, and the growthrate was not affected by the addition of chromate; it reduced chromateeven faster at temperatures between 35 and 39oC. At low temperatures(<25oC), acidic (pH<6.5), or alkaline (pH>8.5) conditions, 100mu M Cr(VI) strongly inhibited growth and chromate reduction. Themini-bioreactor system enabled the rapid determination of theseparameters reproducibly and easily by performing very few experiments.Besides its use for examining parameters of interest to environmentalremediation, the device will also allow one to quickly assess parametersfor optimal production of recombinant proteins or secondarymetabolites« less

  4. Optimal production of 7,10-dihydroxy-8(E)-hexadecenoic acid from palmitoleic acid by Pseudomonas aeruginosa PR3.

    PubMed

    Bae, Jae-Han; Suh, Min-Jung; Kim, Beom-Soo; Hou, Ching T; Lee, In-Jung; Kim, In-Hwan; Kim, Hak-Ryul

    2010-09-30

    The hydroxylation of unsaturated fatty acids by bacterial strains is one type of value-adding bioconversion processes. This process generates new hydroxy fatty acids (HFA) carrying special properties such as higher viscosity and reactivity compared with normal fatty acids. Among microbial strains tested for HFA production, Pseudomonas aeruginosa PR3 is well known to utilize various unsaturated fatty acids to produce mono-, di- and tri-hydroxy fatty acids. Previously we reported that strain PR3 could produce a novel value-added hydroxy fatty acid 7,10-dihydroxy-8(E)-hexadecenoic acid (DHD) from palmitoleic acid (Bae et al. (2007) Appl. Microbiol. Biotechnol. 75, 435-440). In this study, we focused on the development of the optimal nutritional and environmental conditions for DHD production from palmitoleic acid by PR3. Optimal carbon and nitrogen sources for DHD production were fructose and yeast extract, respectively. Optimal initial medium pH and incubation temperature were pH 8.0 and 30 degrees C and magnesium ion was essentially required for DHD production. Substrate concentration and time of substrate addition were also optimized. Under optimized conditions, maximal DHD production was 1600mg/l representing 26.7% conversion yield. Copyright 2009 Elsevier B.V. All rights reserved.

  5. Function of a recombinant Chitinase derived from a virulent Aeromonas hydrophila isolated from diseased channel catfish

    USDA-ARS?s Scientific Manuscript database

    A chitinase was identified in extracellular products of a virulent Aeromonas hydrophila isolated from diseased channel catfish (Ictalurus punctatus). Bioactive recombinant chitinase (rChi-Ah) was produced in Escherichia coli. Purified rChi-Ah had optimal activity at temperature of 42°C and pH 6.5. T...

  6. Investigation on mercury reemission from limestone-gypsum wet flue gas desulfurization slurry.

    PubMed

    Chen, Chuanmin; Liu, Songtao; Gao, Yang; Liu, Yongchao

    2014-01-01

    Secondary atmospheric pollutions may result from wet flue gas desulfurization (WFGD) systems caused by the reduction of Hg(2+) to Hg(0) and lead to a damping of the cobenefit mercury removal efficiency by WFGD systems. The experiment on Hg(0) reemission from limestone-gypsum WFGD slurry was carried out by changing the operating conditions such as the pH, temperature, Cl(-) concentrations, and oxygen concentrations. The partitioning behavior of mercury in the solid and liquid byproducts was also discussed. The experimental results indicated that the Hg(0) reemission rate from WFGD slurry increased as the operational temperatures and pH values increased. The Hg(0) reemission rates decreased as the O2 concentration of flue gas and Cl(-) concentration of WFGD slurry increased. The concentrations of O2 in flue gas have an evident effect on the mercury retention in the solid byproducts. The temperature and Cl(-) concentration have a slight effect on the mercury partitioning in the byproducts. No evident relation was found between mercury retention in the solid byproducts and the pH. The present findings could be valuable for industrial application of characterizing and optimizing mercury control in wet FGD systems.

  7. Investigation on Mercury Reemission from Limestone-Gypsum Wet Flue Gas Desulfurization Slurry

    PubMed Central

    Liu, Songtao; Liu, Yongchao

    2014-01-01

    Secondary atmospheric pollutions may result from wet flue gas desulfurization (WFGD) systems caused by the reduction of Hg2+ to Hg0 and lead to a damping of the cobenefit mercury removal efficiency by WFGD systems. The experiment on Hg0 reemission from limestone-gypsum WFGD slurry was carried out by changing the operating conditions such as the pH, temperature, Cl− concentrations, and oxygen concentrations. The partitioning behavior of mercury in the solid and liquid byproducts was also discussed. The experimental results indicated that the Hg0 reemission rate from WFGD slurry increased as the operational temperatures and pH values increased. The Hg0 reemission rates decreased as the O2 concentration of flue gas and Cl− concentration of WFGD slurry increased. The concentrations of O2 in flue gas have an evident effect on the mercury retention in the solid byproducts. The temperature and Cl− concentration have a slight effect on the mercury partitioning in the byproducts. No evident relation was found between mercury retention in the solid byproducts and the pH. The present findings could be valuable for industrial application of characterizing and optimizing mercury control in wet FGD systems. PMID:24737981

  8. Preparation and Properties of Melamine Urea-Formaldehyde Microcapsules for Self-Healing of Cementitious Materials

    PubMed Central

    Li, Wenting; Zhu, Xujing; Zhao, Nan; Jiang, Zhengwu

    2016-01-01

    Self-healing microcapsules were synthesized by in situ polymerization with a melamine urea-formaldehyde resin shell and an epoxy resin adhesive. The effects of the key factors, i.e., core–wall ratio, reaction temperature, pH and stirring rate, were investigated by characterizing microcapsule morphology, shell thickness, particle size distribution, mechanical properties and chemical nature. Microcapsule healing mechanisms in cement paste were evaluated based on recovery strength and healing microstructure. The results showed that the encapsulation ability, the elasticity modulus and hardness of the capsule increased with an increase of the proportion of shell material. Increased polymerization temperatures were beneficial to the higher degree of shell condensation polymerization, higher resin particles deposition on microcapsule surfaces and enhanced mechanical properties. For relatively low pH values, the less porous three-dimensional structure led to the increased elastic modulus of shell and the more stable chemical structure. Optimized microcapsules were produced at a temperature of 60 °C, a core-wall ratio of 1:1, at pH 2~3 and at a stirring rate of 300~400 r/min. The best strength restoration was observed in the cement paste pre-damaged by 30% fmax and incorporating 4 wt % of capsules. PMID:28773280

  9. A novel cold-adapted and glucose-tolerant GH1 β-glucosidase from Exiguobacterium antarcticum B7.

    PubMed

    Crespim, Elaine; Zanphorlin, Letícia M; de Souza, Flavio H M; Diogo, José A; Gazolla, Alex C; Machado, Carla B; Figueiredo, Fernanda; Sousa, Amanda S; Nóbrega, Felipe; Pellizari, Vivian H; Murakami, Mário T; Ruller, Roberto

    2016-01-01

    A novel GH1 β-glucosidase (EaBgl1A) from a bacterium isolated from Antarctica soil samples was recombinantly overexpressed in Escherichia coli cells and characterized. The enzyme showed unusual pH dependence with maximum activity at neutral pH and retention of high catalytic activity in the pH range 6 to 9, indicating a catalytic machinery compatible with alkaline conditions. EaBgl1A is also a cold-adapted enzyme, exhibiting activity in the temperature range from 10 to 40°C with optimal activity at 30°C, which allows its application in industrial processes using low temperatures. Kinetic characterization revealed an enzymatic turnover (Kcat) of 6.92s(-1) (cellobiose) and 32.98s(-1) (pNPG) and a high tolerance for product inhibition, which is an extremely desirable feature for biotechnological purposes. Interestingly, the enzyme was stimulated by up to 200 mM glucose, whereas the commercial cocktails tested were found fully inhibited at this concentration. These properties indicate EaBgl1A as a promising biocatalyst for biotechnological applications where low temperatures are required. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Isolation and identification of Trichoderma harzianum from groundwater: An effective biosorbent for defluoridation of groundwater.

    PubMed

    Koshle, Shalini; Mahesh, S; Swamy, S Nanjunda

    2016-01-01

    The ability of non-viable form of Trichoderma harzianum, isolated from fluoride rich groundwater, was investigated as biosorbent for defluoridation of groundwater. Biosorption experiments were carried out at laboratory scale for removal of fluoride from groundwater. Significant effect of operational parameters on fluoride biosorption using Trichoderma harzianum as biosorbent was evaluated by varying operational parameters such as: initial fluoride concentration (2-8 mgl(-1)), biosorbent dose (0.4-1.6g/100ml), groundwater pH (6-10), temperature (30-50 degrees C) and biosorption time (30-120 min). The fluoride adsorption isotherms were modeled by Langmuir and Freundlich isotherms. Our result showed that fluoride biosorption, significantly increased with increase in groundwater pH, biosorbent dose, temperature and biosorption time, whereas increase in initial fluoride concentration reduced fluoride removal. The fluoride biosorption was rapid and maximum fluoride uptake was attained with 1.6g 100ml(-1) biosorbent within 60 min. Optimal pH 10 and temperature 50 degrees C gave maximum defluoridation efficiency. Freundlich isotherm fits well for defluoridation of groundwater using Trichoderma harzianum as biosorbent which indicated that biosorbent surface sites were heterogeneous in nature and fitted into heterogeneous site binding model.

  11. New Optical Sensing Materials for Application in Marine Research

    NASA Astrophysics Data System (ADS)

    Borisov, S.; Klimant, I.

    2012-04-01

    Optical chemosensors are versatile analytical tools which find application in numerous fields of science and technology. They proved to be a promising alternative to electrochemical methods and are applied increasingly often in marine research. However, not all state-of-the- art optical chemosensors are suitable for these demanding applications since they do not fully fulfil the requirements of high luminescence brightness, high chemical- and photochemical stability or their spectral properties are not adequate. Therefore, development of new advanced sensing materials is still of utmost importance. Here we present a set of novel optical sensing materials recently developed in the Institute of Analytical Chemistry and Food Chemistry which are optimized for marine applications. Particularly, we present new NIR indicators and sensors for oxygen and pH which feature high brightness and low level of autofluorescence. The oxygen sensors rely on highly photostable metal complexes of benzoporphyrins and azabenzoporphyrins and enable several important applications such as simultaneous monitoring of oxygen and chlorophyll or ultra-fast oxygen monitoring (Eddy correlation). We also developed ulta-sensitive oxygen optodes which enable monitoring in nM range and are primary designed for investigation of oxygen minimum zones. The dynamic range of our new NIR pH indicators based on aza-BODIPY dyes is optimized for the marine environment. A highly sensitive NIR luminescent phosphor (chromium(III) doped yttrium aluminium borate) can be used for non-invasive temperature measurements. Notably, the oxygen, pH sensors and temperature sensors are fully compatible with the commercially available fiber-optic readers (Firesting from PyroScience). An optical CO2 sensor for marine applications employs novel diketopyrrolopyrrol indicators and enables ratiometric imaging using a CCD camera. Oxygen, pH and temperature sensors suitable for lifetime and ratiometric imaging of analytes distribution are also realized. To enable versatility of applications we also obtained a range of nano- and microparticles suitable for intra- and extracellular imaging of the above analytes. Bright ratiometric 2-photon-excitable probes were also developed. Magnetic microparticles are demonstrated to be very promising tools for imaging of oxygen, temperature and other parameters in biofilms, corals etc. since they combine the sensing function with the possibility of external manipulation.

  12. The acid-tolerant L-arabinose isomerase from the mesophilic Shewanella sp. ANA-3 is highly active at low temperatures

    PubMed Central

    2011-01-01

    Background L-arabinose isomerases catalyse the isomerization of L-arabinose into L-ribulose at insight biological systems. At industrial scale of this enzyme is used for the bioconversion of D-galactose into D-tagatose which has many applications in pharmaceutical and agro-food industries. The isomerization reaction is thermodynamically equilibrated, and therefore the bioconversion rates is shifted towards tagatose when the temperature is increased. Moreover, to prevent secondary reactions it will be of interest to operate at low pH. The profitability of this D-tagatose production process is mainly related to the use of lactose as cheaper raw material. In many dairy products it will be interesting to produce D-tagatose during storage. This requires an efficient L-arabinose isomerase acting at low temperature and pH values. Results The gene encoding the L-arabinose isomerase from Shewanella sp. ANA-3 was cloned and overexpressed in Escherichia coli. The purified protein has a tetrameric arrangement composed by four identical 55 kDa subunits. The biochemical characterization of this enzyme showed that it was distinguishable by its maximal activity at low temperatures comprised between 15-35°C. Interestingly, this biocatalyst preserves more than 85% of its activity in a broad range of temperatures from 4.0 to 45°C. Shewanella sp. ANA-3 L-arabinose isomerase was also optimally active at pH 5.5-6.5 and maintained over 80% of its activity at large pH values from 4.0 to 8.5. Furthermore, this enzyme exhibited a weak requirement for metallic ions for its activity evaluated at 0.6 mM Mn2+. Stability studies showed that this protein is highly stable mainly at low temperature and pH values. Remarkably, T268K mutation clearly enhances the enzyme stability at low pH values. Use of this L-arabinose isomerase for D-tagatose production allows the achievement of attractive bioconversion rates of 16% at 4°C and 34% at 35°C. Conclusions Here we reported the purification and the biochemical characterization of the novel Shewanella sp. ANA-3 L-arabinose isomerase. Determination of the biochemical properties demonstrated that this enzyme was highly active at low temperatures. The generated T268K mutant displays an increase of the enzyme stability essentially at low pH. These features seem to be very attractive for the bioconversion of D-galactose into D-tagatose at low temperature which is very interesting from industrial point of view. PMID:22074172

  13. Carrier free immobilization and characterization of trypsin.

    PubMed

    Menfaatli, Esra; Zihnioglu, Figen

    2015-04-01

    Pancreatic trypsin was immobilized by cross-linked enzyme aggregates (CLEA) which is a carrier free immobilization method. Ammonium sulfate was chosen for enzyme precipitation which was followed by cross linking of formed aggregates via glutaraldehyde. Concentrations of precipitant and cross linker were respectively optimized as 60% ammonium sulfate and 1% glutaraldehyde. Optimum pH and temperature for CLEA was increased compared to free enzyme. Furthermore, pH, thermal and storage stability were improved. Presence of additives had no effects on enzyme activity. Prepared cross-linked trypsin aggregates are convenient for in situ protein fragmentation and can be used for protein identification.

  14. Characterization of an L-arabinose isomerase from Bacillus thermoglucosidasius for D-tagatose production.

    PubMed

    Seo, Myung-Ji

    2013-01-01

    L-Arabinose isomerase from Bacillus thermoglucosidasius KCTC 1828 (BTAI) was expressed in Escherichia coli. The optimal temperature and pH for the activity of the purified BTAI were 40 °C and pH 7.0. The Mn(2+) ion was an activator of BTAI activity. The kinetic parameters of BTAI for D-galactose were a K(m) of 175 mM and a k(cat)/K(m) of 2.8 mM(-1)min(-1). The conversion ratio by BTAI to D-tagatose reached 45.6% at 40 °C.

  15. [Production and partial characterization of beta-galactosidase from Kluyveromyces lactis grown in deproteinized whey].

    PubMed

    Ramírez Matheus, Alejandra O; Rivas, Nilo

    2003-06-01

    The purpose of this work was to optimize the beta-galactosidase production by Kluyveromyces lactis, applying the Surface Response Methodology (SRM) and using deproteinized whey as fermentation medium. An Orthogonal Central Compound Design (OCCD) was used without repetition, with four factors: temperature, pH, agitation speed and fermentation time. Then, enzyme activity (U/ml) as response variable was used. Thirty trials in twenty-five treatments, with six repetitions at the central point, were carried out, in a New Brunswick Bioflo 2000 fermentor with a volume of 2 liters. The deproteinized whey obtained by thermocoagulation was chemically analyzed. The results were: moisture 93.83%, total solids 6.17%, protein 0.44%, lactose 4.85%, acidity 0.43% and pH 4.58. The best conditions in the enzyme production were: temperature 30.3 degrees C, pH 4.68, agitation speed 191 r.p.m. and fermentation time 18.5 h. with an enzyme production of 8.3 U/ml. The degree of purification obtained was 7.4 times and the yield was 50.8%. The purified enzyme had an optimum temperature of 60 degrees C and a pH of 6.2. This work shows that the yeast Kluyveromyces lactis grown in deproteinized whey is able to produce the enzyme beta-galactosidase and SRM can be used in the fermentology processes, specifically in determining the best suitable operation conditions.

  16. Enzymatic activities in different strains isolated from healthy and brittle leaf disease affected date palm leaves: study of amylase production conditions.

    PubMed

    Mouna, Jrad; Imen, Fendri; Choba Ines, Ben; Nourredine, Drira; Adel, Kadri; Néji, Gharsallah

    2015-02-01

    The present study aimed to investigate and compare the enzymatic production of endophytic bacteria isolated from healthy and brittle leaf disease affected date palm leaves (pectinase, cellulase, lipase, and amylase). The findings revealed that the enzymatic products from the bacterial isolates of healthy date palm leaves were primarily 33% amylolytic enzyme, 33 % cellulase, 25 % pectinase, and 25 % lipase. The isolates from brittle leaf disease date palm leaves, on the other hand, were noted to produce 16 % amylolytic enzyme, 20 % cellulose, 50 % pectinase, and 50 % lipase. The effects of temperature and pH on amylase, pectinase, and cellulose activities were investigated. The Bacillus subtilis JN934392 strain isolated from healthy date palm leaves produced higher levels of amylase activity at pH 7. A Box Behnken Design (BBD) was employed to optimize amylase extraction. Maximal activity was observed at pH and temperature ranges of pH 6-6.5 and 37-39 °C, respectively. Under those conditions, amylase activity was noted to be attained 9.37 U/ml. The results showed that the enzyme was able to maintain more than 50 % of its activity over a temperature range of 50-80 °C, with an optimum at 70 °C. This bacterial amylase showed high activity compared to other bacteria, which provides support for its promising candidacy for future industrial application.

  17. Optimization of Levan Production by Cold-Active Bacillus licheniformis ANT 179 and Fructooligosaccharide Synthesis by Its Levansucrase.

    PubMed

    Xavier, Janifer Raj; Ramana, Karna Venkata

    2017-03-01

    Fructooligosaccharides (FOS) and levan attract much attention due to a wide range of applications in food technology and pharmaceutical and cosmetic industry. Bacillus licheniformis ANT 179, isolated from Antarctica soil, produced levansucrase and levan in a medium containing sucrose as carbon substrate. In this study, characterization of levansucrase and production of short-chain FOS and levan were investigated. Temperature and pH optimum of the enzyme were found to be 60 °C and pH 6.0, respectively. The optimization of fermentation conditions for levan production using sugarcane juice by response surface methodology (RSM) was carried out. Central composite rotatable design was used to study the main and the interactive effects of medium components: sugarcane juice and casein peptone concentration on levan production by the bacterium. The optimized medium with sugarcane juice at 20 % (v/v) and casein peptone at 2 % (w/v) was found to be optimal at an initial pH of 7.0 and incubation temperature of 35 °C for 48 h. Under these conditions, the maximum levan concentration was 50.25 g/L on wet weight basis and 16.35 g/L on dry weight basis. The produced inulin type FOS (kestose and neokestose) and levan were characterized by Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR) analysis. The study revealed that the levansucrase could form FOS from sucrose. The locally available low-cost substrate such as sugarcane juice in the form of a renewable substrate is proposed to be suitable even for scale-up production of enzyme and FOS for industrial applications. The levan and FOS synthesized by the bacterium are suitable for food applications and biomedical uses as the bacterium has GRAS status and devoid of endotoxin as compared to other Gram-negative bacteria.

  18. pH-Dependent, Thermosensitive Polymeric Nanocarriers for Drug Delivery to Solid Tumors

    PubMed Central

    Chen, Ching-Yi; Kim, Tae Hee; Wu, Wen-Chung; Huang, Chi-Ming; Wei, Hua; Mount, Christopher W.; Tian, Yanqing; Jang, Sei-Hum; Pun, Suzie H.; Jen, Alex K-Y

    2013-01-01

    Polymeric micelles are promising carriers for anticancer agents due to their small size, ease of assembly, and versatility for functionalization. A current challenge in the use of polymeric micelles is the sensitive balance that must be achieved between stability during prolonged blood circulation and release of active drug at the tumor site. Stimuli-responsive materials provide a mechanism for triggered drug release in the acidic tumor and intracellular microenvironments. In this work, we synthesized a series of dual pH- and temperature-responsive block copolymers containing a poly(ε-caprolactone) (PCL) hydrophobic block with a poly(triethylene glycol) block that were copolymerized with an amino acid-functionalized monomer. The block copolymers formed micellar structures in aqueous solutions. An optimized polymer that was functionalized with 6-aminocaproic acid (ACA) possessed pH-sensitive phase transitions at mildly acidic pH and body temperature. Doxorubicin-loaded micelles formed from these polymers were stable at blood pH (~7.4) and showed increased drug release at acidic pH. In addition, these micelles displayed more potent anti-cancer activity than free doxorubicin when tested in a tumor xenograft model in mice. PMID:23498892

  19. Reinforcement of thermoplastic chitosan hydrogel using chitin whiskers optimized with response surface methodology.

    PubMed

    Sun, Guohui; Zhang, Xin; Bao, Zixian; Lang, Xuqian; Zhou, Zhongzheng; Li, Yang; Feng, Chao; Chen, Xiguang

    2018-06-01

    To strengthen the mechanical strength of thermo-sensitive hydroxybutyl chitosan (HBC) hydrogel, chitin whiskers were used as sticker to fabricate reinforced HBC (HBCW) hydrogel by using response surface methodology. Unlike the intrinsic network of HBC hydrogel, HBCW hydrogel showed a laminar shape with firm structure. The preparation condition was optimized by three-factor-three-level Box-Behnken design. The maximum mechanical strength (1011.11 Pa) was achieved at 50 °C, when the concentrations of HBC and chitin whiskers were 5.1 wt% and 2.0 wt%, respectively. The effects of temperature, pH value and NaCl concentration on mechanical strength of HBCW hydrogels were investigated via the oscillatory stress sweeps. The results showed that HBCW hydrogel could reach the maximum stiffness (∼1126 Pa) at 37 °C pH 12.0. Low pH and high salty ions could decrease the stability of hydrogel, while chitin whiskers could increase the stress tolerance and related ruptured strain of HBCW hydrogels. Copyright © 2018. Published by Elsevier Ltd.

  20. Separation, concentration and determination of trace chloramphenicol in shrimp from different waters by using polyoxyethylene lauryl ether-salt aqueous two-phase system coupled with high-performance liquid chromatography.

    PubMed

    Lu, Yang; Yao, Hui; Li, Chuang; Han, Juan; Tan, Zhenjiang; Yan, Yongsheng

    2016-02-01

    Polyoxyethylene lauryl ether (POELE10)-NaH2PO4 aqueous two-phase extraction system (ATPES) is coupled with HPLC to analyze chloramphenicol (CAP) in aquatic product. Response surface methodology (RSM) was adopted in the multi-factor experiment to determine the optimized conditions. The extraction efficiency of CAP (E%) is up to 99.42% under the optimal conditions, namely, the concentration of NaH2PO4, the concentration of POELE10, pH and temperature were 0.186 g · mL(-1), 0.033 g · mL(-1), 3.8 and 25 °C respectively. The optimal value of enrichment factor of CAP (F) was 22.56 when the concentration of NaH2PO4 was 0.192 g · mL(-1), the concentration of POELE10 was 0.024 g/ml, pH was 4.2 and temperature was 30 °C. The limit of detection (LOD) and limit of quantification (LOQ) of this method are 0.8 μg · kg(-1) and 1 μg · kg(-1), which meet the needs of determining trace or ultratrace CAP in food. The E% and F of this technique are much better than other extraction methods. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Biodegradation of bispyribac sodium by a novel bacterial consortium BDAM: Optimization of degradation conditions using response surface methodology.

    PubMed

    Ahmad, Fiaz; Anwar, Samina; Firdous, Sadiqa; Da-Chuan, Yin; Iqbal, Samina

    2018-05-05

    Bispyribac sodium (BS), is a selective, systemic and post emergent herbicide used to eradicate grasses and broad leaf weeds. Extensive use of this herbicide has engendered serious environmental concerns. Hence it is important to develop strategies for bioremediation of BS in a cost effective and environment friendly way. In this study a bacterial consortium named BDAM, comprising three novel isolates Achromobacter xylosoxidans (BD1), Achromobacter pulmonis (BA2), and Ochrobactrum intermedium (BM2), was developed by virtue of its potential for degradation of BS. Different culture conditions (temperature, pH and inoculum size) were optimized for degradation of BS by the consortium BDAM and the mutual interactions of these parameters were analysed using a 2 3 full factorial central composite design (CCD) based on Response Surface Methodology (RSM). The optimal values for temperature, pH and inoculum size were found to be 40 °C, 8 and 0.4 g/L respectively to achieve maximum degradation of BS (85.6%). Moreover, the interactive effects of these parameters were investigated using three dimensional surface plots in terms of maximum fitness function. Importantly, it was concluded that the newly developed consortium is a potential candidate for biodegradation of BS in a safe, cost-effective and environmentally friendly manner. Copyright © 2017. Published by Elsevier B.V.

  2. Separation of 20 coumarin derivatives using the capillary electrophoresis method optimized by a series of Doehlert experimental designs.

    PubMed

    Woźniakiewicz, Michał; Gładysz, Marta; Nowak, Paweł M; Kędzior, Justyna; Kościelniak, Paweł

    2017-05-15

    The aim of this study was to develop the first CE-based method enabling separation of 20 structurally similar coumarin derivatives. To facilitate method optimization a series of three consequent Doehlert experimental designs with the response surface methodology was employed, using number of peaks and the adjusted time of analysis as the selected responses. Initially, three variables were examined: buffer pH, ionic strength and temperature (No. 1 Doehlert design). The optimal conditions provided only partial separation, on that account, several buffer additives were examined at the next step: organic cosolvents and cyclodextrin (No. 2 Doehlert design). The optimal cyclodextrin type was also selected experimentally. The most promising results were obtained for the buffers fortified with methanol, acetonitrile and heptakis(2,3,6-tri-O-methyl)-β-cyclodextrin. Since these additives may potentially affect acid-base equilibrium and ionization state of analytes, the third Doehlert design (No. 3) was used to reconcile concentration of these additives with optimal pH. Ultimately, the total separation of all 20 compounds was achieved using the borate buffer at basic pH 9.5 in the presence of 10mM cyclodextrin, 9% (v/v) acetonitrile and 36% (v/v) methanol. Identity of all compounds was confirmed using the in-lab build UV-VIS spectra library. The developed method succeeded in identification of coumarin derivatives in three real samples. It demonstrates a huge resolving power of CE assisted by addition of cyclodextrins and organic cosolvents. Our unique optimization approach, based on the three Doehlert designs, seems to be prospective for future applications of this technique. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Construction and evaluation of an exopolysaccharide-producing engineered bacterial strain by protoplast fusion for microbial enhanced oil recovery.

    PubMed

    Sun, Shanshan; Luo, Yijing; Cao, Siyuan; Li, Wenhong; Zhang, Zhongzhi; Jiang, Lingxi; Dong, Hanping; Yu, Li; Wu, Wei-Min

    2013-09-01

    Enterobacter cloacae strain JD, which produces water-insoluble biopolymers at optimal temperature of 30°C, and a thermophilic Geobacillus strain were used to construct an engineered strain for exopolysaccharide production at high temperatures by protoplast fusion. The obtained fusant strain ZR3 produced exopolysaccharides at up to 45°C with optimal growth temperature at 35°C. The fusant produced exopolysaccharides of approximately 7.5 g/L or more at pH between 7.0 and 9.0. The feasibility of the enhancement of crude oil recovery with the fusant was tested in a sand-packed column at 40°C. The results demonstrated that bioaugmentation of the fusant was promising approach for MEOR. Mass growth of the fusant was confirmed in fermentor tests. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Optimization study for Pb(II) and COD sequestration by consortium of sulphate-reducing bacteria

    NASA Astrophysics Data System (ADS)

    Verma, Anamika; Bishnoi, Narsi R.; Gupta, Asha

    2017-09-01

    In this study, initial minimum inhibitory concentration (MIC) of Pb(II) ions was analysed to check optimum concentration of Pb(II) ions at which the growth of sulphate-reducing consortium (SRC) was found to be maximum. 80 ppm of Pb(II) ions was investigated as minimum inhibitory concentration for SRC. Influence of electron donors such as lactose, sucrose, glucose and sodium lactate was examined to investigate best carbon source for growth and activity of sulphate-reducing bacteria. Sodium lactate was found to be the prime carbon source for SRC. Later optimization of various parameters was executed using Box-Behnken design model of response surface methodology to explore the effectiveness of three independent operating variables, namely, pH (5.0-9.0), temperature (32-42 °C) and time (5.0-9.0 days), on dependent variables, i.e. protein content, precipitation of Pb(II) ions, and removal of COD by SRC biomass. Maximum removal of COD and Pb(II) was observed to be 91 and 98 %, respectively, at pH 7.0 and temperature 37 °C and incubation time 7 days. According to response surface analysis and analysis of variance, the experimental data were perfectly fitted to the quadratic model, and the interactive influence of pH, temperature and time on Pb(II) and COD removal was highly significant. A high regression coefficient between the variables and response ( r 2 = 0.9974) corroborate eminent evaluation of experimental data by second-order polynomial regression model. SEM and Fourier transform infrared analysis was performed to investigate morphology of PbS precipitates, sorption mechanism and involved functional groups in metal-free and metal-loaded biomass of SRC for Pb(II) binding.

  5. Optimization of Maillard Reaction in Model System of Glucosamine and Cysteine Using Response Surface Methodology

    PubMed Central

    Arachchi, Shanika Jeewantha Thewarapperuma; Kim, Ye-Joo; Kim, Dae-Wook; Oh, Sang-Chul; Lee, Yang-Bong

    2017-01-01

    Sulfur-containing amino acids play important roles in good flavor generation in Maillard reaction of non-enzymatic browning, so aqueous model systems of glucosamine and cysteine were studied to investigate the effects of reaction temperature, initial pH, reaction time, and concentration ratio of glucosamine and cysteine. Response surface methodology was applied to optimize the independent reaction parameters of cysteine and glucosamine in Maillard reaction. Box-Behnken factorial design was used with 30 runs of 16 factorial levels, 8 axial levels and 6 central levels. The degree of Maillard reaction was determined by reading absorption at 425 nm in a spectrophotometer and Hunter’s L, a, and b values. ΔE was consequently set as the fifth response factor. In the statistical analyses, determination coefficients (R2) for their absorbance, Hunter’s L, a, b values, and ΔE were 0.94, 0.79, 0.73, 0.96, and 0.79, respectively, showing that the absorbance and Hunter’s b value were good dependent variables for this model system. The optimum processing parameters were determined to yield glucosamine-cysteine Maillard reaction product with higher absorbance and higher colour change. The optimum estimated absorbance was achieved at the condition of initial pH 8.0, 111°C reaction temperature, 2.47 h reaction time, and 1.30 concentration ratio. The optimum condition for colour change measured by Hunter’s b value was 2.41 h reaction time, 114°C reaction temperature, initial pH 8.3, and 1.26 concentration ratio. These results can provide the basic information for Maillard reaction of aqueous model system between glucosamine and cysteine. PMID:28401086

  6. Optimization of Maillard Reaction in Model System of Glucosamine and Cysteine Using Response Surface Methodology.

    PubMed

    Arachchi, Shanika Jeewantha Thewarapperuma; Kim, Ye-Joo; Kim, Dae-Wook; Oh, Sang-Chul; Lee, Yang-Bong

    2017-03-01

    Sulfur-containing amino acids play important roles in good flavor generation in Maillard reaction of non-enzymatic browning, so aqueous model systems of glucosamine and cysteine were studied to investigate the effects of reaction temperature, initial pH, reaction time, and concentration ratio of glucosamine and cysteine. Response surface methodology was applied to optimize the independent reaction parameters of cysteine and glucosamine in Maillard reaction. Box-Behnken factorial design was used with 30 runs of 16 factorial levels, 8 axial levels and 6 central levels. The degree of Maillard reaction was determined by reading absorption at 425 nm in a spectrophotometer and Hunter's L, a, and b values. ΔE was consequently set as the fifth response factor. In the statistical analyses, determination coefficients (R 2 ) for their absorbance, Hunter's L, a, b values, and ΔE were 0.94, 0.79, 0.73, 0.96, and 0.79, respectively, showing that the absorbance and Hunter's b value were good dependent variables for this model system. The optimum processing parameters were determined to yield glucosamine-cysteine Maillard reaction product with higher absorbance and higher colour change. The optimum estimated absorbance was achieved at the condition of initial pH 8.0, 111°C reaction temperature, 2.47 h reaction time, and 1.30 concentration ratio. The optimum condition for colour change measured by Hunter's b value was 2.41 h reaction time, 114°C reaction temperature, initial pH 8.3, and 1.26 concentration ratio. These results can provide the basic information for Maillard reaction of aqueous model system between glucosamine and cysteine.

  7. Biodegradation of methyl red by Bacillus sp. strain UN2: decolorization capacity, metabolites characterization, and enzyme analysis.

    PubMed

    Zhao, Ming; Sun, Peng-Fei; Du, Lin-Na; Wang, Guan; Jia, Xiao-Ming; Zhao, Yu-Hua

    2014-05-01

    Azo dyes are recalcitrant and refractory pollutants that constitute a significant menace to the environment. The present study is focused on exploring the capability of Bacillus sp. strain UN2 for application in methyl red (MR) degradation. Effects of physicochemical parameters (pH of medium, temperature, initial concentration of dye, and composition of the medium) were studied in detail. The suitable pH and temperature range for MR degradation by strain UN2 were respectively 7.0-9.0 and 30-40 °C, and the optimal pH value and temperature were respectively 8.0 and 35 °C. Mg(2+) and Mn(2+) (1 mM) were found to significantly accelerate the MR removal rate, while the enhancement by either Fe(3+) or Fe(2+) was slight. Under the optimal degradation conditions, strain UN2 exhibited greater than 98 % degradation of the toxic azo dye MR (100 ppm) within 30 min. Analysis of samples from decolorized culture flasks confirmed biodegradation of MR into two prime metabolites: N,N'dimethyl-p-phenyle-nediamine and 2-aminobenzoic acid. A study of the enzymes responsible for the biodegradation of MR, in the control and cells obtained during (10 min) and after (30 min) degradation, showed a significant increase in the activities of azoreductase, laccase, and NADH-DCIP reductase. Furthermore, a phytotoxicity analysis demonstrated that the germination inhibition was almost eliminated for both the plants Triticum aestivum and Sorghum bicolor by MR metabolites at 100 mg/L concentration, yet the germination inhibition of parent dye was significant. Consequently, the high efficiency of MR degradation enables this strain to be a potential candidate for bioremediation of wastewater containing MR.

  8. Study on micro-hardness of electroless composite plating of Ni-P with SiC Nano-particles

    NASA Astrophysics Data System (ADS)

    Sun, Yong; Zhang, Zhaoguo; Li, Jiamin; Xu, Donghui

    2007-07-01

    In this paper, a Ni-P electroless composite coating containing nano SiC particles was produced. The wearability of the composite coating was studied. Temperature, PH of the plating liquid and the concentration of SiC nanoparticles in the plating liquid were taken as parameters and the experiment with three factors and five levels was designed through the method of quadratic orthogonal rotation combination. SiC nanoparticles were dispersed by ultrasonic. The influence of the testing parameters on the hardness of the coating was studied intensively. The optimal parameters were obtained when the temperature is 86+/-1°C, PH is 6+/-0.5 and the concentration of SiC nanoparticles is 6g/L. The maximal hardness of the coating is over 1700HV after heat treatment.

  9. Enrichment of copper and recycling of cyanide from copper-cyanide waste by solvent extraction

    NASA Astrophysics Data System (ADS)

    Gao, Teng-yue; Liu, Kui-ren; Han, Qing; Xu, Bin-shi

    2016-11-01

    The enrichment of copper from copper-cyanide wastewater by solvent extraction was investigated using a quaternary ammonium salt as an extractant. The influences of important parameters, e.g., organic-phase components, aqueous pH values, temperature, inorganic anion impurities, CN/Cu molar ratio, and stripping reagents, were examined systematically, and the optimal conditions were determined. The results indicated that copper was effectively concentrated from low-concentration solutions using Aliquat 336 and that the extraction efficiency increased linearly with increasing temperature. The aqueous pH value and concentrations of inorganic anion impurities only weakly affected the extraction process when varied in appropriate ranges. The CN/Cu molar ratio affected the extraction efficiency by changing the distribution of copper-cyanide complexes. The difference in gold leaching efficiency between using raffinate and fresh water was negligible.

  10. Kinetic analysis of simultaneous denitrification and biomineralization of novel Acinetobacter sp. CN86.

    PubMed

    Su, Jun-Feng; Shi, Jing-Xin; Huang, Ting-Lin; Ma, Fang

    2016-08-15

    A novel aerobic denitrification and biomineralization strain CN86 was isolated from the Qu Jiang artificial lake. Based on phylogenetic characteristics, the isolated strain was identified as Acinetobacter species. Strain CN86 was confirmed to have the ability to perform simultaneous denitrification and biomineralization. Exponential decay equation was used for the matching of kinetic processes on denitrification and biomineralization. A highest nitrate removal rate was achieved at the pH7.0, organic concentration of 1.5g/L and temperature of 30°C. An optimal hardness removal rate was obtained at the pH9.0, organic concentration of 2.0g/L and temperature of 30°C. Strain CN86 is a suitable candidate for the simultaneous removal of nitrate and hardness in groundwater treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Separation of no-carrier-added rhenium from bulk tantalum by the sodium malonate-PEG aqueous biphasic system.

    PubMed

    Dutta, Binita; Lahiri, Susanta; Tomar, B S

    2014-02-01

    The aqueous biphasic system (ABS) involving sodium malonate-polyethylene glycol (PEG) phases has been applied for the first time for separation of no-carrier-added (183)Re (T1/2=70 d) from α-particle irradiated bulk tantalum target. The various ABS conditions were applied for investigating the separation by varying pH, temperature, PEG-molecular weight, concentration of salt. The extraction pattern was hardly affected by change in pH and the molecular weight of PEG. One step separation of nca (183)Re from Ta was achieved at the optimal conditions of (i) 50% (w/w) PEG-4000-2 M sodium malonate, 40 °C and (ii) 50% (w/w) PEG-4000-3 M sodium malonate, room temperature (27 °C). © 2013 Published by Elsevier Ltd.

  12. Enrichment of Thermophilic Ammonia-Oxidizing Archaea from an Alkaline Hot Spring in the Great Basin, USA

    NASA Astrophysics Data System (ADS)

    Zhang, C.; Huang, Z.; Jiang, H.; Wiegel, J.; Li, W.; Dong, H.

    2010-12-01

    One of the major advances in the nitrogen cycle is the recent discovery of ammonia oxidation by archaea. While culture-independent studies have revealed occurrence of ammonia-oxidizing archaea (AOA) in nearly every surface niche on earth, most of these microorganisms have resisted isolation and so far only a few species have been identified. The Great Basin contains numerous hot springs, which are characterized by moderately high temperature (40-65 degree C) and circumneutral or alkaline pH. Unique thermophilic archaea have been identified based on molecular DNA and lipid biomarkers; some of which may be ammonia oxidizers. This study aims to isolate some of these archaea from a California hot spring that has pH around 9.0 and temperature around 42 degree C. Mat material was collected from the spring and transported on ice to the laboratory. A synthetic medium (SCM-5) was inoculated with the mat material and the culture was incubated under varying temperature (35-65 degree C) and pH (7.0-10.0) conditions using antibiotics to suppress bacterial growth. Growth of the culture was monitored by microscopy, decrease in ammonium and increase in nitrite, and increases in Crenarchaeota and AOA abundances over time. Clone libraries were constructed to compare archaeal community structures before and after the enrichment experiment. Temperature and pH profiles indicated that the culture grew optimally at pH 9.0 and temperature 45 degree C, which are consistent with the geochemical conditions of the natural environment. Phylogenetic analysis showed that the final OTU was distantly related to all known hyperthermophilic archaea. Analysis of the amoA genes showed two OTUs in the final culture; one of them was closely related to Candidatus Nitrososphaera gargensis. However, the enrichment culture always contained bacteria and attempts to separate them from archaea have failed. This highlights the difficulty in bringing AOA into pure culture and suggests that some of the AOA may require synergy with bacteria for growth.

  13. Quantum mechanics study of repulsive π-π interaction and flexibility of phenyl moiety in the iron azodioxide complex

    NASA Astrophysics Data System (ADS)

    Liu, Yuemin; Liu, Yucheng; Murru, Siva; Tzeng, Nianfeng; Srivastava, Radhey S.

    2015-10-01

    In this study, repulsive π-π interactions within iron azodioxide complex Fe[Ph(O)NN(O)Ph]3 were quantum mechanically characterized using DFT, MP2 and CCSD(T) methods. Flexibility of six phenyl moieties in this complex structure was also investigated by structural optimization approach using the DFT methods. Our MP2 and CCSD(T) calculations of the closest pair provided interaction energy of 6.62 and 8.29 kcal/mol respectively, which indicate a strongest repulsion among these intra-molecular π-π interactions. Interaction energy of the particular π-π pair calculated from 24 hybrid DFT methods ranges from 4.56 kcal/mol from BHandH method to 15.15 kcal/mol from O3LYP method. Cares should be exercised when interpreting interaction energy and geometry optimization from DFT simulation of systems containing π-π interaction. Comparison between the DFT results and the benchmark CCSD(T) results shows that the DFT calculations of π-π interaction are reasonable but still need to be interpreted with caution. Furthermore, MP2 interaction energy of -44.69 kcal/mol between two substituted π systems/phenyl rings Ph(O)N-moieties suggested that above energetically unfavorable π-π interaction can be compensated by the covalent bond N-N in a single ligand Ph(O)NN(O)Ph, which allows for a reasonable stability across the complex molecules. Optimizations of the entire complex molecule using B3LYP and M06HF methods produced a large variation of π-π distances and orientations, which implied that the complex molecule may perform catalysis at room temperature.

  14. Modeling lactose hydrolysis for efficiency and selectivity: Toward the preservation of sialyloligosaccharides in bovine colostrum whey permeate.

    PubMed

    de Moura Bell, Juliana M L N; Aquino, Leticia F M C; Liu, Yan; Cohen, Joshua L; Lee, Hyeyoung; de Melo Silva, Vitor L; Rodrigues, Maria I; Barile, Daniela

    2016-08-01

    Enzymatic hydrolysis of lactose has been shown to improve the efficiency and selectivity of membrane-based separations toward the recovery of bioactive oligosaccharides. Achieving maximum lactose hydrolysis requires intrinsic process optimization for each specific substrate, but the effects of those processing conditions on the target oligosaccharides are not well understood. Response surface methodology was used to investigate the effects of pH (3.25-8.25), temperature (35-55°C), reaction time (6 to 58 min), and amount of enzyme (0.05-0.25%) on the efficiency of lactose hydrolysis by β-galactosidase and on the preservation of biologically important sialyloligosaccharides (3'-siallylactose, 6'-siallylactose, and 6'-sialyl-N-acetyllactosamine) naturally present in bovine colostrum whey permeate. A central composite rotatable design was used. In general, β-galactosidase activity was favored at pH values ranging from 3.25 to 5.75, with other operational parameters having a less pronounced effect. A pH of 4.5 allowed for the use of a shorter reaction time (19 min), lower temperature (40°C), and reduced amount of enzyme (0.1%), but complete hydrolysis at a higher pH (5.75) required greater values for these operational parameters. The total amount of sialyloligosaccharides was not significantly altered by the reaction parameters evaluated, suggesting specificity of β-galactosidase from Aspergillus oryzae toward lactose as well as the stability of the oligosaccharides at pH, temperature, and reaction time evaluated. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  15. Optimisation of low temperature extraction of banana juice using commercial pectinase.

    PubMed

    Sagu, Sorel Tchewonpi; Nso, Emmanuel Jong; Karmakar, Sankha; De, Sirshendu

    2014-05-15

    The objective of this work was to develop a process with optimum conditions for banana juice. The procedure involves hydrolyzing the banana pulp by commercial pectinase followed by cloth filtration. Response surface methodology with Doehlert design was utilised to optimize the process parameters. The temperature of incubation (30-60 °C), time of reaction (20-120 min) and concentration of pectinase (0.01-0.05% v/w) were the independent variables and viscosity, clarity, alcohol insoluble solids (AIS), total polyphenol and protein concentration were the responses. Total soluble sugar, pH, conductivity, calcium, sodium and potassium concentration in the juice were also evaluated. The results showed reduction of AIS and viscosity with reaction time and pectinase concentration and reduction of polyphenol and protein concentration with temperature. Using numerical optimization, the optimum conditions for the enzymatic extraction of banana juice were estimated. Depectinization kinetics was also studied at optimum temperature and variation of kinetic constants with enzyme dose was evaluated. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Preparation and characterization of CuO catalyst for the thermolysis treatment of distillery wastewater.

    PubMed

    Sharma, Deepak; Prajapati, Abhinesh Kumar; Choudhary, Rumi; Kaushal, Rajesh Kumar; Pal, Dharm; Sawarkar, Ashish N

    2017-08-16

    CuO catalyst was prepared from copper sulfate by alkali precipitation method followed by drying and calcination. Characterization of CuO catalyst using X-ray diffraction, Brunauer-Emmett-Teller, and Barrett-Joyner-Halenda surface area analysis envisaged the effectiveness of CuO as a catalyst for the treatment of biodigester effluent (BDE) emanated from distilleries. The catalytic thermolysis is an efficient advance treatment method for distillery biodigester effluent (BDE). CT treatment of BDE was carried out in a 0.5 dm 3 thermolytic batch reactor using CuO as a catalyst at different pH (1-9), temperatures (80-110°C), and catalyst loadings (1-4 kg/m 3 ). With CuO catalyst, a temperature of 110°C, catalyst loading of 4 kg/m 3 , and pH of 2 was found to be optimal, providing a maximum reduction in chemical oxygen demand of 65%. The settling characteristics at different temperatures of CT-treated sludge were also presented.

  17. Bioleaching of arsenic from highly contaminated mine tailings using Acidithiobacillus thiooxidans.

    PubMed

    Lee, Eunseong; Han, Yosep; Park, Jeonghyun; Hong, Jeongsik; Silva, Rene A; Kim, Seungkon; Kim, Hyunjung

    2015-01-01

    The behavior of arsenic (As) bioleaching from mine tailings containing high amount of As (ca. 34,000 mg/kg) was investigated using Acidithiobacillus thiooxidans to get an insight on the optimal conditions that would be applied to practical heap and/or tank bioleaching tests. Initial pH (1.8-2.2), temperature (25-40 °C), and solid concentration (0.5-4.0%) were employed as experimental parameters. Complementary characterization experiments (e.g., XRD, SEM-EDS, electrophoretic mobility, cell density, and sulfate production) were also carried out to better understand the mechanism of As bioleaching. The results showed that final As leaching efficiency was similar regardless of initial pH. However, greater initial As leaching rate was observed at initial pH 1.8 than other conditions, which could be attributed to greater initial cell attachment to mine tailings. Unlike the trend observed when varying the initial pH, the final As leaching efficiency varied with the changes in temperature and solid concentration. Specifically, As leaching efficiency tended to decrease with increasing temperature due to the decrease in the bacterial growth rate at higher temperature. Meanwhile, As leaching efficiency tended to increase with decreasing solid concentration. The results for jarosite contents in mine tailings residue after bioleaching revealed that much greater amount of the jarosite was formed during the bioleaching reaction at higher solid concentration, suggesting that the coverage of the surface of the mine tailings by jarosite and/or the co-precipitation of the leached As with jarosite could be a dominant factor reducing As leaching efficiency. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Degradation of chitin and chitosan by a recombinant chitinase derived from a virulent Aeromonas hydrophila isolated from diseased channel catfish

    USDA-ARS?s Scientific Manuscript database

    A chitinase was identified in extracellular products of a virulent Aeromonas hydrophila isolated from diseased channel catfish (Ictalurus punctatus). Bioactive recombinant chitinase (rChi-Ah) was produced in Escherichia coli. Purified rChi-Ah had optimal activity at temperature of 42°C and pH 6.5. T...

  19. [Optimization of lime milk precipitation process of Lonicera Japonica aqueous extract based on quality by design concept].

    PubMed

    Shen, Jin-Jing; Gong, Xing-Chu; Pan, Jian-Yang; Qu, Hai-Bin

    2017-03-01

    Design space approach was applied in this study to optimize the lime milk precipitation process of Lonicera Japonica (Jinyinhua) aqueous extract. The evaluation indices for this process were total organic acid purity and amounts of 6 organic acids obtained from per unit mass of medicinal materials. Four critical process parameters (CPPs) including drop speed of lime milk, pH value after adding lime milk, settling time and settling temperature were identified by using the weighted standardized partial regression coefficient method. Quantitative models between process evaluation indices and CPPs were established by a stepwise regression analysis. A design space was calculated by a Monte-Carlo simulation method, and then verified. The verification test results showed that the operation within the design space can guarantee the stability of the lime milk precipitation process. The recommended normal operation space is as follows: drop speed of lime milk of 1.00-1.25 mL•min⁻¹, pH value of 11.5-11.7, settling time of 1.0-1.2 h, and settling temperature of 10-20 ℃.. Copyright© by the Chinese Pharmaceutical Association.

  20. Conditions Optimizing and Application of Laccase-mediator System (LMS) for the Laccase-catalyzed Pesticide Degradation

    PubMed Central

    Jin, Xiaoting; Yu, Xiangyang; Zhu, Guangyan; Zheng, Zuntao; Feng, Fayun; Zhang, Zhiyong

    2016-01-01

    A high capacity of laccase from Trametes versicolor capable of degrading pesticides has been revealed. The conditions for degrading of five selected pesticides including chlorpyrifos, chlorothalonil, pyrimethanil, atrazine and isoproturon with the purified laccases from Trametes versicolor were optimized. The results showed that the optimum conditions for the highest activity were pH at 5.0 and temperature at 25 °C. The best mediators were violuric acid for pyrimethanil and isoproturon, vanillin for chlorpyrifos, and acetosyringone and HBT for chlorothalonil and atrazine, respectively. The laccase was found to be stable at a pH range from 5.0 to 7.0 and temperature from 25 to 30 °C. It was observed that each pesticide required a different laccase mediator concentration typically between 4.0–6.0 mmol/L. In the experiment, the degradation rates of pyrimethanil and isoproturon were significantly faster than those of chlorpyrifos, chlorothalonil and atrazine. For example, it was observed that pyrimethanil and isoproturon degraded up to nearly 100% after 24 hours while the other three pesticides just reached up 90% of degradation after 8 days of incubation. PMID:27775052

  1. Conditions Optimizing and Application of Laccase-mediator System (LMS) for the Laccase-catalyzed Pesticide Degradation.

    PubMed

    Jin, Xiaoting; Yu, Xiangyang; Zhu, Guangyan; Zheng, Zuntao; Feng, Fayun; Zhang, Zhiyong

    2016-10-24

    A high capacity of laccase from Trametes versicolor capable of degrading pesticides has been revealed. The conditions for degrading of five selected pesticides including chlorpyrifos, chlorothalonil, pyrimethanil, atrazine and isoproturon with the purified laccases from Trametes versicolor were optimized. The results showed that the optimum conditions for the highest activity were pH at 5.0 and temperature at 25 °C. The best mediators were violuric acid for pyrimethanil and isoproturon, vanillin for chlorpyrifos, and acetosyringone and HBT for chlorothalonil and atrazine, respectively. The laccase was found to be stable at a pH range from 5.0 to 7.0 and temperature from 25 to 30 °C. It was observed that each pesticide required a different laccase mediator concentration typically between 4.0-6.0 mmol/L. In the experiment, the degradation rates of pyrimethanil and isoproturon were significantly faster than those of chlorpyrifos, chlorothalonil and atrazine. For example, it was observed that pyrimethanil and isoproturon degraded up to nearly 100% after 24 hours while the other three pesticides just reached up 90% of degradation after 8 days of incubation.

  2. Optimization of Bromelain-Aided Production of Angiotensin I-Converting Enzyme Inhibitory Hydrolysates from Stone Fish Using Response Surface Methodology

    PubMed Central

    Auwal, Shehu Muhammad; Zarei, Mohammad; Abdul-Hamid, Azizah; Saari, Nazamid

    2017-01-01

    The stone fish is an under-utilized sea cucumber with many nutritional and ethno-medicinal values. This study aimed to establish the conditions for its optimum hydrolysis with bromelain to generate angiotensin I-converting enzyme (ACE)-inhibitory hydrolysates. Response surface methodology (RSM) based on a central composite design was used to model and optimize the degree of hydrolysis (DH) and ACE-inhibitory activity. Process conditions including pH (4–7), temperature (40–70 °C), enzyme/substrate (E/S) ratio (0.5%–2%) and time (30–360 min) were used. A pH of 7.0, temperature of 40 °C, E/S ratio of 2% and time of 240 min were determined using a response surface model as the optimum levels to obtain the maximum ACE-inhibitory activity of 84.26% at 44.59% degree of hydrolysis. Hence, RSM can serve as an effective approach in the design of experiments to improve the antihypertensive effect of stone fish hydrolysates, which can thus be used as a value-added ingredient for various applications in the functional foods industries. PMID:28362352

  3. Biological oxidation of hydrogen sulfide in mineral media using a biofilm airlift suspension reactor.

    PubMed

    Moghanloo, G M Mojarrad; Fatehifar, E; Saedy, S; Aghaeifar, Z; Abbasnezhad, H

    2010-11-01

    Hydrogen sulfide (H(2)S) removal in mineral media using Thiobacillus thioparus TK-1 in a biofilm airlift suspension reactor (BAS) was investigated to evaluate the relationship between biofilm formation and changes in inlet loading rates. Aqueous sodium sulfide was fed as the substrate into the continuous BAS-reactor. The reactor was operated at a constant temperature of 30 degrees C and a pH of 7, the optimal temperature and pH for biomass growth. The startup of the reactor was performed with basalt carrier material. Optimal treatment performance was obtained at a loading rate of 4.8 mol S(2-) m(-3) h(-1) at a conversion efficiency as high as 100%. The main product of H(2)S oxidation in the BAS-reactor was sulfate because of high oxygen concentrations in the airlift reactor. The maximum sulfide oxidation rate was 6.7 mol S(2-) m(-3) h(-1) at a hydraulic residence time of 3.3 h in the mineral medium. The data showed that the BAS-reactor with this microorganism can be used for sulfide removal from industrial effluent. Copyright 2010 Elsevier Ltd. All rights reserved.

  4. An optimized method for elution of enteroviral RNA from a cellulose-based substrate.

    PubMed

    Li, Yan; Yoshida, Hiromu; Wang, Lu; Tao, Zexin; Wang, Haiyan; Lin, Xiaojuan; Xu, Aiqiang

    2012-12-01

    The Flinders Technology Australia (FTA) Elute Card is a commercial product that facilitates the collection, transport, archiving and processing of nucleic acids from a wide variety of biological samples at room temperature. While the cards have been designed so that sterile/deionized water can elute DNA easily, they are not suitable for some less stable RNAs. This study was undertaken to determine the optimal conditions such as the buffer type, buffer pH and incubation temperature for the elution of enteroviral RNA from FTA Elute Cards prior to quantitative analysis using real-time PCR (qPCR) or consensus degenerate hybrid oligonucleotide primer VP1 RT-semi nested PCR (CODEHOP VP1 RT-snPCR). TE-1 (pH 8.0), rather than sterile water, was the best buffer for high efficiency elution of enteroviral RNA at 95°C. However, as the estimated recovery rate of viral RNA eluted from the cards averaged to be only 6.1%, enterovirus assays using FTA elution should be considered qualitative, especially at low virus titers, and therefore the results of the assay should be interpreted carefully. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Production of L-asparaginase, an anticancer agent, from Aspergillus niger using agricultural waste in solid state fermentation.

    PubMed

    Mishra, Abha

    2006-10-01

    This article reports the production of high levels of L-asparaginase from a new isolate of Aspergillus niger in solid state fermentation (SSF) using agro-wastes from three leguminous crops (bran of Cajanus cajan, Phaseolus mungo, and Glycine max). When used as the sole source for growth in SSF, bran of G. max showed maximum enzyme production followed by that of P. mungo and C. cajan. A 96-h fermentation time under aerobic condition with moisture content of 70%, 30 min of cooking time and 1205-1405 micro range of particle size in SSF appeared optimal for enzyme production. Enzyme yield was maximum (40.9 +/- 3.35 U/g of dry substrate) at pH 6.5 and temperature 30 +/- 2 degrees C. The optimum temperature and pH for enzyme activity were 40 degrees C and 6.5, respectively. The study suggests that choosing an appropriate substrate when coupled with process level optimization improves enzyme production markedly. Developing an asparaginase production process based on bran of G. max as a substrate in SSF is economically attractive as it is a cheap and readily available raw material in agriculture-based countries.

  6. Bioleaching of Gold and Silver from Waste Printed Circuit Boards by Pseudomonas balearica SAE1 Isolated from an e-Waste Recycling Facility.

    PubMed

    Kumar, Anil; Saini, Harvinder Singh; Kumar, Sudhir

    2018-02-01

    Indigenous bacterial strain Pseudomonas balearica SAE1, tolerant to e-waste toxicity was isolated from an e-waste recycling facility Exigo Recycling Pvt. Ltd., India. Toxicity tolerance of bacterial strain was analyzed using crushed (particle size ≤150 µm) waste computer printed circuit boards (PCBs)/liter (L) of culture medium. The EC 50 value for SAE1 was 325.7 g/L of the e-waste pulp density. Two-step bioleaching was then applied to achieve the dissolution of gold (Au) and silver (Ag) from the e-waste. To maximize precious metal dissolution, factors including pulp density, glycine concentration, pH level, and temperature were optimized. The optimization resulted in 68.5 and 33.8% of Au and Ag dissolution, respectively, at a pH of 9.0, a pulp density of 10 g/L, a temperature of 30 °C, and a glycine concentration of 5 g/L. This is the first study of Au and Ag bioleaching using indigenous e-waste bacteria and its analysis to determine e-waste toxicity tolerance.

  7. Conditions Optimizing and Application of Laccase-mediator System (LMS) for the Laccase-catalyzed Pesticide Degradation

    NASA Astrophysics Data System (ADS)

    Jin, Xiaoting; Yu, Xiangyang; Zhu, Guangyan; Zheng, Zuntao; Feng, Fayun; Zhang, Zhiyong

    2016-10-01

    A high capacity of laccase from Trametes versicolor capable of degrading pesticides has been revealed. The conditions for degrading of five selected pesticides including chlorpyrifos, chlorothalonil, pyrimethanil, atrazine and isoproturon with the purified laccases from Trametes versicolor were optimized. The results showed that the optimum conditions for the highest activity were pH at 5.0 and temperature at 25 °C. The best mediators were violuric acid for pyrimethanil and isoproturon, vanillin for chlorpyrifos, and acetosyringone and HBT for chlorothalonil and atrazine, respectively. The laccase was found to be stable at a pH range from 5.0 to 7.0 and temperature from 25 to 30 °C. It was observed that each pesticide required a different laccase mediator concentration typically between 4.0-6.0 mmol/L. In the experiment, the degradation rates of pyrimethanil and isoproturon were significantly faster than those of chlorpyrifos, chlorothalonil and atrazine. For example, it was observed that pyrimethanil and isoproturon degraded up to nearly 100% after 24 hours while the other three pesticides just reached up 90% of degradation after 8 days of incubation.

  8. Chiral recognition ability of an (S)-naproxen- imprinted monolith by capillary electrochromatography.

    PubMed

    Xu, Yan-Li; Liu, Zhao-Sheng; Wang, He-Fang; Yan, Chao; Gao, Ru-Yu

    2005-02-01

    The racemic naproxen was selectively recognized by capillary electrochromatography (CEC) on an (S)-naproxen-imprinted monolith, which was prepared by an in situ thermal-initiated polymerization. The recognition selectivity of a selected monolith strictly relied on the CEC conditions involved. The factors that influence the imprinting selectivity as well as the electroosmotic flow (EOF), including the applied voltage, organic solvent, salt concentration and pH value of the buffer, column temperature, and surfactant modifiers were systematically studied. Once the column was prepared, the experiment results showed that the successful chiral recognition was dependent on CEC variables. For example: the recognition could be observed in acetonitrile and ethanol electrolytes, while methanol and dimethyl sulfoxide (DMSO) electrolytes had no chiral recognition ability. The buffer with pH values of 2.6 or 3.0 at a higher salt concentration had chiral recognition ability. Column temperatures of 25-35 degrees C were optimal. Three surfactants, sodium dodecyl sulfate (SDS), cetyltrimethylammonium bromide (CTAB), and polyoxyethylene sorbitan monolaurate (Tween 20), can improve the recognition. Baseline resolution was obtained under optimized conditions and the column efficiency of the later eluent (S)-naproxen was 90 000 plates/m.

  9. [Extraction and analysis of the essential oil in Pogostemon cablin by enzymatic hydrolysis and inhibitory activity against Hela cell proliferation].

    PubMed

    Yu, Jing; Qi, Yue; Luo, Gang; Duan, Hong-quan; Zhou, Jing

    2012-05-01

    To optimize the extraction method of essential oil in Pogostemon cablin and analyze its inhibitory activity against Hela cell proliferation. The Pogostemon cablin was treated by hemicellulase before steam distillation. The enzyme dosage, treatment time, treatment temperature, pH were optimized through orthogonal experimental design. The components of essential oil were identified by gas chromatography-mass spectrometry (GC-MS). Inhibitory activity of patchouli oil against Hela cell proliferation was determined by MTP method. The optimum extraction process was as follows: pH 4.5, temperature 45 degrees C, the ratio of hemicellulase to Pogostemon cablin was 1% and enzymatic hydrolysis for 1.0 hour. Extraction ratio of the patchouli oil in steam distillation and hemicellulase extraction method was 2.2220 mg/g, 3.1360 mg/g respectively. Patchouli oil could inhibit Hela cell proliferation. IC50 of the patchouli oil in steam distillation and hemicellulase extraction method was 12.2 +/- 0.46 microg/mL and 0.36 +/- 0.03 microg/mL respectively. In comparison with steam distillation method, extraction ratios of essential oil and the inhibitory activity against Hela cell proliferation can be increased by the hemicellulase extraction method.

  10. Immobilization of ammonia-oxidizing bacteria by polyvinyl alcohol and sodium alginate.

    PubMed

    Dong, Yuwei; Zhang, Yanqiu; Tu, Baojun

    Ammonia-oxidizing bacteria were immobilized by polyvinyl alcohol (PVA) and sodium alginate. The immobilization conditions and ammonia oxidation ability of the immobilized bacteria were investigated. The following immobilization conditions were observed to be optimal: PVA, 12%; sodium alginate, 1.1%; calcium chloride, 1.0%; inoculum concentration, 1.3 immobilized balls/mL of immobilized medium; pH, 10; and temperature, 30°C. The immobilized ammonia-oxidizing bacteria exhibited strong ammonia oxidation ability even after being recycled four times. The ammonia nitrogen removal rate of the immobilized ammonia-oxidizing bacteria reached 90.30% under the optimal immobilization conditions. When compared with ammonia-oxidizing bacteria immobilized by sodium alginate alone, the bacteria immobilized by PVA and sodium alginate were superior with respect to pH resistance, the number of reuses, material cost, heat resistance, and ammonia oxidation ability. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  11. Characterization of Purified Staphylococcal Lipase1

    PubMed Central

    Vadehra, D. V.; Harmon, L. G.

    1967-01-01

    Purified staphylococcal lipase had an optimal pH of 8.3 for activity at 37 C, and an optimal temperature of 45 C at pH 8.0. During storage, the enzyme lost less than 10% of the activity over a period of 21 days at 4 and -23 C. The enzyme retained 93% of the activity when heated for 30 min at 50 C and was 95% destroyed in 30 min at 70 C. The purified lipase was capable of hydrolyzing a variety of natural fats and oils. However, the enzyme was three times more active on nonhydrogenated soybean oil than on hydrogenated soybean oil with an iodine value of <3.0. The enzyme was also capable of hydrolyzing fatty acids on the α, β, and α′ positions of a synthetic mixed triglyceride. In general, the presence of oxidizing agents increased the activity and the presence of reducing agents decreased the activity of the lipase enzyme. PMID:6035042

  12. Adsorption performance of mixed dyes on alkalization loofah fibers

    NASA Astrophysics Data System (ADS)

    Wang, Yongli; Liu, Jinyan; Li, Xingxing

    2018-02-01

    When the polyporous structures of loofah fiber is adequately exposed after alkali treatment,lignin, hemicellulose and pectin are removed. Specific surface area is increased to maximum, which means the efficiency of absorptivity is highest. In this paper, by using alkalization loofah (AL) as adsorbent, the effect of loofah fiber on waste water treatment is studied under the efficiency of loofah fiber which contain acridine yellow, methylene blue, mixed solution of the two dyes. The optimum treatment conditions of loofah fiber were studied from five aspects which include dosage, temperature, mixing time, pH and concentration. The results showed that the optimal conditions are 30°C, pH 8.0, 20mg dosage of loofah fiber in 40ml solution and mixing time 25min. The optimal treatment conditions of mixed dyes were studied from the aspects of mixing time, the dosage of AL and the molar ratio of the two components in the mixed dyes.

  13. Production, optimization and characterization of fibrinolytic enzyme by Bacillus subtilis RJAS19.

    PubMed

    Kumar, D J Mukesh; Rakshitha, R; Vidhya, M Annu; Jennifer, P Sharon; Prasad, Sandip; Kumar, M Ravi; Kalaichelvan, P T

    2014-04-01

    The present study aimed at the production, purification and characterization of fibrinolytic nattokinase enzyme from the bacteria isolated from natto food. For the purpose, a fibrinolytic bacterium was isolated and identified as Bacillus subtilis based on 16S rDNA sequence analysis. The strain was employed for the production and optimization of fibrinolytic enzyme. The strain showed better enzyme production during 72nd h of incubation time with 50 degrees C at the pH 9. The lactose and peptone were found to be increasing the enzyme production rate. The enzyme produced was purified and also characterized with the help of SDS-PAGE analysis. The activity and stability profile of the purified enzyme was tested against different temperature and pH. The observations suggesting that the potential of fibrinolytic enzyme produced by Bacillus subtilis RJAS 19 for its applications in preventive medicines.

  14. Approach to method development and validation in capillary electrophoresis for enantiomeric purity testing of active basic pharmaceutical ingredients.

    PubMed

    Sokoliess, Torsten; Köller, Gerhard

    2005-06-01

    A chiral capillary electrophoresis system allowing the determination of the enantiomeric purity of an investigational new drug was developed using a generic method development approach for basic analytes. The method was optimized in terms of type and concentration of both cyclodextrin (CD) and electrolyte, buffer pH, temperature, voltage, and rinsing procedure. Optimal chiral separation of the analyte was obtained using an electrolyte with 2.5% carboxymethyl-beta-CD in 25 mM NaH2PO4 (pH 4.0). Interchanging the inlet and outlet vials after each run improved the method's precision. To assure the method's suitability for the control of enantiomeric impurities in pharmaceutical quality control, its specificity, linearity, precision, accuracy, and robustness were validated according to the requirements of the International Conference on Harmonization. The usefulness of our generic method development approach for the validation of robustness was demonstrated.

  15. Optimization of ultrasound extraction of phenolic compounds from coconut (Cocos nucifera) shell powder by response surface methodology.

    PubMed

    Rodrigues, Sueli; Pinto, Gustavo A S; Fernandes, Fabiano A N

    2008-01-01

    Coconut is a tropical fruit largely consumed in many countries. In some areas of the Brazilian coast, coconut shell represents more than 60% of the domestic waste volume. The coconut shell is composed mainly of lignin and cellulose, having a chemical composition very similar to wood and suitable for phenolic extraction. In this work, the use of ultrasound to extract phenolic compounds from coconut shell was evaluated. The effect of temperature, solution to solid ratio, pH and extraction time were evaluated through a 2(4) experimental planning. The extraction process was also optimized using surface response methodology. At the optimum operating condition (30 degrees C, solution to solid ratio of 50, 15 min of extraction and pH 6.5) the process yielded 22.44 mg of phenolic compounds per gram of coconut shell.

  16. [Biosorption of heavy metals in fluoritum decoction by fungal mycelium].

    PubMed

    Cui, Pei-wu; Hu, Wei; Hu, Ya-qiang; Tan, Zhao-yang

    2014-09-01

    To explore the biosorption technology of heavy metals in Fluoritum decoction by fungal mycelium. Four factors including fungal mycelium amount, adsorption time, pH value and temperature were employed to estimate the fungal biomass adsorption conditions for removing the heavy metals in Fluoritum decoction. Then an orthogonal experimental design was taken to optimize the biosorption process, and the removal efficiency was also evaluated. Under the optimized conditions of 1.0 g/50 mL Fluoritum decoction, 3 hours adsorption time, pH 5.0 and 40 degrees C, a result of 70.12% heavy metals removal rate was accomplished with 35.99% calcium ion loss. The study indicates that removing of heavy metals in Fluoritum decoction through fungal mycelium is feasible, and the experiment results can also provide a basis for further research on biosorption of heavy metals in traditional Chinese medicine

  17. Production of a novel bioflocculant and its flocculation performance in aluminum removal.

    PubMed

    Li, Lixin; Ma, Fang; Zuo, Huimin

    2016-04-02

    A novel bioflocculant CBF with high flocculating activity, produced by mixed culture of Rhizobium radiobacter F2 and Bacillus sphaericus F6 from soil, was investigated with regard to its production and flocculation performance in Al(III) removal. The most preferred carbon source, nitrogen source and C/N ratio (w/w) for strains F2 and F6 to produce CBF were glucose, urea and 20, respectively. The optimal inoculum size for CBF production was 10 % (v/v). The optimal initial pH, culture temperature and shaking speed were 7-8, 30°C and 140 r/min for 24 h, respectively, under which the flocculating activity of the bioflocculant reached 98.52 %. According to literature review, flocculant dosage, coagulant aid dosage, pH, hydraulic condition of coagulation and sedimentation time are considered as influencing parameters for CBF flocculation performance in Al(III) removal by L16(4(5)) orthogonal design. The optimal conditions for Al(III) removal obtained through analysis and verification experiments were as follows: CBF, 28 mg/L; coagulant aid, 1.5 mL/L; initial pH, 8.0; and hydraulic conditions of coagulation: stir speed, 160 r/min; stir time, 40 s; and sedimentation time, 30 min. Under the optimal conditions, the removal efficiency of Al(III) was 92.95 %. Overall, these findings indicate that bioflocculant CBF offers an effective alternative method of decreasing Al(III) during drinking water treatment.

  18. Fabrication of the tea saponin functionalized reduced graphene oxide for fast adsorptive removal of Cd(II) from water

    NASA Astrophysics Data System (ADS)

    Li, Zhigang; Liu, Zhifeng; Wu, Zhibin; Zeng, Guangming; Shao, Binbin; Liu, Yujie; Jiang, Yilin; Zhong, Hua; Liu, Yang

    2018-05-01

    A novel graphene-based material of tea saponin functionalized reduced graphene oxide (TS-RGO) was synthesized via a facil thermal method, and it was characterized as the absorbent for Cd(II) removal from aqueous solutions. The factors on adsorption process including solution pH, contact time, initial concentration of Cd(II) and background electrolyte cations were studied to optimize the conditions for maximum adsorption at room temperature. The results indicated that Cd(II) adsorption was strongly dependent on pH and could be strongly affected by background electrolytes and ionic strength. The optimal pH and required equilibrium time was 6.0 and 10 min, respectively. The Cd(II) removal decreased with the presence of background electrolyte cations (Na+ < Ca2+ < Al3+). The adsorption kinetics of Cd(II) followed well with the pseudo-second-order model. The adsorption isotherm fitted well to the Langmuir model, indicating that the adsorption was a monolayer adsorption process occurred on the homogeneous surfaces of TS-RGO. The maximum monolayer adsorption capacity was 127 mg/g at 313 K and pH 6.0. Therefore, the TS-RGO was considered to be a cost-effective and promising material for the removal of Cd(II) from wastewater.

  19. Heterologous expression, biochemical characterization, and overproduction of alkaline α-amylase from Bacillus alcalophilus in Bacillus subtilis

    PubMed Central

    2011-01-01

    Background Alkaline α-amylases have potential applications for hydrolyzing starch under high pH conditions in the starch and textile industries and as ingredients in detergents for automatic dishwashers and laundries. While the alkaline α-amylase gains increased industrial interest, the yield of alkaline α-amylases from wild-type microbes is low, and the combination of genetic engineering and process optimization is necessary to achieve the overproduction of alkaline α-amylase. Results The alkaline α-amylase gene from Bacillus alcalophilus JN21 (CCTCC NO. M 2011229) was cloned and expressed in Bacillus subtilis strain WB600 with vector pMA5. The recombinant alkaline α-amylase was stable at pH from 7.0 to 11.0 and temperature below 40°C. The optimum pH and temperature of alkaline α-amylase was 9.0 and 50°C, respectively. Using soluble starch as the substrate, the Km and Vmax of alkaline α-amylase were 9.64 g/L and 0.80 g/(L·min), respectively. The effects of medium compositions (starch, peptone, and soybean meal) and temperature on the recombinant production of alkaline α-amylase in B. subtilis were investigated. Under the optimal conditions (starch concentration 0.6% (w/v), peptone concentration 1.45% (w/v), soybean meal concentration 1.3% (w/v), and temperature 37°C), the highest yield of alkaline α-amylase reached 415 U/mL. The yield of alkaline α-amylase in a 3-L fermentor reached 441 U/mL, which was 79 times that of native alkaline α-amylase from B. alcalophilus JN21. Conclusions This is the first report concerning the heterologous expression of alkaline α-amylase in B. subtilis, and the obtained results make it feasible to achieve the industrial production of alkaline α-amylase with the recombinant B. subtilis. PMID:21978209

  20. Modelling the effect of temperature, water activity and pH on the growth of Serpula lacrymans.

    PubMed

    Maurice, S; Coroller, L; Debaets, S; Vasseur, V; Le Floch, G; Barbier, G

    2011-12-01

    To predict the risk factors for building infestation by Serpula lacrymans, which is one of the most destructive fungi causing timber decay in buildings. The growth rate was assessed on malt extract agar media at temperatures between 1.5 and 45°C, at water activity (a(w)) over the range of 0.800-0.993 and at pH ranges from 1.5 to 11.0. The radial growth rate (μ) and the lag phase (λ) were estimated from the radial growth kinetics via the plots radius vs time. These parameters were then modelled as a function of the environmental factors tested. Models derived from the cardinal model (CM) were used to fit the experimental data and allowed an estimation of the optimal and limit values for fungal growth. Optimal growth rate occurred at 20°C, at high a(w) level (0.993) and at a pH range between 4.0 and 6.0. The strain effect on the temperature parameters was further evaluated using 14 strains of S. lacrymans. The robustness of the temperature model was validated on data sets measured in two different wood-based media (Quercus robur L. and Picea abies). The two-step procedure of exponential model with latency followed by the CM with inflection gives reliable predictions for the growth conditions of a filamentous fungus in our study. The procedure was validated for the study of abiotic factors on the growth rate of S. lacrymans. This work describes the usefulness of evaluating the effect of physico-chemical factors on fungal growth in predictive building mycology. Consequently, the developed mathematical models for predicting fungal growth on a macroscopic scale can be used as a tool for risk assessment of timber decay in buildings. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  1. Parameter optimization of electrolytic process of obtaining sodium hypochlorite for disinfection of water

    NASA Astrophysics Data System (ADS)

    Bogoslovskii, S. Yu; Kuznetsov, N. N.; Boldyrev, V. S.

    2017-11-01

    Electrochlorination parameters were optimized in flowing and non-flowing modes for a cell with a volume of 1 l. At a current density of 0.1 A/cm2 in the range of flow rates from 0.8 to 6.0 l/h with a temperature of the initial solution below 20°C the outlet temperature is maintained close to the optimal 40°C. The pH of the solution during electrolysis increases to 8.8 ÷ 9.4. There was studied a process in which a solution with a temperature of 7-8°C and a concentration of sodium chloride of 25 and 35 g/l in non-flowing cell was used. The dependence of the concentration of active chlorine on the electrolysis time varies with the concentration of the initial solution of sodium chloride. In case of chloride concentration of 25 g/l virtually linear relationship makes it easy to choose the time of electrolysis with the aim of obtaining the needed concentration of the product.

  2. Precise Tuning of Facile One-Pot Gelatin Methacryloyl (GelMA) Synthesis

    NASA Astrophysics Data System (ADS)

    Shirahama, Hitomi; Lee, Bae Hoon; Tan, Lay Poh; Cho, Nam-Joon

    2016-08-01

    Gelatin-methacryloyl (GelMA) is one of the most commonly used photopolymerizable biomaterials in bio-applications. However, GelMA synthesis remains suboptimal, as its reaction parameters have not been fully investigated. The goal of this study is to establish an optimal route for effective and controllable GelMA synthesis by systematically examining reaction parameters including carbonate-bicarbonate (CB) buffer molarity, initial pH adjustment, MAA concentration, gelatin concentration, reaction temperature, and reaction time. We employed several analytical techniques in order to determine the degree of substitution (DS) and conducted detailed structural analysis of the synthesized polymer. The results enabled us to optimize GelMA synthesis, showing the optimal conditions to balance the deprotonation of amino groups with minimizing MAA hydrolysis, which led to nearly complete substitution. The optimized conditions (low feed ratio of MAA to gelatin (0.1 mL/g), 0.25 M CB buffer at pH 9, and a gelatin concentration of 10-20%) enable a simplified reaction scheme that produces GelMA with high substitution with just one-step addition of MAA in one pot. Looking forward, these optimal conditions not only enable facile one-pot GelMA synthesis but can also guide researchers to explore the efficient, high methacrylation of other biomacromolecules.

  3. Precise Tuning of Facile One-Pot Gelatin Methacryloyl (GelMA) Synthesis

    PubMed Central

    Shirahama, Hitomi; Lee, Bae Hoon; Tan, Lay Poh; Cho, Nam-Joon

    2016-01-01

    Gelatin-methacryloyl (GelMA) is one of the most commonly used photopolymerizable biomaterials in bio-applications. However, GelMA synthesis remains suboptimal, as its reaction parameters have not been fully investigated. The goal of this study is to establish an optimal route for effective and controllable GelMA synthesis by systematically examining reaction parameters including carbonate-bicarbonate (CB) buffer molarity, initial pH adjustment, MAA concentration, gelatin concentration, reaction temperature, and reaction time. We employed several analytical techniques in order to determine the degree of substitution (DS) and conducted detailed structural analysis of the synthesized polymer. The results enabled us to optimize GelMA synthesis, showing the optimal conditions to balance the deprotonation of amino groups with minimizing MAA hydrolysis, which led to nearly complete substitution. The optimized conditions (low feed ratio of MAA to gelatin (0.1 mL/g), 0.25 M CB buffer at pH 9, and a gelatin concentration of 10–20%) enable a simplified reaction scheme that produces GelMA with high substitution with just one-step addition of MAA in one pot. Looking forward, these optimal conditions not only enable facile one-pot GelMA synthesis but can also guide researchers to explore the efficient, high methacrylation of other biomacromolecules. PMID:27503340

  4. Construction of the yeast whole-cell Rhizopus oryzae lipase biocatalyst with high activity.

    PubMed

    Chen, Mei-ling; Guo, Qin; Wang, Rui-zhi; Xu, Juan; Zhou, Chen-wei; Ruan, Hui; He, Guo-qing

    2011-07-01

    Surface display is effectively utilized to construct a whole-cell biocatalyst. Codon optimization has been proven to be effective in maximizing production of heterologous proteins in yeast. Here, the cDNA sequence of Rhizopus oryzae lipase (ROL) was optimized and synthesized according to the codon bias of Saccharomyces cerevisiae, and based on the Saccharomyces cerevisiae cell surface display system with α-agglutinin as an anchor, recombinant yeast displaying fully codon-optimized ROL with high activity was successfully constructed. Compared with the wild-type ROL-displaying yeast, the activity of the codon-optimized ROL yeast whole-cell biocatalyst (25 U/g dried cells) was 12.8-fold higher in a hydrolysis reaction using p-nitrophenyl palmitate (pNPP) as the substrate. To our knowledge, this was the first attempt to combine the techniques of yeast surface display and codon optimization for whole-cell biocatalyst construction. Consequently, the yeast whole-cell ROL biocatalyst was constructed with high activity. The optimum pH and temperature for the yeast whole-cell ROL biocatalyst were pH 7.0 and 40 °C. Furthermore, this whole-cell biocatalyst was applied to the hydrolysis of tributyrin and the resulted conversion of butyric acid reached 96.91% after 144 h.

  5. The use of artificial neural network (ANN) for the prediction and simulation of oil degradation in wastewater by AOP.

    PubMed

    Mustafa, Yasmen A; Jaid, Ghydaa M; Alwared, Abeer I; Ebrahim, Mothana

    2014-06-01

    The application of advanced oxidation process (AOP) in the treatment of wastewater contaminated with oil was investigated in this study. The AOP investigated is the homogeneous photo-Fenton (UV/H2O2/Fe(+2)) process. The reaction is influenced by the input concentration of hydrogen peroxide H2O2, amount of the iron catalyst Fe(+2), pH, temperature, irradiation time, and concentration of oil in the wastewater. The removal efficiency for the used system at the optimal operational parameters (H2O2 = 400 mg/L, Fe(+2) = 40 mg/L, pH = 3, irradiation time = 150 min, and temperature = 30 °C) for 1,000 mg/L oil load was found to be 72%. The study examined the implementation of artificial neural network (ANN) for the prediction and simulation of oil degradation in aqueous solution by photo-Fenton process. The multilayered feed-forward networks were trained by using a backpropagation algorithm; a three-layer network with 22 neurons in the hidden layer gave optimal results. The results show that the ANN model can predict the experimental results with high correlation coefficient (R (2) = 0.9949). The sensitivity analysis showed that all studied variables (H2O2, Fe(+2), pH, irradiation time, temperature, and oil concentration) have strong effect on the oil degradation. The pH was found to be the most influential parameter with relative importance of 20.6%.

  6. Effects of environmental conditions on growth and survival of Salmonella in pasteurized whole egg.

    PubMed

    Jakočiūnė, Džiuginta; Bisgaard, Magne; Hervé, Gaëlle; Protais, Jocelyne; Olsen, John Elmerdahl; Chemaly, Marianne

    2014-08-01

    This study investigated the influence of three parameters (time, temperature and NaCl concentration) on survival and four parameters (temperature, NaCl and lysozyme concentrations and pH) on growth of Salmonella enterica serovar Enteritidis (S. Enteritidis) in pasteurized whole egg (PWE). Doehlert uniform shell design was employed to choose conditions for trials and data was fitted to polynomial models and were presented as estimated response surfaces. A model for prediction of reduction of S. Enteritidis in PWE within temperatures between 50 and 58°C, NaCl concentrations of 0-12%, and heating times between 30 and 210s and a model for prediction of growth rate of S. Enteritidis in PWE in the temperature range of 1-25°C, NaCl concentration of 0-12%, pH between 5 and 9, and lysozyme concentrations of 107-1007 U/mg proteins were developed. The maximum reduction condition was 58°C, 0% of NaCl at a fixed heating time of 120s, while maximum growth rate was estimated at 25°C and 0% of NaCl. pH and lysozyme concentration were shown not to influence growth performance significantly in the range of values studied. Results inform industry of the optimal pasteurization and storage parameters for liquid whole egg. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Removal of Nitrate in Simulated Water at Low Temperature by a Novel Psychrotrophic and Aerobic Bacterium, Pseudomonas taiwanensis Strain J.

    PubMed

    He, Tengxia; Ye, Qing; Sun, Quan; Cai, Xi; Ni, Jiupai; Li, Zhenlun; Xie, Deti

    2018-01-01

    Low temperatures and high pH generally inhibit the biodenitrification. Thus, it is important to explore the psychrotrophic and alkali-resisting microorganism for degradation of nitrogen. This research was mainly focused on the identification of a psychrotrophic strain and preliminary explored its denitrification characteristics. The new strain J was isolated using the bromothymol blue solid medium and identified as Pseudomonas taiwanensis on the basis of morphology and phospholipid fatty acid as well as 16S rRNA gene sequence analyses, which is further testified to work efficiently for removing nitrate from wastewater at low temperature circumstances. This is the first report that Pseudomonas taiwanensis possessed excellent tolerance to low temperature, with 15°C as its optimum and 5°C as viable. The Pseudomonas taiwanensis showed unusual ability of aerobic denitrification with the nitrate removal efficiencies of 100% at 15°C and 51.61% at 5°C. Single factor experiments showed that the optimal conditions for denitrification were glucose as carbon source, 15°C, shaking speed 150 r/min, C/N 15, pH ≥ 7, and incubation quantity 2.0 × 10 6  CFU/mL. The nitrate and total nitrogen removal efficiencies were up to 100% and 93.79% at 15°C when glucose is served as carbon source. These results suggested that strain J had aerobic denitrification ability, as well as the notable ability to tolerate the low temperature and high pH.

  8. Amylolytic Enzymes Acquired from L-Lactic Acid Producing Enterococcus faecium K-1 and Improvement of Direct Lactic Acid Production from Cassava Starch.

    PubMed

    Unban, Kridsada; Kanpiengjai, Apinun; Takata, Goro; Uechi, Keiko; Lee, Wen-Chien; Khanongnuch, Chartchai

    2017-09-01

    An amylolytic lactic acid bacterium isolate K-1 was isolated from the wastewater of a cassava starch manufacturing factory and identified as Entercoccus faecium based on 16S rRNA gene sequence analysis. An extracellular α-amylase was purified to homogeneity and the molecular weight of the purified enzyme was approximately 112 kDa with optimal pH value and temperature measured of 7.0 and 40 °C, respectively. It was stable at a pH range of 6.0-7.0, but was markedly sensitive to high temperatures and low pH conditions, even at a pH value of 5. Ba 2+ , Al 3+ , and Co 2+ activated enzyme activity. This bacterium was capable of producing 99.2% high optically pure L-lactic acid of 4.3 and 8.2 g/L under uncontrolled and controlled pH at 6.5 conditions, respectively, in the MRS broth containing 10 g/L cassava starch as the sole carbon source when cultivated at 37 °C for 48 h. A control pH condition of 6.5 improved and stabilized the yield of L-lactic acid production directly from starch even at a high concentration of starch at up to 150 g/L. This paper is the first report describing the properties of purified α-amylase from E. faecium. Additionally, pullulanase and cyclodextrinase activities were also firstly recorded from E. faecium K-1.

  9. A Novel Acid-Stable Endo-Polygalacturonase from Penicillium oxalicum CZ1028: Purification, Characterization, and Application in the Beverage Industry.

    PubMed

    Cheng, Zhong; Chen, Dong; Lu, Bo; Wei, Yutuo; Xian, Liang; Li, Yi; Luo, Zhenzhen; Huang, Ribo

    2016-06-28

    Acidic endo-polygalacturonases are the major part of pectinase preparations and extensively applied in the clarification of fruits juice, vegetables extracts, and wines. However, most of the reported fungal endo-polygalacturonases are active and stable under narrow pH range and low temperatures. In this study, an acidic endo-polygalacturonase (EPG4) was purified and characterized from a mutant strain of Penicillium oxalicum. The N-terminal amino acid sequence of EPG4 (ATTCTFSGSNGAASASKSQT) was different from those of reported endopolygalacturonases. EPG4 displayed optimal pH and temperature at 5.0 and 60-70°C towards polygalacturonic acid (PGA), respectively, and was notably stable at pH 2.2-7.0. When tested against pectins, EPG4 showed enzyme activity over a broad acidic pH range (>15.0% activity at pH 2.2-6.0 towards citrus pectin; and >26.6% activity at pH 2.2-7.0 towards apple pectin). The Km and Vmax values were determined as 1.27 mg/ml and 5,504.6 U/mg, respectively. The enzyme hydrolyzed PGA in endo-manner, releasing oligo-galacturonates from PGA, as determined by TLC. Addition of EPG4 (3.6 U/ml) significantly reduced the viscosity (by 42.4%) and increased the light transmittance (by 29.5%) of the papaya pulp, and increased the recovery (by 24.4%) of the papaya extraction. All of these properties make the enzyme a potential application in the beverage industry.

  10. Optimisation of shock absorber process parameters using failure mode and effect analysis and genetic algorithm

    NASA Astrophysics Data System (ADS)

    Mariajayaprakash, Arokiasamy; Senthilvelan, Thiyagarajan; Vivekananthan, Krishnapillai Ponnambal

    2013-07-01

    The various process parameters affecting the quality characteristics of the shock absorber during the process were identified using the Ishikawa diagram and by failure mode and effect analysis. The identified process parameters are welding process parameters (squeeze, heat control, wheel speed, and air pressure), damper sealing process parameters (load, hydraulic pressure, air pressure, and fixture height), washing process parameters (total alkalinity, temperature, pH value of rinsing water, and timing), and painting process parameters (flowability, coating thickness, pointage, and temperature). In this paper, the process parameters, namely, painting and washing process parameters, are optimized by Taguchi method. Though the defects are reasonably minimized by Taguchi method, in order to achieve zero defects during the processes, genetic algorithm technique is applied on the optimized parameters obtained by Taguchi method.

  11. Two growth mechanisms in one-step fabrication of the oxide matrix for FeSiAl soft magnetic composites

    NASA Astrophysics Data System (ADS)

    Wu, Chen; Gao, Xinwei; Zhao, Guoliang; Jiang, Yinzhu; Yan, Mi

    2018-04-01

    Hydrolysis precipitation as a new method was used in the preparation of oxide insulation matrix for FeSiAl soft magnetic composites (SMCs). The growth and composition of the matrix can be tuned by the concentration of the Al(NO3)3 solution, reaction temperature and pH value during the hydrolysis. With optimized Al(NO3)3 concentration of 0.6 mol/L and hydrolysis temperature of 75 °C, two mechanisms have been revealed in the formation of the insulation coating depending on the pH of the Al(NO3)3 solution. When pH = 3, the coating layer contains a mixture of Al2O3 and Fe2O3, while Al2O3 and SiO2 form as the coating for pH = 8. Despite that the Al2O3 dominates for both conditions, it grows via different routes. The Al(OH)3 as the precursor forms through Al3+ hydrolysis and heterogeneous nucleation for pH = 3. With increased pH to 8, the Al3+ directly reacts with OH- to form Al(OH)3 colloidal particles which adsorb onto the surface of FeSiAl powders via electrostatic attraction. Both mechanisms give rise to satisfactory magnetic performance with high effective permeability (μe = 103.5 and 113.4) and low core loss (Pcv = 278.4 mW·cm-3 and 237.8 mW·cm-3) for pH = 3 and 8 measured at 100 mT, 50 kHz.

  12. Exploring the Limits of Cell Adhesion under Shear Stress within Physiological Conditions and beyond on a Chip.

    PubMed

    Stamp, Melanie E M; Jötten, Anna M; Kudella, Patrick W; Breyer, Dominik; Strobl, Florian G; Geislinger, Thomas M; Wixforth, Achim; Westerhausen, Christoph

    2016-10-21

    Cell adhesion processes are of ubiquitous importance for biomedical applications such as optimization of implant materials. Here, not only physiological conditions such as temperature or pH, but also topographical structures play crucial roles, as inflammatory reactions after surgery can diminish osseointegration. In this study, we systematically investigate cell adhesion under static, dynamic and physiologically relevant conditions employing a lab-on-a-chip system. We screen adhesion of the bone osteosarcoma cell line SaOs-2 on a titanium implant material for pH and temperature values in the physiological range and beyond, to explore the limits of cell adhesion, e.g., for feverish and acidic conditions. A detailed study of different surface roughness R q gives insight into the correlation between the cells' abilities to adhere and withstand shear flow and the topography of the substrates, finding a local optimum at R q = 22 nm. We use shear stress induced by acoustic streaming to determine a measure for the ability of cell adhesion under an external force for various conditions. We find an optimum of cell adhesion for T = 37 °C and pH = 7.4 with decreasing cell adhesion outside the physiological range, especially for high T and low pH. We find constant detachment rates in the physiological regime, but this behavior tends to collapse at the limits of 41 °C and pH 4.

  13. Cerebral interstitial tissue oxygen tension, pH, HCO3, CO2.

    PubMed

    Charbel, F T; Hoffman, W E; Misra, M; Hannigan, K; Ausman, J I

    1997-10-01

    There are many techniques for monitoring the injured brain following trauma, subarachnoid hemorrhage, or surgery. It is thought that the major determinants for recovery of injured cerebral tissue are oxygen, glucose delivery, and the clearance of metabolites. These factors, at optimal levels, are probably responsible for the regaining of neuronal functions. These parameters are in turn dependent on the tissue's blood flow and metabolism. We have been using a single, compact, polyethylene sensor, the Paratrend 7 for the measurement of cerebral oxygen tension, CO2, pH, and temperature. This sensor is designed for continuous blood gas analysis to aid in monitoring neurosurgical patients, both during surgery and in the intensive care unit. Using the Paratrend 7 sensor, we found the normal range of values to be: PO2 33 +/- 11 mm Hg; PCO2 48 +/- 7 mm Hg; pH 7.19 +/- 0.11. Critical measurements are considered to be tissue PO2 < 10 mm Hg; PCO2 > 60 mm Hg, and pH < 6.8. We have had no complications with this device; the risks are similar to those of placing a parenchymal intracranial pressure monitor. We believe that assessment of interstitial cerebral oxygen saturation can be of great value both intraoperatively and postoperatively. In our experience, the Paratrend 7 system is an effective method of measuring tissue cerebral oxygen tension, along with carbon dioxide levels, pH, and temperature.

  14. A feasible enzymatic process for D-tagatose production by an immobilized thermostable L-arabinose isomerase in a packed-bed bioreactor.

    PubMed

    Kim, Hye-Jung; Ryu, Se-Ah; Kim, Pil; Oh, Deok-Kun

    2003-01-01

    To develop a feasible enzymatic process for d-tagatose production, a thermostable l-arabinose isomerase, Gali152, was immobilized in alginate, and the galactose isomerization reaction conditions were optimized. The pH and temperature for the maximal galactose isomerization reaction were pH 8.0 and 65 degrees C in the immobilized enzyme system and pH 7.5 and 60 degrees C in the free enzyme system. The presence of manganese ion enhanced galactose isomerization to tagatose in both the free and immobilized enzyme systems. The immobilized enzyme was more stable than the free enzyme at the same pH and temperature. Under stable conditions of pH 8.0 and 60 degrees C, the immobilized enzyme produced 58 g/L of tagatose from 100 g/L galactose in 90 h by batch reaction, whereas the free enzyme produced 37 g/L tagatose due to its lower stability. A packed-bed bioreactor with immobilized Gali152 in alginate beads produced 50 g/L tagatose from 100 g/L galactose in 168 h, with a productivity of 13.3 (g of tagatose)/(L-reactor.h) in continuous mode. The bioreactor produced 230 g/L tagatose from 500 g/L galactose in continuous recycling mode, with a productivity of 9.6 g/(L.h) and a conversion yield of 46%.

  15. Concentrating phenolic acids from Lonicera japonica by nanofiltration technology

    NASA Astrophysics Data System (ADS)

    Li, Cunyu; Ma, Yun; Li, Hongyang; Peng, Guoping

    2017-03-01

    Response surface analysis methodology was used to optimize the concentrate process of phenolic acids from Lonicera japonica by nanofiltration technique. On the basis of the influences of pressure, temperature and circulating volume, the retention rate of neochlorogenic acid, chlorogenic acid and 4-dicaffeoylquinic acid were selected as index, molecular weight cut-off of nanofiltration membrane, concentration and pH were selected as influencing factors during concentrate process. The experiment mathematical model was arranged according to Box-Behnken central composite experiment design. The optimal concentrate conditions were as following: nanofiltration molecular weight cut-off, 150 Da; solutes concentration, 18.34 µg/mL; pH, 4.26. The predicted value of retention rate was 97.99% under the optimum conditions, and the experimental value was 98.03±0.24%, which was in accordance with the predicted value. These results demonstrate that the combination of Box-Behnken design and response surface analysis can well optimize the concentrate process of Lonicera japonica water-extraction by nanofiltration, and the results provide the basis for nanofiltration concentrate for heat-sensitive traditional Chinese medicine.

  16. Fast Synthesis of Gibbsite Nanoplates and Process Optimization using Box-Behnken Experimental Design

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang, Xin; Zhang, Xianwen; Graham, Trent R.

    Developing the ability to synthesize compositionally and morphologically well-defined gibbsite particles at the nanoscale with high yield is an ongoing need that has not yet achieved the level of rational design. Here we report optimization of a clean inorganic synthesis route based on statistical experimental design examining the influence of Al(OH)3 gel precursor concentration, pH, and aging time at temperature. At 80 oC, the optimum synthesis conditions of gel concentration at 0.5 M, pH at 9.2, and time at 72 h maximized the reaction yield up to ~87%. The resulting gibbsite product is composed of highly uniform euhedral hexagonal nanoplatesmore » within a basal plane diameter range of 200-400 nm. The independent roles of key system variables in the growth mechanism are considered. On the basis of these optimized experimental conditions, the synthesis procedure, which is both cost-effective and environmentally friendly, has the potential for mass production scale-up of high quality gibbsite material for various fundamental research and industrial applications.« less

  17. Response surface methodology as an approach to determine the optimal activities of xylose reductase and xylitol dehydrogenase enzymes from Candida Mogii.

    PubMed

    Mayerhoff, Zea D V L; Roberto, Inês C; Franco, Telma T

    2006-05-01

    A central composite experimental design leading to a set of 16 experiments with different combinations of pH and temperature was performed to attain the optimal activities of xylose reductase (XR) and xylitol dehydrogenase (XDH) enzymes from Candida mogii cell extract. Under optimized conditions (pH 6.5 and 38 degrees C), the XR and XDH activities were found to be 0.48 U/ml and 0.22 U/ml, respectively, resulting in an XR to XDH ratio of 2.2. Stability, cofactor specificity and kinetic parameters of the enzyme XR were also evaluated. XR activity remained stable for 3 h under 4 and 38 degrees C and for 4 months of storage at -18 degrees C. Studies on cofactor specificity showed that only NADPH-dependent XR was obtained under the cultivation conditions employed. The XR present in C. mogii extracts showed a superior Km value for xylose when compared with other yeast strains. Besides, this parameter was not modified after enzyme extraction by aqueous two-phase system.

  18. [United Plackett-Burman and Box-Behnken design to control formation of indirubin in process of preparing indigo naturalis].

    PubMed

    Liu, Zeyu; Su, Zhetong; Yang, Ming; Zou, Wenquan

    2010-10-01

    To screen the factors that affect indirubin-generated significantly in the process of preparing indigo naturalis, optimize level combination and determine the optimum technology for indirubin-generated. Using concentration of indirubin (mg x g(-1)) that generated by fresh leaf as an index, Plackett-Burman design, Box-Behnken design response surface analysis as the statistical method, we screened the significantly influencing factors and the optimal level combination. The soaking and making indirubin process in preparing indigo naturalis was identified as the wax is not removed before immersion with immersion pH 7, solvent volume-leaf weight (mL: g)15, soaked not avoided light, soaking 48 h, temperature 60 degrees C, ventilation time of 180 min, and added ammonia water to adjust pH to 10.5. The soaking and making indirubin process in preparing indigo naturalis is optimized systematically. It clarify the various factors on the impact of the active ingredient indirubin which controlled by industrialized production become reality in the process of preparing indigo naturalis, at the same time, it lay the foundation for processing principle of indigo naturalis.

  19. Laccase production by Coriolopsis caperata RCK2011: Optimization under solid state fermentation by Taguchi DOE methodology

    PubMed Central

    Nandal, Preeti; Ravella, Sreenivas Rao; Kuhad, Ramesh Chander

    2013-01-01

    Laccase production by Coriolopsis caperata RCK2011 under solid state fermentation was optimized following Taguchi design of experiment. An orthogonal array layout of L18 (21 × 37) was constructed using Qualitek-4 software with eight most influensive factors on laccase production. At individual level pH contributed higher influence, whereas, corn steep liquor (CSL) accounted for more than 50% of the severity index with biotin and KH2PO4 at the interactive level. The optimum conditions derived were; temperature 30°C, pH 5.0, wheat bran 5.0 g, inoculum size 0.5 ml (fungal cell mass = 0.015 g dry wt.), biotin 0.5% w/v, KH2PO4 0.013% w/v, CSL 0.1% v/v and 0.5 mM xylidine as an inducer. The validation experiments using optimized conditions confirmed an improvement in enzyme production by 58.01%. The laccase production to the level of 1623.55 Ugds−1 indicates that the fungus C. caperata RCK2011 has the commercial potential for laccase. PMID:23463372

  20. Optimization of the Medium for the Production of Extracellular Amylase by the Pseudomonas stutzeri ISL B5 Isolated from Municipal Solid Waste

    PubMed Central

    Dutta, Prajesh; Deb, Akash

    2016-01-01

    The management of municipal solid waste is one of the major problems of the present world. The use of microbial enzymes for sustainable management of the solid waste is the need of the time. In the present study, we have isolated a potent amylase producing strain (ISL B5) from municipal solid waste. The strain was identified as Pseudomonas stutzeri (P. stutzeri) both biochemically and by 16S rDNA sequencing. The optimization studies revealed that the strain ISL B5 exhibited maximum activity in the liquid media containing 2% starch (2.77 U/ml), 0.8% peptone (2.77 U/ml), and 0.001% Ca2+ ion (2.49 U/ml) under the pH 7.5 (2.59 U/ml), temperature 40°C (2.63 U/ml), and 25 h of incubation period (2.49 U/ml). The highest activity of crude enzyme has also been optimized at the pH 8 (2.49 U/ml). PMID:28096816

  1. Potential of Immobilized Whole-Cell Methylocella tundrae as a Biocatalyst for Methanol Production from Methane.

    PubMed

    Mardina, Primata; Li, Jinglin; Patel, Sanjay K S; Kim, In-Won; Lee, Jung-Kul; Selvaraj, Chandrabose

    2016-07-28

    Methanol is a versatile compound that can be biologically synthesized from methane (CH4) by methanotrophs using a low energy-consuming and environment-friendly process. Methylocella tundrae is a type II methanotroph that can utilize CH4 as a carbon and energy source. Methanol is produced in the first step of the metabolic pathway of methanotrophs and is further oxidized into formaldehyde. Several parameters must be optimized to achieve high methanol production. In this study, we optimized the production conditions and process parameters for methanol production. The optimum incubation time, substrate, pH, agitation rate, temperature, phosphate buffer and sodium formate concentration, and cell concentration were determined to be 24 h, 50% CH4, pH 7, 150 rpm, 30°C, 100 mM and 50 mM, and 18 mg/ml, respectively. The optimization of these parameters significantly improved methanol production from 0.66 to 5.18 mM. The use of alginate-encapsulated cells resulted in enhanced methanol production stability and reusability of cells after five cycles of reuse under batch culture conditions.

  2. Optimization of macropore evolution towards high photocatalytic activity enhancement in meso/macro porous Anatase TiO2

    NASA Astrophysics Data System (ADS)

    Nair, Radhika V.; Gayathri, P. K.; Siva Gummaluri, Venkata; Vijayan, C.

    2017-01-01

    We report on an optimization strategy for macro pore evolution leading to the design of highly photocatalytic 3D hierarchical meso/macroporous TiO2 via much simpler, faster and cost effective synthesis scheme. Meso/macro porous TiO2 is an excellent candidate material for photocatalytic applications owing to the availability of internal surfaces as active sites for redox reactions. The current research scenario focuses on the design of highly efficient photocatalytic systems as well as rapid, facile and cost effective methods of synthesis and optimization of parameters. The present report is on the gradual evolution of macropores in anatase TiO2 by the effective control of pH of the solvent, reaction time, temperature, solvent ratio and reactant concentration via a facile hydrothermal method in this regard. 3D hierarchical macroporous structures are obtained at pH 7 within a comparatively short reaction time of 5 h and demonstrated to be highly photocatalytic (with rate constant four times that of P25 nanoparticles) through photodegradation of Rhodamine B dye.

  3. Optimization of preparation of antioxidative peptides from pumpkin seeds using response surface method.

    PubMed

    Fan, Sanhong; Hu, Yanan; Li, Chen; Liu, Yanrong

    2014-01-01

    Protein isolates of pumpkin (Cucurbita pepo L) seeds were hydrolyzed by acid protease to prepare antioxidative peptides. The hydrolysis conditions were optimized through Box-Behnken experimental design combined with response surface method (RSM). The second-order model, developed for the DPPH radical scavenging activity of pumpkin seed hydrolysates, showed good fit with the experiment data with a high value of coefficient of determination (0.9918). The optimal hydrolysis conditions were determined as follows: hydrolyzing temperature 50°C, pH 2.5, enzyme amount 6000 U/g, substrate concentration 0.05 g/ml and hydrolyzing time 5 h. Under the above conditions, the scavenging activity of DPPH radical was as high as 92.82%.

  4. Tolerance to winemaking stress conditions of Patagonian strains of Saccharomyces eubayanus and Saccharomyces uvarum.

    PubMed

    Origone, A C; Del Mónaco, S M; Ávila, J R; González Flores, M; Rodríguez, M E; Lopes, C A

    2017-08-01

    Evaluating the winemaking stress tolerance of a set of both Saccharomyces eubayanus and Saccharomyces uvarum strains from diverse Patagonian habitats. Yeast strains growth was analysed under increasing ethanol concentrations; all of them were able to grow until 8% v/v ethanol. The effect of different temperature and pH conditions as well as at SO 2 and hexose concentrations was evaluated by means of a central composite experimental design. Only two S. uvarum strains (NPCC 1289 and 1321) were able to grow in most stress conditions. Kinetic parameters analysed (μ max and λ) were statistically affected by temperature, pH and SO 2 , but not influenced by sugar concentration. The obtained growth model was used for predicting optimal growth conditions for both strains: 20°C, 0% w/v SO 2 and pH 4·5. Strains from human-associated environments (chichas) presented the highest diversity in the response to different stress factors. Two S. uvarum strains from chichas demonstrated to be the most tolerant to winemaking conditions. This work evidenced the potential use of two S. uvarum yeast strains as starter cultures in wines fermented at low temperatures. Saccharomyces eubayanus was significantly affected by winemaking stress conditions, limiting its use in this industry. © 2017 The Society for Applied Microbiology.

  5. A xylanase with broad pH and temperature adaptability from Streptomyces megasporus DSM 41476, and its potential application in brewing industry.

    PubMed

    Qiu, Zhenhua; Shi, Pengjun; Luo, Huiying; Bai, Yingguo; Yuan, Tiezheng; Yang, Peilong; Liu, Suchun; Yao, Bin

    2010-05-05

    A xylanase gene, xynAM6, was isolated from the genomic DNA library of Streptomyces megasporus DSM 41476 using colony PCR screening method. The 1440-bp full-length gene encodes a 479-amino acid peptide consisting of a putative signal peptide of 36 residues, a family 10 glycoside hydrolase domain and a family 2 carbohydrate-binding module. The mature peptide of xynAM6 was successfully expressed in Pichia pastoris GS115. The optimal pH and temperature were pH 5.5 and 70°C, respectively. The enzyme showed broad temperature adaptability (>60% of the maximum activity at 50-80°C), had good thermostability at 60°C and 70°C, remained stable at pH 4.0-11.0, and was resistant to most proteases. The Km and Vmax values for oat spelt xylan were 1.68mgml(-1) and 436.76μmolmin(-1)mg(-1), respectively, and 2.33mgml(-1) and 406.93μmolmin(-1)mg(-1) for birchwood xylan, respectively. The hydrolysis products of XYNAM6 were mainly xylose and xylobiose. Addition of XYNAM6 (80U) to the brewery mash significantly reduced the filtration rate and viscosity by 36.33% and 35.51%, respectively. These favorable properties probably make XYNAM6 a good candidate for application in brewing industry. Copyright © 2010 Elsevier Inc. All rights reserved.

  6. Factors promoting in vitro excystation of Giardia muris cysts.

    PubMed

    Schaefer, F W; Rice, E W; Hoff, J C

    1984-01-01

    Giardia muris cysts, isolated from mouse faeces, excysted routinely at levels greater than 90%, when induced in 1X Hanks' supplemented with 17 mM glutathione, 29 mM L-cysteine-HCl, and 50 mM NaHCO3 for 30 minutes at 35 degrees C, followed by washing and suspension in trypsin-Tyrode's solution at pH 8.0. Although trypsin was not required in this final step, it enhanced the escape of the trophozoites from their cysts. G. muris excystation was dependent upon the length of the induction period, pH, oxidation-reduction potential and temperature. Optimal induction conditions for excystation were: an induction period of 5 to 30 min; pH of 2; 120 mV oxidation-reduction potential; and a temperature around 35 degrees C. A gradual decline in excystation occurred as pH and oxidation-reduction potential were changed to 7 and 57 mV, respectively. There was a pronounced increase in excystation percentages with increasing temperatures between 0 and 37 degrees C. At 40 degrees C and above, the G. muris cysts showed signs of inactivation. The thermal death point of G. muris cysts was determined to be about 54 degrees C. G. muris cysts showed no polarity; however, the tail or posterior trophozoite portion always emerged through one end of the cyst first. Cytokinensis began within the first hour after excystation. This method always produced extremely active, normal-looking G. muris trophozoites.

  7. Effect of pH and pulsed electric field process parameters on the aflatoxin reduction in model system using response surface methodology: Effect of pH and PEF on Aflatoxin Reduction.

    PubMed

    Vijayalakshmi, Subramanian; Nadanasabhapathi, Shanmugam; Kumar, Ranganathan; Sunny Kumar, S

    2018-03-01

    The presence of aflatoxin, a carcinogenic and toxigenic secondary metabolite produced by Aspergillus species, in food matrix has been a major worldwide problem for years now. Food processing methods such as roasting, extrusion, etc. have been employed for effective destruction of aflatoxins, which are known for their thermo-stable nature. The high temperature treatment, adversely affects the nutritive and other quality attributes of the food, leading to the necessity of application of non-thermal processing techniques such as ultrasonication, gamma irradiation, high pressure processing, pulsed electric field (PEF), etc. The present study was focused on analysing the efficacy of the PEF process in the reduction of the toxin content, which was subsequently quantified using HPLC. The process parameters of different pH model system (potato dextrose agar) artificially spiked with aflatoxin mix standard was optimized using the response surface methodology. The optimization of PEF process effects on the responses aflatoxin B1 and total aflatoxin reduction (%) by pH (4-10), pulse width (10-26 µs) and output voltage (20-65%), fitted 2FI model and quadratic model respectively. The response surface plots obtained for the processes were of saddle point type, with the absence of minimum or maximum response at the centre point. The implemented numerical optimization showed that the predicted and actual values were similar, proving the adequacy of the fitted models and also proved the possible application of PEF in toxin reduction.

  8. Systematic optimization of ethyl glucuronide extraction conditions from scalp hair by design of experiments and its potential effect on cut-off values appraisal.

    PubMed

    Alladio, Eugenio; Biosa, Giulia; Seganti, Fabrizio; Di Corcia, Daniele; Salomone, Alberto; Vincenti, Marco; Baumgartner, Markus R

    2018-05-11

    The quantitative determination of ethyl glucuronide (EtG) in hair samples is consistently used throughout the world to assess chronic excessive alcohol consumption. For administrative and legal purposes, the analytical results are compared with cut-off values recognized by regulatory authorities and scientific societies. However, it has been recently recognized that the analytical results depend on the hair sample pretreatment procedures, including the crumbling and extraction conditions. A systematic evaluation of the EtG extraction conditions from pulverized scalp hair was conducted by design of experiments (DoE) considering the extraction time, temperature, pH, and solvent composition as potential influencing factors. It was concluded that an overnight extraction at 60°C with pure water at neutral pH represents the most effective conditions to achieve high extraction yields. The absence of differential degradation of the internal standard (isotopically-labeled EtG) under such conditions was confirmed and the overall analytical method was validated according to SGWTOX and ISO17025 criteria. Twenty real hair samples with different EtG content were analyzed with three commonly accepted procedures: (a) hair manually cut in snippets and extracted at room temperature; (b) pulverized hair extracted at room temperature; (c) hair treated with the optimized method. Average increments of EtG concentration around 69% (from a to c) and 29% (from b to c) were recorded. In light of these results, the authors urge the scientific community to undertake an inter-laboratory study with the aim of defining more in detail the optimal hair EtG detection method and verifying the corresponding cut-off level for legal enforcements. This article is protected by copyright. All rights reserved.

  9. [Characterization of a thermophilic Geobacillus strain DM-2 degrading hydrocarbons].

    PubMed

    Liu, Qing-kun; Wang, Jun; Li, Guo-qiang; Ma, Ting; Liang, Feng-lai; Liu, Ru-lin

    2008-12-01

    A thermophilic Geobacillus strain DM-2 from a deep-subsurface oil reservoir was investigated on its capability of degrading crude oil under various conditions as well as its characters on degrading hydrocarbons in optimal conditions. The results showed that Geobacillus strain DM-2 was able to degrade crude oil under anoxic wide-range conditions with pH ranging from 4.0 to 10.0, high temperature in the range of 45-70 degrees C and saline concentration ranging from 0.2% to 3.0%. Furthermore, the optimal temperature and pH value for utilizing hydrocarbons by the strain were 60 degrees C and 7.0, respectively. Under such optimal conditions, the strain utilized liquid paraffine emulsified by itself as its carbon source for growth; further analysis by gas chromatography (GC) and infrared absorption spectroscopy demonstrated that it was able to degrade n-alkanes (C14-C30), branched-chain alkanes and aromatic hydrocarbons in crude oil and could also utilize long-chain n-alkanes from C16 to C36, among of which the degradation efficiency of C28 was the highest, up to 88.95%. One metabolite of the strain oxidizing alkanes is fatty acid.While utilizing C16 as carbon source for 5 d, only one fatty acid-acetic acid was detected by HPLC and MS as the product, with the amount of 0.312 g/L, which indicated that it degraded n-alkanes with pathway of inferior terminal oxidation,and then followed by a beta-oxidation pathway. Due to its characters of efficient emulsification, high-performance degradation of hydrocarbons and fatty-acid production under high temperature and anoxic condition, the strain DM-2 may be potentially applied to oil-waste treatment and microbial enhanced heavy oil recovery in extreme conditions.

  10. Studies on the production of alkaline α-amylase from Bacillus subtilis CB-18.

    PubMed

    Nwokoro, Ogbonnaya; Anthonia, Odiase

    2015-01-01

    Amylases are among the main enzymes used in food and other industries. They hydrolyse starch molecules into polymers composing glucose units. Amylases have potential applications in a number of industrial processes including foods and pharmaceutical industries. Alkaline α-amylase has the potential of hydrolysing starch under alkaline pH and is useful in the starch and textile industries and as an ingredient of detergents. Amylases are produced from plants, however, microbial production processes have dominated applications in the industries. Optimization of microbial production processes can result in improved enzyme yields. Amylase activity was assayed by incubating the enzyme solution (0.5 ml) with 1% soluble starch (0.5 ml) in 0.1 M Tris/HCl buffer (pH 8.5). After 30 minutes, the reaction was stopped by the addition of 4 mL of 3,5-dinitrosalicylic acid (DNS) reagent then heated for 10 min in boiling water bath and cooled in a refrigerator. Absorbance readings were used to estimate the units of enzyme activity from glucose standard curve. Hydrolysed native starches from cassava, rice, corn, coco yam, maize and potato and soluble starch were adjusted to pH 8.5 prior to incubation with crude enzyme solution. Reducing sugars produced were therefore determined. The effect of pH on enzyme activity of the alkaline α-amylase was determined by using buffer solutions of different pH (potassium phosphate buffer, 6.0-7.0; Tris-HCl buffer 7.5 to 9.0 and carbonate/bicarbonate buffer, pH 9.5-11) for enzyme assay. The pH stability profile of the enzyme was determined by incubating 0.5 ml of α-amylase enzyme in 0.1 M Tris/HCl buffer (pH 8.5) and 0.5 ml of 1% (w/v) soluble starch (Merck) in 0.1 M Tris/HCl buffer (pH 8.5) for 3 h in various buffers. The effect of temperature on enzyme activity was studied by incubating 0.5 mL of the enzyme solution contained in the test tube and 0.5 mL of 1% soluble starch (Merck) solution prepared in 0.1 M Tris/HCl buffer (pH 8.5) for 3 h at various temperatures (25, 30, 35, 40, 45, 50, 55 and 60°C) in a thermo static water bath. The reactions were stopped by adding DNS reagent. The enzyme activity was therefore determined. Thermal stability was studied by incubating 0.5 ml of enzyme solution in 0.1 M Tris/HCl buffer (pH 8.5) and 0.5 ml of 1% (w/v) soluble starch (Merck) in 0.1 M Tris/HCl buffer (pH 8.5) for 3 h at various temperatures (20, 30, 40, 50, 60 and 70°C) for 60 min. The enzyme displayed optimal activity at pH 8.0 at which it produced maximum specific activity of 34.3 units/mg protein. Maximum stability was at pH 8.0 to 9.0. Maximum activity was observed at temperature of 50°C while thermo stability of the enzyme was observed at 40-50°C. The enzyme displayed a wide range of activities on starch and caused the release of 5.86, 4.75, 5.98, 3.44, 3.96, 8.84 mg/mL reducing sugar from cassava, potato, cocoyam, corn, rice and soluble starch respectively. This investigation reports some biochemical characterization of alkaline α-amylase from Bacillus subtilis CB-18. The substrate specificities of this enzyme on various starches suggested that the alkaline α-amylase enzyme had combined activities on raw and soluble starch.

  11. Effect of variable hydrothermal conditions on sulfur speciation and isotopic compositions mediated by two Thiomicrospira strains

    NASA Astrophysics Data System (ADS)

    Houghton, J.; Wills, E.; Fike, D. A.

    2012-12-01

    Microbially mediated reactions involving elemental sulfur in low temperature hydrothermal environments are a critical component of the net hydrothermal flux of sulfur to the global oceans. We assess here the physiological impact on sulfur speciation and isotopic composition of two microbial strains at a range of pH conditions consistent with the sharp gradients found in seafloor hydrothermal environments. Thiomicrospira thermophila and T. crunogena, both isolated from hydrothermal vents at East Pacific Rise, were grown with thiosulfate as the electron donor under aerobic, closed system conditions at controlled pH and optimal temperature (35°C). T. thermophila at pH 8 produced sulfate at a 1:1 ratio with thiosulfate consumption during exponential growth, with the ratio decreasing as pH decreases. This stoichiometric ratio decreases more steeply as a function of pH during metabolism by T. crunogena. Sulfate:thiosulfate ratios less than one indicate the production of alternative oxidized sulfur compounds such as polythionates. The rate of sulfate production is comparable in both strains and is dependent on pH, decreasing from 0.8mM/hr at pH 8 to 0.2mM/hr at pH 5.6. Fractionation of 34S expressed as Δ34S between reactant and product range from 0‰ to 3‰ for both sulfate and elemental sulfur produced, with no difference between products in pH buffered experiments (pH 5.6 and 8.0). However, in unbuffered experiments during which growth causes pH to decrease from 7 to below 4.5, Δ34S(S2O3-SO4) is consistently larger than Δ34S(S2O3-S) in both strains by a factor of 2. The metabolic activity of these (and similar) strains indicate that complex and cryptic sulfur cycling may be occurring in the subsurface, associated with only minimal variation in the δ34S isotopic composition of sulfate and elemental sulfur.

  12. Phosphonate removal from discharged circulating cooling water using iron-carbon micro-electrolysis.

    PubMed

    Zhou, Zhen; Qiao, Weimin; Lin, Yangbo; Shen, Xuelian; Hu, Dalong; Zhang, Jianqiao; Jiang, Lu-Man; Wang, Luochun

    2014-01-01

    Phosphonate is a commonly used corrosion and scale inhibitor for a circulating cooling water (CCW) system. Its discharge could cause eutrophication of receiving waters. The iron-carbon (Fe/C) micro-electrolysis technology was used to degrade and remove phosphonate from discharged CCW. The influences of initial pH, Fe/C ratio (FCR) and temperature on phosphonate removal were investigated in a series of batch tests and optimized by response surface methodology. The quadratic model of phosphonate removal was obtained with satisfactory degrees of fitness. The optimum conditions with total phosphorus removal efficiency of 95% were obtained at pH 7.0, FCR of 1.25, and temperature of 45 °C. The phosphonate removal mechanisms were also studied. Phosphonate removal occurred predominantly via two consecutive reactive phases: the degradation of phosphonate complexes (Ca-phosphonate) and the precipitation of Fe/C micro-electrolysis products (PO₄(3-), Ca²⁺ and Fe³⁺).

  13. Radiolabelling of isopeptide N epsilon-(gamma-glutamyl)-L-lysine by conjugation with N-succinimidyl-4-[18F]fluorobenzoate.

    PubMed

    Wüst, F; Hultsch, C; Bergmann, R; Johannsen, B; Henle, T

    2003-07-01

    The isopeptide N(epsilon)-(gamma-glutamyl)-L-lysine 4 was labelled with 18F via N-succinimidyl-4-[18F]fluorobenzoate ([18F]SFB). A modified approach for the convenient synthesis of [18F]SFB was used, and [18F]SFB could be obtained in decay-corrected radiochemical yields of 44-53% (n = 20) and radiochemical purity >95% within 40 min after EOB. For labelling N(epsilon)-(gamma-glutamyl)-L-lysine with [18F]SFB the effects of isopeptide concentration, temperature, and pH were studied to determine the optimum reaction conditions. The coupling reaction was shown to be temperature and pH independent while being strongly affected by the isopeptide concentration. Using the optimized labelling conditions, in a typical experiment 1.3GBq of [18F]SFB could be converted into 447MBq (46%, decay-corrected) of [18F]fluorobenzoylated isopeptide within 45 min, including HPLC purification.

  14. Culture conditions affect cytotoxin production by Serratia marcescens.

    PubMed

    Carbonell, G V; Fonseca, B A; Figueiredo, L T; Darini, A L; Yanaguita, R M

    1996-12-31

    Cytotoxins have been implicated in the pathogenesis of bacterial infections. In this study, the influence of different culture conditions was evaluated on cytotoxin production of Serratia marcescens. Parameters such as culture media, incubation temperature, starting pH of culture medium, aeration, anaerobiosis, carbon sources, iron concentration in he culture media, and release of cell-bond toxin by polymyxin B were investigated. The data suggest that this cytotoxin is predominantly extracellular and is not induced by iron limitation. Aerobic culture with shaking resulted in higher cytotoxicity than static aerobic or anaerobic culture. Bacteria grown in glucose, sucrose or galactose were more cytotoxic than those grown in inositol or maltose. The culture conditions that were identified as optimal for cytotoxin production by Serratia marcescens were incubation temperature ranging from 30 to 37 degrees C, in medium adjusted pH 8.5, with shaking. This work will contribute to further studies on the identification of this cytotoxic activity.

  15. [Study on preparation of sagittatoside B with epimedin B converted from cellulase].

    PubMed

    Xu, Feng-Juan; Sun, E; Zhang, Zhen-Hai; Cui, Li; Jia, Xiao-Bin

    2014-01-01

    To prepare sagittatoside B with epimedin B Hydrolyzed from cellulase. With the conversion ratio as the index, the effects of pH value, temperature, reaction time, dosage of enzyme and concentration of substrates on the conversion ratio were detected. L9 (3(4)) orthogonal design was adopted to optimize the preparation process. Hydrolyzed products were identified by MS, 1H-NMR, and 13C-NMR. The results showed that the optimum reaction conditions for the enzymatic hydrolysis were that the temperature was 50 degrees C, the reaction medium was pH 5.6 acetic acid-sodium acetate buffer solution, the concentration of substrates was 20 g x L(-1), the mass ratio between enzyme and substrate was 3: 5, and the relative molecular mass of the reaction product was 646.23. NMR data proved that the product was sagittatoside B. The process is simple and reliable under mild reaction conditions, thus suitable for industrial production.

  16. Vegetative Growth of Four Strains of Hericium erinaceus Collected from Different Habitats

    PubMed Central

    Imtiaj, Ahmed; Jayasinghe, Chandana; Lee, Geon Woo; Shim, Mi Ja; Rho, Hyun-Su; Lee, Hyun Sook; Hur, Hyun; Lee, Min Woong; Lee, U-Youn

    2008-01-01

    Vegetative growth of four different strains of Hericium erinaceus was observed. The temperature suitable for optimal mycelial growth was determined to be 25℃, with growth observed in the extend temperature range of 20~30℃. The different strains of this mushroom showed distinct pH requirements for their optimum vegetative growth, with the most favorable growth observed at pH 6. Considering vegetative mycelial growth, PDA, YM, Hennerberg, Hamada, and Glucose peptone were the most favorable media, and Czapek Dox, Hoppkins, Glucose tryptone, and Lilly were the most unfavorable media for these mushroom strains. With the exception of lactose, most of the carbon sources assayed demonstrated favorable vegetative growth of H. erinaceus. For mycelial growth, the most suitable nitrogen source was alanine and the most unsuitable was histidine. Oak sawdust medium supplemented with 10~20% rice bran was the best for mycelial growth of the mushroom. PMID:23990739

  17. Utilization of Diamine Oxidase Enzyme from Mung Bean Sprouts (Vigna radiata L) for Histamine biosensors

    NASA Astrophysics Data System (ADS)

    Karim, Abdul; Wahab, A. W.; Raya, I.; Natsir, H.; Arif, A. R.

    2018-03-01

    This research is aimed to utilize the diamine oxidase enzyme (DAO) which isolated from mung bean sprouts (Vigna radiata L) to develop histamine biosensors based on electode enzyme with the amperometric method (cyclic voltammetry).The DAO enzyme is trapped inside the membrane of chitin-cellulose acetate 2:1 and glutaraldehyde which super imposed on a Pt electrode. Histamine will be oxidized by DAO enzyme to produce aldehydes and H2O2 that acting as electron transfer mediators.The performance of biosensors will be measured at various concentrations of glutaraldehyde, temperature changes and different range of pH. Recently, it has been found that the optimal conditions obtained from the paramaters as follows; at 25% of glutaraldehyde, temperature of 37°C and pH of 7.4. Eventually, the results provided an expectation for applying histamine biosensors in determining the freshness and safety of fish specifically skombroidae families.

  18. Bacillus sphaericus: the highest bacterial tannase producer with potential for gallic acid synthesis.

    PubMed

    Raghuwanshi, Shailendra; Dutt, Kakoli; Gupta, Pritesh; Misra, Swati; Saxena, Rajendra Kumar

    2011-06-01

    An indigenously isolated strain of Bacillus sphaericus was found to produce 1.21 IU/ml of tannase under unoptimized conditions. Optimizing the process one variable at a time resulted in the production of 7.6 IU/ml of tannase in 48 h in the presence of 1.5% tannic acid. A 9.26-fold increase in tannase production was achieved upon further optimization using response surface methodology (RSM), a statistical approach. This increase led to a production level of 11.2I U/ml in medium containing 2.0% tannic acid, 2.5% galactose, 0.25% ammonium chloride, and 0.1% MgSO(4) pH 6.0 incubated at 37°C and 100 rpm for 48 h with a 2.0% inoculum level. Scaling up tannase production in a 30-l bioreactor resulted in the production of 16.54 IU/ml after 36 h. Thus far, this tannase production is the highest reported in this bacterial strain. Partially purified tannase exhibited an optimum pH of 5.0 with activity in the pH range of 3 to 8; 50°C was the optimal temperature for activity. Efficient conversion of tannic acid to purified gallic acid (90.80%) was achieved through crystallization. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  19. Expression, purification and characterization of a phyAm-phyCs fusion phytase*

    PubMed Central

    Zou, Li-kou; Wang, Hong-ning; Pan, Xin; Tian, Guo-bao; Xie, Zi-wen; Wu, Qi; Chen, Hui; Xie, Tao; Yang, Zhi-rong

    2008-01-01

    The phyAm gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichia pastoris in order to expand the pH profile of phytase and decrease the cost of production. The fusion phytase phyAm-phyCs gene was successfully overexpressed in P. pastoris as an active and extracellular phytase. The yield of total extracellular fusion phytase activity is (25.4±0.53) U/ml at the flask scale and (159.1±2.92) U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 °C and an optimal pH at 5.5~6.0 and its relative activity remains at a relatively high level of above 70% in the range of pH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 °C to 95 °C for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After deglycosylation by endoglycosidase H (EndoHf), the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those of phyCs and phyAm. PMID:18600783

  20. Degradation of dichlorvos using hydrodynamic cavitation based treatment strategies.

    PubMed

    Joshi, Ravi K; Gogate, Parag R

    2012-05-01

    The degradation of an aqueous solution of dichlorvos, a commonly used pesticide in India, has been systematically investigated using hydrodynamic cavitation reactor. All the experiments have been carried out using a 20 ppm solution of commercially available dichlorvos. The effect of important operating parameters such as inlet pressure (over a range 3-6 bar), temperature (31 °C, 36 °C and 39 °C) and pH (natural pH = 5.7 and acidic pH = 3) on the extent of degradation has been investigated initially. It has been observed that an optimum value of pressure gives maximum degradation whereas low temperature and pH of 3 are favorable. Intensification studies have been carried out using different additives such as hydrogen peroxide, carbon tetrachloride, and Fenton's reagent. Use of hydrogen peroxide and carbon tetrachloride resulted in the enhancement of the extent of degradation at optimized conditions but significant enhancement was obtained with the combined use of hydrodynamic cavitation and Fenton's chemistry. The maximum extent of degradation as obtained by using a combination of hydrodynamic cavitation and Fenton's chemistry was 91.5% in 1h of treatment time. The present work has conclusively established that hydrodynamic cavitation in combination with Fenton's chemistry can be effectively used for the degradation of dichlorvos. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. Enhancing Degradation of Low Density Polyethylene Films by Curvularia lunata SG1 Using Particle Swarm Optimization Strategy.

    PubMed

    Raut, Sangeeta; Raut, Smita; Sharma, Manisha; Srivastav, Chaitanya; Adhikari, Basudam; Sen, Sudip Kumar

    2015-09-01

    In the present study, artificial neural network (ANN) modelling coupled with particle swarm optimization (PSO) algorithm was used to optimize the process variables for enhanced low density polyethylene (LDPE) degradation by Curvularia lunata SG1. In the non-linear ANN model, temperature, pH, contact time and agitation were used as input variables and polyethylene bio-degradation as the output variable. Further, on application of PSO to the ANN model, the optimum values of the process parameters were as follows: pH = 7.6, temperature = 37.97 °C, agitation rate = 190.48 rpm and incubation time = 261.95 days. A comparison between the model results and experimental data gave a high correlation coefficient ([Formula: see text]). Significant enhancement of LDPE bio-degradation using C. lunata SG1by about 48 % was achieved under optimum conditions. Thus, the novelty of the work lies in the application of combination of ANN-PSO as optimization strategy to enhance the bio-degradation of LDPE.

  2. Optimization of Enzymatic Saccharification of Alkali Pretreated Parthenium sp. Using Response Surface Methodology

    PubMed Central

    Pandiyan, K.; Tiwari, Rameshwar; Singh, Surender; Nain, Pawan K. S.; Rana, Sarika; Arora, Anju; Singh, Shashi B.; Nain, Lata

    2014-01-01

    Parthenium sp. is a noxious weed which threatens the environment and biodiversity due to its rapid invasion. This lignocellulosic weed was investigated for its potential in biofuel production by subjecting it to mild alkali pretreatment followed by enzymatic saccharification which resulted in significant amount of fermentable sugar yield (76.6%). Optimization of enzymatic hydrolysis variables such as temperature, pH, enzyme, and substrate loading was carried out using central composite design (CCD) in response to surface methodology (RSM) to achieve the maximum saccharification yield. Data obtained from RSM was validated using ANOVA. After the optimization process, a model was proposed with predicted value of 80.08% saccharification yield under optimum conditions which was confirmed by the experimental value of 85.80%. This illustrated a good agreement between predicted and experimental response (saccharification yield). The saccharification yield was enhanced by enzyme loading and reduced by temperature and substrate loading. This study reveals that under optimized condition, sugar yield was significantly increased which was higher than earlier reports and promises the use of Parthenium sp. biomass as a feedstock for bioethanol production. PMID:24900917

  3. Study on Power Ultrasound Optimization and Its Comparison with Conventional Thermal Processing for Treatment of Raw Honey

    PubMed Central

    2017-01-01

    Summary The present study was done to optimize the power ultrasound processing for maximizing diastase activity of and minimizing hydroxymethylfurfural (HMF) content in honey using response surface methodology. Experimental design with treatment time (1-15 min), amplitude (20-100%) and volume (40-80 mL) as independent variables under controlled temperature conditions was studied and it was concluded that treatment time of 8 min, amplitude of 60% and volume of 60 mL give optimal diastase activity and HMF content, i.e. 32.07 Schade units and 30.14 mg/kg, respectively. Further thermal profile analyses were done with initial heating temperatures of 65, 75, 85 and 95 ºC until temperature of honey reached up to 65 ºC followed by holding time of 25 min at 65 ºC, and the results were compared with thermal profile of honey treated with optimized power ultrasound. The quality characteristics like moisture, pH, diastase activity, HMF content, colour parameters and total colour difference were least affected by optimized power ultrasound treatment. Microbiological analysis also showed lower counts of aerobic mesophilic bacteria and in ultrasonically treated honey than in thermally processed honey samples complete destruction of coliforms, yeasts and moulds. Thus, it was concluded that power ultrasound under suggested operating conditions is an alternative nonthermal processing technique for honey. PMID:29540991

  4. Evaluation and optimization of the conditions for an improved ferulic acid intercalation into a synthetic lamellar anionic clay.

    PubMed

    Schoubben, Aurélie; Blasi, Paolo; Giovagnoli, Stefano; Nocchetti, Morena; Ricci, Maurizio; Perioli, Luana; Rossi, Carlo

    2006-03-01

    The aim of the study is to optimize the intercalation conditions of ferulic acid (FERH), an antioxidant compound, into Mg-Al-hydrotalcite for a safe skin photoprotection. The intercalation products were prepared incubating hydrotalcite (HTlc) in aqueous solutions of FERH sodium salt at different temperatures over 4 and 8 days. Quantitative determination of intercalated FERH was performed by thermogravimetric analysis and morphology by scanning electron microscopy (SEM). FERH stability study was carried out at different pHs and temperatures. FERH was analyzed by reversed phase-high-performance liquid chromatography. Response surface methods (RSMs) were used to assess optimal intercalation conditions and FERH stability. In all intercalation products, FERH content was found to be about 48% w/w except when the intercalation process was carried out at 52 degrees C for 8 days and at 60 degrees C for both 4 and 8 days, which resulted to be 40.39, 39.99, and 34.99%, respectively. The RSM designs showed that intercalation improvement can be achieved by working at pH 6, at temperatures below 40 degrees C, and over 4 days of incubation. The optimal conditions for a proper FERH intercalation were assessed. The development of a new optimized protocol may improve HTlc-FER complex performances and safety by augmenting dosage and reducing the presence of harmful reactive species in the final formulation.

  5. Research on Influencing Factors of Biological Filtration Tower Treating Toluene Gas

    NASA Astrophysics Data System (ADS)

    Zhang, Changping; Cao, Ziqing; Lu, Yuqi; Du, Linggai

    2017-05-01

    Through the orthogonal experimental design, the optimal combination of Triton X-100, nitrogen source, Fe2+, temperature, concentration of antibiotics, pH and spray quantity was determined with surfactants, nitrogen and iron elements as additive, by which the key influencing factors were determined. In the test, the removal efficiency of the second groups was higher than that of the eighth groups, which were 89% and 87%, respectively. The best combination of a group of removal was as follows: nitrogen source concentration was 2 g ·L-1, antibiotic concentration was 300 U·mL-1, the concentration of Triton X-100 was 0.05 mL·L-1, Fe2+ concentration was 14 mL·L-1, pH was 7, the temperature was 34°C, spray amount was 6 L ·h-1. The antibiotic concentration was the most important factor on the removal efficiency of the toluene. The concentration of gas in each layer of toluene was detected; the curve of the outlet concentration in the optimal combination and the average state was obtained. The removal efficiency of the optimal combination was much better than the average, and it was found that the removal rate decreased with the increase of the height of the filling layer. The change of oxygen content in each layer was detected which was no significant change. It showed that oxygen was not the limiting factor of the removal of toluene by microorganisms. Keywords: surfactants; biological filtration tower; toluene; orthogonal test

  6. Optimization of papain hydrolysis conditions for release of glycosaminoglycans from the chicken keel cartilage

    NASA Astrophysics Data System (ADS)

    Le Vien, Nguyen Thi; Nguyen, Pham Bao; Cuong, Lam Duc; An, Trinh Thi Thua; Dao, Dong Thi Anh

    2017-09-01

    Glycosaminoglycans (GAGs) are natural biocompounds which join to construct cartilage tissuses, it can be extracted from cartilage of sharks, pigs, cows, chickens, etc. GAGs contain a Chondroitin sulfate (CS) content which is a supplement of functional food used for preventing and supporting treatment of arthritis and eye diseases. Therefore, the GAGs extraction from byproducts of the industry of cattle and poultry slaughter to identify the CS content by papain enzyme is necessary. In this study, the optimal hydrolysis conditions were obtained by response surface methodology (RSM). The independent variables were coded as: pH (x1), enzyme concentration (x2), incubation temperature (x3) and hydrolysis time (x4). The results of the analysis of variance (ANOVA) shown that the variables actively affected GAGs content. The optimal conditions of hydrolysis were derived at pH of 7.1, ratio of enzyme per substances of 0.62% w/wpo, temperature of 65°C and hydrolysis time of 230 minutes, GAGs content reached 14.3% of the dry matter of raw material. Analyzes by HPLC revealed that 56.17% of the dry preparations of GAGs were CS compound, were equivalent to 8.11% of the dry matter of chicken keel cartilage. Molecular weight of the dry preparations GAGs was 259.6 kDa. The dry preparations included the contents of moisture 12.2%, protein 8.42%, lipid 0%, ash 10.03% and extracted GAGs 69.35%.

  7. Optimization of tannase production by Aspergillus niger in solid-state packed-bed bioreactor.

    PubMed

    Rodríguez-Durán, Luis V; Contreras-Esquivel, Juan C; Rodríguez, Raúl; Prado-Barragán, L Arely; Aguilar, Cristóbal N

    2011-09-01

    Tannin acyl hydrolase, also known as tannase, is an enzyme with important applications in the food, feed, pharmaceutical, and chemical industries. However, despite a growing interest in the catalytic properties of tannase, its practical use is very limited owing to high production costs. Several studies have already demonstrated the advantages of solid-state fermentation (SSF) for the production of fungal tannase, yet the optimal conditions for enzyme production strongly depend on the microbial strain utilized. Therefore, the aim of this study was to improve the tannase production by a locally isolated A. niger strain in an SSF system. The SSF was carried out in packed-bed bioreactors using polyurethane foam as an inert support impregnated with defined culture media. The process parameters influencing the enzyme production were identified using a Plackett–Burman design, where the substrate concentration, initial pH, and incubation temperature were determined as the most significant. These parameters were then further optimized using a Box-Behnken design. The maximum tannase production was obtained with a high tannic acid concentration (50 g/l), relatively low incubation temperature (30°C), and unique low initial pH (4.0). The statistical strategy aided in increasing the enzyme activity nearly 1.97-fold, from 4,030 to 7,955 U/l. Consequently, these findings can lead to the development of a fermentation system that is able to produce large amounts of tannase in economical, compact, and scalable reactors.

  8. Enhancement of tannase production by Lactobacillus plantarum CIR1: validation in gas-lift bioreactor.

    PubMed

    Aguilar-Zarate, Pedro; Cruz-Hernandez, Mario A; Montañez, Julio C; Belmares-Cerda, Ruth E; Aguilar, Cristobal N

    2014-11-01

    The optimization of tannase production by Lactobacillus plantarum CIR1 was carried out following the Taguchi methodology. The orthogonal array employed was L18 (2(1) × 3(5)) considering six important factors (pH and temperature, also phosphate, nitrogen, magnesium, and carbon sources) for tannase biosynthesis. The experimental results obtained from 18 trials were processed using the software Statistical version 7.1 using the character higher the better. Optimal culture conditions were pH, 6; temperature, 40 °C; tannic acid, 15.0 g/L; KH2PO4, 1.5 g/L; NH4Cl, 7.0 g/L; and MgSO4, 1.5 g/L which were obtained and further validated resulting in an enhance tannase yield of 2.52-fold compared with unoptimized conditions. Tannase production was further carried out in a 1-L gas-lift bioreactor where two nitrogen flows (0.5 and 1.0 vvm) were used to provide anaerobic conditions. Taguchi methodology allowed obtaining the optimal culture conditions for the production of tannase by L. plantarum CIR1. At the gas-lift bioreactor the tannase productivity yields increase 5.17 and 8.08-fold for the flow rates of 0.5 and 1.0 vvm, respectively. Lactobacillus plantarum CIR1 has the capability to produce tannase at laboratory-scale. This is the first report for bacterial tannase production using a gas-lift bioreactor.

  9. [Enzymatic characteristics of peroxidase from Chrysanthemum morifolium cv. Bo-ju].

    PubMed

    Zhu, Yu-Yun; Lyu, Xin-Lin; Li, Xiang-Wei; Zhang, Dong; Dong, Li-Hua; Zhu, Jing-Jing; Wang, Zhi-Min; Zhang, Jin-Zhen

    2018-04-01

    The enzymatic browning is one of the main reasons for affecting the quality of medicinal flowers. In the process of chrysanthemum harvesting and processing, improper treatment will lead to the browning and severely impact the appearance and quality of chrysanthemum. Peroxidase enzyme is one of the oxidoreductases that cause enzymatic browning of fresh chrysanthemum. The enzymatic characteristics of peroxidase (POD) in chrysanthemum were studied in this paper. In this experiment, the effects of different reaction substrates and their concentrations, PH value of buffer and reaction temperatures on the activity of POD enzyme were investigated. The results showed that the optimal substrate of POD was guaiacol, and the optimal concentration of POD was 50 mmol·L⁻¹. The optimal pH value and reaction temperature were 4.4 and 30-35 °C, respectively. Michaelis-Menten equation was obtained to express the kinetics of enzyme-catalyzed reaction of POD, Km=0.193 mol·L⁻¹, Vmax=0.329 D·min⁻¹. In addition, the results of POD enzyme thermal stability test showed that the POD enzyme activity was inhibited when being treated at 80 °C for 4 min or at 100 °C for 2 min. The above results were of practical significance to reveal the enzymatic browning mechanism, control the enzymatic browning and improve the quality of chrysanthemum, and can also provide the basis for the harvesting and processing of medicinal materials containing polyphenols. Copyright© by the Chinese Pharmaceutical Association.

  10. Acetone photophysics at 282 nm excitation at elevated pressure and temperature. II: Fluorescence modeling

    NASA Astrophysics Data System (ADS)

    Hartwig, Jason; Raju, Mandhapati; Sung, Chih-Jen

    2017-07-01

    This is the second in a series of two papers that presents an updated fluorescence model and compares with the new experimental data reported in the first paper, as well as the available literature data, to extend the range of acetone photophysics to elevated pressure and temperature conditions. This work elucidates the complete acetone photophysical model in terms of each and every competing radiative and non-radiative rate. The acetone fluorescence model is then thoroughly examined and optimized based on disparity with recently conducted elevated pressure and temperature photophysical calibration experiments. The current work offers insight into the competition between non-radiative and vibrational energy decay rates at elevated temperature and pressure and proposes a global optimization of model parameters from the photophysical model developed by Thurber (Acetone Laser-Induced Fluorescence for Temperature and Multiparameter Imaging in Gaseous Flows. PhD thesis, Stanford University Mechanical Engineering Department, 1999). The collisional constants of proportionality, which govern vibrational relaxation, are shown to be temperature dependent at elevated pressures. A new oxygen quenching rate is proposed which takes into account collisions with oxygen as well as the oxygen-assisted intersystem crossing component. Additionally, global trends in ketone photophysics are presented and discussed.

  11. Enhanced production of leech hyaluronidase by optimizing secretion and cultivation in Pichia pastoris.

    PubMed

    Kang, Zhen; Zhang, Na; Zhang, Yunfeng

    2016-01-01

    Leech hyaluronidase (LHAase) was recently cloned and successfully expressed in Pichia pastoris. To increase its secretory expression level, four signal peptides (nsB, YTP1, SCS3, and HKR1) and six amphipathic peptides (APs) were comparatively investigated. After substitution with nsB and fusion with AP2, the production of LHAase was significantly increased, from 8.42 × 10(5) to 1.24 × 10(6) U/ml. Compared with the parental LHAase, the variant AP2-LHAase showed a lower optimum pH (5.0), higher optimum temperature (50 °C), and a broader range of thermal stability (20-60 °C). To further promote fermentative production of the variant AP2-LHAase, the cultivation temperature was systematically optimized according to cell viability and alcohol oxidase activity. Eventually, through a combination of N-terminal engineering and optimization of cultivation, the production of LHAase was improved to 1.68 × 10(6) U/ml, with a high productivity of 1.87 × 10(4) U/ml/h.

  12. Removal of Nitrate in Simulated Water at Low Temperature by a Novel Psychrotrophic and Aerobic Bacterium, Pseudomonas taiwanensis Strain J

    PubMed Central

    He, Tengxia; Ye, Qing; Sun, Quan; Cai, Xi; Ni, Jiupai

    2018-01-01

    Low temperatures and high pH generally inhibit the biodenitrification. Thus, it is important to explore the psychrotrophic and alkali-resisting microorganism for degradation of nitrogen. This research was mainly focused on the identification of a psychrotrophic strain and preliminary explored its denitrification characteristics. The new strain J was isolated using the bromothymol blue solid medium and identified as Pseudomonas taiwanensis on the basis of morphology and phospholipid fatty acid as well as 16S rRNA gene sequence analyses, which is further testified to work efficiently for removing nitrate from wastewater at low temperature circumstances. This is the first report that Pseudomonas taiwanensis possessed excellent tolerance to low temperature, with 15°C as its optimum and 5°C as viable. The Pseudomonas taiwanensis showed unusual ability of aerobic denitrification with the nitrate removal efficiencies of 100% at 15°C and 51.61% at 5°C. Single factor experiments showed that the optimal conditions for denitrification were glucose as carbon source, 15°C, shaking speed 150 r/min, C/N 15, pH ≥ 7, and incubation quantity 2.0 × 106 CFU/mL. The nitrate and total nitrogen removal efficiencies were up to 100% and 93.79% at 15°C when glucose is served as carbon source. These results suggested that strain J had aerobic denitrification ability, as well as the notable ability to tolerate the low temperature and high pH. PMID:29789796

  13. The effect of operational parameters on the biodegradation of bisphenols by Trametes versicolor laccase immobilized on Hippospongia communis spongin scaffolds.

    PubMed

    Zdarta, Jakub; Antecka, Katarzyna; Frankowski, Robert; Zgoła-Grześkowiak, Agnieszka; Ehrlich, Hermann; Jesionowski, Teofil

    2018-02-15

    Due to the rapid growth in quantities of phenolic compounds in wastewater, the development of efficient and environmentally friendly methods for their removal becomes a necessity. Thus, in a presented work, for the first time, a novel material, Hippospongia communis spongin-based scaffold, was used as a biopolymeric support for the immobilization of laccase from Trametes versicolor. The resulting biocatalytic systems were used for the biodegradation of three bisphenols: bisphenol A (BPA), bisphenol F (BPF) and bioremoval-resistant bisphenol S (BPS). Optimization of the immobilization and biodegradation methodologies was performed to increase bisphenols removal. The effect of temperature, pH and initial pollutant concentration was evaluated. It was shown that under optimal conditions, almost 100% of BPA (pH5, 30°C) and BPF (pH5, 40°C), and over 40% of BPS (pH4, 30°C) was removed from the solution at a concentration of 2mg/mL. Furthermore, the immobilized laccase exhibited good reusability and storage stability, retaining over 80% of its initial activity after 50days of storage. In addition, the main biodegradation products of BPA and BPF were identified. It was shown that mainly dimers and trimers were formed following the oxidation of bisphenols by the immobilized laccase. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Production and Partial Characterization of α-Amylase Enzyme from Bacillus sp. BCC 01-50 and Potential Applications

    PubMed Central

    Qureshi, Abdul Sattar; Khushk, Imrana; Ali, Chaudhry Haider; Lashari, Safia; Bhutto, Muhammad Aqeel; Mangrio, Ghulam Sughra; Lu, Changrui

    2017-01-01

    Amylase is an industrially important enzyme and applied in many industrial processes such as saccharification of starchy materials, food, pharmaceutical, detergent, and textile industries. This research work deals with the optimization of fermentation conditions for α-amylase production from thermophilic bacterial strain Bacillus sp. BCC 01-50 and characterization of crude amylase. The time profile of bacterial growth and amylase production was investigated in synthetic medium and maximum enzyme titer was observed after 60 h. In addition, effects of different carbon sources were tested as a substrate for amylase production and molasses was found to be the best. Various organic and inorganic compounds, potassium nitrate, ammonium chloride, sodium nitrate, urea, yeast extract, tryptone, beef extract, and peptone, were used and beef extract was found to be the best among the nitrogen sources used. Temperature, pH, agitation speed, and size of inoculum were also optimized. Highest enzyme activity was obtained when the strain was cultured in molasses medium for 60 h in shaking incubator (150 rpm) at 50°C and pH 8. Crude amylase showed maximal activity at pH 9 and 65°C. Enzyme remained stable in alkaline pH range 9-10 and 60–70°C. Crude amylase showed great potential for its application in detergent industry and saccharification of starchy materials. PMID:28168200

  15. Production and Partial Characterization of α-Amylase Enzyme from Bacillus sp. BCC 01-50 and Potential Applications.

    PubMed

    Simair, Altaf Ahmed; Qureshi, Abdul Sattar; Khushk, Imrana; Ali, Chaudhry Haider; Lashari, Safia; Bhutto, Muhammad Aqeel; Mangrio, Ghulam Sughra; Lu, Changrui

    2017-01-01

    Amylase is an industrially important enzyme and applied in many industrial processes such as saccharification of starchy materials, food, pharmaceutical, detergent, and textile industries. This research work deals with the optimization of fermentation conditions for α -amylase production from thermophilic bacterial strain Bacillus sp. BCC 01-50 and characterization of crude amylase. The time profile of bacterial growth and amylase production was investigated in synthetic medium and maximum enzyme titer was observed after 60 h. In addition, effects of different carbon sources were tested as a substrate for amylase production and molasses was found to be the best. Various organic and inorganic compounds, potassium nitrate, ammonium chloride, sodium nitrate, urea, yeast extract, tryptone, beef extract, and peptone, were used and beef extract was found to be the best among the nitrogen sources used. Temperature, pH, agitation speed, and size of inoculum were also optimized. Highest enzyme activity was obtained when the strain was cultured in molasses medium for 60 h in shaking incubator (150 rpm) at 50°C and pH 8. Crude amylase showed maximal activity at pH 9 and 65°C. Enzyme remained stable in alkaline pH range 9-10 and 60-70°C. Crude amylase showed great potential for its application in detergent industry and saccharification of starchy materials.

  16. Quality by design approach of a pharmaceutical gel manufacturing process, part 1: determination of the design space.

    PubMed

    Rosas, Juan G; Blanco, Marcel; González, Josep M; Alcalá, Manel

    2011-10-01

    This work was conducted in the framework of a quality by design project involving the production of a pharmaceutical gel. Preliminary work included the identification of the quality target product profiles (QTPPs) from historical values for previously manufactured batches, as well as the critical quality attributes for the process (viscosity and pH), which were used to construct a D-optimal experimental design. The experimental design comprised 13 gel batches, three of which were replicates at the domain center intended to assess the reproducibility of the target process. The viscosity and pH models established exhibited very high linearity and negligible lack of fit (LOF). Thus, R(2) was 0.996 for viscosity and 0.975 for pH, and LOF was 0.53 for the former parameter and 0.84 for the latter. The process proved reproducible at the domain center. Water content and temperature were the most influential factors for viscosity, and water content and acid neutralized fraction were the most influential factors for pH. A desirability function was used to find the best compromise to optimize the QTPPs. The body of information was used to identify and define the design space for the process. A model capable of combining the two response variables into a single one was constructed to facilitate monitoring of the process. Copyright © 2011 Wiley-Liss, Inc.

  17. Enhancing the auto-flocculation of photosynthetic bacteria to realize biomass recovery in brewery wastewater treatment.

    PubMed

    Lu, Haifeng; Dong, Shan; Zhang, Guangming; Han, Ting; Zhang, Yuanhui; Li, Baoming

    2018-02-15

    Photosynthetic bacteria (PSB) wastewater treatment technology can simultaneously realize wastewater purification and biomass production. The produced biomass contains high value-added products, which can be used in medical and agricultural industry. However, because of the small size and high electronegativity, PSB are hard to be collected from wastewater, which hampers the commercialization of PSB-based industrial processes. Auto-flocculation is a low cost, energy saving, non-toxic biomass collection method for microbiology. In this work, the influence factors with their optimal levels and mechanism for enhancing the auto-flocculation of PSB were investigated in pure cultivation medium. Then PSB auto-flocculation performance in real brewery wastewater was probed. Results showed that Na + concentration, pH and light intensity were three crucial factors except the initial inoculum sizes and temperature. In the pure medium cultivation system, the optimal condition for PSB auto-flocculation was as follows: pH was 9.5, inoculum size was 420 mg l -1 , Na + concentration was 0.067 mol l -1 , light intensity was 5000 lux, temperature was 30°C. Under the optimal condition, the auto-flocculation ratio and biomass recovery reached 85.0% and 1488 mg l -1 , which improved by 1.67-fold and 2.14-fold compared with the PSB enrichment cultivation conditions, respectively. Mechanism analysis showed that the protein/polysaccharides ratio and absolute Zeta potential value had a liner relationship. For the brewery wastewater treatment, under the above optimal condition, the chemical oxygen demand removal reached 94.3% with the auto-flocculation ratio and biomass recovery of 89.6% and 1510 mg l -1 , which increased 2.75-fold and 2.77-fold, respectively.

  18. Efficient Accumulation and In Vitro Antitumor Activities of Triterpene Acids from Submerged Batch--Cultured Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes).

    PubMed

    Wang, Xiao-Ling; Ding, Zhong-Yang; Zhao, Yan; Liu, Gao-Qiang; Zhou, Guo-Ying

    2017-01-01

    Triterpene acids are among the major bioactive constituents of lucidum. However, submerged fermentation techniques for isolating triterpene acids from G. lucidum have not been optimized for commercial use, and the antitumor activity of the mycelial triterpene acids needs to be further proven. The aim of this work was to optimize the conditions for G. lucidum culture with respect to triterpene acid production, scaling up the process, and examining the in vitro antitumor activity of mycelial triterpene acids. The key conditions (i.e., initial pH, fermentation temperature, and rotation speed) were optimized using response surface methodology, and the in vitro antitumor activity was evaluated using the MTT method. The optimum key fermentation conditions for triterpene acid production were pH 6.0; rotation speed, 161.9 rpm; and temperature, 30.1°C, resulting in a triterpene acid yield of 291.0 mg/L in the validation experiment in a 5-L stirred bioreactor; this yield represented a 70.8% increase in titer compared with the nonoptimized conditions. Furthermore, the optimized conditions were then successfully scaled up to a production scale of 200 L, and a triterpene productivity of 47.9 mg/L/day was achieved, which is, to our knowledge, the highest reported in the large-scale fermentation of G. lucidum. In addition, the mycelial triterpene acids were found to be cytotoxic to the SMMC-7721 and SW620 cell lines in vitro. Chemical analysis showed that the key active triterpene acid compounds, ganoderic acids T and Me, predominated in the extract, at 69.2 and 41.6 mg/g, respectively. Thus, this work develops a simple and feasible batch fermentation technique for the large-scale production of antitumor triterpene acids from G. lucidum.

  19. Biodiesel production from algae grown on food industry wastewater.

    PubMed

    Mureed, Khadija; Kanwal, Shamsa; Hussain, Azhar; Noureen, Shamaila; Hussain, Sabir; Ahmad, Shakeel; Ahmad, Maqshoof; Waqas, Rashid

    2018-04-10

    Algae have an ample potential to produce biodiesel from spent wash of food industry. In addition, it is cheaper and presents an environment friendly way to handle food industry wastewater. This study was conducted to optimize the growth of microalgal strains and to assess biodiesel production potential of algae using untreated food industry wastewater as a source of nutrients. The food industry wastewater was collected and analyzed for its physicochemical characteristics. Different dilutions (10, 20, 40, 80, and 100%) of this wastewater were made with distilled water, and growth of two microalgal strains (Cladophora sp. and Spyrogyra sp.) was recorded. Each type of wastewater was inoculated with microalgae, and biomass was harvested after 7 days. The growth of both strains was also evaluated at varying temperatures, pH and light periods to optimize the algal growth for enhanced biodiesel production. After optimization, biodiesel production by Spyrogyra sp. was recorded in real food industry wastewater. The algal biomass increased with increasing level of food industry wastewater and was at maximum with 100% wastewater. Moreover, statistically similar results were found with algal growth on 100% wastewater and also on Bristol's media. The Cladophora sp. produced higher biomass than Spyrogyra sp. while growing on food industry wastewater. The optimal growth of both microalgal strains was observed at temperature 30 °C, pH: 8, light 24 h. Cladophora sp. was further evaluated for biodiesel production while growing on 100% wastewater and found that this strain produced high level of oil and biodiesel. Algae have an ample potential to produce biodiesel from spent wash of food industry. In addition, it is cheaper and presents an environment friendly way to handle food industry wastewater.

  20. Improved production of tannase by Klebsiella pneumoniae using Indian gooseberry leaves under submerged fermentation using Taguchi approach.

    PubMed

    Kumar, Mukesh; Singh, Amrinder; Beniwal, Vikas; Salar, Raj Kumar

    2016-12-01

    Tannase (tannin acyl hydrolase E.C 3.1.1.20) is an inducible, largely extracellular enzyme that causes the hydrolysis of ester and depside bonds present in various substrates. Large scale industrial application of this enzyme is very limited owing to its high production costs. In the present study, cost effective production of tannase by Klebsiella pneumoniae KP715242 was studied under submerged fermentation using different tannin rich agro-residues like Indian gooseberry leaves (Phyllanthus emblica), Black plum leaves (Syzygium cumini), Eucalyptus leaves (Eucalyptus glogus) and Babul leaves (Acacia nilotica). Among all agro-residues, Indian gooseberry leaves were found to be the best substrate for tannase production under submerged fermentation. Sequential optimization approach using Taguchi orthogonal array screening and response surface methodology was adopted to optimize the fermentation variables in order to enhance the enzyme production. Eleven medium components were screened primarily by Taguchi orthogonal array design to identify the most contributing factors towards the enzyme production. The four most significant contributing variables affecting tannase production were found to be pH (23.62 %), tannin extract (20.70 %), temperature (20.33 %) and incubation time (14.99 %). These factors were further optimized with central composite design using response surface methodology. Maximum tannase production was observed at 5.52 pH, 39.72 °C temperature, 91.82 h of incubation time and 2.17 % tannin content. The enzyme activity was enhanced by 1.26 fold under these optimized conditions. The present study emphasizes the use of agro-residues as a potential substrate with an aim to lower down the input costs for tannase production so that the enzyme could be used proficiently for commercial purposes.

  1. Optimization of biogenic methane production from coal

    DOE PAGES

    Fuertez, John; Nguyen, Van; McLennan, John D.; ...

    2017-09-29

    Given continuously increasing global energy needs, diversified efforts have been made to find and exploit new natural gas resources. These include coalbed methane (CBM), which represents an important global, unconventional source of natural gas. Efforts have been underway for some time to more effectively generate methane in-situ in coal plays by introduction of nutrients and/or microbial consortia. However, much is still to be learned about the limitations and environmental conditions that support microbial growth and are conducive to biogenic methane production from coal. Here we evaluated environmental conditions that led to increased methane production from subbituminous coal by introducing amore » foreign methanogenic consortium that included Methanobacterium sp. Furthermore, we used a central composite design (CCD) to explore a broad range of operational conditions, examine the effects of the important environmental factors, such as temperature, pH and salt concentration, and query a feasible region of operation to maximize methane production from coal. An anticipated detrimental effect of NaCl concentration on methane production was observed for the consortium assessed. The range of feasible operational conditions comprised initial pH values between 4.2 and 6.8, temperatures between 23 °C and 37 °C, and NaCl concentrations between 3.7 mg/cm 3 and 9.0 mg/cm 3. Coal biogasification was optimal for this consortium at an initial pH value of 5.5, at 30 °C, and at a NaCl concentration 3.7 mg/cm 3 (i.e., 145,165 ppm, which is 25.6 sft 3/ton).« less

  2. Optimization of biogenic methane production from coal

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fuertez, John; Nguyen, Van; McLennan, John D.

    Given continuously increasing global energy needs, diversified efforts have been made to find and exploit new natural gas resources. These include coalbed methane (CBM), which represents an important global, unconventional source of natural gas. Efforts have been underway for some time to more effectively generate methane in-situ in coal plays by introduction of nutrients and/or microbial consortia. However, much is still to be learned about the limitations and environmental conditions that support microbial growth and are conducive to biogenic methane production from coal. Here we evaluated environmental conditions that led to increased methane production from subbituminous coal by introducing amore » foreign methanogenic consortium that included Methanobacterium sp. Furthermore, we used a central composite design (CCD) to explore a broad range of operational conditions, examine the effects of the important environmental factors, such as temperature, pH and salt concentration, and query a feasible region of operation to maximize methane production from coal. An anticipated detrimental effect of NaCl concentration on methane production was observed for the consortium assessed. The range of feasible operational conditions comprised initial pH values between 4.2 and 6.8, temperatures between 23 °C and 37 °C, and NaCl concentrations between 3.7 mg/cm 3 and 9.0 mg/cm 3. Coal biogasification was optimal for this consortium at an initial pH value of 5.5, at 30 °C, and at a NaCl concentration 3.7 mg/cm 3 (i.e., 145,165 ppm, which is 25.6 sft 3/ton).« less

  3. A halotolerant thermostable lipase from the marine bacterium Oceanobacillus sp. PUMB02 with an ability to disrupt bacterial biofilms

    PubMed Central

    Seghal Kiran, George; Nishanth Lipton, Anuj; Kennedy, Jonathan; Dobson, Alan DW; Selvin, Joseph

    2014-01-01

    A halotolerant thermostable lipase was purified and characterized from the marine bacterium Oceanobacillus sp. PUMB02. This lipase displayed a high degree of stability over a wide range of conditions including pH, salinity, and temperature. It was optimally active at 30 °C and pH 8.0 respectively and was stable at higher temperatures (50–70 °C) and alkaline pH. The molecular mass of the lipase was approximately 31 kDa based on SDS-PAGE and MALDI-ToF fingerprint analysis. Conditions for enhanced production of lipase by Oceanobacillus sp. PUMB02 were attained in response surface method-guided optimization with factors such as olive oil, sucrose, potassium chromate, and NaCl being evaluated, resulting in levels of 58.84 U/ml being achieved. The biofilm disruption potential of the PUMB02 lipase was evaluated and compared with a marine sponge metagenome derived halotolerant lipase Lpc53E1. Good biofilm disruption activity was observed with both lipases against potential food pathogens such as Bacillus cereus MTCC1272, Listeria sp. MTCC1143, Serratia sp. MTCC4822, Escherichia coli MTCC443, Pseudomonas fluorescens MTCC1748, and Vibrio parahemolyticus MTCC459. Phase contrast microscopy, scanning electron microscopy, and confocal laser scanning microscopy showed very effective disruption of pathogenic biofilms. This study reveals that marine derived hydrolytic enzymes such as lipases may have potential utility in inhibiting biofilm formation in a food processing environment and is the first report of the potential application of lipases from the genus Oceanobacillus in biofilm disruption strategies. PMID:25482232

  4. Autolytic degradation of skipjack tuna during heating as affected by initial quality and processing conditions.

    PubMed

    Stagg, Nicola J; Amato, Penny M; Giesbrecht, Francis; Lanier, Tyre C

    2012-02-01

    Several factors were studied as affecting protein degradation and texture of skipjack tuna muscle following ambient pressure thermal processing (precooking). These included degree of mushy tuna syndrome (MTS) evidenced in the raw meat, raw meat pH, abusive thawing/holding, and precooking temperature/time. Slurries and intact pieces from frozen skipjack tuna, either tempered for 2 h or thawed and held at 25 °C for 22 h (abusive treatment) were heated at temperatures ranging from 40 to 80 °C for up to 2 h, and also at 90 °C for 1 h, with or without prior adjustment of pH to 5 or 7 to favor cathepsin or calpain activity, respectively. Proteolysis of precooked samples was monitored by Lowry assay and SDS-PAGE; cooked texture of intact meat was measured using a Kramer shear press and by sensory profile analysis. Proteolysis maximally occurred in slurries of skipjack tuna muscle that had been abusively stored (22 h at 25 °C) and adjusted to pH 5 prior to heating at 55 °C. Intact pieces of tuna abusively thawed/held for 22 h with subsequent heating at 55 °C also evidenced the most proteolysis and were the least firm in texture. Raw fish that evidenced higher severity of MTS when raw displayed higher levels of proteolysis prior to cooking, which were further increased after cooking at 55 °C. The kinetic data presented here can be used to optimize processing conditions for skipjack tuna canning to minimize textural degradation and optimize quality. © 2012 Institute of Food Technologists®

  5. Enhancement stability and catalytic activity of immobilized α-amylase using bioactive phospho-silicate glass as a novel inorganic support.

    PubMed

    Ahmed, Samia A; Mostafa, Faten A; Ouis, Mona A

    2018-06-01

    α-Amylase enzyme was immobilized on bioactive phospho-silicate glass (PS-glass) as a novel inorganic support by physical adsorption and covalent binding methods using glutaraldehyde and poly glutaraldehyde as a spacer. Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) studies confirmed the glass-enzyme linkage. Dissolution of PS-glass in acidic and neutral pH is higher than that of alkaline pH. Some immobilization variables were optimized using statistical factorial design (Central Composite Design). Optimized immobilization variables enhanced the immobilization yield (IY) from 27.9 to 79.9% (2.9-fold). It was found that the immobilized enzyme had higher optimum temperature, higher half-life time (t 1/2 ), lower activation energy (E a ), lower deactivation constant rate (k d ) and higher decimal reduction time (D-values) within the temperature range of 40-60°C. Differential scanning calorimetry analysis (DSC) confirmed the thermalstability of the immobilized enzyme. The immobilized enzyme was stable at a wide pH range (5.0-8.0). Kinetic studies of starch hydrolysis demonstrated that immobilized enzyme had lower Michaelis constant (K m ), maximum velocity (V max ) and catalytic efficiency (V max /K m ) values. The storage stability and reusability of the immobilized enzyme were found to be about 74.7 and 62.5% of its initial activity after 28days and 11cycles, respectively. Enhanced α-amylase stabilities upon immobilization make it suitable for industrial application. Copyright © 2018 Elsevier B.V. All rights reserved.

  6. Development and Optimization of HPLC Analysis of Metronidazole, Diloxanide, Spiramycin and Cliquinol in Pharmaceutical Dosage Forms Using Experimental Design.

    PubMed

    Elkhoudary, Mahmoud M; Abdel Salam, Randa A; Hadad, Ghada M

    2016-11-01

    A new simple, sensitive, rapid and accurate gradient reversed-phase high-performance liquid chromatography with photodiode array detector (RP-HPLC-DAD) was developed and validated for simultaneous analysis of Metronidazole (MNZ), Spiramycin (SPY), Diloxanidefuroate (DIX) and Cliquinol (CLQ) using statistical experimental design. Initially, a resolution V fractional factorial design was used in order to screen five independent factors: the column temperature (°C), pH, phosphate buffer concentration (mM), flow rate (ml/min) and the initial fraction of mobile phase B (%). pH, flow rate and initial fraction of mobile phase B were identified as significant, using analysis of variance. The optimum conditions of separation determined with the aid of central composite design were: (1) initial mobile phase concentration: phosphate buffer/methanol (50/50, v/v), (2) phosphate buffer concentration (50 mM), (3) pH (4.72), (4) column temperature 30°C and (5) mobile phase flow rate (0.8 ml min -1 ). Excellent linearity was observed for all of the standard calibration curves, and the correlation coefficients were above 0.9999. Limits of detection for all of the analyzed compounds ranged between 0.02 and 0.11 μg ml -1 ; limits of quantitation ranged between 0.06 and 0.33 μg ml -1 The proposed method showed good prediction ability. The optimized method was validated according to ICH guidelines. Three commercially available tablets were analyzed showing good % recovery and %RSD. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  7. Biodegradation of kerosene: Study of growth optimization and metabolic fate of P. janthinellum SDX7

    PubMed Central

    Khan, Shamiyan R.; Nirmal, J.I. Kumar; Kumar, Rita N.; Patel, Jignasha G.

    2015-01-01

    Penicillum janthinellum SDX7 was isolated from aged petroleum hydrocarbon-affected soil at the site of Anand, Gujarat, India, and was tested for different pH, temperature, agitation and concentrations for optimal growth of the isolate that was capable of degrading upto 95%, 63% and 58% of 1%, 3% and 5% kerosene, respectively, after a period of 16 days, at optimal growth conditions of pH 6.0, 30 °C and 180 rpm agitation. The GC/MS chromatograms revealed that then-alkane fractions are easily degraded; however, the rate might be lower for branched alkanes, n-alkylaromatics, cyclic alkanes and polynuclear aromatics. The test doses caused a concentration-dependent depletion of carbohydrates of P. janthinellum SDX7 by 3% to 80%, proteins by 4% to 81% and amino acids by 8% to 95% upto 16 days of treatment. The optimal concentration of 3% kerosene resulted in the least reduction of the metabolites of P. janthinellum such as carbohydrates, proteins and amino acids with optimal growth compared to 5% and 1% (v/v) kerosene doses on the 12th and 16th day of exposure. Phenols were found to be mounted by 43% to 66% at lower and higher concentrations during the experimental period. Fungal isolate P. janthinellum SDX7 was also tested for growth on various xenobiotic compounds. PMID:26273254

  8. Biodegradation of kerosene: Study of growth optimization and metabolic fate of P. janthinellum SDX7.

    PubMed

    Khan, Shamiyan R; Nirmal, J I Kumar; Kumar, Rita N; Patel, Jignasha G

    2015-06-01

    Penicillum janthinellum SDX7 was isolated from aged petroleum hydrocarbon-affected soil at the site of Anand, Gujarat, India, and was tested for different pH, temperature, agitation and concentrations for optimal growth of the isolate that was capable of degrading upto 95%, 63% and 58% of 1%, 3% and 5% kerosene, respectively, after a period of 16 days, at optimal growth conditions of pH 6.0, 30 °C and 180 rpm agitation. The GC/MS chromatograms revealed that then-alkane fractions are easily degraded; however, the rate might be lower for branched alkanes, n-alkylaromatics, cyclic alkanes and polynuclear aromatics. The test doses caused a concentration-dependent depletion of carbohydrates of P. janthinellum SDX7 by 3% to 80%, proteins by 4% to 81% and amino acids by 8% to 95% upto 16 days of treatment. The optimal concentration of 3% kerosene resulted in the least reduction of the metabolites of P. janthinellum such as carbohydrates, proteins and amino acids with optimal growth compared to 5% and 1% (v/v) kerosene doses on the 12(th) and 16(th) day of exposure. Phenols were found to be mounted by 43% to 66% at lower and higher concentrations during the experimental period. Fungal isolate P. janthinellum SDX7 was also tested for growth on various xenobiotic compounds.

  9. First report of the production of a potent biosurfactant with α,β-trehalose by Fusarium fujikuroi under optimized conditions of submerged fermentation.

    PubMed

    Reis, Cristiane Bianchi Loureiro Dos; Morandini, Liziane Maria Barassuol; Bevilacqua, Caroline Borges; Bublitz, Fabricio; Ugalde, Gustavo; Mazutti, Marcio Antonio; Jacques, Rodrigo Josemar Seminoti

    2018-04-24

    Biosurfactants have many advantages over synthetic surfactants but have higher production costs. Identifying microorganisms with high production capacities for these molecules and optimizing their growth conditions can reduce cost. The present work aimed to isolate and identify a fungus with high biosurfactant production capacity, optimize its growth conditions in a low cost culture medium, and characterize the chemical structure of the biosurfactant molecule. The fungal strain UFSM-BAS-01 was isolated from soil contaminated with hydrocarbons and identified as Fusarium fujikuroi. To optimize biosurfactant production, a Plackett-Burman design and a central composite rotational design were used. The variables evaluated were pH, incubation period, temperature, agitation and amount of inoculum in a liquid medium containing glucose. The partial structure of the biosurfactant molecule was identified by nuclear magnetic resonance spectrometry. F. fujikuroi reduced surface tension from 72 to 20mNm -1 under the optimized conditions of pH 5.0, 37°C and 7 days of incubation with 190rpm agitation. The partial identification of the structure of the biosurfactant demonstrated the presence of an α,β-trehalose. The present study is the first report of the biosynthesis of this compound by F. fujikuroi, suggesting that the biosurfactant produced belongs to the class of trehalolipids. Copyright © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  10. Novel cyanine dyes with vinylsulfone group for labeling biomolecules.

    PubMed

    Park, Jin Woo; Kim, YoungSoo; Lee, Kee-Jung; Kim, Dong Jin

    2012-03-21

    Novel vinylsulfone cyanine dyes (em. 550-850 nm) were designed and synthesized for fluorescent labeling of biomolecules via 1,2-addition reaction in aqueous conditions. Due to the virtue of chemical structures of both fluorophore and reactive group, these dyes could be significantly stable and reactive in various aqueous/organic conditions. A wide variety of pH, temperature, buffer concentration, and protein were tested for the optimal labeling condition.

  11. Characterization and optimization of carboxylesterase-catalyzed esterification between capric acid and glycerol for the production of 1-monocaprin in reversed micellar system.

    PubMed

    Park, Kyung Min; Kwon, Oh Taek; Ahn, Seon Min; Lee, JaeHwan; Chang, Pahn-Shick

    2010-02-28

    Calotropis procera R. Br. carboxylesterase (EC 3.1.1.1) solubilized in reversed micellar glycerol droplets containing a very small amount of water (less than 5ppm) and stabilized by a surfactant effectively catalyzed the esterification between glycerol and capric acid to produce 1-monocaprin. Reaction variables including surfactant types, organic solvent media, reaction time, G-value ([glycerol]/[capric acid]), R-value ([water]/[surfactant]), pH, temperature, and types of metal ion inhibitors on the carboxylesterase-catalyzed esterification were characterized and optimized to efficiently produce 1-monocaprin. Bis(2-ethylhexyl) sodium sulfosuccinate (AOT) and isooctane were the most effective surfactant and organic solvent medium, respectively, for 1-monocaprin formation in reversed micelles. The optimum G- and R-values were 3.0 and 0.05, respectively, and the optimum pH and temperature were determined to be 10.0 and 60 degrees C, respectively. K(m,app.) and V(max,app.) were calculated from a Hanes-Woolf plot, and the values were 9.64 mM and 2.45 microM/min mg protein, respectively. Among various metal ions, Cu(2+) and Fe(2+) severely inhibited carboxylesterase-catalyzed esterification activity (less than 6.0% of relative activity). Copyright 2009 Elsevier B.V. All rights reserved.

  12. Optimization of Enterobacter cloacae (KU923381) for diesel oil degradation using response surface methodology (RSM).

    PubMed

    Ramasamy, Sugumar; Arumugam, Arumugam; Chandran, Preethy

    2017-02-01

    Efficiency of Enterobacter cloacae KU923381 isolated from petroleum hydrocarbon contaminated soil was evaluated in batch culture and bioreactor mode. The isolate were screened for biofilm formation using qualitative and quantitative assays. Response surface methodology (RSM) was used to study the effect of pH, temperature, glucose concentration, and sodium chloride on diesel degradation. The predicted values for diesel oil degradation efficiency by the statistical designs are in a close agreement with experimental data (R 2 = 99.66%). Degradation efficiency is increased by 36.78% at pH = 7, temperature = 35°C, glucose = 5%, and sodium chloride concentration = 5%. Under the optimized conditions, the experiments were performed for diesel oil degradation by gas chromatographic mass spectrometric analysis (GC-MS). GC-MS analysis confirmed that E. cloacae had highly degrade hexadecane, heptadecane, tridecane, and docosane by 99.71%, 99.23%, 99.66%, and 98.34% respectively. This study shows that rapid bioremoval of hydrocarbons in diesel oil is acheived by E. cloacae with abet of biofilm formation. The potential use of the biofilms for preparing trickling filters (gravel particles) for the degradation of hydrocarbons from petroleum wastes before their disposal in the open environment is highly suggested. This is the first successful attempt for artificially establishing petroleum hydrocarbon degrading bacterial biofilm on solid substrates in bioreactor.

  13. Chitosan production by psychrotolerant Rhizopus oryzae in non-sterile open fermentation conditions.

    PubMed

    Tasar, Ozden Canli; Erdal, Serkan; Taskin, Mesut

    2016-08-01

    A new chitosan producing fungus was locally isolated from soil samples collected around Erzurum, Turkey and identified as Rhizopus oryzae PAS 17 (GenBank accession number KU318422.1). Cultivation in low cost non-sterile conditions was achieved by exploiting its ability to grow at low temperature and pH, thus, undesired microbial contamination could be eliminated when appropriate culture conditions (incubation temperature as 15°C and initial pH of the medium as 4.5) were selected. Medium composition and culture conditions were optimized using Taguchi orthogonal array (OA) design of experiment (DOE). An OA layout of L16 (4(5)) was constructed with five most influensive factors at four levels on chitosan production like, carbon source (molasses), metal ion (Mg(2+)), inoculum amount, agitation speed and incubation time. The optimal combinations of factors (molasses, 70ml/l; MgSO4·7H2O, 0.5g/l; inoculum, 6.7×10(6) spores/disc; agitation speed, 150rpm and incubation time, 8days) obtained from the proposed DOE methodology was further validated by analysis of variance (ANOVA) test and the results revealed the increment of chitosan and biomass yields of 14.45 and 8.58 folds from its unoptimized condition, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Experimental design data for the biosynthesis of citric acid using Central Composite Design method.

    PubMed

    Kola, Anand Kishore; Mekala, Mallaiah; Goli, Venkat Reddy

    2017-06-01

    In the present investigation, we report that statistical design and optimization of significant variables for the microbial production of citric acid from sucrose in presence of filamentous fungi A. niger NCIM 705. Various combinations of experiments were designed with Central Composite Design (CCD) of Response Surface Methodology (RSM) for the production of citric acid as a function of six variables. The variables are; initial sucrose concentration, initial pH of medium, fermentation temperature, incubation time, stirrer rotational speed, and oxygen flow rate. From experimental data, a statistical model for this process has been developed. The optimum conditions reported in the present article are initial concentration of sucrose of 163.6 g/L, initial pH of medium 5.26, stirrer rotational speed of 247.78 rpm, incubation time of 8.18 days, fermentation temperature of 30.06 °C and flow rate of oxygen of 1.35 lpm. Under optimum conditions the predicted maximum citric acid is 86.42 g/L. The experimental validation carried out under the optimal values and reported citric acid to be 82.0 g/L. The model is able to represent the experimental data and the agreement between the model and experimental data is good.

  15. Mineralization of hetero bi-functional reactive dye in aqueous solution by Fenton and photo-Fenton reactions.

    PubMed

    Torrades, Francesc; García-Hortal, José Antonio; García-Montaño, Julia

    2015-01-01

    This study focused on the advanced oxidation of the hetero bi-functional reactive dye Sumifix Supra Yellow 3RF (CI Reactive Yellow 145) using dark Fenton and photo-Fenton conditions in a lab-scale experiment. A 2(3) factorial design was used to evaluate the effects of the three key factors: temperature, Fe(II) and H2O2 concentrations, for a dye concentration of 250 mg L(-1) with chemical oxygen demand (COD) of 172 mg L(-1) O2 at pH=3. The response function was the COD reduction. This methodology lets us find the effects and interactions of the studied variables and their roles in the efficiency of the treatment process. In the optimization, the correlation coefficients for the model (R2) were 0.948 and 0.965 for Fenton and photo-Fenton treatments, respectively. Under optimized reaction conditions: pH=3, temperature=298 K, [H2O2]=11.765 mM and [Fe(II)]=1.075 mM; 60 min of treatment resulted in a 79% and 92.2% decrease in COD, for the dye taken as the model organic compound, after Fenton and photo-Fenton treatments, respectively.

  16. Photo Degradation of Methyl Orange by Persulfate Activated with Zero Valent Iron

    NASA Astrophysics Data System (ADS)

    Munkoeva, V. A.; Sizykh, M. R.; Batoeva, A. A.

    2017-11-01

    The oxidative degradation of Methyl Orange (MO) subjected to direct photolysis (Solar) and various oxidative systems was studied. The comparative experiments have shown that MO conversion and mineralization increases in the following order: Solar ∼ Solar/Fe0 ∼ Solar/S2O82- < S2O82-/Fe0 < Solar/S2O82-/Fe0. The influence of the main factors (duration of exposure, the ratio of initial concentrations of MO:S2O82-:Fe0, pH and temperature of the reaction medium) on the degree of MO conversion and mineralization was studied. The optimal pH and temperature of the reaction medium were 5.8 and 25°C, respectively. The rate of MO decomposition and mineralization increased proportionally to the initial concentration of the oxidant at the molar ratios [S2O82-] :[MO] ≤ 12. Judging by the nature of the kinetic curves, a further increase of this ratio is impractical. However, an increase in the oxidant concentration had a positive effect on the degrees of conversion and mineralization of total organic carbon (TOC). Thus, at the ratios of 12:1 and 48:1, the conversion efficiency of TOC was 23 and 60 %, respectively. The optimal concentration of Fe0 was 100 mg/l.

  17. Optimization of Exopolysaccharide Production by Lactobacillus delbrueckii subsp. bulgaricus RR Grown in a Semidefined Medium

    PubMed Central

    Kimmel, Stacy A.; Roberts, Robert F.; Ziegler, Gregory R.

    1998-01-01

    The optimal fermentation temperature, pH, and Bacto-casitone (Difco Laboratories, Detroit, Mich.) concentration for production of exopolysaccharide by Lactobacillus delbrueckii subsp. bulgaricus RR in a semidefined medium were determined by using response surface methods. The design consisted of 20 experiments, 15 unique combinations, and five replications. All fermentations were conducted in a fermentor with a 2.5-liter working volume and were terminated when 90% of the glucose in the medium had been consumed. The population of L. delbrueckii subsp. bulgaricus RR and exopolysaccharide content were measured at the end of each fermentation. The optimum temperature, pH, and Bacto-casitone concentration for exopolysaccharide production were 38°C, 5, and 30 g/liter, respectively, with a predicted yield of 295 mg of exopolysaccharide/liter. The actual yield under these conditions was 354 mg of exopolysaccharide/liter, which was within the 95% confidence interval (217 to 374 mg of exopolysaccharide/liter). An additional experiment conducted under optimum conditions showed that exopolysaccharide production was growth associated, with a specific production at the endpoint of 101.4 mg/g of dry cells. Finally, to obtain material for further characterization, a 100-liter fermentation was conducted under optimum conditions. Twenty-nine grams of exopolysaccharide was isolated from centrifuged, ultrafiltered fermentation broth by ethanol precipitation. PMID:9464404

  18. Optimization of enzymes-microwave-ultrasound assisted extraction of Lentinus edodes polysaccharides and determination of its antioxidant activity.

    PubMed

    Yin, Chaomin; Fan, Xiuzhi; Fan, Zhe; Shi, Defang; Gao, Hong

    2018-05-01

    Enzymes-microwave-ultrasound assisted extraction (EMUE) method had been used to extract Lentinus edodes polysaccharides (LEPs). The enzymatic temperature, enzymatic pH, microwave power and microwave time were optimized by response surface methodology. The yields, properties and antioxidant activities of LEPs from EMUE and other extraction methods including hot-water extraction, enzymes-assisted extraction, microwave-assisted extraction and ultrasound-assisted extraction were evaluated. The results showed that the highest LEPs yield of 9.38% was achieved with enzymatic temperature of 48°C, enzymatic pH of 5.0, microwave power of 440W and microwave time of 10min, which correlated well with the predicted value of 9.79%. Additionally, LEPs from different extraction methods possessed typical absorption peak of polysaccharides, which meant different extraction methods had no significant effects on type of glycosidic bonds and sugar ring of LEPs. However, SEM images of LEPs from different extraction methods were significantly different. Moreover, the different LEPs all showed antioxidant activities, but LEPs from EMUE showed the highest reducing power when compared to other LEPs. The results indicated LEPs from EMUE can be used as natural antioxidant component in the pharmaceutical and functional food industries. Copyright © 2018 Elsevier B.V. All rights reserved.

  19. Optimization of edible coating formulations for improving postharvest quality and shelf life of pear fruit using response surface methodology.

    PubMed

    Nandane, A S; Dave, Rudri K; Rao, T V Ramana

    2017-01-01

    The effect of composite edible films containing soy protein isolate (SPI) in combination with additives like hydroxypropyl methylcellulose (HPMC) and olive oil on 'Babughosha' pear ( Pyrus communis L.) stored at ambient temperature (28 ± 5 °C and 60 ± 10% RH) was evaluated using Response surface methodology (RSM). A total of 30 edible coating formulations comprising of SPI (2-6%, w/v), olive oil (0.7-1.1%, v/v), HPMC (0.1-0.5%, w/v) and potassium sorbate (0-0.4% w/v) were evaluated for optimizing the most suitable combination. Quality parameters like weight loss%, TSS, pH and titrable acidity of the stored pears were selected as response variables for optimization. The optimization procedure was carried out using RSM. It was observed that the response variables were mainly effected by concentration of SPI and olive oil in the formulation. Edible coating comprising of SPI 5%, HPMC 0.40%, olive oil 1% and potassium sorbate 0.22% was found to be most suitable combination for pear fruit with predicted values of response variables indicated as weight loss% 3.50, pH 3.41, TSS 11.13 and TA% 0.513.

  20. Optimization of Preparation of Antioxidative Peptides from Pumpkin Seeds Using Response Surface Method

    PubMed Central

    Fan, Sanhong; Hu, Yanan; Li, Chen; Liu, Yanrong

    2014-01-01

    Protein isolates of pumpkin (Cucurbita pepo L) seeds were hydrolyzed by acid protease to prepare antioxidative peptides. The hydrolysis conditions were optimized through Box-Behnken experimental design combined with response surface method (RSM). The second-order model, developed for the DPPH radical scavenging activity of pumpkin seed hydrolysates, showed good fit with the experiment data with a high value of coefficient of determination (0.9918). The optimal hydrolysis conditions were determined as follows: hydrolyzing temperature 50°C, pH 2.5, enzyme amount 6000 U/g, substrate concentration 0.05 g/ml and hydrolyzing time 5 h. Under the above conditions, the scavenging activity of DPPH radical was as high as 92.82%. PMID:24637721

  1. CO2 induced pHi changes in the brain of polar fish: a TauCEST application.

    PubMed

    Wermter, Felizitas C; Maus, Bastian; Pörtner, Hans-O; Dreher, Wolfgang; Bock, Christian

    2018-06-22

    Chemical exchange saturation transfer (CEST) from taurine to water (TauCEST) can be used for in vivo mapping of taurine concentrations as well as for measurements of relative changes in intracellular pH (pH i ) at temperatures below 37°C. Therefore, TauCEST offers the opportunity to investigate acid-base regulation and neurological disturbances of ectothermic animals living at low temperatures, and in particular to study the impact of ocean acidification (OA) on neurophysiological changes of fish. Here, we report the first in vivo application of TauCEST imaging. Thus, the study aimed to investigate the TauCEST effect in a broad range of temperatures (1-37°C) and pH (5.5-8.0), motivated by the high taurine concentration measured in the brains of polar fish. The in vitro data show that the TauCEST effect is especially detectable in the low temperature range and strictly monotonic for the relevant pH range (6.8-7.5). To investigate the specificity of TauCEST imaging for the brain of polar cod (Boreogadus saida) at 1.5°C simulations were carried out, indicating a taurine contribution of about 65% to the in vivo expected CEST effect, if experimental parameters are optimized. B. saida was acutely exposed to three different CO 2 concentrations in the sea water (control normocapnia; comparatively moderate hypercapnia OA m  = 3300 μatm; high hypercapnia OA h  = 4900 μatm). TauCEST imaging of the brain showed a significant increase in the TauCEST effect under the different CO 2 concentrations of about 1.5-3% in comparison with control measurements, indicative of changes in pH i or metabolite concentration. Consecutive recordings of 1 H MR spectra gave no support for a concentration induced change of the in vivo observed TauCEST effect. Thus, the in vivo application of TauCEST offers the possibility of mapping relative changes in pH i in the brain of polar cod during exposure to CO 2 . © 2018 John Wiley & Sons, Ltd.

  2. Influence of pH on optoelectronic properties of zinc sulphide thin films prepared using hydrothermal and spin coating method

    NASA Astrophysics Data System (ADS)

    Choudapur, V. H.; Bennal, A. S.; Raju, A. B.

    2018-04-01

    The ZnS nanomaterial is synthesized by hydrothermal method under optimized conditions using Zinc acetate and sodium sulphide as precursors. The Zinc Sulphide thin films are obtained by simple spin coating method with high optical transmittance. The prepared thin films are adhesive and uniform. The x-ray diffraction analysis showed that the films are polycrystalline in cubic phase with the preferred orientation along (111) direction. Current-voltage curves were recorded at room temperature using Keithley 617 programmable electrometer and conductivity is calculated for the film coated on ITO by two probe method. The pH of the solution is varied by using ammonia and hydrochloric acid. The comparative studies of effect of pH on the morphology, crystallanity and optoelectronic properties of the films are studied. It is observed that the pH of the solution has large influence on optoelectronic properties. The thin film prepared with neutral pH has higher crystallanity, bandgap and conductivity as compared to the samples prepared in acidic or basic solutions.

  3. Optimization of L-asparaginase production from novel Enterobacter sp., by submerged fermentation using response surface methodology.

    PubMed

    Erva, Rajeswara Reddy; Goswami, Ajgebi Nath; Suman, Priyanka; Vedanabhatla, Ravali; Rajulapati, Satish Babu

    2017-03-16

    The culture conditions and nutritional rations influencing the production of extra cellular antileukemic enzyme by novel Enterobacter aerogenes KCTC2190/MTCC111 were optimized in shake-flask culture. Process variables like pH, temperature, incubation time, carbon and nitrogen sources, inducer concentration, and inoculum size were taken into account. In the present study, finest enzyme activity achieved by traditional one variable at a time method was 7.6 IU/mL which was a 2.6-fold increase compared to the initial value. Further, the L-asparaginase production was optimized using response surface methodology, and validated experimental result at optimized process variables gave 18.35 IU/mL of L-asparaginase activity, which is 2.4-times higher than the traditional optimization approach. The study explored the E. aerogenes MTCC111 as a potent and potential bacterial source for high yield of antileukemic drug.

  4. Optimization to the Culture Conditions for Phellinus Production with Regression Analysis and Gene-Set Based Genetic Algorithm

    PubMed Central

    Li, Zhongwei; Xin, Yuezhen; Wang, Xun; Sun, Beibei; Xia, Shengyu; Li, Hui

    2016-01-01

    Phellinus is a kind of fungus and is known as one of the elemental components in drugs to avoid cancers. With the purpose of finding optimized culture conditions for Phellinus production in the laboratory, plenty of experiments focusing on single factor were operated and large scale of experimental data were generated. In this work, we use the data collected from experiments for regression analysis, and then a mathematical model of predicting Phellinus production is achieved. Subsequently, a gene-set based genetic algorithm is developed to optimize the values of parameters involved in culture conditions, including inoculum size, PH value, initial liquid volume, temperature, seed age, fermentation time, and rotation speed. These optimized values of the parameters have accordance with biological experimental results, which indicate that our method has a good predictability for culture conditions optimization. PMID:27610365

  5. Investigation on solubility of hydroxy dibasic acids in alkanolamine solutions

    NASA Astrophysics Data System (ADS)

    Du, M.

    2017-12-01

    Solubilities of three hydroxy dibasic (adipic, suberic, and sebacic) acids in alkanolamine solutions were measured within the 30-90℃ temperature range. It is found that solubility of these acids sharply grows with temperature and concentration of alkanolamine solvent. In addition, the study substantiates the adjustment of pH to optimize the CO2 absorption and desorption processes. The precipitation of added acids from alkanolamine solvents by cooling is found to be quite problematic, which makes the recovery of residual acids from lean alkanolamine solvents non-feasible and requires the application of alternative methods.

  6. Evaluation of a new polymeric stationary phase with reversed-phase properties for high temperature liquid chromatography.

    PubMed

    Vanhoenacker, Gerd; Dos Santos Pereira, Alberto; Kotsuka, Takashi; Cabooter, Deirdre; Desmet, Gert; Sandra, Pat

    2010-05-07

    The performance of a polymeric stationary phase with reversed-phase properties (ET-RP1) was evaluated for LC separations at elevated temperature. The most significant observation was that the reduced plate height (h) decreased from 3.4 at 25 degrees C (optimal flow 0.5 mL/min) to 2.4 at 150 degrees C (optimal flow 2.5 mL/min) which is comparable to the efficiency obtained with silica-based reversed-phase columns of 4.6mm ID operated at 0.8 mL/min. The phase showed no deterioration after long use at 150 degrees C within the pH range 1-9. Catalytic activity originating from the stationary phase material, e.g. as experienced on zirconium columns operated at elevated temperature, was absent. The performance of ET-RP1 is illustrated with the analysis of some pharmaceutical samples by LC and LC-MS. Operation at elevated temperature also allows to reduce the amount of organic modifier or to replace acetonitrile and methanol by the biodegradable ethanol. Copyright (c) 2009 Elsevier B.V. All rights reserved.

  7. Biochemical characterization of a recombinant plant class III chitinase from the pitcher of the carnivorous plant Nepenthes alata.

    PubMed

    Ishisaki, Kana; Arai, Sachiko; Hamada, Tatsuro; Honda, Yuji

    2012-11-01

    A class III chitinase belonging to the GH18 family from Nepenthes alata (NaCHIT3) was expressed in Escherichia coli. The enzyme exhibited hydrolytic activity toward colloidal chitin, ethylene glycol chitin, and (GlcNAc)(n) (n=5 and 6). The enzyme hydrolyzed the fourth glycosidic linkage from the non-reducing end of (GlcNAc)(6). The anomeric form of the products indicated it was a retaining enzyme. The colloidal chitin hydrolytic reaction displayed high activity between pH 3.9 and 6.9, but the pH optimum of the (GlcNAc)(6) hydrolytic reaction was 3.9 at 37 °C. The optimal temperature for activity was 65 °C in 50 mM sodium acetate buffer (pH 3.9). The pH optima of NaCHIT3 and NaCHIT1 might be related to their roles in chitin degradation in the pitcher fluid. Copyright © 2012 Elsevier Ltd. All rights reserved.

  8. Properties of lactate dehydrogenase from the isopod, Saduria entomon.

    PubMed

    Mulkiewicz, E; Zietara, M S; Stachowiak, K; Skorkowski, E F

    2000-07-01

    Saduria entomon lactate dehydrogenase (LDH-A4*) from thorax muscle was purified about 89 fold to specific activity 510 micromol NADH/min/mg using Cibacron Blue 3GA Agarose and Oxamate-Agarose chromatographies. The enzyme is a tetramer, with molecular weight of 140 kDa for the native enzyme and 36 kDa for the subunit. The isoelectric point was at pH 5.7. The enzyme possesses high heat stability (T50 = 71.5 degrees C). The optimum pH for pyruvate reduction reaction was 6.5, while for lactate oxidation one, the maximum activity was at pH 9.1. The Km for pyruvate was minimal at 5 degrees C, the average environmental temperature of the isopod. The Km values determined at 30 degrees C and optimal pH for pyruvate reduction and lactate oxidation were 0.18 and 90.04 mM, respectively. Amino acid compositional analyses showed the strongest resemblance of the isopod isoenzyme to cod (Gadus morhua) LDH-C4.

  9. Acid-catalysed xylose dehydration into furfural in the presence of kraft lignin.

    PubMed

    Lamminpää, Kaisa; Ahola, Juha; Tanskanen, Juha

    2015-02-01

    In this study, the effects of kraft lignin (Indulin AT) on acid-catalysed xylose dehydration into furfural were studied in formic and sulphuric acids. The study was done using D-optimal design. Three variables in both acids were included in the design: time (20-80 min), temperature (160-180°C) and initial lignin concentration (0-20 g/l). The dependent variables were xylose conversion, furfural yield, furfural selectivity and pH change. The results showed that the xylose conversion and furfural yield decreased in sulphuric acid, while in formic acid the changes were minor. Additionally, it was showed that lignin has an acid-neutralising capacity, and the added lignin increased the pH of reactant solutions in both acids. The pH rise was considerably lower in formic acid than in sulphuric acid. However, the higher pH did not explain all the changes in conversion and yield, and thus lignin evidently inhibits the formation of furfural. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Production and Characterization of Lipases by Two New Isolates of Aspergillus through Solid-State and Submerged Fermentation

    PubMed Central

    Colla, Luciane Maria; Ficanha, Aline M. M.; Rizzardi, Juliana; Bertolin, Telma Elita; Reinehr, Christian Oliveira; Costa, Jorge Alberto Vieira

    2015-01-01

    Due to the numerous applications of lipases in industry, there is a need to study their characteristics, because lipases obtained from different sources may present different properties. The aim of this work was to accomplish the partial characterization of lipases obtained through submerged fermentation and solid-state fermentation by two species of Aspergillus. Fungal strains were isolated from a diesel-contaminated soil and selected as good lipases producers. Lipases obtained through submerged fermentation presented optimal activities at 37°C and pH 7.2 and those obtained through solid-state fermentation at 35°C and pH 6.0. The enzymes produced by submerged fermentation were more temperature-stable than those obtained by solid-state fermentation, presenting 72% of residual activity after one hour of exposition at 90°C. Lipases obtained through submerged fermentation had 80% of stability in acidic pH and those obtained through solid-state fermentation had stability greater than 60% in alkaline pH. PMID:26180809

  11. Thermo-controlled rheology of electro-assembled polyanionic polysaccharide (alginate) and polycationic thermo-sensitive polymers.

    PubMed

    Niang, Pape Momar; Huang, Zhiwei; Dulong, Virginie; Souguir, Zied; Le Cerf, Didier; Picton, Luc

    2016-03-30

    Several thermo-sensitive polyelectrolyte complexes were prepared by ionic self-association between an anionic polysaccharide (alginate) and a monocationic copolymer (polyether amine, Jeffamine®-M2005) with a 'Low Critical Solubility Temperature' (LCST). We show that electro-association must be established below the aggregation temperature of the free Jeffamine®, after which the organization of the system is controlled by the thermo-association of Jeffamine® that was previously electro-associated with the alginate. Evidence for this comes primarily from the rheology in the semi-dilute region. Electro- and thermo-associative behaviours are optimal at a pH corresponding to maximum ionization of both compounds (around pH 7). High ionic strength could prevent the electro-association. The reversibility of the transition is possible only at temperatures lower than the LCST of Jeffamine®. Similar behaviour has been obtained with carboxymethyl cellulose (CMC), which suggests that this behaviour can be observed using a range of anionic polyelectrolytes. In contrast, no specific properties have been found for pullulan, which is a neutral polysaccharide. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Effects of pH, temperature, and osmolarity on the morphology and survival rate of primary hemocyte cultures from the mitten crab, Eriocheir sinensis.

    PubMed

    Hong, Yuhang; Yang, Xiaozhen; Cheng, Yongxu; Liang, Pan; Zhang, Jinbiao; Li, Meng; Shen, Cheng; Yang, Zhigang; Wang, Chun

    2013-10-01

    Hemocytes are the main immune defense cells in crustacean, and its in vitro culture can be a useful tool for the study of host and pathogen interaction. In the present study, the primary hemocyte culture of Chinese mitten crab (Eriocheir sinensis), including mixed and single hemocyte, was set up for the first time. In this study, different pH (6.4, 6.8, 7.2, 7.6, and 8.0), temperature (26, 28, and 30°C), and osmolarity (500, 700, 900, 1,100, and 1,300 mOsm kg(-1)) values were tested. Moreover, the effects of two types of medium (1× L-15 and 3× L-15) with the same osmolarity on hemocyte culture were evaluated. After incubation at different culture conditions, the morphological changes (degranulation, lysis, shrinkage, and detachment) and survival rate of hemocytes were taken into account in order to evaluate the culture condition effect. Our results showed that the total hemocyte counts of Chinese mitten crab were about 2.5 × 10(7) cells ml(-1), and three subpopulations of hemocytes were distinguished as granulocytes (43.46 ± 4.98%), semigranulocytes (31.04 ± 1.95%), and hyalinocytes (25.50 ± 4.89%). The optimal culture condition for primary hemocytes of Chinese mitten crab was 3× L-15 medium, 1,100 mOsm kg(-1), pH 6.8 at 28°C. Hemocytes at optimal culture condition could retain a better morphology and higher survival rate: hemocytes retained a survival rate >60% after 5 d and >40% after 7 d. Furthermore, the hemocyte subpopulations were isolated by Percoll step gradient centrifugation and cultured in optimized hemocyte culture conditions. The results showed that hyalinocytes and semigranulocytes could maintain a survival rate of >50% after 15 d, while granulocytes only retained a survival rate of 26% after 5 d.

  13. Hexavalent Chromate Reductase Activity in Cell Free Extracts of Penicillium sp.

    PubMed Central

    Arévalo-Rangel, Damaris L.; Cárdenas-González, Juan F.; Martínez-Juárez, Víctor M.; Acosta-Rodríguez, Ismael

    2013-01-01

    A chromium-resistant fungus isolated from contaminated air with industrial vapors can be used for reducing toxic Cr(VI) to Cr(III). This study analyzes in vitro reduction of hexavalent chromium using cell free extract(s) of the fungus that was characterized based on optimal temperature, pH, use of electron donors, metal ions and initial Cr(VI) concentration in the reaction mixture. This showed the highest activity at 37°C and pH 7.0; there is an increase in Cr(VI) reductase activity with addition of NADH as an electron donor, and it was highly inhibited by Hg2+, Ca2+ and Mg2+, and azide, EDTA, and KCN. PMID:24027493

  14. Functionalized vertically aligned ZnO nanorods for application in electrolyte-insulator-semiconductor based pH sensors and label-free immuno-sensors

    NASA Astrophysics Data System (ADS)

    Kumar, Narendra; Senapati, Sujata; Kumar, Satyendra; Kumar, Jitendra; Panda, Siddhartha

    2016-04-01

    Vertically aligned ZnO nanorods were grown on a SiO2/Si surface by optimization of the temperature and atmosphere for annealing of the seed. The seed layer annealed at 500 °C in vacuum provided well separated and uniform seeds which also provided the best condition to get densely packed, uniformly distributed, and vertically aligned nanorods. These nanorods grown on the substrates were used to fabricate electrolyte-insulator-semiconductor (EIS) devices for pH sensing. Etching of ZnO at acidic pH prevents the direct use of nanorods for pH sensing. Therefore, the nanorods functionalised with 3-aminopropyltriethoxysilane (APTES) were utilized for pH sensing and showed the pH sensitivity of 50.1 mV/pH. APTES is also known to be used as a linker to immobilize biomolecules (such as antibodies). The EIS device with APTES functionalized nanorods was used for the label free detection of prostate-specific antigen (PSA). Finally, voltage shifts of 23 mV and 35 mV were observed with PSA concentrations of 1 ng/ml and 100 ng/ml, respectively.

  15. OPTIMIZATION OF ALKALINE Α-AMYLASE PRODUCTION BY THERMOPHILIC BACILL US SUBTILIS.

    PubMed

    Al-Johani, Nuha Bakeet; Al-Seeni, Madeha N; Ahmed, Youssri Mohamed

    2017-01-01

    Starch-degrading amylase enzyme is important in biotechnological applications as food, fermentation, textile, paper and pharmaceutical purposes. The aim of current study to isolate alkaline thermostable α-amylase bacteria and then study the composition of medium and culture conditions to optimize cells growth and a-amylase production. Thermophilic amylase producing bacterium was isolated from local hot water-springs in Gazan city Saudi Arabia. Phylogenetic analysis of 16 S rRNA sequence for the strain revealed that the strain have the same sequence of Bacillus subtilis . Maximum amylase production was observed, when B. subtilis cultured in medium containing starch at concentration 0.5%, and 10 g/L peptones as nitrogen source at pH 8.5 in when it was incubated for 48 h at 45°C. An amylase-producing bacterium were isolated from hot-spring water and was identified as B. subtilis . Amylase produced from B.subtilis had optimum temperature 45°C and pH 8.5 in shaking media.

  16. Immobilization of Paecilomyces variotii tannase and properties of the immobilized enzyme.

    PubMed

    Schons, Patrícia Fernanda; Lopes, Fernanda Cristina Rezende; Battestin, Vania; Macedo, Gabriela Alves

    2011-01-01

    Tannase produced by Paecilomyces variotii was encapsulated in sodium alginate beads and used for the effective hydrolysis of tannic acid; the efficiency of hydrolysis was comparable to that of the free enzyme. The alginate beads retained 100% of their efficiency in the first three rounds of successive use and 60% in rounds 4 and 5. The response surface methodology showed that the best conditions to hydrolysis of tannic acid by immobilized tannase were: sodium alginate 5.2%, CaCl₂ 0.55 M and 9 h to curing time. The optimized process resulted in 2.4 times more hydrolysed tannic acid than that obtained before optimization. The optimum pH for the actions of both the encapsulated and the free enzymes was 5.5. The optimum temperature of the reaction was determined to be 40 °C for the free enzyme and 60 °C for the immobilized form. The immobilization process improved the stability at low pH.

  17. Optimization of antioxidant activity by response surface methodology in hydrolysates of jellyfish (Rhopilema esculentum) umbrella collagen.

    PubMed

    Zhuang, Yong-liang; Zhao, Xue; Li, Ba-fang

    2009-08-01

    To optimize the hydrolysis conditions to prepare hydrolysates of jellyfish umbrella collagen with the highest hydroxyl radical scavenging activity, collagen extracted from jellyfish umbrella was hydrolyzed with trypsin, and response surface methodology (RSM) was applied. The optimum conditions obtained from experiments were pH 7.75, temperature (T) 48.77 degrees C, and enzyme-to-substrate ratio ([E]/[S]) 3.50%. The analysis of variance in RSM showed that pH and [E]/[S] were important factors that significantly affected the process (P<0.05 and P<0.01, respectively). The hydrolysates of jellyfish umbrella collagen were fractionated by high performance liquid chromatography (HPLC), and three fractions (HF-1>3000 Da, 1000 Da

  18. Optimization of antioxidant activity by response surface methodology in hydrolysates of jellyfish (Rhopilema esculentum) umbrella collagen*

    PubMed Central

    Zhuang, Yong-liang; Zhao, Xue; Li, Ba-fang

    2009-01-01

    To optimize the hydrolysis conditions to prepare hydrolysates of jellyfish umbrella collagen with the highest hydroxyl radical scavenging activity, collagen extracted from jellyfish umbrella was hydrolyzed with trypsin, and response surface methodology (RSM) was applied. The optimum conditions obtained from experiments were pH 7.75, temperature (T) 48.77 °C, and enzyme-to-substrate ratio ([E]/[S]) 3.50%. The analysis of variance in RSM showed that pH and [E]/[S] were important factors that significantly affected the process (P<0.05 and P<0.01, respectively). The hydrolysates of jellyfish umbrella collagen were fractionated by high performance liquid chromatography (HPLC), and three fractions (HF-1>3000 Da, 1000 Da

  19. First evidence of bioflocculant from Shinella albus with flocculation activity on harvesting of Chlorella vulgaris biomass.

    PubMed

    Li, Yi; Xu, Yanting; Liu, Lei; Jiang, Xiaobing; Zhang, Kun; Zheng, Tianling; Wang, Hailei

    2016-10-01

    Bioflocculant from Shinella albus xn-1 could be used to harvest energy-producing microalga Chlorella vulgaris biomass for the first time. In this study, we investigated the flocculation activity and mode of strain xn-1, the characteristics of bioflocculant, the effect of flocculation conditions and optimized the flocculation efficiency. The results indicated that strain xn-1 exhibited flocculation activity through secreting bioflocculant; the bioflocculant with high thermal stability, pH stability and low molecular weight was proved to be not protein and polysaccharide, and flocculation active component was confirmed to contain triple bond and cumulated double bonds; algal pH, temperature and metal ions showed great impacts on the flocculation efficiency of bioflocculant; the maximum flocculation activity of bioflocculant reached 85.65% after the response surface optimization. According to the results, the bioflocculant from S. albus xn-1 could be a good potential in applications for high-efficiency harvesting of microalgae. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Immobilization of an enzyme from a Fusarium fungus WZ-I for chlorpyrifos degradation.

    PubMed

    Xie, Hui; Zhu, Lusheng; Ma, Tingting; Wang, Jun; Wang, Jinhua; Su, Jun; Shao, Bo

    2010-01-01

    The free enzyme extracted from WZ-I, which was identified as Fusarium LK. ex Fx, could effectively degrade chlorpyrifos, an organophosphate insecticide. The methods of immobilizing this free enzyme and determined its degradation-related characteristics were investigated. The properties of the immobilized enzyme were compared with those of the free enzyme. The optimal immobilization of the enzyme was achieved in a solution of 30 g/L sodium alginate at 4 degrees C for 4-12 hr. The immobilized enzyme showed the maximal activity at pH 8.0, 45 degrees C. The maximum initial rate and the substrate concentration of the immobilized enzyme were less than that of the free enzyme. The immobilized enzyme, therefore, had a higher capacity to withstand a broader range of temperatures and pH conditions than the free enzyme. With varying pH and temperatures, the immobilized enzyme was more active than the free enzyme in the degradation reaction. In addition, the immobilized enzyme exhibited only a slight loss in its initial activity, even after three repeated uses. The results showed that the immobilized enzyme was more resistant to different environmental conditions, suggesting that it was viable for future practical use.

  1. Bacillus sphaericus LMG 22257 is physiologically suitable for self-healing concrete.

    PubMed

    Wang, Jianyun; Jonkers, Henk M; Boon, Nico; De Belie, Nele

    2017-06-01

    The suitability of using a spore-forming ureolytic strain, Bacillus sphaericus, was evaluated for self-healing of concrete cracks. The main focus was on alkaline tolerance, calcium tolerance, oxygen dependence, and low-temperature adaptability. Experimental results show that B. sphaericus had a good tolerance. It can grow and germinate in a broad range of alkaline pH. The optimal pH range is 7 ∼ 9. High alkaline conditions (pH 10 ∼ 11) slow down but not stop the growth and germination. Oxygen was strictly needed during bacterial growth and germination, but not an essential factor during bacterial urea decomposition. B. sphaericus also had a good Ca tolerance, especially at a high bacterial concentration of 10 8  cells/mL; no significant influence was observed on bacterial ureolytic activity of the presence of 0.9M Ca 2+ . Furthermore, at a low temperature (10 °C), bacterial spores germinated and revived ureolytic activity with some retardation. However, this retardation can be counteracted by using a higher bacterial concentration and by supplementing yeast extract. It can be concluded that B. sphaericus is a suitable bacterium for application in bacteria-based self-healing concrete.

  2. Banana peel: a green and economical sorbent for the selective removal of Cr(VI) from industrial wastewater.

    PubMed

    Memon, Jamil R; Memon, Saima Q; Bhanger, Muhammad I; El-Turki, Adel; Hallam, Keith R; Allen, Geoffrey C

    2009-05-01

    This study describes the use of banana peel, a commonly produced fruit waste, for the removal of Cr(VI) from industrial wastewater. The parameters pH, contact time, initial metal ion concentration, and temperature were investigated and the conditions resulting in rapid and efficient adsorption (95% within 10 min) were determined. The binding of metal ions was found to be pH dependent with the optimal sorption occurring at pH 2. The retained species were eluted with 5 mL of 2M H(2)SO(4). To elucidate the mechanism of the process, total amounts of chromium and Cr(VI) were analyzed using flame atomic absorption and ultraviolet-visible (UV-vis) spectroscopic techniques, respectively. The Langmuir and Dubinin-Radushkevich (D-R) isotherms were used to describe the partitioning behavior for the system at different temperatures. Kinetics and thermodynamics of Cr(VI) removal by banana peel were also studied. The influence of diverse ions on the sorption behavior revealed that only Fe(II) ions (of those tested) suppressed the sorption of Cr(VI) ions to some extent. The method was applied for the removal of Cr(VI) from industrial wastewater.

  3. Characterization of a bi-functional cellulase produced by a gut bacterial resident of Rosaceae branch borer beetle, Osphranteria coerulescens (Coleoptera: Cerambycidae).

    PubMed

    Hatefi, Atousa; Makhdoumi, Ali; Asoodeh, Ahmad; Mirshamsi, Omid

    2017-10-01

    A cellulolytic bacterium was obtained from the digestive tract of Osphranteria coerulescens. The breakdown of woody and cellulosic substances by this insect may be relative in part to its symbiont bacteria. Under optimal cultural conditions the novel isolate produced 5.35U/ml cellulase after 72h. The enzyme was purified to 36 fold with a 0.59% yield and showed a specific activity of 9.0U/mg. It presented its maximum activity at 60°C and pH 5, while it was stable in a wide range of temperature from 20 to 60°C and pH from 5 to 10. The purified enzyme had a molecular weight of 42.50kDa based on SDS-PAGE and zymogram analyses. It demonstrated high ions and solvent stability and its activity was stimulated by Mn 2+ , Na + , DMSO and chloroform. The enzyme could hydrolyze CMC, avicel, cellulose and sawdust. TLC analysis represented the cellobiose as the hydrolytic product of CMC. With regard to endo/exo glucanase activity and wide pH, temperature and solvent stability, it has potential for industrial application. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. A novel biochar from Manihot esculenta Crantz waste: application for the removal of Malachite Green from wastewater and optimization of the adsorption process.

    PubMed

    Beakou, Buscotin Horax; El Hassani, Kaoutar; Houssaini, Mohammed Amine; Belbahloul, Mounir; Oukani, Elhassan; Anouar, Abdellah

    2017-09-01

    The adsorptive removal of Malachite Green (MG) by a novel biochar namely Cassava Rind Carbon (CRC) was studied in a batch system. Moreover, Box-Behnken Response Surface Methodology was used to optimize operating conditions of the adsorption process. Characterization was done by Thermo Gravimetric Analysis (TGA), Attenuated Total Reflectance Fourier Transform Infra-Red Spectroscopy (ATR/FTIR), Brunauer-Emmett-Teller (BET) surface area, Scanning Electron Microscopy (SEM), X-Ray Diffraction (XRD) and pH zero charge point (pH ZCP ). The pseudo-second-order model and Langmuir model provided the best fit for kinetic and isotherm, respectively. The maximum capacity of dye adsorbed was 932.98 mg/g at 25 °C. The influence of temperature, the mass of adsorbent and the concentration of dye was studied. The optimal amount of adsorbed MG was 1,363.58 mg/g corresponding to 50 °C, 5 mg of CRC and 150 mg/L of dye. According to the high performance exhibited by CRC in this study, Manihot esculenta Crantz waste can be used as a better and low-cost biomass for wastewater decolourization.

  5. Optimization of the Production of 1-Phenylethanol Using Enzymes from Flowers of Tea (Camellia sinensis) Plants.

    PubMed

    Dong, Fang; Zhou, Ying; Zeng, Lanting; Watanabe, Naoharu; Su, Xinguo; Yang, Ziyin

    2017-01-13

    1-Phenylethanol (1PE) can be used as a fragrance in food flavoring and cosmetic industries and as an intermediate in the pharmaceutical industry. 1PE can be synthesized from acetophenone, and the cost of 1PE is higher than the cost of acetophenone. Therefore, it is important to establish an effective and low-cost approach for producing 1PE. Our previous studies found that tea ( Camellia sinensis ) flowers, which are an abundant and waste resource, contained enzymes that could transform acetophenone to 1PE. In the present study, we extracted crude enzymes from tea flowers and optimized the production conditions of 1PE using response surface methodology. The optimized conditions were an extraction pH of 7.0, a reaction pH of 5.3, a reaction temperature of 55 °C, a reaction time of 100 min, a coenzyme NADPH concentration of 3.75 μmol/mL in the reaction assay, and a substrate acetophenone concentration of 1.25 μmol/mL in the reaction assay. The results provide essential information for future industrial 1PE production using plant-derived enzymes.

  6. Bioprocessing of wheat bran for the production of lignocellulolytic enzyme cocktail by Cotylidia pannosa under submerged conditions.

    PubMed

    Sharma, Deepika; Garlapat, Vijay Kumar; Goel, Gunjan

    2016-04-02

    Characterization and production of efficient lignocellulytic enzyme cocktails for biomass conversion is the need for biofuel industry. The present investigation reports the modeling and optimization studies of lignocellulolytic enzyme cocktail production by Cotylidia pannosa under submerged conditions. The predominant enzyme activities of cellulase, xylanase and laccase were produced in the cocktail through submerged conditions using wheat bran as a substrate. A central composite design approach was utilized to model the production process using temperature, pH, incubation time and agitation as input variables with the goal of optimizing the output variables namely cellulase, xylanase and laccase activities. The effect of individual, square and interaction terms on cellulase, xylanase and laccase activities were depicted through the non-linear regression equations with significant R(2) and P-values. An optimized value of 20 U/ml, 17 U/ml and 13 U/ml of cellulase, xylanase and laccase activities, respectively, were obtained with a media pH of 5.0 in 77 h at 31C, 140 rpm using wheatbran as a substrate. Overall, the present study introduces a fungal strain, capable of producing lignocellulolytic enzyme cocktail for subsequent applications in biofuel industry.

  7. Bioprocessing of wheat bran for the production of lignocellulolytic enzyme cocktail by Cotylidia pannosa under submerged conditions

    PubMed Central

    Sharma, Deepika; Garlapat, Vijay Kumar; Goel, Gunjan

    2016-01-01

    ABSTRACT Characterization and production of efficient lignocellulytic enzyme cocktails for biomass conversion is the need for biofuel industry. The present investigation reports the modeling and optimization studies of lignocellulolytic enzyme cocktail production by Cotylidia pannosa under submerged conditions. The predominant enzyme activities of cellulase, xylanase and laccase were produced in the cocktail through submerged conditions using wheat bran as a substrate. A central composite design approach was utilized to model the production process using temperature, pH, incubation time and agitation as input variables with the goal of optimizing the output variables namely cellulase, xylanase and laccase activities. The effect of individual, square and interaction terms on cellulase, xylanase and laccase activities were depicted through the non-linear regression equations with significant R2 and P-values. An optimized value of 20 U/ml, 17 U/ml and 13 U/ml of cellulase, xylanase and laccase activities, respectively, were obtained with a media pH of 5.0 in 77 h at 31C, 140 rpm using wheatbran as a substrate. Overall, the present study introduces a fungal strain, capable of producing lignocellulolytic enzyme cocktail for subsequent applications in biofuel industry. PMID:26941214

  8. Seawater Acidification and Elevated Temperature Affect Gene Expression Patterns of the Pearl Oyster Pinctada fucata

    PubMed Central

    Liu, Wenguang; Huang, Xiande; Lin, Jianshi; He, Maoxian

    2012-01-01

    Oceanic uptake of anthropogenic carbon dioxide results in decrease in seawater pH and increase in temperature. In this study, we demonstrated the synergistic effects of elevated seawater temperature and declined seawater pH on gene expression patterns of aspein, calmodulin, nacrein, she-7-F10 and hsp70 in the pearl oyster Pinctada fucata. Under ‘business-as-usual’ scenarios, four treatments were examined: (1) ambient pH (8.10) and ambient temperature (27°C) (control condition), (2) ambient pH and elevated temperature (+3°C), (3) declined pH (7.70) and ambient temperature, (4) declined pH and elevated temperature. The results showed that under warming and acidic seawater conditions, expression of aspein and calmodulin showed no significant differences among different time point in condition 8.10 T. But the levels of aspein and calmodulin in conditions 8.10 T+3, 7.70 T and 7.70 T+3, and levels of nacrein, she-7-F10 in all the four treatments changed significantly. Low pH and pH×temperature interaction influenced the expression of aspein and calmodulin significantly after hours 48 and 96. Significant effects of low pH and pH×temperature interaction on the expression of nacrein were observed at hour 96. The expression level of she-7-F10 was affected significantly by pH after hours 48 and 96. The expression of hsp70 was significantly affected by temperature, pH, temperature×pH interaction at hour 6, and by temperature×pH interaction at hour 24. This study suggested that declined pH and pH×temperature interaction induced down regulation of calcification related genes, and the interaction between declined seawater pH and elevated temperature caused up regulation of hsp70 in P. facata. These results demonstrate that the declined seawater pH and elevated temperature will impact the physiological process, and potentially the adaptability of P. fucata to future warming and acidified ocean. PMID:22438983

  9. Variations of thiaminase I activity pH dependencies among typical Great Lakes forage fish and Paenibacillus thiaminolyticus.

    USGS Publications Warehouse

    Zajicek, J.L.; Brown, L.; Brown, S.B.; Honeyfield, D.C.; Fitzsimons, J.D.; Tillitt, D.E.

    2009-01-01

    The source of thiaminase in the Great Lakes food web remains unknown. Biochemical characterization of the thiaminase I activities observed in forage fish was undertaken to provide insights into potential thiaminase sources and to optimize catalytic assay conditions. We measured the thiaminase I activities of crude extracts from five forage fish species and one strain of Paenibacillus thiaminolyticus over a range of pH values. The clupeids, alewife Alosa pseudoharengus and gizzard shad Dorosoma cepedianum, had very similar thiaminase I pH dependencies, with optimal activity ranges (> or = 90% of maximum activity) between pH 4.6 and 5.5. Rainbow smelt Osmerus mordax and spottail shiner Notropis hudsonius had optimal activity ranges between pH 5.5-6.6. The thiaminase I activity pH dependence profile of P. thiaminolyticus had an optimal activity range between pH 5.4 and 6.3, which was similar to the optimal range for rainbow smelt and spottail shiners. Incubation of P. thiaminolyticus extracts with extracts from bloater Coregonus hoyi (normally, bloaters have little or no detectable thiaminase I activity) did not significantly alter the pH dependence profile of P. thiaminolyticus-derived thiaminase I, such that it continued to resemble that of the rainbow smelt and spottail shiner, with an apparent optimal activity range between pH 5.7 and 6.6. These data are consistent with the hypothesis of a bacterial source for thiaminase I in the nonclupeid species of forage fish; however, the data also suggest different sources of thiaminase I enzymes in the clupeid species.

  10. Predictive microbiology in a dynamic environment: a system theory approach.

    PubMed

    Van Impe, J F; Nicolaï, B M; Schellekens, M; Martens, T; De Baerdemaeker, J

    1995-05-01

    The main factors influencing the microbial stability of chilled prepared food products for which there is an increased consumer interest-are temperature, pH, and water activity. Unlike the pH and the water activity, the temperature may vary extensively throughout the complete production and distribution chain. The shelf life of this kind of foods is usually limited due to spoilage by common microorganisms, and the increased risk for food pathogens. In predicting the shelf life, mathematical models are a powerful tool to increase the insight in the different subprocesses and their interactions. However, the predictive value of the sigmoidal functions reported in the literature to describe a bacterial growth curve as an explicit function of time is only guaranteed at a constant temperature within the temperature range of microbial growth. As a result, they are less appropriate in optimization studies of a whole production and distribution chain. In this paper a more general modeling approach, inspired by system theory concepts, is presented if for instance time varying temperature profiles are to be taken into account. As a case study, we discuss a recently proposed dynamic model to predict microbial growth and inactivation under time varying temperature conditions from a system theory point of view. Further, the validity of this methodology is illustrated with experimental data of Brochothrix thermosphacta and Lactobacillus plantarum. Finally, we propose some possible refinements of this model inspired by experimental results.

  11. Purification and Characterization of a Highly Efficient Calcium-Independent α-Amylase from Talaromyces pinophilus 1-95

    PubMed Central

    Xian, Liang; Wang, Fei; Luo, Xiang; Feng, Yu-Liang; Feng, Jia-Xun

    2015-01-01

    Alpha-amylase is a very important enzyme in the starch conversion process. Most of the α-amylases are calcium-dependent and exhibit poor performance in the simultaneous saccharification and fermentation process of industrial bioethanol production that uses starch as feedstock. In this study, an extracellular amylolytic enzyme was purified from the culture broth of newly isolated Talaromyces pinophilus strain 1-95. The purified amylolytic enzyme, with an apparent molecular weight of 58 kDa on SDS-PAGE, hydrolyzed maltopentaose, maltohexaose, and maltoheptaose into mainly maltose and maltotriose and minor amount of glucose, confirming the endo-acting mode of the enzyme, and hence, was named Talaromyces pinophilus α-amylase (TpAA). TpAA was most active at pH 4.0–5.0 (with the temperature held at 37°C) and 55°C (at pH 5.0), and stable within the pH range of 5.0–9.5 (at 4°C) and below 45°C (at pH 5.0). Interestingly, the Ca2+ did not improve its enzymatic activity, optimal temperature, or thermostability of the enzyme, indicating that the TpAA was Ca2+-independent. TpAA displayed higher enzyme activity toward malto-oligosaccharides and dextrin than other previously reported α-amylases. This highly active Ca2+-independent α-amylase may have potential applications in starch-to-ethanol conversion process. PMID:25811759

  12. SynTec Final Technical Report: Synthetic biology for Tailored Enzyme cocktails

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lin, Janine; Teter, Sarah

    Using a novel enzyme screening method inspired by synthetic biology, Novozymes developed new technology under SynTec which allows for more rapidly tailoring of enzyme cocktails. The methodology can be applied to specific feedstocks, and or coupled to address a specific hydrolytic conversion process context. Using combinatorial high throughput screening of libraries of enzyme domains, we can quickly assess which combination of catalytic modules delivers the best performance for a specific condition. To demonstrate the effectiveness of the screening process, we measured performance of the output catalytic cocktail compared to CTec3/HTec3. SynTec benchmark cocktail - blend of Cellic® CTec3 and HTec3.more » The test substrate was - ammonia fiber expansion pretreated corn stover (AFEX™ PCS).CTec3/HTec3 was assayed at the optimal pH and temperature, and also in the absence of any pH adjustment. The new enzyme cocktail discovered under SynTec was assayed in the absence of any pH adjustment and at the optimal temperature. Conversion is delivered by SynTec enzyme at significant dose reduction relative to CTec3/HTec3 at the controlled pH optimum, and without titrant required to maintain pH, which delivers additional cost savings relative to current state of the art process. In this 2.5 year $4M project, the team delivered an experimental cocktail that significantly outperformed CTec3/HTec3 for a specific substrate, and for specific hydrolysis conditions. As a means of comparing performance improvement delivered per research dollar spent, we note that SynTec delivered a similar performance improvement to the previous award, in a shorter time and with fewer resources than for the previously successful DOE project DECREASE, a 3.5 year, $25M project, though this project focused on a different substrate and used different hydrolysis conditions. The newly implemented technology for rapid sourcing of new cellulases and hemicellulases from nature is an example of Novozymes' continued innovation that results in more effective products for the advanced biofuel market.« less

  13. Optimization of Maillard Reaction between Glucosamine and Other Precursors by Measuring Browning with a Spectrophotometer.

    PubMed

    Ogutu, Benrick; Kim, Ye-Joo; Kim, Dae-Wook; Oh, Sang-Chul; Hong, Dong-Lee; Lee, Yang-Bong

    2017-09-01

    The individual Maillard reactions of glucose, glucosamine, cyclohexylamine, and benzylamine were studied at a fixed temperature of 120°C under different durations by monitoring the absorbance of the final products at 425 nm. Glucosamine was the most individually reactive compound, whereas the reactions of glucose, cyclohexylamine, and benzylamine were not significantly different from each other. Maillard reactions of reaction mixtures consisting of glucosamine-cyclohexylamine, glucosamine-benzylamine, glucose-cyclohexylamine, and glucose-benzylamine were also studied using different concentration ratios under different durations at a fixed temperature of 120°C and pH 9. Maillard reactions in the pairs involving glucosamine were observed to be more intense than those of the pairs involving glucose. Finally, with respect to the concentration ratios, it was observed that in most instances, optimal activity was realized, when the reaction mixtures were in the ratio of 1:1.

  14. Optimization of Maillard Reaction between Glucosamine and Other Precursors by Measuring Browning with a Spectrophotometer

    PubMed Central

    Ogutu, Benrick; Kim, Ye-Joo; Kim, Dae-Wook; Oh, Sang-Chul; Hong, Dong-Lee; Lee, Yang-Bong

    2017-01-01

    The individual Maillard reactions of glucose, glucosamine, cyclohexylamine, and benzylamine were studied at a fixed temperature of 120°C under different durations by monitoring the absorbance of the final products at 425 nm. Glucosamine was the most individually reactive compound, whereas the reactions of glucose, cyclohexylamine, and benzylamine were not significantly different from each other. Maillard reactions of reaction mixtures consisting of glucosamine-cyclohexylamine, glucosamine-benzylamine, glucose-cyclohexylamine, and glucose-benzylamine were also studied using different concentration ratios under different durations at a fixed temperature of 120°C and pH 9. Maillard reactions in the pairs involving glucosamine were observed to be more intense than those of the pairs involving glucose. Finally, with respect to the concentration ratios, it was observed that in most instances, optimal activity was realized, when the reaction mixtures were in the ratio of 1:1. PMID:29043219

  15. Optimization of pH sensing using silicon nanowire field effect transistors with HfO2 as the sensing surface.

    PubMed

    Zafar, Sufi; D'Emic, Christopher; Afzali, Ali; Fletcher, Benjamin; Zhu, Y; Ning, Tak

    2011-10-07

    Silicon nanowire field effect transistor sensors with SiO(2)/HfO(2) as the gate dielectric sensing surface are fabricated using a top down approach. These sensors are optimized for pH sensing with two key characteristics. First, the pH sensitivity is shown to be independent of buffer concentration. Second, the observed pH sensitivity is enhanced and is equal to the Nernst maximum sensitivity limit of 59 mV/pH with a corresponding subthreshold drain current change of ∼ 650%/pH. These two enhanced pH sensing characteristics are attributed to the use of HfO(2) as the sensing surface and an optimized fabrication process compatible with silicon processing technology.

  16. Ultrasound-assisted three-phase partitioning of polyphenol oxidase from potato peel (Solanum tuberosum).

    PubMed

    Niphadkar, Sonali S; Rathod, Virendra K

    2015-01-01

    Conventional three phase partitioning (TPP) and ultrasound assisted three phase partitioning (UATPP) were optimized for achieving the maximum extraction and purification of polyphenol oxidase (PPO) from waste potato peels. Different process parameters such as ammonium sulfate (NH4)2SO4 concentration, crude extract to t-butanol ratio, time, temperature and pH were studied for conventional TPP. Except agitation speed, the similar parameters were also optimized for UATPP. Further additional parameters were also studied for UATPP viz. irradiation time at different frequencies, duty cycle and, rated power in order to obtain the maximum purification factor and recovery of PPO. The optimized conditions for conventional TPP were (NH4)2SO4 0-40% (w/v), extract to t-butanol ratio 1:1 (v/v), time 40 min and pH 7 at 30°C. These conditions provided 6.3 purification factor and 70% recovery of PPO from bottom phase. On the other hand, UATPP gives maximum purification fold of 19.7 with 98.3% recovery under optimized parameters which includes (NH4)2SO4 0-40% (w/v), crude extract to t-butanol ratio 1: 1 (v/v) pH 7, irradiation time 5 min with 25 kHz, duty cycle 40% and rated power 150W at 30°C. UATPP delivers higher purification factor and % recovery of PPO along with reduced operation time from 40 min to 5 min when compared with TPP. SDS PAGE showed partial purification of PPO enzyme with UATPP with molecular weight in the range of 26-36 kDa. Results reveal that UATPP would be an attractive option for the isolation and purification of PPO without need of multiple steps. © 2015 American Institute of Chemical Engineers.

  17. Hydrophilic interaction liquid chromatography in analysis of granisetron HCl and its related substances. Retention mechanisms and method development.

    PubMed

    Maksić, Jelena; Tumpa, Anja; Stajić, Ana; Jovanović, Marko; Rakić, Tijana; Jančić-Stojanović, Biljana

    2016-05-10

    In this paper separation of granisetron and its two related substances in HILIC mode is presented. Separation was done on silica column derivatized with sulfoalkylbetaine groups (ZIC-HILIC). Firstly, retention mechanisms were assessed whereby retention factors of substances were followed in wide range of acetonitrile content (80-97%), at constant concentration of aqueous buffer (10mM) as well as at constant pH value of 3.0. Further, in order to developed optimal HILIC method, Design of Experiments (DoE) methodology was applied. For optimization full factorial design 3(2) was employed. Influence of acetonitrile content and ammonium acetate concentration were investigated while pH of the water phase was kept at 3.3. Adequacy of obtained mathematical models was confirmed by ANOVA. Optimization goals (α>1.15 and minimal run time) were accomplished with 94.7% of acetonitrile in mobile phase and 70 mM of ammonium acetate in water phase. Optimal point was in the middle of defined Design Space. In the next phase, robustness was experimetally tested by Rechtschaffen design. The investigated factors and their levels were: acetonitrile content (±1%), ammonium acetate molarity in water phase (±2 mM), pH value of water phase (±0.2) and column temperature (±4 °C). The validation scope included selectivity, linearity, accuracy and precision as well as determination of limit of detection (LOD) and limit of quantification (LOQ) for the related substances. Additionally, the validation acceptance criteria were met in all cases. Finally, the proposed method could be successfully utilized for estimation of granisetron HCl and its related substances in tablets and parenteral dosage forms, as well as for monitoring degradation under various stress conditions. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Effects of medium components and culture conditions on mycelial biomass and the production of bioactive ingredients in submerged culture of Xylaria nigripes (Ascomycetes), a Chinese medicinal fungus.

    PubMed

    Chen, Jian-Zhi; Lo, Hui-Chen; Lin, Fang-Yi; Chang, Shih-Liang; Hsieh, Changwei; Liang, Zeng-Chin; Ho, Wai-Jane; Hsu, Tai-Hao

    2014-01-01

    The optimal culture conditions were investigated to maximize the production of mycelial biomass and bioactive ingredients in submerged cultivation of Xylaria nigripes, a Chinese medicinal fungus. The one-factor-at-a-time method was used to explore the effects of medium components, including carbon, nitrogen, mineral sources, and initial pH of the medium and environmental factors, such as culture temperature and rotation speed, on mycelial growth and production of bioactive ingredients. The results indicated that the optimal culture temperature and rotation speed were 25°C and 100 rpm in a medium with 20 g fructose, 6 g yeast extract, and 2 g magnesiun sulfate heptahydrate as carbon, nitrogen, and mineral sources, respectively, in 1 L distilled water with an initial medium pH of 5.5. With optimal medium components and conditions of cultivation, the maximal production of mycelial biomass was 6.64 ± 0.88 g/L, with maximal production of bioactive ingredients such as extracellular polysaccharides (2.36 ± 0.18 mg/mL), intracellular polysaccharides (2.38 ± 0.07 mg/g), adenosine (43.27 ± 2.37 mg/g), total polyphenols (36.57 ± 1.36 mg/g), and triterpenoids (31.29 ± 1.17 mg/g) in a shake flask culture. These results suggest that different bioactive ingredients including intracellular polysaccharides, adenosine, total polyphenols and triterpenoids in mycelia and extracellular polysaccharides in broth can be obtained from one simple medium for submerged cultivation of X. nigripes.

  19. Optimization of Culture Parameters for Maximum Polyhydroxybutyrate Production by Selected Bacterial Strains Isolated from Rhizospheric Soils.

    PubMed

    Lathwal, Priyanka; Nehra, Kiran; Singh, Manpreet; Jamdagni, Pragati; Rana, Jogender S

    2015-01-01

    The enormous applications of conventional non-biodegradable plastics have led towards their increased usage and accumulation in the environment. This has become one of the major causes of global environmental concern in the present century. Polyhydroxybutyrate (PHB), a biodegradable plastic is known to have properties similar to conventional plastics, thus exhibiting a potential for replacing conventional non-degradable plastics. In the present study, a total of 303 different bacterial isolates were obtained from soil samples collected from the rhizospheric area of three crops, viz., wheat, mustard and sugarcane. All the isolates were screened for PHB (Poly-3-hydroxy butyric acid) production using Sudan Black staining method, and 194 isolates were found to be PHB positive. Based upon the amount of PHB produced, the isolates were divided into three categories: high, medium and low producers. Representative isolates from each category were selected for biochemical characterization; and for optimization of various culture parameters (carbon source, nitrogen source, C/N ratio, different pH, temperature and incubation time periods) for maximizing PHB accumulation. The highest PHB yield was obtained when the culture medium was supplemented with glucose as the carbon source, ammonium sulphate at a concentration of 1.0 g/l as the nitrogen source, and by maintaining the C/N ratio of the medium as 20:1. The physical growth parameters which supported maximum PHB accumulation included a pH of 7.0, and an incubation temperature of 30 degrees C for a period of 48 h. A few isolates exhibited high PHB accumulation under optimized conditions, thus showing a potential for their industrial exploitation.

  20. Processed dairy beverages pH evaluation: consequences of temperature variation.

    PubMed

    Ferreira, Fabiana Vargas; Pozzobon, Roselaine Terezinha

    2009-01-01

    This study assessed the pH from processed dairy beverages as well as eventual consequences deriving from different ingestion temperatures. 50 adults who accompanied children attended to at the Dentistry School were randomly selected and they answered a questionnaire on beverages. The beverages were divided into 4 groups: yogurt (GI) fermented milk (GII), chocolate-based products (GIII) and fermented dairy beverages (GIV). They were asked which type, flavor and temperature. The most popular beverages were selected, and these made up the sample. A pH meter Quimis 400A device was used to verify pH. The average pH from each beverage was calculated and submitted to statistical analysis (Variance and Tukey test with a 5% significance level). for groups I, II and III beverages, type x temperature interaction was significant, showing the pH averages were influenced by temperature variation. At iced temperatures, they presented lower pH values, which were considered statistically significant when compared to the values found for the same beverages at room temperature. All dairy beverages, with the exception of the chocolate-based type presented pH below critical level for enamel and present corrosive potential; as to ingestion temperature, iced temperature influenced pH reducing its values, in vitro.

  1. Bioactive characteristics and optimization of tamarind seed protein hydrolysate for antioxidant-rich food formulations.

    PubMed

    Bagul, Mayuri B; Sonawane, Sachin K; Arya, Shalini S

    2018-04-01

    Tamarind seed has been a source of valuable nutrients such as protein (contains high amount of many essential amino acids), essential fatty acids, and minerals which are recognized as additive to develop perfect balanced functional foods. The objective of present work was to optimize the process parameters for extraction and hydrolysis of protein from tamarind seeds. Papain-derived hydrolysates showed a maximum degree of hydrolysis (39.49%) and radical scavenging activity (42.92 ± 2.83%) at optimized conditions such as enzyme-to-substrate ratio (1:5), hydrolysis time (3 h), hydrolysis temperature (65 °C), and pH 6. From this study, papain hydrolysate can be considered as good source of natural antioxidants in developing food formulations.

  2. Combined ultrasound and Fenton (US-Fenton) process for the treatment of ammunition wastewater.

    PubMed

    Li, Yangang; Hsieh, Wen-Pin; Mahmudov, Rovshan; Wei, Xiaomei; Huang, C P

    2013-01-15

    A wastewater collected from a regional ammunition process site was treated with combined US-Fenton process. Factors such as pH, temperature, reaction time, US energy intensity, initial TOC concentration, and the molar ratio of iron to hydrogen peroxide that might affect the treatment efficiency were investigated. The removal of TOC, COD, and color increased with decreasing pH and increasing temperature and US intensity. Color was removed rapidly reaching 85% in 10 min; whereas TOC and COD were removed slowly, only about 20% for both in 10 min and approaching 65 and 92% removal in 120 min, respectively. The optimal molar ratio of Fe(II) to H(2)O(2) for TOC and COD removal was 500. The results showed that the change in the average carbon oxidation number (ACON) was parallel to that of the removal efficiency of TOC, COD, and color. The toxicity of treated wastewater was reduced as assessed by the respiration rate of Escherichia coli. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Immobilization of an L-aminoacylase-producing strain of Aspergillus oryzae into gelatin pellets and its application in the resolution of D,L-methionine.

    PubMed

    Yuan Yj, Ying-jin; Wang Sh, Shu-hao; Song Zx, Zheng-xiao; Gao Rc, Rui-chang

    2002-04-01

    The conditions for immobilization of an l-aminoacylase-producing strain of Aspergillus oryzae in gelatin and the enzymic characteristics of the immobilized pellets were studied. The optimal concentrations of gelatin, glutaraldehyde and ethyldiamine and time of immobilization were determined. Scanning electron micrographs reveal the cross-linked structure differences between the native and immobilized pellets. Optimum pH and temperature of the native and immobilized pellets were determined. Effects of ionic strength and substrate concentration on relative activity of the native and immobilized pellets were investigated in detail. The immobilized pellets were more stable over broader temperature and pH ranges. In addition, the immobilized pellets showed stable activity under operational and storage conditions. The immobilized pellets lost about 20% of their initial activity after five cycles of reuse. The results reported in this paper show the potential for using the immobilized A. oryzae pellets to resolve d,l-methionine.

  4. Remediation of lead from lead electroplating industrial effluent using sago waste.

    PubMed

    Jeyanthi, G P; Shanthi, G

    2007-01-01

    Heavy metals are known toxicants, which inflict acute disorders to the living beings. Electroplating industries pose great threat to the environment through heavy load of metals in the wastewater discharged on land and water sources. In the present study, sago processing waste, which is both a waste and a pollutant, was used to adsorb lead ions from lead electroplating industrial effluent. Two types of sago wastes, namely, coarse sago waste and fine sago waste were used to study their adsorption capacity with the batch adsorption and Freundlich adsorption isotherm. The parameters that were considered for batch adsorption were pH (4, 5 and 6), time of contact (1, 2 and 3 hrs), temperature (30, 37 and 45 degrees C) and dosage of the adsorbent (2,4 and 6 g/L). The optimal condition for the effective removal of lead was found to be pH 5, time of contact 3 hrs, temperature 30 degrees C and dosage 4 g/L with coarse sago waste than fine sago waste.

  5. Green synthesis and characterisation of platinum nanoparticles using quail egg yolk

    NASA Astrophysics Data System (ADS)

    Nadaroglu, Hayrunnisa; Gungor, Azize Alayli; Ince, Selvi; Babagil, Aynur

    2017-02-01

    Nanotechnology is extensively used in all parts today. Therefore, nano synthesis is also significant in all explored areas. The results of studies conducted have revealed that nanoparticle synthesis is performed by using both chemical and physical methods. It is well known that these syntheses are carried out at high charge, pressure and temperature in harsh environments. Therefore, this study investigated green synthesis method that sustains more mild conditions. In this study, quail egg yolk having high vitamin and protein content was prepared for green synthesis reaction and used for the synthesis of platinum nanoparticles in the reaction medium. Reaction situations were optimised as a function of pH, temperature, time and concentration by using quail egg yolk. The results showed that the highest platinum nanoparticles were synthesised at 20 °C and pH 6.0 for 4 h. Also, optimal concentration of metal ions was established as 0.5 mM. The synthesised platinum nanoparticles were characterised by using UV spectrum, X-ray diffraction and scanning electron microscope.

  6. Biological indicators for steam sterilization: characterization of a rapid biological indicator utilizing Bacillus stearothermophilus spore-associated alpha-glucosidase enzyme.

    PubMed

    Albert, H; Davies, D J; Woodson, L P; Soper, C J

    1998-11-01

    The alpha-glucosidase enzyme was isolated from vegetative cells and spores of Bacillus stearothermophilus, ATCC 7953. Spore-associated enzyme had a molecular weight of approximately 92,700, a temperature optimum of 60 degrees C, and a pH optimum of 7.0-7.5. The enzyme in crude aqueous spore extract was stable for 30 min up to a temperature of 65 degrees C, above which the enzyme was rapidly denatured. The optimal pH for stability of the enzyme was approximately 7.2. The alpha-glucosidase in crude vegetative cell extract had similar characteristics to the spore-associated enzyme but its molecular weight was 86,700. The vegetative cell and spore-associated enzymes were cross-reactive. The enzymes are postulated to derive from a single gene product, which undergoes modification to produce the spore-associated form. The location of alpha-glucosidase in the spore coats (outside the spore protoplast) is consistent with the location of most enzymes involved in activation, germination and outgrowth.

  7. Statistical analysis and isotherm study of uranium biosorption by Padina sp. algae biomass.

    PubMed

    Khani, Mohammad Hassan

    2011-06-01

    The application of response surface methodology is presented for optimizing the removal of U ions from aqueous solutions using Padina sp., a brown marine algal biomass. Box-Wilson central composite design was employed to assess individual and interactive effects of the four main parameters (pH and initial uranium concentration in solutions, contact time and temperature) on uranium uptake. Response surface analysis showed that the data were adequately fitted to second-order polynomial model. Analysis of variance showed a high coefficient of determination value (R (2)=0.9746) and satisfactory second-order regression model was derived. The optimum pH and initial uranium concentration in solutions, contact time and temperature were found to be 4.07, 778.48 mg/l, 74.31 min, and 37.47°C, respectively. Maximized uranium uptake was predicted and experimentally validated. The equilibrium data for biosorption of U onto the Padina sp. were well represented by the Langmuir isotherm, giving maximum monolayer adsorption capacity as high as 376.73 mg/g.

  8. Immobilization of pectinase on oxidized pulp fiber and its application in whitewater treatment.

    PubMed

    Wu, Rina; He, Bei-Hai; Zhao, Guang-Lei; Qian, Li-Ying; Li, Xiao-Feng

    2013-09-12

    Modified pulp fiber was originally used as a new type of carrier for pectinase immobilization. Pulp fiber was oxidized by sodium periodate to produce aldehyde groups for covalently binding with amino groups of pectinase. Results showed that the enzymatic activity of immobilized pectinase on pulp fiber reached 65 μgg(-1)min(-1) when immobilization pH value, temperature and time were of 7.0, 20 °C and 15 min, respectively. The immobilized pectinase showed higher thermo stability in a wider temperature range of 40-70 °C than its free type and its optimal pH shifted from 8.0 to 8.8. Furthermore, the immobilized pectinase exhibited good operational stability. When employed in whitewater treatment of papermaking industry, it still efficiently decreased the cationic demand after operating repeatedly for six batches. The results obtained demonstrate a promising route to prepare available, cheap and biodegradable carrier for immobilizing enzymes with potential application in wastewater treatment in papermaking industry. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Production, characterization, and immobilization of partially purified surfactant-detergent and alkali-thermostable protease from newly isolated Aeromonas caviae.

    PubMed

    Datta, Sumitra; Menon, Gopalakrishnan; Varughese, Bincy

    2017-04-21

    Proteolytic Aeromonas caviae P-1-1 growing at wide-ranging pH (7.0-11.0) and moderate salinity (0-5% NaCl) was isolated from cattle shed of Thanjavur, India. It produced lipase, gelatinase, and polyhydroxybutyrate. Different culture conditions, incubation time, carbon and nitrogen sources, vitamins, amino acids, surfactants, and metal ions for optimal growth and protease production of P-1-1 were examined. Maximum protease (0.128 U/mL) production was achieved with 1% fructose, 1% yeast extract, 0.1% ammonium sulfate, 3% NaCl, 0.1% CaCl 2  · 2H 2 O, 1% glycine, 0.1% vitamin E, and 0.1% Tween-40 at pH 8.0 after 42 hr of incubation at 37°C. It was active over broad range of pH (7.0-12.0), temperature (15-100°C), and salinity (0-9% NaCl) with optima at pH 10.0, 55°C, and 3% NaCl. It retained 65 and 48% activities at pH 12.0 and 100°C, respectively. Partially purified protease was highly stable (100%) within pH range 7.0-12.0 and salinities of 0-5% NaCl for 48 hr. Cu 2+ , Mn 2+ , Co 2+ , and Ca 2+ did not inhibit its activity. Its stability at extreme pHs, temperatures, and in the presence of surfactants and commercial detergents suggests its possible application in laundry detergents. Partially purified protease was immobilized and reused. This is the first report of alkali-thermotolerant, surfactant-detergent-stable partially purified extracellular protease from A. caviae.

  10. Thermal inactivation kinetics of Bacillus coagulans spores in tomato juice.

    PubMed

    Peng, Jing; Mah, Jae-Hyung; Somavat, Romel; Mohamed, Hussein; Sastry, Sudhir; Tang, Juming

    2012-07-01

    The thermal characteristics of the spores and vegetative cells of three strains of Bacillus coagulans (ATCC 8038, ATCC 7050, and 185A) in tomato juice were evaluated. B. coagulans ATCC 8038 was chosen as the target microorganism for thermal processing of tomato products due to its spores having the highest thermal resistance among the three strains. The thermal inactivation kinetics of B. coagulans ATCC 8038 spores in tomato juice between 95 and 115°C were determined independently in two different laboratories using two different heating setups. The results obtained from both laboratories were in general agreement, with z-values (z-value is defined as the change in temperature required for a 10-fold reduction of the D-value, which is defined as the time required at a certain temperature for a 1-log reduction of the target microorganisms) of 8.3 and 8.7°C, respectively. The z-value of B. coagulans 185A spores in tomato juice (pH 4.3) was found to be 10.2°C. The influence of environmental factors, including cold storage time, pH, and preconditioning, upon the thermal resistance of these bacterial spores is discussed. The results obtained showed that a storage temperature of 4°C was appropriate for maintaining the viability and thermal resistance of B. coagulans ATCC 8038 spores. Acidifying the pH of tomato juice decreased the thermal resistance of these spores. A 1-h exposure at room temperature was considered optimal for preconditioning B. coagulans ATCC 8038 spores in tomato juice.

  11. Preparation of dual-stimuli-responsive liposomes using methacrylate-based copolymers with pH and temperature sensitivities for precisely controlled release.

    PubMed

    Sugimoto, Takumi; Yamazaki, Naoko; Hayashi, Takaaki; Yuba, Eiji; Harada, Atsushi; Kotaka, Aki; Shinde, Chiharu; Kumei, Takayuki; Sumida, Yasushi; Fukushima, Mitsuhiro; Munekata, Yuki; Maruyama, Keiichi; Kono, Kenji

    2017-07-01

    Dual-signal-sensitive copolymers were synthesized by copolymerization of methoxy diethylene glycol methacrylate, methacrylic acid, and lauroxy tetraethylene glycol methacrylate, which respectively provide temperature sensitivity, pH sensitivity, and anchoring to liposome surfaces. These novel copolymers, with water solubility that differs depending on temperature and pH, are soluble in water under neutral pH and low-temperature conditions, but they become water-insoluble and form aggregates under acidic pH and high-temperature conditions. Liposomes modified with these copolymers exhibited enhanced content release at weakly acidic pH with increasing temperature, although no temperature-dependent content release was observed in neutral conditions. Interaction between the copolymers and the lipid monolayer at the air-water interface revealed that the copolymer chains penetrate more deeply into the monolayer with increasing temperature at acidic pH than at neutral pH, where the penetration of copolymer chains was moderate and temperature-independent at neutral pH. Interaction of the copolymer-modified liposomes with HeLa cells demonstrated that the copolymer-modified liposomes were adsorbed quickly and efficiently onto the cell surface and that they were internalized more gradually than the unmodified liposomes through endocytosis. Furthermore, the copolymer-modified liposomes enhanced the content release in endosomes with increasing temperature, but no such temperature-dependent enhancement of content release was observed for unmodified liposomes. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Modeling and optimization of fermentation variables for enhanced production of lactase by isolated Bacillus subtilis strain VUVD001 using artificial neural networking and response surface methodology.

    PubMed

    Venkateswarulu, T C; Prabhakar, K Vidya; Kumar, R Bharath; Krupanidhi, S

    2017-07-01

    Modeling and optimization were performed to enhance production of lactase through submerged fermentation by Bacillus subtilis VUVD001 using artificial neural networks (ANN) and response surface methodology (RSM). The effect of process parameters namely temperature (°C), pH, and incubation time (h) and their combinational interactions on production was studied in shake flask culture by Box-Behnken design. The model was validated by conducting an experiment at optimized process variables which gave the maximum lactase activity of 91.32 U/ml. Compared to traditional activity, 3.48-folds improved production was obtained after RSM optimization. This study clearly shows that both RSM and ANN models provided desired predictions. However, compared with RSM (R 2  = 0.9496), the ANN model (R 2  = 0.99456) gave a better prediction for the production of lactase.

  13. Nematicidal activity of three novel extracellular proteases of the nematophagous fungus Monacrosporium sinense.

    PubMed

    Soares, Filippe E F; Braga, Fabio R; Araújo, Jackson V; Geniêr, Hugo L A; Gouveia, Angélica S; Queiroz, José H

    2013-04-01

    Extracellular proteases are an important virulence factor for the nematophagous fungi Monacrosporium. The objective of this study was to optimize, purify, partially characterize, and to evaluate the nematicidal activity of the proteases produced by the nematophagous fungus Monacrosporium sinense (SF53) by solid-state fermentation. Wheat bran was used as substrate for protease production. The variables moisture, pH, incubation time, temperature, glucose, yeast extract, and the number of conidia were tested for their influences on protease production by SF53. To determine the optimal level of the selected variables the central composite design was applied. The crude extract obtained was purified in two steps, an ion exchange chromatography and a gel excision. SDS-PAGE and zymogram were performed for analysis of the purification process. Proteolytic activity was also tested at different pHs and temperatures. In the in vitro assay, the nematicidal activity of the three proteases was evaluated. pH and incubation time showed a significant effect (p<0.05) on production of protease. The highest value of activity was 38.0 (U/ml) under the conditions of pH 5.0 and incubation time of 211 h. SF53 produced three different proteases (Ms1, Ms2, and Ms3) which were directly purified from the zymogram. Ms1, Ms2, and Ms3 showed the following percentage of reduction (p<0.05) on the number of Panagrellus redivivus compared to control after 24 h: 76.8, 68.1, and 92.1%. This is the first report of the use of proteases of the isolate SF53 on a phytonematode, which may be a research tool in future works.

  14. Optimization of the production and characterization of lipase from Candida rugosa and Geotrichum candidum in soybean molasses by submerged fermentation.

    PubMed

    de Morais, Wilson Galvão; Kamimura, Eliana Setsuko; Ribeiro, Eloízio Júlio; Pessela, Benevides Costa; Cardoso, Vicelma Luiz; de Resende, Miriam Maria

    2016-07-01

    This present work describes the production and biochemical characterization of lipase by Candida rugosa and Geotrichum candidum in a culture supplemented with soybean molasses. After optimizing the fermentation times for both microorganisms, the effects of changing the soybean molasses concentration, the fermentative medium pH and the fermentation temperature were evaluated using the Central Composite Planning. When soybean molasses was used at a concentration of 200 g/L at 27 ± 1 °C and pH 3.5, the lipolytic activity measured in the broth was 12.3 U/mL after 12 h for C. rugosa and 11.48 U/mL after 24 h for G. candidum. The molecular masses were 38.3 kDa to G. candidum lipase and 59.7 kDa to C. rugosa lipase, determined by SDS-PAGE. The lipase from both microorganisms exhibited maximal hydrolytic activity at a temperature of 40 °C and were inhibited at pH 10.0. Using different concentration of p-nitrophenylbutyrate (p-NPB), the kinetic parameters were calculated, as follows: the Km of lipase from G. candidum was 465.44 μM and the Vmax 0.384 μmol/min; the Km and Vmax of lipase from C. rugosa were 129.21 μM and 0.034 μmol/min, respectively. Lipases activity were increased by metallic ions Mg(2+) and Na(+) and inhibited by metallic ion Cu(3+). Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Application of response surface methodology for optimization of biosorption of fluoride from groundwater using Shorea robusta flower petal

    NASA Astrophysics Data System (ADS)

    Biswas, G.; Kumari, M.; Adhikari, K.; Dutta, S.

    2017-12-01

    Fluoride pollution in groundwater is a major concern in rural areas. The flower petal of Shorea robusta, commonly known as sal tree, is used in the present study both in its native form and Ca-impregnated activated form to eradicate excess fluoride from simulated wastewater. Response surface methodology (RSM) was used for experimental designing and analyzing optimum condition for carbonization vis-à-vis calcium impregnation for preparation of adsorbent. During carbonization, temperature, time and weight ratio of calcium chloride to sal flower petal (SFP) have been considered as input factors and percentage removal of fluoride as response. Optimum condition for carbonization has been obtained as temperature, 500 °C; time, 1 h and weight ratio, 2.5 and the sample prepared has been termed as calcium-impregnated carbonized sal flower petal (CCSFP). Optimum condition as analyzed by one-factor-at-a-time (OFAT) method is initial fluoride concentration, 2.91 mg/L; pH 3 and adsorbent dose, 4 g/L. CCSFP shows maximum removal of 98.5% at this condition. RSM has also been used for finding out optimum condition for defluoridation considering initial concentration, pH and adsorbent dose as input parameters. The optimum condition as analyzed by RSM is: initial concentration, 5 mg/L; pH 3.5 and adsorbent dose, 2 g/L. Kinetic and equilibrium data follow Ho pseudo-second-order kinetic model and Freundlich isotherm model, respectively. Adsorption capacity of CCSFP has been found to be 5.465 mg/g. At optimized condition, CCSFP has been found to remove fluoride (80.4%) efficiently from groundwater collected from Bankura district in West Bengal, a fluoride-contaminated province in India.

  16. Optimization of maltodextrin production from avocado seed starch by response surface methodology

    NASA Astrophysics Data System (ADS)

    Nguyen, Thanh Viet; Ma, Tuyen-Hoang Nguyen; Nguyen, Tha Thi; Ho, Vinh-Nghi Kim; Vo, Hau Tan

    2018-04-01

    A process for maltodextrin production from avocado seed starch was reported in this study. Response surface methodology was used to investigate the effects of three independent variables for hydrolysis of the starch using a commercial food-grade α-amylase, Termamyl SC. These variables included enzyme concentration (0.05 - 0.15% starch), pH (5.0 - 6.0) and hydrolysis time (1.0 - 3.0 h), while the temperature fixed at 95°C. The result showed that the optimum conditions were using enzyme concentration at 0.12%, pH at 5.5 and 2.75 h of the incubation time. Under the optimum conditions, the recovered starch yield was 79.8% and the maltodextrin powder had 15.8 of dextrose equivalent.

  17. Crystal Structure and Superconductivity of PH 3 at High Pressures

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Liu, Hanyu; Li, Yinwei; Gao, Guoying

    2016-02-04

    We have performed a systematic structure search on solid PH3 at high pressures using the particle swarm optimization method. At 100–200 GPa, the search led to two structures which along with others have P–P bonds. These structures are structurally and chemically distinct from those predicted for the high-pressure superconducting H2S phase, which has a different topology (i.e., does not contain S–S bonds). Phonon and electron–phonon coupling calculations indicate that both structures are dynamically stable and superconducting. The pressure dependence and critical temperature for the monoclinic (C2/m) phase of 83 K at 200 GPa are in excellent agreement with a recentmore » experimental report.« less

  18. Relationship between fumonisin production and FUM gene expression in Fusarium verticillioides under different environmental conditions.

    PubMed

    Fanelli, Francesca; Iversen, Anita; Logrieco, Antonio F; Mulè, Giuseppina

    2013-01-01

    Fusarium verticillioides is the main source of fumonisins, a group of mycotoxins that can contaminate maize-based food and feed and cause diseases in humans and animals. The study of the effect of different environmental conditions on toxin production should provide information that can be used to develop strategies to minimize the risk. This study analysed the effect of temperature (15°C-35°C), water activity (a(w): 0.999-0.93), salinity (0-125 g l(-1) NaCl) and pH (5-8) on the growth and production of fumonisins B(1) (FB1), B(2) (FB2) and B(3) (FB3) and the expression of FUM1 and FUM21 in F. verticillioides. The highest growth rate was measured at 25°C, a(w) of 0.998-0.99 and 0-25 g l(-1) of NaCl. Optimal conditions for fumonisin production were 30°C, a(w) of 0.99, 25 g l(-1) of NaCl and pH 5; nevertheless, the strain showed a good adaptability and was able to produce moderate levels of fumonisins under a wide range of conditions. Gene expression mirrored fumonisin production profile under all conditions with the exception of temperature: FUM1 and FUM21 expression was highest at 15°C, while maximum fumonisin production was at 30°C. These data indicate that a post-transcriptional regulation mechanism could account for the different optimal temperatures for FUM gene expression and fumonisin production.

  19. Optimization of the ultrasound-assisted extraction of anthocyanins and total phenolic compounds in mulberry (Morus nigra) pulp.

    PubMed

    Espada-Bellido, Estrella; Ferreiro-González, Marta; Carrera, Ceferino; Palma, Miguel; Barroso, Carmelo G; Barbero, Gerardo F

    2017-03-15

    New ultrasound-assisted extraction methods for the determination of anthocyanins and total phenolic compounds present in mulberries have been developed. Several extraction variables, including methanol composition (50-100%), temperature (10-70°C), ultrasound amplitude (30-70%), cycle (0.2-0.7s), solvent pH (3-7) and solvent-solid ratio (10:1.5-20:1.5) were optimized. A Box-Behnken design in conjunction with a response surface methodology was employed to optimize the conditions for the maximum response based on 54 different experiments. Two response variables were considered: total anthocyanins and total phenolic compounds. Extraction temperature and solvent composition were found to be the most influential parameters for anthocyanins (48°C and 76%) and phenolic compounds (64°C and 61%). The developed methods showed high reproducibility and repeatability (RSD<5%). Finally, the new methods were successfully applied to real samples in order to investigate the presence of anthocyanins and total phenolic compounds in several mulberry jams. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Pork Quality Traits According to Postmortem pH and Temperature in Berkshire

    PubMed Central

    Kim, Tae Wan; Kim, Chul Wook; Yang, Mi Ra; No, Gun Ryoung; Kim, Il-Suk

    2016-01-01

    This study was performed to investigate the role of pH and temperature postmortem, and to demonstrate the importance of these factors in determining meat quality. Postmortem pH45min (pH at 45 min postmortem or initial pH) via analysis of Pearson’s correlation showed high positive correlation with pH change pHc24 (pH change from pH45min to pH24h postmortem). However, postmortem pH after 24 h (pH24h or ultimate pH) had a high negative correlation with pH change, pHc24, CIE L*, and protein content. Initial temperature postmortem (T1h ) was positively associated with a change in temperature from 45 min to 24 h postmortem (Tc24) and cooking loss, but negatively correlated with water holding capacity. Temperature at 24 h postmortem (T24h) was negatively associated with Tc24. Collectively, these results indicate that higher initial pH was associated with higher pHc24, T1h, and Tc24. However, higher initial pH was associated with a reduction in carcass weight, backfat thickness, CIE a* and b*, water holding capacity, collagen and fat content, drip loss, and cooking loss as well as decreased shear force. In contrast, CIE a* and b*, drip loss, cooking loss, and shear force in higher ultimate pH was showed by a similar pattern to higher initial pH, whereas pHc24, carcass weight, backfat thickness, water holding capacity, fat content, moisture content, protein content, T1h, T24h, and Tc24 were exhibited by completely differential patterns (p<0.05). Therefore, we suggest that initial pH, ultimate pH, and temperatures postmortem are important factors in determining the meat quality of pork. PMID:27499661

  1. Optimization of pectin extraction and antioxidant activities from Jerusalem artichoke

    NASA Astrophysics Data System (ADS)

    Liu, Shengyi; Shi, Xuejie; Xu, Lanlan; Yi, Yuetao

    2016-03-01

    Jerusalem artichoke is an economic crop widely planted in saline-alkaline soil. The use of Jerusalem artichoke is of great significance. In this study, the response surface method was employed to optimize the effects of processing variables (extraction temperature, pH, extraction time, and liquid-to-solid ratio) on the yield of Jerusalem artichoke pectin. Under the optimal extraction conditions: pH 1.52, 63.62 min, 100°C and a liquid-to-solid ratio of 44.4 mL/g, the maximum pectin yield was predicted to be 18.76%. Experiments were conducted under these optimal conditions and a pectin yield of 18.52±0.90% was obtained, which validated the model prediction. The effects of diff erent drying methods (freeze drying, spray drying and vacuum drying) on the properties of Jerusalem artichoke pectin were evaluated and they were compared with apple pectin. FTIR spectral analysis showed no major structural diff erences in Jerusalem artichoke pectin samples produced by various drying treatments. The antioxidant activities of pectin dried by diff erent methods were investigated using in vitro hydroxyl and DPPH radical scavenging systems. The results revealed that the activities of spray dried pectin (SDP) and apple pectin (AP) were stronger than those of vacuum oven dried pectin (ODP) and vacuum freeze dried pectin (FDP). Therefore compared with the other two drying methods, the spray drying method was the best.

  2. Determination of the Optimal Ammonium Sulfate Concentration for the Fractionation of Rabbit, Sheep, Horse, and Goat Antisera

    PubMed Central

    Hebert, G. Ann; Pelham, Patricia L.; Pittman, Bertie

    1973-01-01

    Various ammonium sulfate concentrations and reaction conditions were employed in the fractionation of sera from rabbits, sheep, horses, and goats. Precipitates and supernatant fluids were analyzed by electrophoresis to study the effects of the controlled variables. At room temperature, the third precipitate in 35% saturated (NH4)2SO4 was the best fraction from both rabbit and sheep sera; 80 to 90% of the gamma globulins were recovered. The second and third precipitates of horse sera proteins in 30% saturated (NH4)2SO4 were both satisfactory, but only 44% of the gamma globulin was recovered after three precipitations. Goat sera yielded a very satisfactory fraction; 80% of the gamma globulin was recovered after two precipitations—the first in 30% and the second in 45% saturated (NH4)2SO4. The composition of these fractions was not influenced by the pH of the sulfate solutions (pH 5.8 and 7.2), by a range of normal room temperatures (20 to 30 C), or by diluting the sera before fractionation. Crude globulins and fluorescein isothiocyanate-labeled globulins were successfully refractionated by one precipitation in the optimal sulfate concentration for the appropriate animal species. The refractionated products contained considerably less beta and alpha globulins than did the original crude fractions and little or no albumin. PMID:4119831

  3. Optimization of cellulase-assisted extraction process and antioxidant activities of polysaccharides from Tricholoma mongolicum Imai.

    PubMed

    Zhao, Yong-Ming; Song, Jin-Hui; Wang, Jin; Yang, Jian-Ming; Wang, Zhi-Bao; Liu, Ying-Hui

    2016-10-01

    Tricholoma mongolicum Imai is a well-known edible and medicinal mushroom which in recent years has attracted increasing attention because of its bioactivities. In this study, water-soluble polysaccharides were extracted from T. mongolicum Imai by cellulase-assisted extraction and their antioxidant activities were investigated. In order to improve the yield of polysaccharides, four variables, cellulase amount (X1 ), pH (X2 ), temperature (X3 ) and extraction time (X4 ), were investigated with a Box-Behnken design. The optimal conditions were predicted to be cellulase amount of 20 g kg(-1) , pH of 4.0, temperature of 50 °C and extraction time of 127 min, with a predicted polysaccharide yield of 190.1 g kg(-1) . The actual yield of polysaccharides under these conditions was 189.6 g kg(-1) , which matched the predicted value well. The crude polysaccharides were purified to obtain four fractions, and characterization of each was carried out. In addition, antioxidant properties of four polysaccharides assessed by 1,1-diphenyl-2-picryldydrazyl (DPPH) and hydroxyl radical-scavenging assays indicated that polysaccharides from T. mongolicum Imai (TMIPs) possessed antioxidant activity in a dose-dependent manner. TMIPs show moderate antioxidant activities in vitro. Therefore it is suggested that TMIPs are potential natural antioxidants for use in functional foods. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  4. Biological and physiological characteristics of Neotyphodium gansuense symbiotic with Achnatherum inebrians.

    PubMed

    Li, Chunjie; Nan, Zhibiao; Li, Fei

    2008-01-01

    Biological and physiological characteristics of Neotyphodium gansuense were compared with Neotyphodium coenophialum and Epichloë festucae at a range of temperatures and pH values, and on carbon and nitrogen amended media. N. gansuense was able to grow at 10-30 degrees C, but not at 5 degrees C, and slowly at 35 degrees C. The optimal temperature for both N. gansuense and N. coenophialum was 25 degrees C, but that of E. festucae was 20-25 degrees C. The optimal pH ranges for mycelial growth of N. gansuense, N. coenophialum and E. festucae were 5-9, 5-9 and 5-7, respectively. The Neotyphodium and Epichloë endophytes varied in their ability to grow on media containing different carbon and nitrogen nutrients. The preference of N. gansuense for carbon source was sucrose>glucose, lactose, sorbitol, inulin, maltose, mannitol, starch, fructose>xylose. Growth of all three endophytes tested was significantly improved by peptone, tryptone, casein, yeast extract and l-proline. Yeast extract, peptone, casein, tryptone, l-proline, potassium nitrate, ammonium oxalic acid and l-leucine significantly improved growth of N. gansuense. However, ammonium nitrite was not utilized at all by any tested endophyte. N. gansuense grew significantly better on potato dextrose agar (PDA) and oat meal agar (OMA) than on corn meal agar (CMA) and drunken-horse-grass agar (DA), and most slowly on water agar (WA) and saltwater nutrient agar (SNA).

  5. Lipase production in lipolytic yeast from Wonorejo mangrove area

    NASA Astrophysics Data System (ADS)

    Alami, Nur Hidayatul; Nasihah, Liziyatin; Umar, Rurin Luswidya Artaty; Kuswytasari, Nengah Dwianita; Zulaika, Enny; Shovitri, Maya

    2017-06-01

    Lipase is an enzyme that is often used in industry and become a commercial enzyme. One group of microorganisms capable of producing lipase is a yeast. This study aims to screen yeast from Wonorejo mangrove that potential to produce lipase and to optimize the production of these enzymes. Screening test include the measurement of lipolytic index and value of fatty acid. Yeast with the best value of fatty acid will be continued to the measurement of lipase activity. It is affected by several environmental factors, such as pH, temperature, and incubation time. This research was conducted to observe the optimization variation on environmental factors combination to produce lipase. Lipase activity was tested by using p-Nitrophenyl Palmitate (pNPP). Absorbency was measured by spectrofotometer on wavelength of 410 nm. Measurement of the enzyme activity was done by interpolating the absorbance values on the p-nitrophenol standard curve then calculated by the formula. All data were analyzed by using descriptive quantitative method. The results show that the highest lypolityc index was 2.08. The highest value of fatty acid was 0.49 that was reached on 168 hours of incubation. Candida W3.8 expressed the highest lypolylitic potential. The optimum environment to produce lipase by Candida W 3.8 was on 120 hours of incubation time, in temperature range of 27°C - 45°C and pH range of 4,5 - 7.

  6. Nanoparticle titanium dioxide aqueous interfacial energy can be modified to control phase stability, coarsening, and morphology

    NASA Astrophysics Data System (ADS)

    Finnegan, Michael Patrick

    The effect of solution chemistry on the phase stability, coarsening kinetics and morphology of titanium dioxide (TiO2) nanoparticles is investigated in order to attain efficient production pathways to desired nano-structures with optimal properties. To obtain sample, TiO2 was synthesized via hydrolysis of titanium isopropoxide producing an 85% anatase/15% brookite mixture. The titania was hydrothermally heated in an array of temperatures and pH values for various times. There are distinct phase stability fields for nanoscale titania based on pH alone due to slight interface charging behavior differences among the polymorphs. The mixture transforms to rutile below the pH of zero point of charge (ZPC) and remains anatase above the ZPC. This phenomenon is partially reversible. The solution chemistry also dictates the hydrothermal coarsening mechanism of the anatase polymorph. Ostwald ripening (OR) takes place in basic pH where titania solubility is elevated relative to neutral pH where lower solubility prevents rapid OR but allows for coarsening via oriented attachment (OA) of nanoparticles. This OA event can alter the symmetry of anatase causing unexpected and perhaps technically useful morphologies such as straight and curved nanorods during coarsening.

  7. [Establishment of optimal conditions at laboratory and pilot plant levels for the preparation of a protein isolated from Lupinus mutabilis].

    PubMed

    Rodríguez Pacheco, T; Aliaga, T; Schoeneberger, H; Gross, R

    1981-12-01

    Laboratory conditions were first investigated to determine are optimum processing parameters for the preparation of a protein isolate from the ground, defatted, commercial flakes of Lupinus mutabilis. The extraction variables were: pH (2-10); solvent to lupine ratio (5:1 to 40:1); temperature (28 degrees C - 60 degrees C) and time (10-50 min). The isoelectric point of the lupine protein was found to be pH 4.5 with a protein solubility higher than 80% above pH 8.0. Using 70-100 mesh, ground defatted flakes, and extracting at pH 8.7 for 60 min, a protein isolate was obtained on acidification to pH 4.5 which was 99.8 protein (dry basis), compared to 55.25% protein for the original material. This protein isolate represented 32% of the initial material and 57.6% of the initial nitrogen. When making pilot plant assays we found that the yield of protein isolate decreased to 20.4% of the original material and 36.4% of the initial nitrogen. The protein efficiency ratio for the protein isolate was 2.15 when supplemented with methionine, and had a digestibility of 89.33

  8. Optimal Autonomous Spacecraft Resiliency Maneuvers Using Metaheuristics

    DTIC Science & Technology

    2014-09-15

    Optimization of Interplanetary Trajectories,” Journal of Spacecraft and Rockets, Vol. 46, No. 2, March-April 2009, pp. 365–372. [23] Subbarao , K . and...Black, PhD (Chairman) /signed/ Darryl K . Ahner, PhD (Member) /signed/ Raymond R. Hill, PhD (Member) /signed/ William E. Wiesel, PhD (Member) 29 Jul 2014...124 Appendix A: Derivation of Spherical Equations of Motion . . . . . . . . . . . . . . 126 Appendix B : Equations of

  9. Unique pH dynamics in GABAergic synaptic vesicles illuminates the mechanism and kinetics of GABA loading.

    PubMed

    Egashira, Yoshihiro; Takase, Miki; Watanabe, Shoji; Ishida, Junji; Fukamizu, Akiyoshi; Kaneko, Ryosuke; Yanagawa, Yuchio; Takamori, Shigeo

    2016-09-20

    GABA acts as the major inhibitory neurotransmitter in the mammalian brain, shaping neuronal and circuit activity. For sustained synaptic transmission, synaptic vesicles (SVs) are required to be recycled and refilled with neurotransmitters using an H(+) electrochemical gradient. However, neither the mechanism underlying vesicular GABA uptake nor the kinetics of GABA loading in living neurons have been fully elucidated. To characterize the process of GABA uptake into SVs in functional synapses, we monitored luminal pH of GABAergic SVs separately from that of excitatory glutamatergic SVs in cultured hippocampal neurons. By using a pH sensor optimal for the SV lumen, we found that GABAergic SVs exhibited an unexpectedly higher resting pH (∼6.4) than glutamatergic SVs (pH ∼5.8). Moreover, unlike glutamatergic SVs, GABAergic SVs displayed unique pH dynamics after endocytosis that involved initial overacidification and subsequent alkalization that restored their resting pH. GABAergic SVs that lacked the vesicular GABA transporter (VGAT) did not show the pH overshoot and acidified further to ∼6.0. Comparison of luminal pH dynamics in the presence or absence of VGAT showed that VGAT operates as a GABA/H(+) exchanger, which is continuously required to offset GABA leakage. Furthermore, the kinetics of GABA transport was slower (τ > 20 s at physiological temperature) than that of glutamate uptake and may exceed the time required for reuse of exocytosed SVs, allowing reuse of incompletely filled vesicles in the presence of high demand for inhibitory transmission.

  10. Investigation of antimicrobial activity and statistical optimization of Bacillus subtilis SPB1 biosurfactant production in solid-state fermentation.

    PubMed

    Ghribi, Dhouha; Abdelkefi-Mesrati, Lobna; Mnif, Ines; Kammoun, Radhouan; Ayadi, Imen; Saadaoui, Imen; Maktouf, Sameh; Chaabouni-Ellouze, Semia

    2012-01-01

    During the last years, several applications of biosurfactants with medical purposes have been reported. Biosurfactants are considered relevant molecules for applications in combating many diseases. However, their use is currently extremely limited due to their high cost in relation to that of chemical surfactants. Use of inexpensive substrates can drastically decrease its production cost. Here, twelve solid substrates were screened for the production of Bacillus subtilis SPB1 biosurfactant and the maximum yield was found with millet. A Plackett-Burman design was then used to evaluate the effects of five variables (temperature, moisture, initial pH, inoculum age, and inoculum size). Statistical analyses showed that temperature, inoculum age, and moisture content had significantly positive effect on SPB1 biosurfactant production. Their values were further optimized using a central composite design and a response surface methodology. The optimal conditions of temperature, inoculum age, and moisture content obtained under the conditions of study were 37°C, 14 h, and 88%, respectively. The evaluation of the antimicrobial activity of this compound was carried out against 11 bacteria and 8 fungi. The results demonstrated that this biosurfactant exhibited an important antimicrobial activity against microorganisms with multidrug-resistant profiles. Its activity was very effective against Staphylococcus aureus, Staphylococcus xylosus, Enterococcus faecalis, Klebsiella pneumonia, and so forth.

  11. Ultraviolet irradiation and gradient temperature assisted autolysis for protein recovery from shrimp head waste.

    PubMed

    Cao, Wenhong; Tan, Caiyun; Zhan, Xiaojian; Li, Huiyi; Zhang, Chaohua

    2014-12-01

    A novel autolysis method using ultraviolet (UV) irradiation and gradient temperature was investigated to efficiently recover proteins from the head of the shrimp Penaeus vannamei. The proteolytic activity of shrimp head subjected to 30W UV irradiation for 20 min was increased by 62%, compared with that of untreated samples. After irradiation, the enzymes remained active across a wide range of temperatures (45-60°C) and pH (7-10). An orthogonal design was used to optimize autolysis condition. After 5h autolysis, protein recovery from the UV-heat treated samples was up to 92.1%. These results indicate the potential of using UV irradiation in combination with gradient temperatures to improve recovery of proteins from shrimp head waste. Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

  12. The Influence of pH on Prokaryotic Cell Size and Temperature

    NASA Astrophysics Data System (ADS)

    Sundararajan, D.; Gutierrez, F.; Heim, N. A.; Payne, J.

    2015-12-01

    The pH of a habitat is essential to an organism's growth and success in its environment. Although most organisms maintain a neutral internal pH, their environmental pH can vary greatly. However, little research has been done concerning an organism's environmental pH across a wide range of taxa. We studied pH tolerance in prokaryotes and its relationship with biovolume, taxonomic classification, and ideal temperature. We had three hypotheses: pH and temperature are not correlated; pH tolerance is similar within taxonomic groups; and extremophiles have small cell sizes. To test these hypotheses, we used pH, size, and taxonomic data from The Prokaryotes. We found that the mean optimum external pH was neutral for prokaryotes as a whole and when divided by domain, phylum, and class. Using ANOVA to test for pH within and among group variances, we found that variation of pH in domains, phyla, classes, and families was greater than between them. pH and size did not show much of a correlation, except that the largest and smallest sized prokaryotes had nearly neutral pH. This seems significant because extremophiles need to divert more of their energy from growth to maintain a neutral internal pH. Acidophiles showed a larger range of optimum pH values than alkaliphiles. A similar result was seen with the minimum and maximum pH values of acidophiles and alkaliphiles. While acidophiles were spread out and had some alkaline maximum values, alkaliphiles had smaller ranges, and unlike some acidophiles that had pH minimums close to zero, alkaliphile pH maximums did not go beyond a pH of 12. No statistically significant differences were found between sizes of acidophiles and alkaliphiles. However, optimum temperatures of acidophiles and alkaliphiles did have a statistically significant difference. pH and temperature had a negative correlation. Therefore, pH seems to have a correlation with cell size, temperature, and taxonomy to some extent.

  13. Evaluation of pectinolytic activities for oenological uses from psychrotrophic yeasts.

    PubMed

    Sahay, S; Hamid, B; Singh, P; Ranjan, K; Chauhan, D; Rana, R S; Chaurse, V K

    2013-08-01

    Of the twenty-three morphotypes of yeasts isolated from soil capable of utilizing pectin as sole carbon source at 6°C, two yeast isolates, one psychrotolerant (PT1) and one psychrophilic (SPY11), were selected according to their ability to secrete pectinolytic enzymes under some oenological conditions (temperature 6 and 12°C and pH 3.5) and ability or inability to grow above 20°C, respectively. As compared to their optimal activity, the three pectinolytic enzymes viz., pectin methyl esterase (PME), endopolygalacturonase (endo-PG) and exopolygalacturonase (exo-PG) isolated and assayed at pH 3.5 from PT1 were found to retain 39, 60 and 60% activity at 12°C and 40, 79 and 74% activity at 28°C, respectively. Likewise, the enzymes PME and endo-PG at pH 3.5 from SPY11 displayed 46 and 86% activity at 12°C and 50 and 60% activity at 28°C, respectively. All these enzymes showed 20-90% of residual activity at pH 3.5 and 6°C. The yeast isolates PT1 and SPY11 were identified as Rhodotorula mucilaginosa and Cystofilobasidium capitatum, respectively, on the basis of morphological, physiological and molecular characteristics. This study presents the first report on pectinolytic activities under major oenological conditions from psychrotolerant isolate R. mucilaginosa PT1 and psychrophilic isolate C. capitatum SPY11. The cold-active pectinolytic enzymes (PME, endo-PG and exo-PG) from the newly isolated and identified psychrophilic yeast Cystofilobasidium capitatum SPY11 and psychrotolerant yeast Rhodotorula mucilaginosa PT1that exhibited 50-80% of their optimum activity under some major oenological conditions pH (3.5) and temperatures (6 and 12°C) could be applied to wine production and juice clarification at low temperature. The psychrotrophic yeasts themselves could be applied to cold process for the production of enzymes thus saving cost of energy and protecting process from contamination. © 2013 The Society for Applied Microbiology.

  14. Comparison between the univariate and multivariate analysis on the partial characterization of the endoglucanase produced in the solid state fermentation by Aspergillus oryzae ATCC 10124.

    PubMed

    de Brito, Aila Riany; Santos Reis, Nadabe Dos; Silva, Tatielle Pereira; Ferreira Bonomo, Renata Cristina; Trovatti Uetanabaro, Ana Paula; de Assis, Sandra Aparecida; da Silva, Erik Galvão Paranhos; Aguiar-Oliveira, Elizama; Oliveira, Julieta Rangel; Franco, Marcelo

    2017-11-26

    Endoglucanase production by Aspergillus oryzae ATCC 10124 cultivated in rice husks or peanut shells was optimized by experimental design as a function of humidity, time, and temperature. The optimum temperature for the endoglucanase activity was estimated by a univariate analysis (one factor at the time) as 50°C (rice husks) and 60°C (peanut shells), however, by a multivariate analysis (synergism of factors), it was determined a different temperature (56°C) for endoglucanase from peanut shells. For the optimum pH, values determined by univariate and multivariate analysis were 5 and 5.2 (rice husk) and 5 and 7.6 (peanut shells). In addition, the best half-lives were observed at 50°C as 22.8 hr (rice husks) and 7.3 hr (peanut shells), also, 80% of residual activities was obtained between 30 and 50°C for both substrates, and the pH stability was improved at 5-7 (rice hulls) and 6-9 (peanut shells). Both endoglucanases obtained presented different characteristics as a result of the versatility of fungi in different substrates.

  15. Bacterial navigation in chemical and nonchemical environments

    NASA Astrophysics Data System (ADS)

    Hu, Bo; Tu, Yuhai

    2014-03-01

    Navigation of cells to the optimal environmental niches is critical for their survival and growth. E. coli cells, for example, can detect various chemicals and move up or down those chemical gradients (i.e., chemotaxis). Using the same signaling machinery, they can also sense other external factors such as pH and temperature and navigate from both sides toward some intermediate levels of those stimuli. This mode of precision sensing is more sophisticated than the (unidirectional) chemotaxis strategy and requires distinctive molecular mechanisms. To understand different bacterial taxis behaviors, we develop a theoretical model which incorporates microscopic signaling events in individual cells into macroscopic population dynamics. We find that the equilibrium population distribution is governed by an effective potential, the landscape of which depends on the external environment (chemical stimuli, pH, and temperature). We uncover the key conditions for various taxis behaviors and directly connects the cellular taxis performances with the underlying molecular parameters. This approach is used to examine and predict how background attractants and downstream temperature effects influence the performance and stability of thermotaxis, which can be tested in future experiments. This work is supported by the National Institutes of Health Grant GM081747.

  16. Optimal conditions of mycelia growth of Laetiporus sulphureus sensu lato

    PubMed Central

    Luangharn, Thatsanee; Karunarathna, Samantha C.; Hyde, Kevin D.; Chukeatirote, Ekachai

    2014-01-01

    Laetiporus sulphureus is an edible wood-rotting basidiomycete, growing on decaying logs, stumps, and trunks of many deciduous and coniferous tree species. This fungus produces relatively large striking yellowish or orange-coloured bracket-like fruitbodies. L. sulphureus is widely consumed as a nutritional food because of its fragrance and texture. In this study, two L. sulphureus strains, MFLUCC 12-0546 and MFLUCC 12-0547, isolated from Chiang Rai, Thailand, were investigated for optimal conditions of mycelia growth. Potato dextrose agar and malt extract agar were observed as the favourable medium for mycelia growth. The optimum pH and temperature for the mushroom mycelia were 6–8 and 25–30°C, respectively. PMID:25544934

  17. Physiological and fermentation properties of Bacillus coagulans and a mutant lacking fermentative lactate dehydrogenase activity.

    PubMed

    Su, Yue; Rhee, Mun Su; Ingram, Lonnie O; Shanmugam, K T

    2011-03-01

    Bacillus coagulans, a sporogenic lactic acid bacterium, grows optimally at 50-55 °C and produces lactic acid as the primary fermentation product from both hexoses and pentoses. The amount of fungal cellulases required for simultaneous saccharification and fermentation (SSF) at 55 °C was previously reported to be three to four times lower than for SSF at the optimum growth temperature for Saccharomyces cerevisiae of 35 °C. An ethanologenic B. coagulans is expected to lower the cellulase loading and production cost of cellulosic ethanol due to SSF at 55 °C. As a first step towards developing B. coagulans as an ethanologenic microbial biocatalyst, activity of the primary fermentation enzyme L-lactate dehydrogenase was removed by mutation (strain Suy27). Strain Suy27 produced ethanol as the main fermentation product from glucose during growth at pH 7.0 (0.33 g ethanol per g glucose fermented). Pyruvate dehydrogenase (PDH) and alcohol dehydrogenase (ADH) acting in series contributed to about 55% of the ethanol produced by this mutant while pyruvate formate lyase and ADH were responsible for the remainder. Due to the absence of PDH activity in B. coagulans during fermentative growth at pH 5.0, the l-ldh mutant failed to grow anaerobically at pH 5.0. Strain Suy27-13, a derivative of the l-ldh mutant strain Suy27, that produced PDH activity during anaerobic growth at pH 5.0 grew at this pH and also produced ethanol as the fermentation product (0.39 g per g glucose). These results show that construction of an ethanologenic B. coagulans requires optimal expression of PDH activity in addition to the removal of the LDH activity to support growth and ethanol production.

  18. Recovery of fermented inulin fiber by lactic acid bacteria (LAB) from inulin hydrolysate using fungi inulinase enzymes of Scopulariopsis sp.-CBS1 and class of Deuteromycetes-CBS4 as cholesterol binder

    NASA Astrophysics Data System (ADS)

    Susilowati, Agustine; Melanie, Hakiki; Maryati, Yati; Aspiyanto

    2017-01-01

    Fermentation of Lactobacillus Acid Bacteria (LAB) which are mixtures of Lactobacillus acidophilus, Bifidobacteriumbifidum, Lactobacillus bulgaricus and Streptococcus thermophillus on hydrolysate as a result of inulin hydrolysis using inulinase enzymes obtained from endophytic fungi ofScopulariopsis sp.-CBS1 (inulin hydrolysate of S) and Class of Deuteromycetes-CBS4 (inulin hydrolysate of D) generate potential fermented inulin fiber as cholesterol binder. Fermentation process was conducted under concentrations of inulin hydrolysate 50% (w/v), LAB 15% (v/v) and skim milk 12.5% (w/v) at room temperature and 40°C for 0, 12, 24, 36 and 48 hours, respectively. Result of experimental work showed that longer time of LAB fermentation increased total acids, TPC and CBC at pH 2, but decreased total sugar, reducing, IDF, SDF, CBC pH 2 and CBC pH 7. Based on Cholesterol Binding Capacity (CBC), optimization of fermentation process on inulin hydrolysate of S was achieved by combining treatment at 40°C for 24 hours resulted in CBC pH 2 of 19.11 mg/g TDF and inulin hydrolysate of D was achieved by fermentation at 40 °C for 48 hours resulted in CBC pH 2 of 24.28 mg/g TDF. Inulin hydrolysate of class of Deutrymecetes CBS4 fermented by LAB had better functional property as cholesterol binder than that inulin hydrolysate of S fermented by LAB. This is due to cholesterol binder and cholesterol derivatives as a result of degradation of LAB on digestive system (stomach) when compared to higher colon under optimal process condition.

  19. Influence of temperature, pH and metal ions on guaiacol oxidation of purified laccase from Leptographium qinlingensis.

    PubMed

    Hu, Xia; Wang, Chunyan; Wang, Le; Zhang, Ranran; Chen, Hui

    2014-04-01

    The bark beetle Dendroctonus armandi is able to kill living Pinus armandi and has caused serious damage to pine forest in Northern China. As the most important symbiotic fungus of D. armandi, Leptographium qinlingensis plays an important role in the invasion process of the bark beetle. The laccase secreted by it are involved in lignin degradation to provide utilizable nutrition for D. armandi, and catalyze some biochemical reactions, causing the damages of tree tissue. In present study, the extracellular laccase of L. qinlingensis was purified by using the ammonium sulfate precipitation and DEAE-cellulose (DE-52) column chromatography. Furthermore, the effects of temperature, pH value and metal ions on it were investigated and characterized. The purified enzyme exerted its optimal activity with guaiacol. The catalytic efficiencies K(m) and V(max) determined for substrate guaiacol were 15.4 μM and 372.9 IU mg⁻¹, respectively. The optimum pH and temperature for the purified enzyme was 4.4 and 45 °C, respectively, with the highest enzyme specific activity of 7,000 IU mg⁻¹. Moreover, the metal ions, Co²⁺, Mn²⁺, Ca²⁺, Mg²⁺, Fe²⁺ and Cd²⁺, especially Hg²⁺, showed significantly inhibition effects on its activity. To understand the characteristics of this laccase might provide an opportunity and theoretical basis to promote integrated pest management of D. armandi.

  20. Optimization of supercritical carbon dioxide treatment for the inactivation of the natural microbial flora in cubed cooked ham.

    PubMed

    Ferrentino, Giovanna; Balzan, Sara; Spilimbergo, Sara

    2013-02-15

    This study aims to investigate the effects of supercritical carbon dioxide (SC-CO₂) treatment on the inactivation of the natural microbial flora in cubed cooked ham. Response surface methodology with a central composite design was applied to determine the optimal process conditions and investigate the effect of three independent variables (pressure, temperature and treatment time). Additionally, analyses of texture, pH and color together with a storage study of the product were performed to determine its microbial and qualitative stability. Response surface analysis revealed that 12 MPa, 50 °C, 5 min were the optimal conditions to obtain about 3.0, 1.6, and 2.5 Log(CFU/g) reductions of mesophilic aerobic bacteria, psychrophilic bacteria and lactic acid bacteria respectively. Inactivation to undetectable levels of yeasts and molds and coliforms was also obtained. A storage study of 30 days at 4 °C was carried out on the treated product (12 MPa, 50 °C, 5 min) monitoring microbial growth, pH, texture, and color parameters (L*, a*, b* and ΔE). Microbial loads slightly increased and after 30 days of storage reached the same levels detected in the fresh product. Color parameters (L*, a*, b*) showed slight variations while pH and texture did not change significantly. On the basis of the results obtained, SC-CO₂ can be considered a promising technique to microbiologically stabilize cubed cooked ham and, in general, cut/sliced meat products without affecting its quality attributes. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Purification and characterization of chitinases and chitosanases from a new species strain Pseudomonas sp. TKU015 using shrimp shells as a substrate.

    PubMed

    Wang, San-Lang; Chen, Shin-Jen; Wang, Chuan-Lu

    2008-05-19

    A chitinase (CHT1) and a chitosanase (CHS1) were purified from the culture supernatant of Pseudomonas sp. TKU015 with shrimp shell wastes as the sole carbon and nitrogen source. The optimized conditions of this new species strain (Gen Bank Accession Number EU103629) for the production of chitinases were found to be when the culture was shaken at 30 degrees C for 3 days in 100 mL of medium (pH 8) containing 0.5% shrimp shell powder (SSP) (w/v), 0.1% K2HPO4, and 0.05% MgSO(4).7H2O. The molecular weights of CHT1 and CHS1 determined by SDS-PAGE were approximately 68 kDa and 30 kDa, respectively. The optimum pH, optimum temperature, pH stability, and the thermal stability of CHT1 and CHS1 were pH 6, 50 degrees C, pH 5-7, <50 degrees C and pH 4, 50 degrees C, pH 3-9, <50 degrees C, respectively. CHT1 was inhibited completely by Mn2+ and Fe2+, and CHS1 was inhibited by Mn2+, Cu2+, and PMSF. CHT1 was only specific to chitin substrates, whereas the relative activity of CHS1 increased when the degree of deacetylation of soluble chitosan increased.

  2. Effect of immobilized biosorbents on the heavy metals (Cu2+) biosorption with variations of temperature and initial concentration of waste

    NASA Astrophysics Data System (ADS)

    Siwi, W. P.; Rinanti, A.; Silalahi, M. D. S.; Hadisoebroto, R.; Fachrul, M. F.

    2018-01-01

    The aims of research is to studying the efficiency of copper removal by combining immobilized microalgae with optimizations of temperature and initial Copper concentration. The research was conducted in batch culture with temperature variations of 25°C, 30°C, and 35°C, as well as initial Cu2+ concentrations (mg/l) of 3, 5, 10, 15 and 20 using monoculture of S. cerevisiae, Chlorella sp., and mixed culture of them both as immobilized biosorbents. The optimum adsorption of 83.4% obtained in temperature of 30°C with an initial waste concentration of 17.62 mg/l, initial biomass concentration of 200 mg, pH of 4, and 120 minutes detention time by the immobilized mixed culture biosorbent. The cell morphology examined using Scanning Electron Microscope (SEM) has proved that the biosorbent surface was damaged after being in contact with copper (waste), implying that heavy metals (molecules) attach to different functional cell surfaces and change the biosorbent surface. The adsorption process of this research follows Langmuir Isotherm with the R2 value close to 1. The immobilized mixed culture biosorbent is capable of optimally removing copper at temperature of 30°C and initial Cu2+ concentration of 17.62 mg/l.

  3. Treatment of pulp and paper industry bleaching effluent by electrocoagulant process.

    PubMed

    Sridhar, R; Sivakumar, V; Prince Immanuel, V; Prakash Maran, J

    2011-02-28

    The experiments were carried out in an electrocoagulation reactor with aluminum as sacrificial electrodes. The influence of electrolysis time, current density, pH, NaCl concentration, rotational speed of the stirrer and electrode distance on reduction of color, COD and BOD were studied in detail. From the experimental results, 15 mA/cm(2) current density, pH of 7, 1 g/l NaCl, 100 rpm, 28°C temperature and 3 cm electrode distance were found to be optimum for maximum reduction of color, COD and BOD. The reduction of color, COD and BOD under the optimum condition were found to be 94%, 90% and 87% respectively. The electrode energy consumption was calculated and found to be varied from 10.1 to 12.9 kWh/m(3) depending on the operating conditions. Under optimal operating condition such as 15 mA/cm(2) current density, pH of 7, 1 g/l NaCl, 100 rpm, 28°C temperature and 3 cm electrode distance, the operating cost was found to be 1.56 US $/m(3). The experimental results proved that the electrocoagulation is a suitable method for treating bleaching plant effluents for reuse. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. The use of a natural coagulant (Opuntia ficus-indica) in the removal for organic materials of textile effluents.

    PubMed

    de Souza, Maísa Tatiane Ferreira; Ambrosio, Elizangela; de Almeida, Cibele Andrade; de Souza Freitas, Thábata Karoliny Formicoli; Santos, Lídia Brizola; de Cinque Almeida, Vitor; Garcia, Juliana Carla

    2014-08-01

    The goal of this study was to investigate the activity of the coagulant extracted from the cactus Opuntia ficus-indica (OFI) in the process of coagulation/flocculation of textile effluents. Preliminary tests of a kaolinite suspension achieved maximum turbidity removal of 95 % using an NaCl extraction solution. Optimization assays were conducted with actual effluents using the response surface methodology (RSM) based on the Box-Behnken experimental design. The responses of the variables FeCl3, dosage, cactus dosage, and pH in the removal of COD and turbidity from both effluents were investigated. The optimum conditions determined for jeans washing laundry effluent were the following: FeCl3 160 mg L(-1), cactus dosage 2.60 mg L(-1), and pH 5.0. For the fabric dyeing effluent, the optimum conditions were the following: FeCl3 640 mg L(-1), cactus dosage 160 mg L(-1), and pH 6.0. Investigation of the effects of the storage time and temperature of the cactus O. ficus-indica showed that coagulation efficiency was not significantly affected for storage at room temperature for up to 4 days.

  5. Sorption behavior of nano-TiO2 for the removal of selenium ions from aqueous solution.

    PubMed

    Zhang, Lei; Liu, Na; Yang, Lijun; Lin, Qing

    2009-10-30

    Titanium dioxide nanoparticles were employed for the sorption of selenium ions from aqueous solution. The process was studied in detail by varying the sorption time, pH, and temperature. The sorption was found to be fast, and to reach equilibrium basically within 5.0 min. The sorption has been optimized with respect to the pH, maximum sorption has been achieved from solution of pH 2-6. Sorbed Se(IV) and Se(VI) were desorbed with 2.0 mL 0.1 mol L(-1) NaOH. The kinetics and thermodynamics of the sorption of Se(IV) onto nano-TiO2 have been studied. The kinetic experimental data properly correlate with the second-order kinetic model (k(2)=0.69 g mg(-1) min(-1), 293 K). The overall rate process appears to be influenced by both boundary layer diffusion and intraparticle diffusion. The sorption data could be well interpreted by the Langmuir sorption isotherm. The mean energy of adsorption (14.46 kJ mol(-1)) was calculated from the Dubinin-Radushkevich (D-R) adsorption isotherm at room temperature. The thermodynamic parameters for the sorption were also determined, and the DeltaH(0) and DeltaG(0) values indicate exothermic behavior.

  6. Whole cell immobilization of refractory glucose isomerase using tris(hydroxymethyl)phosphine as crosslinker for preparation of high fructose corn syrup at elevated temperature.

    PubMed

    Jia, Dong-Xu; Wang, Teng; Liu, Zi-Jian; Jin, Li-Qun; Li, Jia-Jia; Liao, Cheng-Jun; Chen, De-Shui; Zheng, Yu-Guo

    2018-04-04

    Glucose isomerase (GI) responsible for catalyzing the isomerization from d-glucose to d-fructose, was an important enzyme for producing high fructose corn syrup (HFCS). In a quest to prepare HFCS at elevated temperature and facilitate enzymatic recovery, an effective procedure for whole cell immobilization of refractory Thermus oshimai glucose isomerase (ToGI) onto Celite 545 using tris(hydroxymethyl)phosphine (THP) as crosslinker was established. The immobilized biocatalyst showed an activity of approximate 127.3 U/(g·immobilized product) via optimization in terms of cells loading, crosslinker concentration and crosslinking time. The pH optimum of the immobilized biocatalyst was displaced from pH 8.0 of native enzyme to neutral pH 7.0. Compared with conventional glutaraldehyde (GLU)-immobilized cells, it possessed the enhanced thermostability with 70.1% residual activity retaining after incubation at 90°C for 72 h. Moreover, the THP-immobilized biocatalyst exhibited superior operational stability, in which it retained 85.8% of initial activity after 15 batches of bioconversion at 85°C. This study paved a way for reducing catalysis cost for upscale preparation of HFCS with higher d-fructose concentration. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  7. Hydrogen production and metal-dye bioremoval by a Nostoc linckia strain isolated from textile mill oxidation pond.

    PubMed

    Mona, Sharma; Kaushik, Anubha; Kaushik, C P

    2011-02-01

    Biohydrogen production by Nostoc linckia HA-46, isolated from a textile-industry oxidation-pond was studied by varying light/dark period, pH, temperature and ratio of carbon-dioxide and argon in the gas-mixture. Hydrogen production rates were maximum under 18 h of light and 6 h of darkness, pH 8.0, 31°C, a CO(2):Ar ratio 2:10. Hydrogen production of the strain acclimatized to 20 mg/L of chromium/cobalt and 100 mg/L of Reactive red 198/crystal violet dye studied in N-supplemented/deficient medium was 6-10% higher in the presence of 1.5 g/L of NaNO(3). Rates of hydrogen production in the presence of dyes/metals by the strain (93-105 μmol/h/mg Chlorophyll) were significantly higher than in medium without metals/dyes serving as control (91.3 μmol/h/mg Chlorophyll). About 58-60% of the two metals and 35-73% of dyes were removed by cyanobacterium. Optimal conditions of temperature, pH and metals/dyes concentration for achieving high hydrogen production and wastewater treatment were found practically applicable as similar conditions are found in the effluent of regional textile-mills. Copyright © 2010 Elsevier Ltd. All rights reserved.

  8. Isolation and molecular characterization of thermostable phytase from Bacillus subtilis (BSPhyARRMK33).

    PubMed

    Reddy, Chinreddy Subramanyam; Achary, V Mohan Murali; Manna, Mrinalini; Singh, Jitender; Kaul, Tanushri; Reddy, Malireddy K

    2015-03-01

    The thermostable phytase gene was isolated from Bacillus subtilis ARRMK33 (BsPhyARRMK33). The gene has an ORF of 1152 bp and that encodes a protein of 383 amino acids. Sequence analysis showed high homology with Bacillus sp. phytase proteins, but no similarity was found with other phytases. SDS-PAGE analysis exhibited a predicted molecular mass of 42 kDa. Homology modeling of BsPhyARRMK33 protein based on Bacillus amyloliquefaciens crystal structure disclosed its β-propeller structure. BsPhyARRMK33 recombinant plasmid in pET-28a(+) was expressed in Rosetta gami B DE3 cells and the maximum phytase activity 15.3 U mg(-1) obtained. The enzyme exhibits high thermostability at various temperatures and broad pH ranges. The recombinant protein retained 74% of its original activity after incubation at 95 °C for 10 min. In the presence of Ca(2+), the recombinant phytase activity was maximal where as it was inhibited by EDTA. The optimal pH and temperature for the recombinant phytase activity is achieved at 7.0 and 55 °C, respectively. Thermostable nature and wide range of pH are promising features of recombinant BsPhyARRMK33 protein that may be employed as an efficient alternative to commercially known phytases and thereby alleviate environmental eutrophication.

  9. Use of Cu/Zn-superoxide dismutase tool for biomonitoring marine environment pollution in the Persian Gulf and the Gulf of Oman.

    PubMed

    Zeinali, Farrokhzad; Homaei, Ahmad; Kamrani, Ehsan; Patel, Seema

    2018-04-30

    Superoxide dismutase (SOD) is the pivotal antioxidant enzyme that defends organisms against the oxidative stresses of superoxide radicals. In this experimental study, purification of SOD from the leaves of Avicennia marina (grey mangrove or white mangrove) from the family Acanthaceae, located in Sirik mangrove forest on the shore of the Gulf of Oman was performed, for the intended characterization of SOD. The Sirik AmSOD (A. marina SOD) expressed optimum activity in the pH range of 6-9 with the maximum activity at pH 8. The optimal temperature for Sirik AmSOD activity was 70°C. Comparison of the pH and temperature optima in two regions (the Persian Gulf and the Gulf of Oman) showed significant differences with P<0.05. The SOD from the Persian Gulf was more resistant against the environmental stressors, because of the biochemical adaption to this environment, which is harsher. The evidence from these results suggests that AmSOD has different characteristics in each place, and mangroves undergo different adaptations and require different protections. The results of the enzymatic research can be useful for ecological management of organisms. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Equilibrium partitioning of organic compounds to OASIS HLB® as a function of compound concentration, pH, temperature and salinity.

    PubMed

    Jeong, Yoonah; Schäffer, Andreas; Smith, Kilian

    2017-05-01

    Oasis hydrophilic lipophilic balance ® (Oasis HLB) is commonly employed in solid phase extraction (SPE) of environmental contaminants and within polar organic chemical integrative passive samplers (POCIS). In this study batch experiments were carried out to evaluate the relative affinity of a range of relevant organic pollutants to Oasis HLB in aqueous systems. The influence of sorbate concentration, temperature, pH, and salinity on the equilibrium sorption was investigated. Equilibrium partition ratios (K D ) of 28 compounds were determined, ranging over three orders of magnitude from 1.16 × 10 3  L/kg (atenolol) to 1.07 × 10 6  L/kg (isoproturon). The Freundlich model was able to describe the equilibrium partitioning to Oasis HLB, and an analysis of the thermodynamic parameters revealed the spontaneous and exothermic nature of the partitioning process. Ionic strength had only a minor effect on the partitioning, whereas pH had a considerable effect but only for ionizable compounds. The results show that apolar interactions between the Oasis HLB and analyte mainly determine the equilibrium partitioning. These research findings can be used to optimize the application of SPE and POCIS for analyses of environmental contaminants even in complex mixtures. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Improving dyeability of modified cotton fabrics by the natural aqueous extract from red cabbage using ultrasonic energy.

    PubMed

    Ben Ticha, Manel; Haddar, Wafa; Meksi, Nizar; Guesmi, Ahlem; Mhenni, M Farouk

    2016-12-10

    The concern regarding sustainable utilization of available resources is growing due to its global importance. In this paper, the dyeability of cotton fabrics with natural colorant extracted from red cabbage was improved by applying cationic groups on cotton fibers. Modification of cotton was carried using acid tannic, Rewin Os, Denitex BC and Sera Fast as cationic agents. The dyeing process was done by ultrasonic energy. The effects of the cationising agent amount, the dye bath pH, the dyeing temperature and duration, on the sonicator dyeing quality were studied. The performances of this process were evaluated by measuring the colour yield (K/S) and the dyeing fastness of the coloured cotton. Besides, modified cotton fibers were characterized by morphology analysis (SEM) and Fourier transform infrared (FTIR) spectra and compared to untreated cotton. Moreover, a two-level full factorial design was employed to optimize the sonicator dyeing process. Mathematical model equation and statistical analysis were derived by computer simulation programming applying the least squares method using Minitab 15. Best dyeing conditions were found to be: 10%, pH 11, 60min and 100°C respectively for the Sera Fast amount, dye bath pH, dyeing duration and temperature. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Characterisation of a novel Bacillus sp. SJ-10 β-1,3-1,4-glucanase isolated from jeotgal, a traditional Korean fermented fish.

    PubMed

    Kim, Yu-Ri; Kim, Eun-Young; Lee, Jong Min; Kim, Joong Kyun; Kong, In-Soo

    2013-06-01

    A novel β-1,3-1,4-glucanase gene was identified in Bacillus sp. SJ-10 (KCCM 90078) isolated from jeotgal, a traditional Korean fermented fish. We analysed the β-1,3-1,4-glucanase gene sequence and examined the recombinant enzyme. The open reading frame of the gene encoded 244 amino acids. The sequence was not identical to any β-glucanases deposited in GenBank. The gene was cloned into pET22b(+) and expressed in Escherichia coli BL21. Purification of recombinant β-1,3-1,4-glucanase was conducted by affinity chromatography using a Ni-NTA column. Enzyme specificity of β-1,3-1,4-glucanase was confirmed based on substrate specificity. The optimal temperature and pH of the purified enzyme towards barley β-glucan were 50 °C and pH 6, respectively. More than 80 % of activity was retained at temperatures of 30-70 °C and pH values of 4-9, which differed from all other bacterial β-1,3-1,4-glucanases. The degradation products of barley β-glucan by β-1,3-1,4-glucanase were analysed using thin-layer chromatography, and ultimately glucose was produced by treatment with cellobiase.

  13. Evaluation of the individuality of white rot macro fungus for the decolorization of synthetic dye.

    PubMed

    Pandey, Priyanka; Singh, Ram Praksh; Singh, Kailash Nath; Manisankar, Paramasivam

    2013-01-01

    A biosorbent was developed by simple dried Agaricus bisporus (SDAB) and effectively used for the biosorption of cationic dyes, Crystal Violet and Brilliant Green. For the evaluation of the biosorbent system, all the batch equilibrium parameters like pH, biomass dose, contact time, and temperature were optimized to determine the decolorization efficiency of the biosorbent. The maximum yields of dye removal were achieved at pH 4.0 for Crystal Violet (CV) and pH 5.0 for Brilliant Green (BG), which are closer to their natural pH also. Equilibrium was established at 60 and 40 min for CV and BG, respectively. Pseudo first-order, pseudo second-order, and intraparticle-diffusion kinetic models were studied at different temperatures. Isotherm models such as Freundlich, Langmuir, and Dubinin-Radushkevich were also studied. Biosorption processes were successfully described by Langmuir isotherm model and the pseudo second-order kinetic model. The biosorption capacity of A. bisporus over CV and BG were found as 21.74 and 12.16 mg gm(-1). Thermodynamic parameters indicated that the CV and BG dye adsorption onto A. bisporus is spontaneous and exothermic in the single and ternary systems. Scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy were used for the surface morphology, crystalline structure of biosorbent, and dye-biosorbent interaction, respectively. This analysis of the biosorption data confirmed that these biosorption processes are ecofriendly and economical. Thus, this biomass system may be useful for the removal of contaminating cationic dyes.

  14. Bioconversion of D-galactose to D-tagatose: continuous packed bed reaction with an immobilized thermostable L-arabinose isomerase and efficient purification by selective microbial degradation.

    PubMed

    Liang, Min; Chen, Min; Liu, Xinying; Zhai, Yafei; Liu, Xian-wei; Zhang, Houcheng; Xiao, Min; Wang, Peng

    2012-02-01

    The continuous enzymatic conversion of D-galactose to D-tagatose with an immobilized thermostable L-arabinose isomerase in packed-bed reactor and a novel method for D-tagatose purification were studied. L-arabinose isomerase from Thermoanaerobacter mathranii (TMAI) was recombinantly overexpressed and immobilized in calcium alginate. The effects of pH and temperature on D-tagatose production reaction catalyzed by free and immobilized TMAI were investigated. The optimal condition for free enzyme was pH 8.0, 60°C, 5 mM MnCl(2). However, that for immobilized enzyme was pH 7.5, 75°C, 5 mM MnCl(2). In addition, the catalytic activity of immobilized enzyme at high temperature and low pH was significantly improved compared with free enzyme. The optimum reaction yield with immobilized TMAI increased by four percentage points to 43.9% compared with that of free TMAI. The highest productivity of 10 g/L h was achieved with the yield of 23.3%. Continuous production was performed at 70°C; after 168 h, the reaction yield was still above 30%. The resultant syrup was then incubated with Saccharomyces cerevisiae L1 cells. The selective degradation of D-galactose was achieved, obtaining D-tagatose with the purity above 95%. The established production and separation methods further potentiate the industrial production of D-tagatose via bioconversion and biopurification processes.

  15. Statistical approach for the enhanced production of cold-active β-galactosidase from Thalassospira frigidphilosprofundus: a novel marine psychrophile from deep waters of Bay of Bengal.

    PubMed

    Pulicherla, K K; Kumar, P Suresh; Manideep, K; Rekha, V P B; Ghosh, Mrinmoy; Sambasiva Rao, K R S

    2013-01-01

    In the present investigation Thalassospira frigidphilosprofundus, a novel species from the deep waters of the Bay of Bengal, was explored for the production of cold-active β-galactosidase by submerged fermentation using marine broth medium as the basal medium. Effects of various medium constituents, namely, carbon, nitrogen source, pH, and temperature, were investigated using a conventional one-factor-at-a-time method. It was found that lactose, yeast extract, and bactopeptones are the most influential components for β-galactosidase production. Under optimal conditions, the production of β-galactosidase was found to be 3,864 U/mL at 20 ± 2°C, pH 6.5 ± 0.2, after 48 hr of incubation. β-Galactosidase production was further optimized by the Taguchi orthogonal array design of experiments and the central composite rotatable design (CCRD) of response surface methodology. Under optimal experimental conditions the cold-active β-galactosidase enzyme production from Thalassospira frigidphilosprofundus was enhanced from 3,864 U/mL to 10,657 U/mL, which is almost three times higher than the cold-active β-galactosidase production from the well-reported psychrophile Pseudoalteromonas haloplanktis.

  16. Deglycosylation of isoflavones in isoflavone-rich soy germ flour by Aspergillus oryzae KACC 40247.

    PubMed

    Lee, Seon-Hwa; Seo, Min-Ho; Oh, Deok-Kun

    2013-12-11

    Aspergillus oryzae KACC 40247 was selected as an efficient daidzein-producing fungus from strains of the genus Aspergillus by using 5% (w/v) soy germ flour (SGF) as an isoflavone-glycoside-rich medium. The culture conditions, including SGF concentration, agitation speed, initial pH, temperature, and time, were optimized as follows: 7% (w/v) SGF, initial pH 6.0, 33 °C, 300 rpm, and 24 h in a 100 mL baffled flask. The determined amount of isoflavone aglycons in SGF using 50% ethyl acetate was the highest among the solvent systems tested and it was 3.7-fold higher than that using 70% methanol. Under the optimized conditions, the content and concentration of daidzein were 134 mg/g of SGF and 9.4 g/L, respectively, with a productivity of 391 ± 2.8 mg/L/h, and those of isoflavone aglycons were 165 mg/g of SGF and 11.5 g/L, respectively, with a productivity of 479 mg/L/h. Optimization of culture conditions increased the content, concentration, and productivity of isoflavone aglycons by 3.1-, 3.0-, and 3.7-fold, respectively. To our knowledge, this is the highest production of isoflavone aglycons reported to date.

  17. Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis.

    PubMed

    Masarik, Michal; Gumulec, Jaromir; Sztalmachova, Marketa; Hlavna, Marian; Babula, Petr; Krizkova, Sona; Ryvolova, Marketa; Jurajda, Michal; Sochor, Jiri; Adam, Vojtech; Kizek, Rene

    2011-12-01

    Prostate cancer with altered zinc(II) cell metabolism is the second most frequently diagnosed cancer in developed countries. The alterations of zinc(II) metabolism can influence metabolism of other metal ions and can also be associated with the expression and translation of metal-binding proteins including metallothioneins. The aim of this article was to optimize immunoseparation protocol based on paramagnetic beads conjugated with protein G for the isolation of metallothionein. Isolated metallothionein was determined by differential pulse voltammetry Brdicka reaction and SDS-PAGE. Optimal conditions: antigen-binding time - 60 min, temperature - 70°C, and buffer composition and pH - acetate buffer, pH 4.3, were determined. Under the optimized conditions, lysates from 22Rv1 prostate cancer cells treated with various concentrations of cadmium(II) and copper(II) ions were analyzed. We observed strong correlation in all experimental groups and all lysate types (r>0.83 at p<0.041) between metallothionein concentration related to viability and concentration of copper(II) ions and cadmium(II) ions in medium. Moreover, the results were compared with standard sample preparation as heat treatment and SDS-PAGE analysis. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Enzymatic Production of Ascorbic Acid-2-phosphate by Recombinant Acid Phosphatase.

    PubMed

    Zheng, Kai; Song, Wei; Sun, Anran; Chen, Xiulai; Liu, Jia; Luo, Qiuling; Wu, Jing

    2017-05-24

    In this study, an environmentally friendly and efficient enzymatic method for the synthesis of l-ascorbic acid-2-phosphate (AsA-2P) from l-ascorbic acid (AsA) was developed. The Pseudomonas aeruginosa acid phosphatase (PaAPase) was expressed in Escherichia coli BL21. The optimal temperature, optimal pH, K m , k cat , and catalytic efficiency of recombinant PaAPase were 50 °C, 5.0, 93 mM, 4.2 s -1 , and 2.7 mM -1 min -1 , respectively. The maximal dry cell weight and PaAPase phosphorylating activity reached 8.5 g/L and 1127.7 U/L, respectively. The highest AsA-2P concentration (50.0 g/L) and the maximal conversion (39.2%) were obtained by incubating 75 g/L intact cells with 88 g/L AsA and 160 g/L sodium pyrophosphate under optimal conditions (0.1 mM Ca 2+ , pH 4.0, 30 °C) for 10 h; the average AsA-2P production rate was 5.0 g/L/h, and the AsA-2P production system was successfully scaled up to a 7.5 L fermenter. Therefore, the enzymatic process showed great potential for production of AsA-2P in industry.

  19. Encapsulation of brewing yeast in alginate/chitosan matrix: lab-scale optimization of lager beer fermentation.

    PubMed

    Naydenova, Vessela; Badova, Mariyana; Vassilev, Stoyan; Iliev, Vasil; Kaneva, Maria; Kostov, Georgi

    2014-03-04

    Two mathematical models were developed for studying the effect of main fermentation temperature ( T MF ), immobilized cell mass ( M IC ) and original wort extract (OE) on beer fermentation with alginate-chitosan microcapsules with a liquid core. During the experiments, the investigated parameters were varied in order to find the optimal conditions for beer fermentation with immobilized cells. The basic beer characteristics, i.e. extract, ethanol, biomass concentration, pH and colour, as well as the concentration of aldehydes and vicinal diketones, were measured. The results suggested that the process parameters represented a powerful tool in controlling the fermentation time. Subsequently, the optimized process parameters were used to produce beer in laboratory batch fermentation. The system productivity was also investigated and the data were used for the development of another mathematical model.

  20. Collagen-Immobilized Lipases Show Good Activity and Reusability for Butyl Butyrate Synthesis.

    PubMed

    Dewei, Song; Min, Chen; Haiming, Cheng

    2016-11-01

    Candida rugosa lipases were immobilized onto collagen fibers through glutaraldehyde cross-linking method. The immobilization process has been optimized. Under the optimal immobilization conditions, the activity of the collagen-immobilized lipase reached 340 U/g. The activity was recovered of 28.3 % by immobilization. The operational stability of the obtained collagen-immobilized lipase for hydrolysis of olive oil emulsion was determined. The collagen-immobilized lipase showed good tolerance to temperature and pH variations in comparison to free lipase. The collagen-immobilized lipase was also applied as biocatalyst for synthesis of butyl butyrate from butyric acid and 1-butanol in n-hexane. The conversion yield was 94 % at the optimal conditions. Of its initial activity, 64 % was retained after 5 cycles for synthesizing butyl butyrate in n-hexane.

  1. Process optimization for the preparation of straw feedstuff for rearing yellow mealworms (Tenebrio molitor L.) in BLSS

    NASA Astrophysics Data System (ADS)

    Li, Leyuan; Liu, lh64. Hong

    2012-07-01

    It has been confirmed in our previous work that in bioregenerative life support systems, feeding yellow mealworms (Tenebrio molitor L.) using fermented straw has the potential to provide good animal protein for astronauts, meanwhile treating with plant wastes. However, since the nitrogen content in straw is very low, T. molitor larvae can not obtain sufficient nitrogen, which results in a relatively low growth efficiency. In this study, wheat straw powder was mixed with simulated human urine before fermentation. Condition parameters, e.g. urine:straw ratio, moisture content, inoculation dose, fermentation time, fermentation temperature and pH were optimized using Taguchi method. Larval growth rate and average individual mass of mature larva increased significantly in the group of T. molitor larvae fed with feedstuff prepared with the optimized process.

  2. Encapsulation of brewing yeast in alginate/chitosan matrix: lab-scale optimization of lager beer fermentation

    PubMed Central

    Naydenova, Vessela; Badova, Mariyana; Vassilev, Stoyan; Iliev, Vasil; Kaneva, Maria; Kostov, Georgi

    2014-01-01

    Two mathematical models were developed for studying the effect of main fermentation temperature (T MF), immobilized cell mass (M IC) and original wort extract (OE) on beer fermentation with alginate-chitosan microcapsules with a liquid core. During the experiments, the investigated parameters were varied in order to find the optimal conditions for beer fermentation with immobilized cells. The basic beer characteristics, i.e. extract, ethanol, biomass concentration, pH and colour, as well as the concentration of aldehydes and vicinal diketones, were measured. The results suggested that the process parameters represented a powerful tool in controlling the fermentation time. Subsequently, the optimized process parameters were used to produce beer in laboratory batch fermentation. The system productivity was also investigated and the data were used for the development of another mathematical model. PMID:26019512

  3. Zinc tetraaminophthalocyanine-Fe3O4 nanoparticle composite for laccase immobilization

    PubMed Central

    Huang, Jun; Liu, Cheng; Xiao, Haiyan; Wang, Juntao; Jiang, Desheng; GU, Erdan

    2007-01-01

    Zinc tetraaminophthalocyanine-Fe3O4 nanoparticle composites were prepared by organic-inorganic complex technology and characterized. It has been proved that the ZnTAPc dispersed randomly onto the surface of Fe3O4 nanoparticles to form molecular dispersion layer and there was a relatively strong bond between central zinc cation and oxygen. The nanoparticle composite took the shape of roundish spheres with the mean diameter of about 15 nm. Active amino groups of magnetic carriers could be used to bind laccase via glutaraldehyde. The optimal pH for the activity of the immobilized laccases and free laccase were the same at pH 3.0 and the optimal temperature for laccase immobilization on ZnTAPc-Fe3O4 nanoparticle composite was 45°. The immobilization yields and Km value of the laccase immobilized on ZnTAPc-Fe3O4 nanoparticle composite were 25% and 20.1 μM, respectively. This kind of immobilized laccase has good thermal, storage and operation stability, and could be used as the sensing biocomponent for the fiber optic biosensor based on enzyme catalysis. PMID:18203444

  4. Production of Fungal Glucoamylase for Glucose Production from Food Waste

    PubMed Central

    Lam, Wan Chi; Pleissner, Daniel; Lin, Carol Sze Ki

    2013-01-01

    The feasibility of using pastry waste as resource for glucoamylase (GA) production via solid state fermentation (SSF) was studied. The crude GA extract obtained was used for glucose production from mixed food waste. Our results showed that pastry waste could be used as a sole substrate for GA production. A maximal GA activity of 76.1 ± 6.1 U/mL was obtained at Day 10. The optimal pH and reaction temperature for the crude GA extract for hydrolysis were pH 5.5 and 55 °C, respectively. Under this condition, the half-life of the GA extract was 315.0 minutes with a deactivation constant (kd) 2.20 × 10−3 minutes−1. The application of the crude GA extract for mixed food waste hydrolysis and glucose production was successfully demonstrated. Approximately 53 g glucose was recovered from 100 g of mixed food waste in 1 h under the optimal digestion conditions, highlighting the potential of this approach as an alternative strategy for waste management and sustainable production of glucose applicable as carbon source in many biotechnological processes. PMID:24970186

  5. Synthesis and characterization of Cadmium selenide nanoparticles loaded on activated carbon and its efficient application for removal of Muroxide from aqueous solution

    NASA Astrophysics Data System (ADS)

    Ghaedi, M.; Amirabad, S. Zamani; Marahel, F.; Nasiri Kokhdan, S.; Sahraei, R.; Nosrati, M.; Daneshfar, A.

    2011-12-01

    In the first, Cadmium selenide Nanoparticle loaded on activated carbon (CdSe-NP-AC) has been synthesized and characterized by different techniques including XRD and SEM. Then, this new adsorbent successfully has been applied for the removal of muroxide (MO) from aqueous solution in batch studies, while the effect of various experimental parameters like initial pH (pH 0), contact time, amount of (CdSe-NP-AC) and initial MO concentration ( C0) on its removal percentage was examined by one at a time optimization method. It was found following optimization of variable, the adsorption of MO onto (CdSe-NP-AC) followed pseudo-second-order kinetics and show Tempkin and Langmuir models for interpretation of experimental data. It was observed that by increasing the temperature the removal percentage was improved and the positive change in entropy (Δ S°) and heat of adsorption (Δ H°) show the endothermic nature of process, while the high negative value in Gibbs free energy change (Δ G°) indicates the feasible nature of adsorption process.

  6. Screening and optimization of some inorganic salts for the production of ergot alkaloids from Penicillium species using surface culture fermentation process.

    PubMed

    Shahid, Memuna Ghafoor; Nadeem, Muhammad; Baig, Shahjehan; Cheema, Tanzeem Akbar; Atta, Saira; Ghafoor, Gul Zareen

    2016-03-01

    The present study deals with the production of ergot alkaloids from Penicillium commune and Penicillium citrinum, using surface culture fermentation process. Impact of various inorganic salts was tested on the production of ergot alkaloids during the optimization studies of fermentation medium such as impact of various concentration levels of succinic acid, ammonium chloride, MgSO4, FeSO4, ZnSO4, pH and the effect of various incubation time periods was also determined on the production of ergot alkaloids from Penicillium commune and Penicillium citrinum. Highest yield of ergot alkaloids was obtained when Penicillium commune and Penicillium citrinum that were grown on optimum levels of ingredients such as 2 g succinic acid, 1.5 and 2 g NH4Cl, 1.5 g MgSO4, 1 g FeSO4, 1 and 1.5 g ZnSO4 after 21 days of incubation time period using pH 5 at 25(°)C incubation temperature in the fermentation medium. Ergot alkaloids were determined using Spectrophotometry and Thin Layer Chromatography (TLC) techniques.

  7. Catalytic characteristics and application of l-asparaginase immobilized on aluminum oxide pellets.

    PubMed

    Agrawal, Sarika; Sharma, Isha; Prajapati, Bhanu Pratap; Suryawanshi, Rahul Kumar; Kango, Naveen

    2018-07-15

    l-asparaginase from Escherichia coli (l-ASNase) was covalently immobilized on aluminum oxide pellets (AlOPs) using a cross-linking agent, glutaraldehyde. Maximum immobilization yield (85.0%) was obtained after optimizing immobilization parameters using response surface methodology (RSM). Both free and immobilized l-ASNase (AlOP-ASNase) were optimally active at 37°C and pH7.5. However, the bioconjugate exhibited enhanced activity and stability at different pH and temperatures. It had higher affinity (low K m ) and was comparatively more stable in presence of some solvents (ethyl acetate, acetone, acetonitrile), metal ions (Ag + , Zn 2+ ) and β-mercaptoethanol. AlOP-ASNase was reused in a glass column reactor for l-asparagine hydrolysis upto nine successive cycles without any loss in activity. The AlOP-ASNase was effective in lowering l-asparagine level in blanched potato chips indicating its potential use in mitigating acrylamide formation in starchy foods. This cost-effective enzyme preparation had shelf-life of more than 30days and can be effectively used in starch based food industries. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. Production of high activity Aspergillus niger BCC4525 β-mannanase in Pichia pastoris and its application for mannooligosaccharides production from biomass hydrolysis.

    PubMed

    Harnpicharnchai, Piyanun; Pinngoen, Waraporn; Teanngam, Wanwisa; Sornlake, Warasirin; Sae-Tang, Kittapong; Manitchotpisit, Pennapa; Tanapongpipat, Sutipa

    2016-12-01

    A cDNA encoding β-mannanase was cloned from Aspergillus niger BCC4525 and expressed in Pichia pastoris KM71. The secreted enzyme hydrolyzed locust bean gum substrate with very high activity (1625 U/mL) and a relatively high k cat /K m (461 mg -1 s -1  mL). The enzyme is thermophilic and thermostable with an optimal temperature of 70 °C and 40% retention of endo-β-1,4-mannanase activity after preincubation at 70 °C. In addition, the enzyme exhibited broad pH stability with an optimal pH of 5.5. The recombinant enzyme hydrolyzes low-cost biomass, including palm kernel meal (PKM) and copra meal, to produce mannooligosaccharides, which is used as prebiotics to promote the growth of beneficial microflora in animals. An in vitro digestibility test simulating the gastrointestinal tract system of broilers suggested that the recombinant β-mannanase could effectively liberate reducing sugars from PKM-containing diet. These characteristics render this enzyme suitable for utilization as a feed additive to improve animal performance.

  9. Purification and Antithrombotic Potential of a Fibrinolytic Enzyme from Shiitake Culinary- Medicinal Mushroom, Lentinus edodes GNA01 (Agaricomycetes).

    PubMed

    Choi, Jun-Hui; Kim, Kyung-Je; Kim, Seung

    2018-01-01

    We purified Lentinus edodes GNA01 fibrinolytic enzyme (LEFE) and identified it as a novel metalloprotease. LEFE was purified to homogeneity through a 2-step procedure, with an 8.28-fold increase in specific activity and 5.3% recovery. The molecular mass of LEFE was approximately 38 kDa, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its optimal pH, optimal temperature, pH stability, and thermal stability were 5, 30°C, 6-7, and 40°C, respectively. LEFE was inhibited by zinc and magnesium ions, and by EDTA and EGTA, indicating that LEFE is a metalloprotease. The protease exhibited fibrinolytic activity and a degradative effect on clot formation and blood clots. The protease prolonged activated partial thromboplastin time, prothrombin time, and coagulation time as induced by platelet aggregators (collagen and epinephrine). Taken together, our results indicate that L. edodes GNA01 produces a metalloprotease/fibrinolytic enzyme and that this enzyme might be applied as a new thrombolytic and antithrombotic agent for thrombosis-related cardiovascular disorders.

  10. The initial freezing point temperature of beef rises with the rise in pH: a short communication.

    PubMed

    Farouk, M M; Kemp, R M; Cartwright, S; North, M

    2013-05-01

    This study tested the hypothesis that the initial freezing point temperature of meat is affected by pH. Sixty four bovine M. longissimus thoracis et lumborum were classified into two ultimate pH groups: low (<5.8) and high pH (>6.2) and their cooling and freezing point temperatures were determined. The initial freezing temperatures for beef ranged from -0.9 to -1.5°C (∆=0.6°C) with the higher and lower temperatures associated with high and low ultimate pH respectively. There was a significant correlation (r=+0.73, P<0.01) between beef pH and freezing point temperature in the present study. The outcome of this study has implications for the meat industry where evidence of freezing (ice formation) in a shipment as a result of high pH meat could result in a container load of valuable chilled product being downgraded to a lower value frozen product. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Biosorption of Cr(VI) by Ceratocystis paradoxa MSR2 Using Isotherm Modelling, Kinetic Study and Optimization of Batch Parameters Using Response Surface Methodology

    PubMed Central

    Ramalingam, Chidambaram

    2015-01-01

    This study is focused on the possible use of Ceratocystis paradoxa MSR2 native biomass for Cr(VI) biosorption. The influence of experimental parameters such as initial pH, temperature, biomass dosage, initial Cr(VI) concentration and contact time were optimized using batch systems as well as response surface methodology (RSM). Maximum Cr(VI) removal of 68.72% was achieved, at an optimal condition of biomass dosage 2g L−1, initial Cr(VI) concentration of 62.5 mg L−1 and contact time of 60 min. The closeness of the experimental and the predicted values exhibit the success of RSM. The biosorption mechanism of MSR2 biosorbent was well described by Langmuir isotherm and a pseudo second order kinetic model, with a high regression coefficient. The thermodynamic study also revealed the spontaneity and exothermic nature of the process. The surface characterization using FT-IR analysis revealed the involvement of amine, carbonyl and carboxyl groups in the biosorption process. Additionally, desorption efficiency of 92% was found with 0.1 M HNO3. The Cr(VI) removal efficiency, increased with increase in metal ion concentration, biomass concentration, temperature but with a decrease in pH. The size of the MSR2 biosorbent material was found to be 80 μm using particle size analyzer. Atomic force microscopy (AFM) visualizes the distribution of Cr(VI) on the biosorbent binding sites with alterations in the MSR2 surface structure. The SEM-EDAX analysis was also used to evaluate the binding characteristics of MSR2 strain with Cr(VI) metals. The mechanism of Cr(VI) removal of MSR2 biomass has also been proposed. PMID:25822726

  12. A novel malic enzyme gene, Mime2, from Mortierella isabellina M6-22 contributes to lipid accumulation.

    PubMed

    Li, Shan; Li, Lingyan; Xiong, Xiangfeng; Ji, Xiuling; Wei, Yunlin; Lin, Lianbing; Zhang, Qi

    2018-05-18

    This study was aimed at cloning and characterizing a novel malic enzyme (ME) gene of Mortierella isabellina M6-22 and identifying its relation with lipid accumulation. Mime2 was cloned from strain M6-22. Plasmid pET32aMIME2 was constructed to express ME of MIME2 in Escherichia coli BL21. After purification, the optimal pH and temperature of MIME2, as well as K m and V max for NADP + were determined. The effects of EDTA or metal ions (Mn 2+ , Mg 2+ , Co 2+ , Cu 2+ , Ca 2+ , or Zn 2+ ) on the enzymatic activity of MIME2 were evaluated. Besides, plasmid pRHMIME2 was created to express MIME2 in Rhodosporidium kratochvilovae YM25235, and its cell lipid content was measured by the acid-heating method. The optimal pH and temperature of MIME2 are 5.8 and 30 °C, respectively. The act ivity of MIME2 was significantly increased by Mg 2+ , Ca 2+ , or Mn 2+ at 0.5 mM but inhibited by Cu 2+ or Zn 2+ (p < 0.05). The optimal enzymatic activity of MIME2 is 177.46 U/mg, and the K m and V max for NADP + are 0.703 mM and 156.25 μg/min, respectively. Besides, Mime2 transformation significantly increased the cell lipid content in strain YM25235 (3.15 ± 0.24 vs. 2.17 ± 0.31 g/L, p < 0.01). The novel ME gene Mime2 isolated from strain M6-22 contributes to lipid accumulation in strain YM25235.

  13. Bioprocess and downstream optimization of recombinant bovine chymosin B in Pichia (Komagataella) pastoris under methanol-inducible AOXI promoter.

    PubMed

    Noseda, Diego Gabriel; Blasco, Martín; Recúpero, Matías; Galvagno, Miguel Ángel

    2014-12-01

    A clone of the methylotrophic yeast Pichia pastoris strain GS115 transformed with the bovine prochymosin B gene was used to optimize the production and downstream of recombinant bovine chymosin expressed under the methanol-inducible AOXI promoter. Cell growth and recombinant chymosin production were analyzed in flask cultures containing basal salts medium with biodiesel-byproduct glycerol as the carbon source, obtaining values of biomass level and milk-clotting activity similar to those achieved with analytical glycerol. The effect of biomass level at the beginning of methanol-induction phase on cell growth and chymosin expression was evaluated, determining that a high concentration of cells at the start of such period generated an increase in the production of chymosin. The impact of the specific growth rate on chymosin expression was studied throughout the induction stage by methanol exponential feeding fermentations in a lab-scale stirred bioreactor, achieving the highest production of heterologous chymosin with a constant specific growth rate of 0.01h(-1). By gel filtration chromatography performed at a semi-preparative scale, recombinant chymosin was purified from exponential fed-batch fermentation cultures, obtaining a specific milk-clotting activity of 6400IMCU/mg of chymosin and a purity level of 95%. The effect of temperature and pH on milk-clotting activity was analyzed, establishing that the optimal temperature and pH values for the purified recombinant chymosin are 37°C and 5.5, respectively. This study reported the features of a sustainable bioprocess for the production of recombinant bovine chymosin in P. pastoris by fermentation in stirred-tank bioreactors using biodiesel-derived glycerol as a low-cost carbon source. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Production and characterization of bioactive metabolites from piezotolerant deep sea fungus Nigrospora sp. in submerged fermentation.

    PubMed

    Arumugam, G K; Srinivasan, S K; Joshi, G; Gopal, D; Ramalingam, K

    2015-01-01

    To produce and characterize bioactive metabolites from piezotolerant marine fungus Nigrospora sp. in submerged fermentation. A distinct marine strain, Nigrospora sp. NIOT has been isolated from a depth of 800 m at the Arabian Sea. The 18S rRNA and internal transcribed spacers (ITS) analysis demonstrates its close association with the genus Nigrospora. Effect of pH, temperature, salinity, carbon source and amino acids was studied to optimize the fermentation conditions. Optimal mycelia growth and secondary metabolites production were observed at 6·0-8·0 pH, 20-30°C temperature, 7·5% salinity, sucrose as carbon source and tryptophan as amino acid source. The extracellular secondary metabolites exhibited high antimicrobial activities against both gram-positive and gram-negative pathogenic bacteria with minimal inhibitory concentration (MIC) values higher than 30 μg ml(-1). Strongest cytotoxicity was observed in all cell lines tested, GI50 (growth inhibition by 50%) was calculated to be 1·35, 3·2, 0·13 and 0·35 μg ml(-1) against U937, MCF-7, A673 and Jurkat, respectively. Fourier transform infrared spectroscopy (FTIR) and gas chromatography-mass spectrometry (GC-MS) analyses of secondary metabolites confirmed the production of antimicrobial and anticancer substances. A piezotolerant fungus Nigrospora sp. NIOT isolated from deep sea environment was successfully cultured under submerged fermentation. The secondary metabolites produced from this organism showed potent antimicrobial and anticancer activities with immediate application to cosmetics and pharmaceutical industries. This is the first study exploring Nigrospora sp. from 800 m in marine environment. This deep sea fungus under optimized culture conditions effectively produced bioactive secondary metabolites such as griseofulvin, spirobenzofuran and pyrone derivatives at higher concentrations. © 2014 The Society for Applied Microbiology.

  15. Preparation and Optimisation of Cross-Linked Enzyme Aggregates Using Native Isolate White Rot Fungi Trametes versicolor and Fomes fomentarius for the Decolourisation of Synthetic Dyes

    PubMed Central

    Vršanská, Martina; Voběrková, Stanislava; Jiménez Jiménez, Ana María; Strmiska, Vladislav; Adam, Vojtěch

    2017-01-01

    The key to obtaining an optimum performance of an enzyme is often a question of devising a suitable enzyme and optimisation of conditions for its immobilization. In this study, laccases from the native isolates of white rot fungi Fomes fomentarius and/or Trametes versicolor, obtained from Czech forests, were used. From these, cross-linked enzyme aggregates (CLEA) were prepared and characterised when the experimental conditions were optimized. Based on the optimization steps, saturated ammonium sulphate solution (75 wt.%) was used as the precipitating agent, and different concentrations of glutaraldehyde as a cross-linking agent were investigated. CLEA aggregates formed under the optimal conditions showed higher catalytic efficiency and stabilities (thermal, pH, and storage, against denaturation) as well as high reusability compared to free laccase for both fungal strains. The best concentration of glutaraldehyde seemed to be 50 mM and higher efficiency of cross-linking was observed at a low temperature 4 °C. An insignificant increase in optimum pH for CLEA laccases with respect to free laccases for both fungi was observed. The results show that the optimum temperature for both free laccase and CLEA laccase was 35 °C for T. versicolor and 30 °C for F. fomentarius. The CLEAs retained 80% of their initial activity for Trametes and 74% for Fomes after 70 days of cultivation. Prepared cross-linked enzyme aggregates were also investigated for their decolourisation activity on malachite green, bromothymol blue, and methyl red dyes. Immobilised CLEA laccase from Trametes versicolor showed 95% decolourisation potential and CLEA from Fomes fomentarius demonstrated 90% decolourisation efficiency within 10 h for all dyes used. These results suggest that these CLEAs have promising potential in dye decolourisation. PMID:29295505

  16. Expression of food-grade phytase in Lactococcus lactis from optimized conditions in milk broth.

    PubMed

    Miao, Yuzhi; Xu, Hui; Fei, Baojin; Qiao, Dairong; Cao, Yi

    2013-07-01

    The major objective of this study was to engineer lactic acid bacteria to produce the enzyme phytase from a gene native to Bacillus subtilis GYPB04. The phytase gene (phyC) of B. subtilis GYPB04 was cloned into the plasmid pMG36e for expression in Lactococcus lactis. The enzyme activity in L. lactis cultured in GM17 broth was 20.25 U/mL at 36°C. The expressed phytase was characterized as active in a pH range of 2.0-9.0 at a temperature range of 20-80°C, with an optimum pH of 5.5-6.5 and temperature of 60°C. When cultured in food-grade milk broth, the transformed L. lactis grew to an OD(600 nm) value of 1.05 and had a phytase yield of 13.58 U/mL. In same broth under optimized conditions for cell growth and phytase production, the transformant reached an OD(600 nm) value of 1.68 and a phytase yield of 42.12 U/mL, representing approximately 1.6-fold and 3.1-fold increases, respectively, compared to growth in natural milk broth. Fermentation was scaled to 5 L under optimized conditions, and product analysis revealed a final OD(600 nm) value of 1.89 and an extracellular enzyme activity of 24.23 U/mL. The results of this study may be used in the dairy fermentation industry for the development of functional, healthy yogurts and other fermented dairy foods that provide both active phytase and viable probiotics to the consumer. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  17. Biodegradation of waste lubricants by a newly isolated Ochrobactrum sp. C1.

    PubMed

    Bhattacharya, Munna; Biswas, Dipa; Sana, Santanu; Datta, Sriparna

    2015-10-01

    A potential degrader of paraffinic and aromatic hydrocarbons was isolated from oil-contaminated soil from steel plant effluent area in Burnpur, India. The strain was investigated for degradation of waste lubricants (waste engine oil and waste transformer oil) that often contain EPA (Environmental Protection Agency, USA) classified priority pollutants and was identified as Ochrobactrum sp. C1 by 16S rRNA gene sequencing. The strain C1 was found to tolerate unusually high waste lubricant concentration along with emulsification capability of the culture broth, and its degradation efficiency was 48.5 ± 0.5 % for waste engine oil and 30.47 ± 0.25 % for waste transformer oil during 7 days incubation period. In order to get optimal degradation efficiency, a three level Box-Behnken design was employed to optimize the physical parameters namely pH, temperature and waste oil concentration. The results indicate that at temperature 36.4 °C, pH 7.3 and with 4.6 % (v/v) oil concentration, the percentage degradation of waste engine oil will be 57 % within 7 days. At this optimized condition, the experimental values (56.7 ± 0.25 %) are in a good agreement with the predicted values with a calculated R 2 to be 0.998 and significant correlation between biodegradation and emulsification activity (E 24  = 69.42 ± 0.32 %) of the culture broth toward engine oil was found with a correlation coefficient of 0.972. This is the first study showing that an Ochrobactrum sp. strain is capable of degrading waste lubricants, which might contribute to the bioremediation of waste lubricating oil-contaminated soil.

  18. Interaction of diazepam with surfactants. Spectrophotometric and spectrofluorometric study

    NASA Astrophysics Data System (ADS)

    De La Guardia, M.; Rodilla, F.

    1986-03-01

    The interaction of diazepam with non-ionic, anionic and cationic surfactants has been studied spectrophotometrically and fluorometrically. It has been verified that the absorption spectrum of diazepam is not modified in micellar medium. However, a dramatic five-fold increase in fluorescence sensitivity is observed in the presence of sodium lauryl sulphate (SDS). The experimental conditions, temperature, pH and surfactant concentration have been optimized to improve the fluorometric determination of diazepam and a detection limit of 0,04 ppmhas been obtained.

  19. [Leaching of Rare Earth Elements from Coal Ashes Using Acidophilic Chemolithotrophic Microbial Communities].

    PubMed

    Muravyov, M I; Bulaev, A G; Melamud, V S; Kondrat'eva, T F

    2015-01-01

    A method for leaching rare earth elements from coal ash in the presence of elemental sulfur using communities of acidophilic chemolithotrophic microorganisms was proposed. The optimal parameters determined for rare element leaching in reactors were as follows: temperature, 45 degrees C; initial pH, 2.0; pulp density, 10%; and the coal ash to elemental sulfur ratio, 10 : 1. After ten days of leaching, 52.0, 52.6, and 59.5% of scandium, yttrium, and lanthanum, respectively, were recovered.

  20. Study on the preparation process of cross-linked porous cassava starch

    NASA Astrophysics Data System (ADS)

    Yin, Xiulian; You, Qinghong; Wan, Miaomiao; Zhang, Xuejuan; Dai, Chunhua

    2017-04-01

    Using cassava starch as raw material, preparation process of porous cross-linked cassava starch was studied. Using TSTP as cross-linking agents, Orthogonal design was applied for the optimization of cross-linked porous starch preparation process. The results showed that the opitmal conditions of cross-linked porous cassava starch were as follows: reaction temperature 45°C, reaction time 20 h, 1% of the amount of the enzyme, the enzyme ratio of 1:5, pH 5.50, substrate concentration of 40%.

  1. Improving the Catalytic Activity of Hyperthermophilic Pyrococcus horikoshii Prolidase for Detoxification of Organophosphorus Nerve Agents over a Broad Range of Temperatures

    DTIC Science & Technology

    2011-01-01

    affinity for metal, and increased thermostability compared to P. furiosus prolidase, Pf prol (PF1343). To obtain a better enzyme for OP nerve agent...decontamination and to investigate the structural factors that may influence protein thermostability and thermoactivity, randomly mutated Ph1prol enzymes ...Introduction Pyrococcus horikoshii and Pyrococcus furiosus are both hyper- thermophilic archaea, growing optimally at 98 –100◦C that were isolated from a

  2. Electroless plating Cu-Co-P polyalloy on UV/ozonolysis irradiated polyethylene terephthalate film and its corrosion resistance

    NASA Astrophysics Data System (ADS)

    Hou, Lei; Bi, Siyi; Zhao, Hang; Xu, Yumeng; Mu, Yuhang; Lu, Yinxiang

    2017-05-01

    High corrosion resistant Cu-Co-P coatings were firstly prepared on polyethylene terephthalate (PET) substrate by electroless plating in combination with UV/ozonolysis irradiation under optimized cobalt sulfate heptahydrate concentration, pH value, plating temperature and time. The copper polyalloy/PET composite can be obtained in three steps, namely: (i) the generation of oxygen-containing functionalities (carboxylic groups) onto PET surface through UV irradiation combined with ozone, (ii) Cu seeding catalysts were obtained after being immersed into cupric citrate and NaBH4 solutions subsequently, and (iii) Cu-Co-P polyalloy metallization using electroless plating bath. Attenuated total reflection fourier transformation infrared spectrometer (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), water contact angle measurement and energy dispersive X-ray analysis (EDAX) were utilized to track the surface changes during the whole process. The electroless plating conditions were optimized by an orthogonal experiment (L9(3)4) for Cu-Co-P coating as follows: CoSO4·7H2O addition of 0.08 M, pH value, plating temperature and time were set on 10.0, 35 °C and 25 min, respectively. Under the optimal conditions, copper polyalloy possessed high adhesive strength and the lowest surface resistance (8.06 Ω/sq), while maintaining reliability even after over 1000 times of bending and mechanical stress. The results of scanning electron microscope (SEM) and atomic force microscope (AFM) measurements showed that Cu-Co-P layer formed on PET surface was imparted with fine uniformity and high compactness. Electrochemical test revealed the optimized Cu-Co-P coatings exhibited high corrosion resistance in NaCl, NaOH and HCl solutions, respectively. The excellent electromagnetic interference shielding effectiveness (EMI SE >99.999% at frequency ranging from 30 MHz to 1000 MHz) of copper polyalloy/PET composites was confirmed by the spectrum analyzer. Therefore, this copper polyalloy will have potential applications in microelectronics packaging and coatings for anti-corrosion and electromagnetic interference shielding.

  3. Enhanced saturated fatty acids accumulation in cultures of newly-isolated strains of Schizochytrium sp. and Thraustochytriidae sp. for large-scale biodiesel production.

    PubMed

    Wang, Qiuzhen; Sen, Biswarup; Liu, Xianhua; He, Yaodong; Xie, Yunxuan; Wang, Guangyi

    2018-08-01

    Heterotrophic marine protists (Thraustochytrids) have received increasingly global attention as a renewable, sustainable and alternative source of biodiesel because of their high ability of saturated fatty acids (SFAs) accumulation. Yet, the influence of extrinsic factors (nutrients and environmental conditions) on thraustochytrid culture and optimal conditions for high SFAs production are poorly described. In the present study, two different thraustochytrid strains, Schizochytrium sp. PKU#Mn4 and Thraustochytriidae sp. PKU#Mn16 were studied for their growth and SFAs production profiles under various conditions (carbon, nitrogen, temperature, pH, KH 2 PO 4 , salinity, and agitation speed). Of the culture conditions, substrates (C and N) source and conc., temperature, and agitation speed significantly influenced the cell growth and SFAs production of both strains. Although both the strains were capable of growth and SFAs production in the broad range of culture conditions, their physiological responses to KH 2 PO 4 , pH, and salinity were dissimilar. Under their optimal batch culture conditions, peak SFAs productions of 3.3g/L and 2.2g/L with 62% and 49% SFAs contents (relative to total fatty acids) were achieved, respectively. The results of 5-L fed-batch fermentation under optimal conditions showed a nearly 4.5-fold increase in SFAs production (i.e., 7.5g/L) by both strains compared to unoptimized conditions. Of the two strains, the quality of biodiesel produced from the fatty acids of PKU#Mn4 met the biodiesel standard defined by ASTM6751. This study, to the knowledge of the authors, is the first comprehensive report of optimal fermentation conditions demonstrating enhanced SFAs production by strains belonging to two different thraustochytrid genera and provides the basis for large-scale biodiesel production. Copyright © 2018. Published by Elsevier B.V.

  4. An Optimized Method for the Measurement of Acetaldehyde by High-Performance Liquid Chromatography

    PubMed Central

    Guan, Xiangying; Rubin, Emanuel; Anni, Helen

    2011-01-01

    Background Acetaldehyde is produced during ethanol metabolism predominantly in the liver by alcohol dehydrogenase, and rapidly eliminated by oxidation to acetate via aldehyde dehydrogenase. Assessment of circulating acetaldehyde levels in biological matrices is performed by headspace gas chromatography and reverse phase high-performance liquid chromatography (RP-HPLC). Methods We have developed an optimized method for the measurement of acetaldehyde by RP-HPLC in hepatoma cell culture medium, blood and plasma. After sample deproteinization, acetaldehyde was derivatized with 2,4-dinitrophenylhydrazine (DNPH). The reaction was optimized for pH, amount of derivatization reagent,, time and temperature. Extraction methods of the acetaldehyde-hydrazone (AcH-DPN) stable derivative and product stability studies were carried out. Acetaldehyde was identified by its retention time in comparison to AcH-DPN standard, using a new chromatography gradient program, and quantitated based on external reference standards and standard addition calibration curves in the presence and absence of ethanol. Results Derivatization of acetaldehyde was performed at pH 4.0 with a 80-fold molar excess of DNPH. The reaction was completed in 40 min at ambient temperature, and the product was stable for 2 days. A clear separation of AcH-DNP from DNPH was obtained with a new 11-min chromatography program. Acetaldehyde detection was linear up to 80 μM. The recovery of acetaldehyde was >88% in culture media, and >78% in plasma. We quantitatively determined the ethanol-derived acetaldehyde in hepatoma cells, rat blood and plasma with a detection limit around 3 μM. The accuracy of the method was <9% for intraday and <15% for interday measurements, in small volume (70 μl) plasma sampling. Conclusions An optimized method for the quantitative determination of acetaldehyde in biological systems was developed using derivatization with DNPH, followed by a short RP-HPLC separation of AcH-DNP. The method has an extended linear range, is reproducible and applicable to small volume sampling of culture media and biological fluids. PMID:21895715

  5. An optimized method for the measurement of acetaldehyde by high-performance liquid chromatography.

    PubMed

    Guan, Xiangying; Rubin, Emanuel; Anni, Helen

    2012-03-01

    Acetaldehyde is produced during ethanol metabolism predominantly in the liver by alcohol dehydrogenase and rapidly eliminated by oxidation to acetate via aldehyde dehydrogenase. Assessment of circulating acetaldehyde levels in biological matrices is performed by headspace gas chromatography and reverse phase high-performance liquid chromatography (RP-HPLC). We have developed an optimized method for the measurement of acetaldehyde by RP-HPLC in hepatoma cell culture medium, blood, and plasma. After sample deproteinization, acetaldehyde was derivatized with 2,4-dinitrophenylhydrazine (DNPH). The reaction was optimized for pH, amount of derivatization reagent, time, and temperature. Extraction methods of the acetaldehyde-hydrazone (AcH-DNP) stable derivative and product stability studies were carried out. Acetaldehyde was identified by its retention time in comparison with AcH-DNP standard, using a new chromatography gradient program, and quantitated based on external reference standards and standard addition calibration curves in the presence and absence of ethanol. Derivatization of acetaldehyde was performed at pH 4.0 with an 80-fold molar excess of DNPH. The reaction was completed in 40 minutes at ambient temperature, and the product was stable for 2 days. A clear separation of AcH-DNP from DNPH was obtained with a new 11-minute chromatography program. Acetaldehyde detection was linear up to 80 μM. The recovery of acetaldehyde was >88% in culture media and >78% in plasma. We quantitatively determined the ethanol-derived acetaldehyde in hepatoma cells, rat blood and plasma with a detection limit around 3 μM. The accuracy of the method was <9% for intraday and <15% for interday measurements, in small volume (70 μl) plasma sampling. An optimized method for the quantitative determination of acetaldehyde in biological systems was developed using derivatization with DNPH, followed by a short RP-HPLC separation of AcH-DNP. The method has an extended linear range, is reproducible and applicable to small-volume sampling of culture media and biological fluids. Copyright © 2011 by the Research Society on Alcoholism.

  6. Removal of Malachite Green Dye by Mangifera indica Seed Kernel Powder

    NASA Astrophysics Data System (ADS)

    Singh, Dilbagh; Sowmya, V.; Abinandan, S.; Shanthakumar, S.

    2017-11-01

    In this study, batch experiments were carried out to study the adsorption of Malachite green dye from aqueous solution by Mangifera indica (mango) seed kernel powder. The mango seed kernel powder was characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. Effect of various parameters including pH, contact time, adsorbent dosage, initial dye concentration and temperature on adsorption capacity of the adsorbent was observed and the optimized condition for maximum dye removal was identified. Maximum percentage removal of 96% was achieved with an adsorption capacity of 22.8 mg/g at pH 6 with an initial concentration of 100 mg/l. The equilibrium data were examined to fit the Langmuir and Freundlich isotherm models. Thermodynamic parameters for the adsorption process were also calculated.

  7. Bioproperties of potent nattokinase from Bacillus subtilis YJ1.

    PubMed

    Yin, Li-Jung; Lin, Hsin-Hung; Jiang, Shann-Tzong

    2010-05-12

    Fibrinolytic enzyme activity was observed during cultivation of Bacillus subtilis YJ1 in a medium containing 1% skim milk, 1% rice husk, 0.5% NaCl, and 0.25% glucose. It was purified to electrophoretical homogeneity after CM-sepharose FF chromatography. The specific activity and yield were 1791.9 FU/mg and 9.5%, respectively. This purified fibrinolytic enzyme had M of 27.5 kDa, optimal temperature and pH at 50 degrees C and 8.5, respectively. It was stable at pH 6.0-10.0 and 10-40 degrees C and inhibited by Fe(3+), Hg(2+), Cu(2+), Zn(2+), and PMSF. Compared the N terminal of amino acids and full DNA sequence with those in NCBI, it was considered to be a nattokinase.

  8. The immobilization of lipase on PVDF-co-HFP membrane

    NASA Astrophysics Data System (ADS)

    Kayhan, Naciye; Eyüpoǧlu, Volkan; Adem, Şevki

    2016-04-01

    Lipase is an enzyme having a lot of different industrial applications such as biodiesel production, biopolymer synthesis, enantiopure pharmaceutical productions, agrochemicals, etc. Its immobilized form on different substances is more conventional and useful than its free form. Supporting material was prepared using PVDF-co-HFP in laboratory conditions and attached 1,4-diaminobutane (DA) and epichlorohydrin (EPI) ligands to the membrane to immobilize lipase enzyme. The immobilization conditions such as enzyme amount, pH, the concentration of salt, thermal stability and activity were stabilized for our experimental setup. Then, biochemical characterizations were performed on immobilized lipase PVDF-co-HFP regarding optimal pH activity, temperature and thermal stability. Also, the desorption ratios of immobilized enzyme in two different pathway were investigated to confirm immobilization stability for 24 hours.

  9. A mathematical model for the generation and control of a pH gradient in an immobilized enzyme system involving acid generation.

    PubMed

    Chen, G; Fournier, R L; Varanasi, S

    1998-02-20

    An optimal pH control technique has been developed for multistep enzymatic synthesis reactions where the optimal pH differs by several units for each step. This technique separates an acidic environment from a basic environment by the hydrolysis of urea within a thin layer of immobilized urease. With this technique, a two-step enzymatic reaction can take place simultaneously, in proximity to each other, and at their respective optimal pH. Because a reaction system involving an acid generation represents a more challenging test of this pH control technique, a number of factors that affect the generation of such a pH gradient are considered in this study. The mathematical model proposed is based on several simplifying assumptions and represents a first attempt to provide an analysis of this complex problem. The results show that, by choosing appropriate parameters, the pH control technique still can generate the desired pH gradient even if there is an acid-generating reaction in the system. Copyright 1998 John Wiley & Sons, Inc.

  10. Self-collimating photonic crystal polarization beam splitter.

    PubMed

    Zabelin, V; Dunbar, L A; Le Thomas, N; Houdré, R; Kotlyar, M V; O'Faolain, L; Krauss, T F

    2007-03-01

    We present theoretical and experimental results of a polarization splitter device that consists of a photonic crystal (PhC) slab, which exhibits a large reflection coefficient for TE and a high transmission coefficient for TM polarization. The slab is embedded in a PhC tile operating in the self-collimation mode. Embedding the polarization-discriminating slab in a PhC with identical lattice symmetry suppresses the in-plane diffraction losses at the PhC-non-PhC interface. The optimization of the PhC-non-PhC interface is thereby decoupled from the optimization of the polarizing function. Transmissions as high as 35% for TM- and 30% for TE-polarized light are reported.

  11. A gold nanocluster-based fluorescent probe for simultaneous pH and temperature sensing and its application to cellular imaging and logic gates

    NASA Astrophysics Data System (ADS)

    Wu, Yun-Tse; Shanmugam, Chandirasekar; Tseng, Wei-Bin; Hiseh, Ming-Mu; Tseng, Wei-Lung

    2016-05-01

    Metal nanocluster-based nanomaterials for the simultaneous determination of temperature and pH variations in micro-environments are still a challenge. In this study, we develop a dual-emission fluorescent probe consisting of bovine serum albumin-stabilized gold nanoclusters (BSA-AuNCs) and fluorescein-5-isothiocyanate (FITC) as temperature- and pH-responsive fluorescence signals. Under single wavelength excitation the FITC/BSA-AuNCs exhibited well-separated dual emission bands at 525 and 670 nm. When FITC was used as a reference fluorophore, FITC/BSA-AuNCs showed a good linear response over the temperature range 1-71 °C and offered temperature-independent spectral shifts, temperature accuracy, activation energy, and reusability. The possible mechanism for high temperature-induced fluorescence quenching of FITC/BSA-AuNCs could be attributed to a weakening of the Au-S bond, thereby lowering the charge transfer from BSA to AuNCs. Additionally, the pH- and temperature-responsive properties of FITC/BSA-AuNCs allow simultaneous temperature sensing from 21 to 41 °C (at intervals of 5 °C) and pH from 6.0 to 8.0 (at intervals of 0.5 pH unit), facilitating the construction of two-input AND logic gates. Three-input AND logic gates were also designed using temperature, pH, and trypsin as inputs. The practicality of using FITC/BSA-AuNCs to determine the temperature and pH changes in HeLa cells is also validated.Metal nanocluster-based nanomaterials for the simultaneous determination of temperature and pH variations in micro-environments are still a challenge. In this study, we develop a dual-emission fluorescent probe consisting of bovine serum albumin-stabilized gold nanoclusters (BSA-AuNCs) and fluorescein-5-isothiocyanate (FITC) as temperature- and pH-responsive fluorescence signals. Under single wavelength excitation the FITC/BSA-AuNCs exhibited well-separated dual emission bands at 525 and 670 nm. When FITC was used as a reference fluorophore, FITC/BSA-AuNCs showed a good linear response over the temperature range 1-71 °C and offered temperature-independent spectral shifts, temperature accuracy, activation energy, and reusability. The possible mechanism for high temperature-induced fluorescence quenching of FITC/BSA-AuNCs could be attributed to a weakening of the Au-S bond, thereby lowering the charge transfer from BSA to AuNCs. Additionally, the pH- and temperature-responsive properties of FITC/BSA-AuNCs allow simultaneous temperature sensing from 21 to 41 °C (at intervals of 5 °C) and pH from 6.0 to 8.0 (at intervals of 0.5 pH unit), facilitating the construction of two-input AND logic gates. Three-input AND logic gates were also designed using temperature, pH, and trypsin as inputs. The practicality of using FITC/BSA-AuNCs to determine the temperature and pH changes in HeLa cells is also validated. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr02341j

  12. Biotherapeutic formulation factors affecting metal leachables from stainless steel studied by design of experiments.

    PubMed

    Zhou, Shuxia; Evans, Brad; Schöneich, Christian; Singh, Satish K

    2012-03-01

    Trace amounts of metals are inevitably present in biotherapeutic products. They can arise from various sources. The impact of common formulation factors such as protein concentration, antioxidant, metal chelator concentration and type, surfactant, pH, and contact time with stainless steel on metal leachables was investigated by a design of experiments approach. Three major metal leachables, iron, chromium, and nickel were monitored by inductively coupled plasma-mass spectrometry. It was observed that among all the tested factors, contact time, metal chelator concentration, and protein concentration were statistically significant factors with higher temperature resulting in higher levels of leached metals. Within a pH range of 5.5-6.5, solution pH played a minor role for chromium leaching at 25°C. No statistically significant difference was observed due to type of chelator, presence of antioxidant, or surfactant. In order to optimize a biotherapeutic formulation to achieve a target drug product shelf life with acceptable quality, each formulation component must be evaluated for its impact.

  13. Production of surfactant and detergent-stable, halophilic, and alkalitolerant alpha-amylase by a moderately halophilic Bacillus sp. Strain TSCVKK.

    PubMed

    Kiran, Kondepudi Kanthi; Chandra, T S

    2008-01-01

    A moderately halophilic alkalitolerant Bacillus sp. Strain TSCVKK, with an ability to produce extracellular halophilic, alkalitolerant, surfactant, and detergent-stable alpha-amylase was isolated from soil samples obtained from a salt-manufacturing industry in Chennai. The culture conditions for higher amylase production were optimized with respect to NaCl, substrate, pH, and temperature. Maximum amylase production of 592 mU/ml was achieved in the medium at 48 h with 10% NaCl, 1% dextrin, 0.4% yeast extract, 0.2% tryptone, and 0.2% CaCl(2) at pH 8.0 at 30 degrees C. The enzyme activity in the culture supernatant was highest with 10% NaCl at pH 7.5 and 55 degrees C. The amylase that was partially purified by acetone precipitation was highly stable in various surfactants and detergents. Glucose, maltose, and maltooligosaccharides were the main end products of starch hydrolysis indicating that it is an alpha-amylase.

  14. Removal of Heavy Metals by Adsorption onto Activated Carbon Derived from Pine Cones of Pinus roxburghii.

    PubMed

    Saif, Muhammad Jawwad; Zia, Khalid Mahmood; Fazal-ur-Rehman; Usman, Muhammad; Hussain, Abdullah Ijaz; Chatha, Shahzad Ali Shahid

    2015-04-01

    Activated carbon derived from cones of Pinus roxburghii (Himalayan Pine) was used as an adsorbent for the removal of copper, nickel and chromium ions from waste water. Surface analysis was carried out to determine the specific surface area and pore size distribution of the pine cone derived activated carbon. Optimal parameters, effect of adsorbent quantity, pH, equilibrium time, agitation speed and temperature were studied. Equilibrium data were evaluated by Langmuir and Freundlich isotherm models. Langmuir isotherm afforded the best fit to the equilibrium data with a maximum adsorption capacity of 14.2, 31.4 and 29.6 mg/g for Cu(II), Ni(II) and Cr(VI) respectively. Maximum adsorption of Cu(II), Ni(II) was observed in the pH range 4.0 to 4.5, whereas the best adsorption of Cr(VI) was observed at pH 2.5. It was found that 180 minutes was sufficient to gain adsorption equilibrium. The adsorption process follows a pseudo-second-order kinetic model.

  15. Facile and green synthesis of highly stable L-cysteine functionalized copper nanoparticles

    NASA Astrophysics Data System (ADS)

    Kumar, Nikhil; Upadhyay, Lata Sheo Bachan

    2016-11-01

    A simple eco-friendly method for L-cysteine capped copper nanoparticles (CCNPs) synthesis in aqueous solution has been developed. Glucose and L-cysteine were used as reducing agent and capping/functionalizing agent, respectively. Different parameters such as capping agent concentration, pH, reaction temperature, and reducing agent concentration were optimized during the synthesis. The L-cysteine capped copper nanoparticle were characterized by ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, Particle size and zeta potential analyser, and high resolution transmission electron microscopy. Spherical shaped cysteine functionalized/capped copper nanoparticles with an average size of 40 nm were found to be highly stable at room temperature (RT) for a period of 1 month

  16. Biosorptive uptake of arsenic(V) by steam activated carbon from mung bean husk: equilibrium, kinetics, thermodynamics and modeling

    NASA Astrophysics Data System (ADS)

    Mondal, Sandip; Aikat, Kaustav; Halder, Gopinath

    2017-12-01

    The present investigation emphasizes on the biosorptive removal of toxic pentavalent arsenic from water using steam activated carbon prepared from mung bean husk (SAC-MBH). Characterization of the synthesized sorbent was done using different instrumental techniques, i.e., SEM, BET and point of zero charge. Sorptive uptake of As(V) over steam activated MBH as a function of pH (3-9), agitation speed (40-200 rpm), dosage (50-1000 mg) and temperature (298-313 K) was studied by batch process at arsenic concentration of 2 mg L-1. Lower pH increases the arsenic removal over the pH range of 3-9. Among three adsorption isotherm models examined, Langmuir model was observed to show superior results over Freundlich model. The mean sorption energy (E) estimated by Dubinin-Radushkevich model suggested that the process of adsorption was chemisorption. Thermodynamic parameters confer that the sorption process was spontaneous, exothermic and feasible in nature. The pseudo-second-order rate kinetics of arsenic gave better correlation coefficients as compared to pseudo-first-order kinetics equation. Three process parameters, viz. adsorbent dosage, agitation speed and pH were opted for optimizing As(V) elimination using central composite design matrix of response surface methodology (RSM). The identical design setup was used for artificial neural network (ANN) for comparing its prediction capability with RSM towards As(V) removal. Maximum arsenic removal was observed to be 98.75% at sorbent dosage 0.75 gm L-1, pH 3.0, agitation speed 160 rpm and temperature 308 K. The study concluded that SAC-MBH could be a competent adsorbent for As(V) removal and ANN model was better in arsenic removal predictability results than RSM model.

  17. Use of oxalic acid as inducer in photocatalytic oxidation of cresol red in aqueous solution under natural and artificial light.

    PubMed

    Seraghni, N; Ghoul, I; Lemmize, I; Reguig, A; Debbache, N; Sehili, T

    2017-08-30

    This work was carried out in the field of water treatment using advanced oxidation processes (AOPs), especially photolysis of carboxylic acid that leads to the formation in situ of hydroxyl radical (·OH). Cresol red (CR) degradation induced by organic acids/UV system was investigated in aqueous solution. The preliminary study of CR-organic acid mixture in the dark and at room temperature allowed confirming the absence of interaction under our experimental conditions. However, upon irradiation at 365 nm, the proportion of elimination of CR was 89% after 5 h of irradiation. Indeed, the CR degradation efficiency depends on the acid concentration and the pH of the medium. The concentration of acid is optimized to the 5 × 10 -3 M. pH 2.39 was the optimal one when C 2 HO - 4 was the most important species at this pH. The use of i-PrOH as · OH confirmed the involvement of · OH in photodegradation of CR induced by Ox. The addition of metal ions including Zn 2+ and Cu 2+ to the CR-organic acid mixture slowed the CR degradation unlike Fe 2+ , hence an improvement of its disappearance was observed. The results showed a faster degradation of the pollutant under excitation by sunlight. This environmentally friendly method appears to be very effective in the treatment of wastewater.

  18. Production and characterization of thermostable alkaline protease of Bacillus subtilis (ATCC 6633) from optimized solid-state fermentation.

    PubMed

    Chatterjee, Joyee; Giri, Sudipta; Maity, Sujan; Sinha, Ankan; Ranjan, Ashish; Rajshekhar; Gupta, Suvroma

    2015-01-01

    Proteases are the most important group of enzymes utilized commercially in various arenas of industries, such as food, detergent, leather, dairy, pharmaceutical, diagnostics, and waste management, accounting for nearly 20% of the world enzyme market. Microorganisms of specially Bacillus genera serve as a vast repository of diverse set of industrially important enzymes and utilized for the large-scale enzyme production using a fermentation technology. Approximately 30%-40% of the cost of industrial enzymes originates from the cost of the growth medium. This study is attempted to produce protease from Bacillus subtilis (ATCC 6633) after optimization of various process parameters with the aid of solid-state fermentation using a cheap nutrient source such as wheat bran. B. subtilis (ATCC 6633) produces proteases of molecular weight 36 and 20 kDa, respectively, in the fermented medium as evident from SDS zymogram. Alkaline protease activity has been detected with optimum temperature at 50 °C and is insensitive to ethylenediaminetetraacetic acid. This thermostable alkaline protease exhibits dual pH optimum at 7 and 10 with moderate pH stability at alkaline pH range. It preserves its activity in the presence of detergent such as SDS, Tween 20, and Triton X-100 and may be considered as an effective additive to detergent formulation with some industrial importance. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

  19. Enzymatic hydrolysis of oleuropein from Olea europea (olive) leaf extract and antioxidant activities.

    PubMed

    Yuan, Jiao-Jiao; Wang, Cheng-Zhang; Ye, Jian-Zhong; Tao, Ran; Zhang, Yu-Si

    2015-02-11

    Oleuropein (OE), the main polyphenol in olive leaf extract, is likely to decompose into hydroxytyrosol (HT) and elenolic acid under the action of light, acid, base, high temperature. In the enzymatic process, the content of OE in olive leaf extract and enzyme are key factors that affect the yield of HT. A selective enzyme was screened from among 10 enzymes with a high OE degradation rate. A single factor (pH, temperature, time, enzyme quantity) optimization process and a Box-Behnken design were studied for the enzymatic hydrolysis of 81.04% OE olive leaf extract. Additionally, enzymatic hydrolysis results with different substrates (38.6% and 81.04% OE) were compared and the DPPH antioxidant properties were also evaluated. The result showed that the performance of hydrolysis treatments was best using hemicellulase as a bio-catalyst, and the high purity of OE in olive extract was beneficial to biotransform OE into HT. The optimal enzymatic conditions for achieving a maximal yield of HT content obtained by the regression were as follows: pH 5, temperature 55 °C and enzyme quantity 55 mg. The experimental result was 11.31% ± 0.15%, and the degradation rate of OE was 98.54%. From the present investigation of the antioxidant activity determined by the DPPH method, the phenol content and radical scavenging effect were both decreased after enzymatic hydrolysis by hemicellulase. However, a high antioxidant activity of the ethyl acetate extract enzymatic hydrolysate (IC50 = 41.82 μg/mL) was demonstated. The results presented in this work suggested that hemicellulase has promising and attractive properties for industrial production of HT, and indicated that HT might be a valuable biological component for use in pharmaceutical products and functional foods.

  20. In vivo oxygen, temperature and pH dynamics in the female reproductive tract and their importance in human conception: a systematic review.

    PubMed

    Ng, Ka Ying Bonnie; Mingels, Roel; Morgan, Hywel; Macklon, Nick; Cheong, Ying

    2018-01-01

    Despite advances in ART, implantation and pregnancy rates per embryo transfer still remain low. IVF laboratories strive to ensure that the process of handling gametes in vitro closely mimics the in vivo environment. However, there remains a lack of knowledge regarding the in vivo regulation and dynamic variation in biophysical parameters such as oxygen concentration, pH and temperature within the reproductive tract. To undertake a systematic review of the current understanding of the physico-chemical parameters of oxygen tension (pO2), pH and temperature within the female reproductive tract, and their potential implications in clinical and pathological processes related to fertility and those pertaining to limited reproductive capacity. A comprehensive literature search was performed using electronic databases including Medline, Embase, Cochrane Library and Pubmed to identify original and review articles addressing the biophysical parameters (pO2, pH and temperature) in the female reproductive tract of any species. The search included all studies published between 1946 and November 2015. Search terms included 'oxygen', 'pH', 'hydrogen ion concentration', 'acid base' and others terms. We also used special features and truncations to identify synonyms and broaden the search. Studies were excluded if they only assessed embryo culture conditions, fetal acid-base status, oxidative stress, outcomes of pregnancy and measurements of these parameters in non-reproductive organs. Our search generated 18 685 records and 60 articles were included. pO2 within the female reproductive tract shows cyclical variation and minute-to-minute oscillations, which may be influenced by uterine contractility, hormones, the autonomic system, cardiac pulsatility, and myometrial and smooth muscle integrity. Fine balanced control of pO2 and avoidance of overwhelming oxidative stress is crucial for embryogenesis and implantation. The pH in the female reproductive tract is graduated, with lowest pH in the vagina (~pH 4.42) increasing toward the Fallopian tubes (FTs) (~pH 7.94), reflecting variation in the site-specific microbiome and acid-base buffering at the tissue/cellular level. The temperature variation in humans is cyclical by day and month. In humans, it is biphasic, increasing in the luteal phase; with the caudal region of the oviduct 1-2 degrees cooler than the cranial portion. Temperature variation is influenced by hormones, density of pelvic/uterine vascular beds and effectiveness of heat exchange locally, crucial for sperm motility and embryo development. We have identified significant deficiencies and inconsistencies in the methods used to assess these biophysical factors within the reproductive tract. We have suggested that the technological solutions including the development of methods and models for real time, in vivo recordings of biophysical parameters. The notion of 'back to nature' in assisted conception suggested 20 years ago has yet to be translated into clinical practice. While the findings from this systematic review do not provide evidence to change current in vitro protocols, it highlights our current inability to assess the in vivo reproductive tract environment in real time. Data made available through future development of sensing technology in utero may help to provide new insights into how best to optimize the in vitro embryo environment and allow for more precise and personalized fertility treatment. © The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com

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