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Sample records for guehomyces pullulans 17-1

  1. Pectinolytic yeasts from cold environments: novel findings of Guehomyces pullulans, Cystofilobasidium infirmominiatum and Cryptococcus adeliensis producing pectinases.

    PubMed

    Cavello, Ivana; Albanesi, Agustín; Fratebianchi, Dante; Garmedia, Gabriela; Vero, Silvana; Cavalitto, Sebastián

    2017-03-01

    One hundred and three yeasts isolated from soil samples from King George Island and Tierra del Fuego province were screened in relation with their capability to produce pectinolytic enzymes. Although all the yeasts showed well-developed colonies at 20 °C, only eight showed a clear halo around the colony, indicative of pectin degradation. A secondary screening demonstrated that only four yeasts were capable to produce pectinases at low temperatures (8 °C). It could be seen that the selected yeasts were able to grow and produce high levels of polygalacturonase activity when submerged fermentations were performed using pectin-containing fruit wastes as substrates. None of the strains produced neither lyase nor rhamnogalacturonan hydrolase activities. Regarding pectin esterase activity, it was only produced in lower amounts by G. pullulans 8E (0.022 U ml(-1)). A TLC analysis of the substrate cleavage pattern of the pectinolytic systems was consistent with an endo-type activity. The clarification of apple juice was only accomplished by G. pullulans pectinolytic system, with a clarification of 80% (%T650) using 4 U/ml of enzyme at 20 °C. As far as we concern this work describes for the first time the production of pectinases by the cold-adapted yeasts species Cystofilobasidium infirmominiatum, Cryptococcus adeliensis and G. pullulans.

  2. Pullulan Elaboration by Aureobasidium pullulans Protoplasts

    PubMed Central

    Finkelman, Malcolm A. J.; Vardanis, Alexander

    1982-01-01

    Protoplasts of Aureobasidium pullulans are capable of producing pullulan. Biosynthesis of the polymer pullulan required induction with kinetics similar to those of whole cells. The protoplasts also produced a heteropolysaccharide component containing mannose, glucose, and galactose. The relative proportions of the pullulan and heteropolysaccharide fractions were a function of glucose concentration, with the pullulan content of the total polysaccharide rising from 20% at 2.5 mM glucose to 45% at 20 mM glucose. Elaboration of pullulan by both cells and protoplasts was sensitive to 0.6 M KCl, which was present as the osmotic stabilizer in protoplast experiments. The presence of KCl resulted in a shift in the pH optimum to a more acidic value. The molecular weight of the protoplast-derived pullulan was sharply reduced from the molecular weight of the whole-cell-derived product. Exposure of the protoplasts to proteolytic enzymes had no effect on polysaccharide elaboration. PMID:16346047

  3. Laccases from Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    Laccases are polyphenol oxidases (EC 1.10.3.2) that have numerous industrial and bioremediation applications. Laccases are well known as lignin-degrading enzymes, but these enzymes can play numerous other roles in fungi. In this study, 41 strains of the fungus Aureobasidium pullulans were examined f...

  4. Intraspecific diversity of Aureobasidium pullulans strains from different marine environments

    NASA Astrophysics Data System (ADS)

    Liu, Jia; Liu, Zhiqiang; Chi, Zhenming; Zhang, Liang; Zhang, Dechao

    2009-09-01

    Totally more than 500 yeast strains were isolated from seawater, sea sediments, mud of sea salterns, marine fish guts and marine algae. The results of routine and molecular biology identification methods show that nine strains among these marine yeasts belong to Aureobasidium pullulans, although the morphologies of their colonies are very different. The marine yeasts isolated from different marine environments indicate that A. pullulans is widely distributed in different environmental conditions. These Aureobasidium pullulans strains include A. pullulans 4#2, A. pullulans N13d, A. pullulans HN3-11, A. pullulans HN2-3, A. pullulans JHSc, A. pullulans HN4.7, A. pullulans HN5.3, A. pullulans HN6.2 and A. pullulans W13a. A. pullulans 4#2 could produce cellulase and single cell protein. A. pullulans N13d could produce protease, lipase, amylase and cellulase. Both A. pullulans HN3-11 and A. pullulans HN2-3 were able to produce protease, lipase and cellulase. A. pullulans JHSc could secrete cellulase and killer toxin. Both A. pullulans HN4.7 and A. pullulans HN5.3 could yield lipase and cellulase. A. pullulans W13a was able to secrete extracellular amylase and cellulase while A. pullulans HN4.7 and A. pullulans N13d could produce siderophores. This means that different A. pullulans strains from different marine environments have different physiological characteristics, which may be applied in many different biotechnological industries.

  5. 77 FR 8731 - Aureobasidium pullulans

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-15

    ... active ingredient is 80% w/w (minimum of x 10\\9\\ colony forming units/grams (unit of measure for bacteria... lymph nodes of one animal sacrificed on day 7 tested positive for the presence of Aureobasidium... was positive for Aureobasidium pullulans in three males and three females from the test material...

  6. Efficient production of pullulan by Aureobasidium pullulans grown on mixtures of potato starch hydrolysate and sucrose.

    PubMed

    An, Chao; Ma, Sai-Jian; Chang, Fan; Xue, Wen-Jiao

    Pullulan is a natural exopolysaccharide with many useful characteristics. However, pullulan is more costly than other exopolysaccharides, which limits its effective application. The purpose of this study was to adopt a novel mixed-sugar strategy for maximizing pullulan production, mainly using potato starch hydrolysate as a low-cost substrate for liquid-state fermentation by Aureobasidium pullulans. Based on fermentation kinetics evaluation of pullulan production by A. pullulans 201253, the pullulan production rate of A. pullulans with mixtures of potato starch hydrolysate and sucrose (potato starch hydrolysate:sucrose=80:20) was 0.212h(-1), which was significantly higher than those of potato starch hydrolysate alone (0.146h(-1)) and mixtures of potato starch hydrolysate, glucose, and fructose (potato starch hydrolysate:glucose:fructose=80:10:10, 0.166h(-1)) with 100gL(-1) total carbon source. The results suggest that mixtures of potato starch hydrolysate and sucrose could promote pullulan synthesis and possibly that a small amount of sucrose stimulated the enzyme responsible for pullulan synthesis and promoted effective potato starch hydrolysate conversion effectively. Thus, mixed sugars in potato starch hydrolysate and sucrose fermentation might be a promising alternative for the economical production of pullulan. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  7. HLC/pullulan and pullulan hydrogels: their microstructure, engineering process and biocompatibility.

    PubMed

    Li, Xian; Xue, Wenjiao; Liu, Yannan; Li, Weina; Fan, Daidi; Zhu, Chenhui; Wang, Yaoyu

    2016-01-01

    New locally injectable biomaterials that are suitable for use as soft tissue fillers are needed to address a significant unmet medical need. In this study, we used pullulan and human-like collagen (HLC) based hydrogels with various molecular weights (MWs) in combination therapy against tissue defects. Briefly, pullulan was crosslinked with NaIO4 to form a pullulan hydrogel and then may coupled with HLC using the reaction between the -NH2 end-group of HLC and the -CHO group present on the aldehyde pullulan to form the HLC/pullulan hydrogel, wherein the NaIO4 acted as the crosslinking and oxidizing agent. The good miscibility of pullulan and HLC in the hydrogels was confirmed via Fourier transform infrared spectroscopy, scanning electron microscopy, compression testing, enzyme degradation testing, cell adhesions, live/dead staining and subcutaneous filling assays. Here, pullulan hydrogels with various MWs were fabricated and physicochemically characterized. Limitations of the pullulan hydrogels included inflammation, poor mechanical strength, and degradation. By contrast, the properties of the HLC/pullulan hydrogels strongly enhanced. The efficacy of these hydrogels was evaluated both in vitro and in vivo. Our results indicate that HLC/pullulan hydrogels may have therapeutic value as efficient soft tissue fillers, with reduced inflammation, improved cell adhesion and delayed hydrogel degradation. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Central metabolic pathways of Aureobasidium pullulans CGMCC1234 for pullulan production.

    PubMed

    Sheng, Long; Liu, Chang; Tong, Qunyi; Ma, Meihu

    2015-12-10

    With the purpose of understanding the metabolic network of Aureobasidium pullulans, the central metabolic pathways were confirmed by the activities of the key enzymes involved in different pathways. The effect of different iodoacetic acid concentrations on pullulan fermentation was also investigated in this paper. The activities of phosphofructokinases and glucose-6-phosphate dehydrogenase existed in A. pullulans CGMCC1234, whereas 2-keto-3-deoxy-6-phosphogluconate aldolase activity was not detected. We proposed that the central metabolic pathways of A. pullulans CGMCC1234 included EMP and PPP, but no ED. Pullulan production declined fast as the iodoacetic acid increased, while cell growth offered upgrade firstly than descending latter tendency. Compared to the control group, the ratio of ATP/ADP of 0.60 mM iodoacetic acid group was lower at different stages of pullulan fermentation. The findings revealed that low concentration of iodoacetic acid might impel carbon flux flow toward the PPP, but reduce the flux of the EMP.

  9. Why sucrose is the most suitable substrate for pullulan fermentation by Aureobasidium pullulans CGMCC1234?

    PubMed

    Sheng, Long; Tong, Qunyi; Ma, Meihu

    2016-10-01

    This paper studies the metabolic pathway of sucrose in pullulan fermentation by Aureobasidium pullulans. Because of its high pullulan production, sucrose proved to be the best carbon source for pullulan synthesis by A. pullulans CGMCC1234 (36.3g/L). Compared to other carbon sources, A. pullulans cells reached the stationary phase more quickly in the presence of sucrose. The specific sugar types and concentrations occurring during pullulan fermentation were detected using High Performance Liquid Chromatography (HPLC). HPLC results revealed that sucrose did not simply break down into glucose and fructose in the medium employed. Kestose (22.69g/L) also accumulated during early stages of fermentation (24h), which reduced the osmotic pressure of the extracellular fluid and diminished the inhibition of cell growth and pullulan production. β-Fructofuranosidase activity strongly depended on the carbon source. Sucrose was the best inducer of β-fructofuranosidase production. However, β-fructofuranosidase production did not directly and/or proportionally correlate with the growth of A. pullulans cells. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Efficient pullulan production by bioconversion using Aureobasidium pullulans as the whole-cell catalyst.

    PubMed

    Ju, Xiao-Min; Wang, Da-Hui; Zhang, Gao-Chuan; Cao, Dan; Wei, Gong-Yuan

    2015-01-01

    In this study, pullulan production was achieved by whole-cell bioconversion with Aureobasidium pullulans CCTCC M 2012259. Response surface methodology was applied to optimize the seed medium for incubating cells with high capability of pullulan bioconversion. Three medium components, namely, yeast extract, MgSO4·7H2O, and glucose were identified by Plackett-Berman design as significant factors affecting the cells' pullulan bioconversion capability. A three-level Box-Behnken design was then employed to determine the optimal levels of the three components. A mathematical model was developed to show the influence of each medium component and its effects on the cells' pullulan bioconversion capability. The model predicted a maximum pullulan bioconversion capability of 32.28 mg/g/h at the optimal yeast extract, MgSO4·7H2O, and glucose concentrations of 3.57, 0.18, and 63.97 g/l, respectively. The validation experiments showed that the cells' pullulan bioconversion capability was improved by 23.1% when the optimal medium was used, as compared with that obtained with the basic medium. Subsequently, the gene expression and activities of the key enzymes involved in pullulan biosynthesis were evaluated. When the optimal medium was employed, the transcriptional levels of pgm1 and fks were up-regulated by 2.5- and 1.2-fold, respectively, and the α-phosphoglucose mutase and glucosyltransferase activities were increased by 17 and 19%, respectively, when compared with those achieved using the basic medium. These results indicated that pullulan bioconversion using A. pullulans CCTCC M 2012259 as the whole-cell catalyst is an attractive approach for efficient pullulan production and can be applied for the production of other polysaccharides.

  11. Aureobasidium pullulans xylanase, gene and signal sequence

    DOEpatents

    Xin-Liang, Li; Ljungdahl, Lars G.

    1997-01-01

    A xylanase from Aureobasidium pullulans having a high specific activity is provided as well as a signal protein for controlling excretion into cell culture medium of proteins to which it is attached. DNA encoding these proteins is also provided.

  12. Aureobasidium pullulans fungal keratitis following LASEK.

    PubMed

    Maverick, Kenneth J; Conners, Michael S

    2007-09-01

    To describe a patient who developed Aureobasidium pullulans keratitis following refractive laser epithelial keratomileusis (LASEK). A 52-year-old woman was referred to a tertiary care center 1 month after LASEK for treatment of a corneal ulcer that was unresponsive to conventional therapy. Mycology culture and fungal stain identified Aureobasidium as the infectious organism. The infection responded well to treatment with topical natamycin and systemic itraconazole. Treatment with topical natamycin and systemic itraconazole is effective against Aureobasidium pullulans keratitis.

  13. Efficient production of pullulan using rice hull hydrolysate by adaptive laboratory evolution of Aureobasidium pullulans.

    PubMed

    Wang, Dahui; Ju, Xiaomin; Zhou, Donghai; Wei, Gongyuan

    2014-07-01

    Pullulan production by Aureobasidium pullulans CCTCC M 2012259 using rice hull hydrolysate as the carbon source was conducted. The acetic acid in the hydrolysate was demonstrated to exert a negative effect on pullulan biosynthesis. Instead of employing expensive methods to remove acetic acid from the hydrolysate, a mutant A. pullulans ARH-1 was isolated following 20 cycles of adaptive laboratory evolution of the parental strain on medium containing acetic acid. The maximum pullulan production achieved by the adapted mutant at 48 h using the hydrolysate of untreated rice hull was 22.2 g L(-1), while that obtained by the parental strain at 60 h was 15.6 g L(-1). The assay of key enzymes associated with pullulan biosynthesis revealed that acetic acid inhibited enzyme activity rather than suppressing enzyme synthesis. These results demonstrated that adaptive evolution highly improved the efficiency of pullulan production by A. pullulans using the hydrolysate of untreated rice hull. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Understanding the influence of Tween 80 on pullulan fermentation by Aureobasidium pullulans CGMCC1234.

    PubMed

    Sheng, Long; Tang, Guiyue; Su, Peng; Zhang, Jinling; Xiao, Qian; Tong, Qunyi; Ma, Meihu

    2016-01-20

    In this paper, several new perspectives concerned with the effect of Tween 80 promoting pullulan production were presented. With the presence of Tween 80, the maximum pullulan yield increased by 41% (53.04 g/L). Meanwhile, the carbon source was consumed faster and the broth viscosity was higher. The lower final pH suggested that Tween 80 could protect the integrity of the mycelia. The dispersed filaments were not easily entangled with each other and less pellets were formed in the Tween 80 culture broth. FT-IR spectrum analysis indicated that the evaluated sample structure was coincided with commercial pullulan. The molecular weight of sample significantly dropped comparing with the control. The above findings indicated that Tween 80 facilitated the uptake of nutrient from surroundings to the microorganism and the release of pullulan into the extracellular fluid. These results were useful in better understanding the regulation and optimization of efficient pullulan fermentation.

  15. Effects of Topical Anesthetics on Pullularia pullulans and Debaryomyces hansenii

    PubMed Central

    Merdinger, Emanuel; Guthmann, Walter S.; Mangine, Francis W.

    1969-01-01

    The inhibitory effects of three topical anesthetics of various concentrations on the growth of Pullularia pullulans, Debaryomyces hansenii, and on pigment production by P. pullulans were investigated. The topical anesthetics were benoxinate hydrochloride, proparacaine hydrochloride, and tetracaine hydrochloride. In decreasing order, the inhibiting effects of the drugs on growth were benoxinate, tetracaine, and proparacaine for P. pullulans, and tetracaine, benoxinate, and proparacaine for D. hansenii. The pigment formation in P. pullulans was inhibited by the three drugs. PMID:5392897

  16. Optimization of high molecular weight pullulan production by Aureobasidium pullulans in batch fermentations.

    PubMed

    Gibson, Larry H; Coughlin, Robert W

    2002-01-01

    Of five strains of Aureobasidium pullulans studied, NRRL Y-2311-1 yielded the highest titer (26.2 g/L) of pullulan and formed the lowest amount of melanin-like pigment. Sucrose was superior to glucose as the carbon and energy source on the basis of yield and titer of pullulan produced. Pullulan titer was higher (26.2 vs 5.1 g/L), biomass concentration was lower (6.9 vs 12.7 g/L), and DO was lower (0 vs 60% of saturation) when the fermenter was agitated by a marine propeller compared to Rushton impellers. Pullulan produced by strain NRRL Y-2311-1 ranged in weight-average molar mass (M(w)) from 486 KDa and number-average molar mass (M(n)) from 220 Da on day 1 of growth to 390 KDa and 690 Da on day 6; M(w) declined by about 35% from day 1 to day 3, the day of maximum pullulan titer. For the other strains, the ranges of molar mass on the day of maximum pullulan titer were 338-614 KDa (M(w)) and 100-6820 Da (M(n)).

  17. Production of Pullulan, Poly(beta-L-malic acid), and Heavy Oil by Fungus Aureobasidium pullulans Isolated from Thailand

    USDA-ARS?s Scientific Manuscript database

    Fungus Aureobasidium pullulans is the main source of a polysaccharide, pullulan, in industrial production. Moreover, it can produce many bioproducts, e.g. xylanase, poly(ß-L-malic acid) (PMA), and heavy oil. In this study, we isolated 15 A. pullulans isolates from various sources and habitats in T...

  18. [Lipids of Aureobasidium (Pullularia) pullulans].

    PubMed

    Elinov, N P; Iurlova, N A; Efimova, T P

    1975-01-01

    Fractional composition of free and bound lipids was studied in Aureobasidium (Pullularia) pullulans 8 by preparative TLC on Silufol. Bound lipids contained a fraction (27.76 +/- 0.5%) of dark brown colour, similar to melanin. The composition of fatty acids was studied by GLC. The following fatty acids were identified and determined quantitatively: C12:0, C14:0, C15:0, C16:0, C18:0, C18:1+C15:2. The following fatty acids predominated in free and bound lipids: C16:0, C18:1+C18:2. The ratio between unsaturated and saturated fatty acids in all fractions of free and bound lipids was more than unity. The following parameters were determined for lipids; ester number (173.89 and 178.53); iodine number (44.1 and 33.10), and saponification number (181.17 and 206.03) (the values are given for free and bound lipids, respectively).

  19. Growth of Aureobasidium pullulans on straw hydrolysate.

    PubMed Central

    Han, Y W; Cheeke, P R; Anderson, A W; Lekprayoon, C

    1976-01-01

    Growth characteristics and cell properties of Aureobasidium (Pullularia) pullulans were studied. The organism grew well on an acid hydrolysate of ryegrass straw over a wide range of pH and temperature. The optimum temperature and pH for the growth of the organism were 32 degrees C and 5.5, respectively. A cell yield of 1.5 g/liter of straw hydrolysate was obtained. The dried cell mass contained 42.6% crude protein, 0.4% crude fat, and 6.4% nucleic acids. The essential amino acid profile of the microbial protein was comparable to that of Candida utilis. A rat feeding study indicated that the A. pullulans cells were not toxic and that the feed intake, weight gain, and protein efficiency ratio values were superior to those obtained with C. utilis. Once the question of mathogenicity is resolved, A. pullulans could be useful for production of single-cell protein from cellulosic wastes. PMID:12721

  20. Self-Assembly Behavior of Pullulan Abietate

    NASA Astrophysics Data System (ADS)

    Gradwell, Sheila; Esker, Alan; Glasser, Wolgang; Heinze, Thomas

    2003-03-01

    Wood is one of nature's most fascinating biological composites due to its toughness and resistance to fracture properties. These properties stem from the self-assembly of cellulose microfibrils in an amorphous matrix of hemicellulose and lignin. In recent years, science has looked to nature for guidance in preparing synthetic materials with desirable physical properties. In order to study the self-assembly process in wood, a model system composed of a polysaccharide, pullulan abietate, and a biomimetic cellulose substrate prepared by the Langmuir-Blodgett technique has been developed. Interfacial tension and surface plasmon resonance measurements used to study the self-assembly process will be discussed for different pullulan derivatives.

  1. Zero birefringence films of pullulan ester derivatives

    PubMed Central

    Danjo, Takahiro; Enomoto, Yukiko; Shimada, Hikaru; Nobukawa, Shogo; Yamaguchi, Masayuki; Iwata, Tadahisa

    2017-01-01

    High-performance films with almost zero-birefringence and zero-wavelength dispersion were succeeded to prepare from pullulan esters derivatives (PLEs) without any additives. Optical transmittance analysis, birefringence measurement of PLE cast film and hot stretched films, and infrared dichroism analysis were conducted to characterize optical properties of PLE films comparing with cellulose triacetate which is commercially used as low-birefringence in optical devices. The aims of this study, characterization of optical properties of pullulan esters, can develop a deep understanding of the fundamental knowing and applicability of polysaccharides. Accordingly, authors believe this paper will open the gate for researches in the application of polysaccharides. PMID:28417955

  2. Mechanism study of Tween 80 enhancing the pullulan production by Aureobasidium pullulans.

    PubMed

    Sheng, Long; Zhu, Guilan; Tong, Qunyi

    2013-08-14

    The influences of Tween 80 on the pullulan production, cell growth, glucosyltransferase activity and fatty acid composition of the cells were studied. The addition of Tween 80 to the culture medium significantly enhanced the pullulan production, but had no impact to biomass accumulation. Glucosyltransferase activity involved in pullulan synthesis was remarkable promoted with an increase of Tween 80 content. The ratio of unsaturated/saturated fatty acids contained in the cells was significantly higher in 0.5% Tween 80 in comparison with the absence of Tween 80. Results indicated that the increase of pullulan yield brought by Tween 80 was highly correlated with glucosyltransferase activity and fatty acid composition. Tween 80 was not degraded to serve as extra carbon source.

  3. Production of pullulan from raw potato starch hydrolysates by a new strain of Auerobasidium pullulans.

    PubMed

    Wu, Shengjun; Lu, Mingsheng; Chen, Jing; Fang, Yaowei; Wu, Leilei; Xu, Yan; Wang, Shujun

    2016-01-01

    In the present study, hydrolysis of potato starch with marine cold-adapted α-amylase and pullulan production from the hydrolysates by a new strain of Auerobasidium pullulans isolated from sea mud were conducted. The hydrolysis conditions were optimized as follows: reaction time 2h, pH 6.5, temperature 20°C, and α-amylase amount 12 U/g. Under these optimum hydrolysis conditions, the DE value of the potato starch hydrolysates reached to 49.56. The potato starch hydrolysates consist of glucose, maltose, isomaltose, maltotriose, and trace of other maltooligosaccharides with degree of polymerization ranged 4-7. The maximum production of pullulan at 96 h from the hydrolysate of potato starch was 36.17 g/L, which was higher than those obtained from glucose (22.07 g/L, p<0.05) and sucrose (31.42 g/L, p<0.05). Analysis of the high performance liquid chromatography of the hydrolysates of the pullulan product with pullulanase indicated that the main composition is maltotriose, thus confirming the pullulan structure of this pullulan product.

  4. Biotechnology of Aureobasidium pullulans: A phylogenetic perspective

    USDA-ARS?s Scientific Manuscript database

    Aureobasidium pullulans is a fungus historically included among the "black yeasts." Although many strains are predominantly yeast-like, the species is actually polymorphic, exhibiting complex forms ranging from blastic conidia and swollen cells to pseudophyphae, hyphae, and chlamydospores. A. pull...

  5. Aureobasidium pullulans xylanase, gene and signal sequence

    DOEpatents

    Li Xinliang; Ljungdahl, L.G.

    1997-01-07

    A xylanase from Aureobasidium pullulans having a high specific activity is provided, as well as a signal protein for controlling excretion into cell culture medium of proteins to which it is attached. DNA encoding these proteins is also provided. 4 figs.

  6. Heavy Oils Produced by Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    From a survey of more than 50 diverse strains of Aureobasidium pullulans, 21 strains were observed to produce extracellular heavy oils. These strains represented at least 6 phylogenetic clades, although more than half fell into clades 9 and 11. Oil colors ranged from bright yellow to malachite. M...

  7. Tubular cationized pullulan hydrogels as local reservoirs for plasmid DNA.

    PubMed

    San Juan, Aurélie; Ducrocq, Grégory; Hlawaty, Hanna; Bataille, Isabelle; Guénin, Erwann; Letourneur, Didier; Feldman, Laurent J

    2007-12-01

    In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain plasmid DNA, and tested the ability of retained plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethylamine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies.

  8. Silk fibroin/pullulan blend films: Preparation and characterization

    SciTech Connect

    Shivananda, C. S.; Rao, B. Lakshmeesha; Madhukumar, R.; Asha, S.; Sarojini, B. K.; Somashekhar, R.; Sangappa, Y.

    2016-05-23

    In this work silk fibroin/pullulan blend films have been prepared by solution casting method. The blend films were examined for structural, and thermal properties using X-ray diffraction (XRD), thermogravimatric (TGA) and differential scanning calorimetry (DSC) analysis. The XRD results indicate that with the introduction of pullulan, the interaction between SF and pullulan in the blend films induced the conformation transition of SF films and amorphous phase increases with increasing pullulan ratio. The thermal properties of the blend films were improved significantly in the blend films.

  9. Control of Molecular Weight Distribution of the Biopolymer Pullulan Produced by the Fungus Aureobasidium Pullulans

    DTIC Science & Technology

    1987-10-01

    Source ( 1 0%) [!H QH (%) (k) Fructose 5.44 3.76 27.0 1122 2.5 Sucrose 5.44 3.69 34.4 895 2.4 Maltose 5. 44 4.08 25.4 881 2. l Corn Syrup 5. 44 3.85...pullulan and found that the uptake of glucose at more acid pH was diverted to the synthesis of extra- cellular pullulan, and that high extracellular...at pH 5.5, pH 6.0, and pH 6.5, Phosphate concentrations of 0.2%-0,4% yielded high MW pullulan at the lower pH level. They reported that using

  10. Redefinition of Aureobasidium pullulans and its varieties

    PubMed Central

    Zalar, P.; Gostinčar, C.; de Hoog, G.S.; Uršič, V.; Sudhadham, M.; Gunde-Cimerman, N.

    2008-01-01

    Using media with low water activity, a large numbers of aureobasidium-like black yeasts were isolated from glacial and subglacial ice of three polythermal glaciers from the coastal Arctic environment of Kongsfjorden (Svalbard, Spitsbergen), as well as from adjacent sea water, sea ice and glacial meltwaters. To characterise the genetic variability of Aureobasidium pullulans strains originating from the Arctic and strains originating pan-globally, a multilocus molecular analysis was performed, through rDNA (internal transcribed spacers, partial 28 S rDNA), and partial introns and exons of genes encoding β-tubulin (TUB), translation elongation factor (EF1α) and elongase (ELO). Two globally ubiquitous varieties were distinguished: var. pullulans, occurring particularly in slightly osmotic substrates and in the phyllosphere; and var. melanogenum, mainly isolated from watery habitats. Both varieties were commonly isolated from the sampled Arctic habitats. However, some aureobasidium-like strains from subglacial ice from three different glaciers in Kongsfjorden (Svalbard, Spitsbergen), appeared to represent a new variety of A. pullulans. A strain from dolomitic marble in Namibia was found to belong to yet another variety. No molecular support has as yet been found for the previously described var. aubasidani. A partial elongase-encoding gene was successfully used as a phylogenetic marker at the (infra-)specific level. PMID:19287524

  11. New Method of Producing Protoplasts of Aureobasidium pullulans

    PubMed Central

    Finkelman, Malcolm A. J.; Zajic, James E.; Vardanis, Alex

    1980-01-01

    A rapid and relatively inexpensive method for producing protoplasts of the black yeast Aureobasidium pullulans is described. The procedure involves anaerobic incubation with the lytic preparation Driselase. Images PMID:16345558

  12. Population biology of Aureobasidium pullulans on apple leaf surfaces.

    PubMed

    Andrews, John H; Spear, Russell N; Nordheim, Erik V

    2002-06-01

    Colonization of apple leaves by the yeast-like fungus Aureobasidium pullulans was studied in the field on eight dates over 2 years. Population densities from adaxial leaf surfaces were approximately log10 0.5-2.6 U higher when enumerated directly along line transects as microscopic counts of A. pullulans cells specifically identified by fluorescence in situ hybridization (FISH) than indirectly as CFU obtained by plating leaf washings from comparable surfaces onto nutrient media. Site-specific mapping of the leaf landscape colonized by A. pullulans was facilitated by use of geographic information system (GIS) software. Colonization was plotted and analyzed both qualitatively (as occupancy) and quantitatively (as density). Overall, when expressed as mean occupancy per date, 22-42% of the microscope fields (each 0.196 mm2) contained > or = 1 A. pullulans cell. Occupancy on a microscope field basis was greater over the midvein (47-89%) or smaller veins (49-65%) than over interveinal regions (11-21%). Intensity of colonization, whether expressed as percentage of total A. pullulans cells associated with a particular leaf feature or as cell density per unit area, was also significantly greater (P < 0.05) over the veinal areas compared with the interveinal areas. The primary fungal morphotypes involved in colonization were blastospores, swollen cells, and chlamydospores; only infrequently were hyphae or pseudohyphae seen. Numbers of microcolonies (> or = 10 clustered cells) and percentage of total A. pullulans cells that occurred as microcolonies increased over the growing season and were significantly greater (P < 0.05) over veinal regions compared with interveinal regions. Locally high concentrations of A. pullulans were associated with naturally occurring micro-wounds in interveinal areas. We conclude that A. pullulans colonizes the phylloplane predominantly as single cells and groups thereof in a highly heterogeneous fashion and that sites exist that are relatively

  13. [Phosphate solubilization of Aureobasidium pullulan F4 and its mechanism].

    PubMed

    Wang, Dan; Zhan, Jing; Sun, Qing-Ye

    2014-07-01

    The Aureobasidium pullulans F4 was isolated from the rhizosphere of Hippochaete ramosissimum in Tongguanshan mine wasteland in Tongling City, Anhui Province. Liquid culture was conducted with four kinds of phosphorus sources, calcium phosphate, aluminum phosphate, ferric phosphate and rock phosphate to determine the pH, dissolved phosphorus, phosphorus in the bacteria and organic acid in the solution. The results showed that the phosphate solubilization by A. pullulans F4 varied with phosphorus sources, which decreased in order of aluminum phosphate > ferric phosphate, calcium phosphate > rock phosphate. The amounts of dissolved phosphorus in the different treatments were all higher than 200 mg x L(-1). The pH of the medium dropped immediately in 48 h, and the aluminum phosphate and ferric phosphate treatments showed a greater decrease in pH than the calcium phosphate and rock phosphate treatments. The organic acid synthesized by A. pullulans F4 included oxalic acid, citric acid and tartaric acid, and oxalic acid, among which oxalic acid was the dominated component. The phosphate dissolving capacity of A. pullulans F4 showed no significant correlation with organic acid, but significantly correlated with the pH. The available phosphorus was significantly improved with the combined application of A. pullulans F4 and glucose, suggesting A. pullulans F4 was a potent candidate for remediation of copper mine wastelands.

  14. The effect of Tween 80 on the polymalic acid and pullulan production by Aureobasidium pullulans CCTCC M2012223.

    PubMed

    Tu, Guangwei; Wang, Yongkang; Ji, Yunchao; Zou, Xiang

    2015-01-01

    The effect of Tween 80 on the fermentative production of polymalic acid (PMA) and pullulan using Aureobasidium pullulans CCTCC M2012223 was investigated. Tween 80 is beneficial for the biosynthesis of PMA and pullulan, and can regulate the ratio of PMA to pullulan in a dose-dependent manner. After adding 0.05 % Tween 80 to the media, the maximal PMA and pullulan production was 46.45 and 28.8 g/L at 60 h in a 5 L fermenter, with an increase of 75.08 and 27.21 % when compared to the control. Tween 80 could regulate and enhance oxygen uptake rate and carbon dioxide evolution rate in the early phase of fermentation, and change the cell morphology. The transcription levels of mitochondrial dicarboxylate transporter and transmembrane transporter were also dramatically upregulated. The present work will be helpful in deeply understanding the mechanism of Tween 80 on the effect of PMA and pullulan production.

  15. Bioconversion of industrial solid waste--cassava bagasse for pullulan production in solid state fermentation.

    PubMed

    Sugumaran, K R; Jothi, P; Ponnusami, V

    2014-01-01

    The purpose of the work was to produce commercially important pullulan using industrial solid waste namely cassava bagasse in solid state fermentation and minimize the solid waste disposal problem. First, influence of initial pH on cell morphology and pullulan yield was studied. Effect of various factors like fermentation time, moisture ratio, nitrogen sources and particle size on pullulan yield was investigated. Various supplementary carbon sources (3%, w/w) namely glucose, sucrose, fructose, maltose, mannose and xylose with cassava bagasse was also studied to improve the pullulan yield. After screening the suitable supplement, effect of supplement concentration on pullulan production was investigated. The pullulan from cassava bagasse was characterized by FTIR, (1)H-NMR and (13)C-NMR. Molecular weight of pullulan from cassava bagasse was determined by gel permeation chromatography. Thus, cassava bagasse emerged to be a cheap and novel substrate for pullulan production.

  16. Liamocins from Aureobasidium pullulans: New and highly selective anti-streptococcal agents

    USDA-ARS?s Scientific Manuscript database

    The ubiquitous, black yeast Aureobasidium pullulans, which is used commercially to produce pullulan polysaccharide, also produces several polyol-lipids that we have named liamocins. A survey of >50 strains of A. pullulans identified 21 diverse strains that produce liamocins. Typically the liamocins ...

  17. Cationized pullulan 3D matrices as new materials for gene transfer.

    PubMed

    San Juan, Aurélie; Hlawaty, Hanna; Chaubet, Frédéric; Letourneur, Didier; Feldman, Laurent J

    2007-08-01

    This study deals with the development of a novel biocompatible cationized pullulan three-dimensional matrix for gene delivery. A water-soluble cationic polysaccharide, diethylaminoethyl-pullulan (DEAE-pullulan), was first synthesized and characterized. Fluorescence quenching and gel retardation assays evidenced the complexation in solution of DNA with DEAE-pullulan, but not with neutral pullulan. On cultured smooth muscle cells (SMCs) incubated with DEAE-pullulan and a plasmid vector expressing a secreted form of alkaline phosphatase (pSEAP), SEAP activity was 150-fold higher than with pSEAP alone or pSEAP with neutral pullulan. DEAE-pullulan was then chemically crosslinked using phosphorus oxychloride. The resulting matrices were obtained in less than a minute and molded as discs of 12 mm diameter and 2 mm thickness. Such DEAE-pullulan 3D matrices were loaded with up to 50 microg of plasmid DNA, with a homogeneous plasmid loading observed with YOYO-1 fluorescence staining. Moreover, the DEAE-pullulan matrix was shown to protect pSEAP from DNase I degradation. Incubation of cultured SMCs with pSEAP-loaded DEAE-pullulan matrices resulted in significant gene transfer without cell toxicity. This study suggests that these cationized pullulan 3D matrices could be useful biomaterials for local gene transfer.

  18. Maturation of dendritic cells by pullulan promotes anti-cancer effect

    PubMed Central

    Xu, Li; Zhang, LiJun; Yu, Qing; Jin, Jun-O

    2016-01-01

    Previous studies have demonstrated that pullulan, a polysaccharide purified from Aureobasidium pullulans, has immune-stimulatory effects on T and B cells. Moreover, pullulan has been used as a carrier in the delivery of the antigen (Ag) peptide to lymphoid tissues. However, the in vivo effect of pullulan on dendritic cells (DC) has not been well characterized. In this study, we assessed the effect of pullulan on DC activation and anti-cancer immunity. The results showed that the pullulan treatment up-regulated co-stimulatory molecule expression and enhanced pro-inflammatory cytokine production in bone marrow-derived DCs (BMDC) in vitro and in spleen DCs in vivo. Moreover, the combination of ovalbumin (OVA) and pullulan induced OVA antigen-specific T cell activations in vivo. In tumor-bearing mice, pullulan induced the maturation of DCs in spleen and tumor draining lymph node (drLN), and promoted the OVA-specific T cell activation and migration of the T cells into the tumor. In addition, the combination of OVA and pullulan inhibited B16-OVA tumor growth and liver metastasis. The combination of tyrosinase-related protein 2 (TRP2) peptide and pullulan treatment also suppressed B16 melanoma growth. Thus, the results demonstrated that pullulan enhanced DC maturation and function, and it acted as an adjuvant in promoting Ag-specific immune responses in mice. Thus, pullulan could be a new and useful adjuvant for use in therapeutic cancer vaccines. PMID:27341129

  19. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing system...

  20. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 3 2012-10-01 2012-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed, the...

  1. 46 CFR 78.17-1 - Application.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 3 2012-10-01 2012-10-01 false Application. 78.17-1 Section 78.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS OPERATIONS Tests, Drills, and Inspections § 78.17-1 Application. (a) Except as specifically noted, the provisions of this subpart shall...

  2. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing system...

  3. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 3 2010-10-01 2010-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed, the...

  4. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing system...

  5. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 3 2014-10-01 2014-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed, the...

  6. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 3 2013-10-01 2013-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed, the...

  7. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing system...

  8. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing system...

  9. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 3 2011-10-01 2011-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed, the...

  10. 28 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 1 2013-07-01 2013-07-01 false Purpose. 17.1 Section 17.1 Judicial Administration DEPARTMENT OF JUSTICE CLASSIFIED NATIONAL SECURITY INFORMATION AND ACCESS TO CLASSIFIED INFORMATION § 17.1 Purpose. The purpose of this part is to ensure that information within the Department...

  11. Pullulan-based nanoparticles as carriers for transmucosal protein delivery.

    PubMed

    Dionísio, Marita; Cordeiro, Clara; Remuñán-López, Carmen; Seijo, Begoña; Rosa da Costa, Ana M; Grenha, Ana

    2013-09-27

    Polymeric nanoparticles have revealed very effective in transmucosal delivery of proteins. Polysaccharides are among the most used materials for the production of these carriers, owing to their structural flexibility and propensity to evidence biocompatibility and biodegradability. In parallel, there is a preference for the use of mild methods for their production, in order to prevent protein degradation, ensure lower costs and easier procedures that enable scaling up. In this work we propose the production of pullulan-based nanoparticles by a mild method of polyelectrolyte complexation. As pullulan is a neutral polysaccharide, sulfated and aminated derivatives of the polymer were synthesized to provide pullulan with a charge. These derivatives were then complexed with chitosan and carrageenan, respectively, to produce the nanocarriers. Positively charged nanoparticles of 180-270 nm were obtained, evidencing ability to associate bovine serum albumin, which was selected as model protein. In PBS pH 7.4, pullulan-based nanoparticles were found to have a burst release of 30% of the protein, which maintained up to 24h. Nanoparticle size and zeta potential were preserved upon freeze-drying in the presence of appropriate cryoprotectants. A factorial design was approached to assess the cytotoxicity of raw materials and nanoparticles by the metabolic test MTT. Nanoparticles demonstrated to not cause overt toxicity in a respiratory cell model (Calu-3). Pullulan has, thus, demonstrated to hold potential for the production of nanoparticles with an application in protein delivery. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Diterpenes and phenolic compounds from Sideritis pullulans.

    PubMed

    Faiella, Laura; Piaz, Fabrizio Dal; Bader, Ammar; Braca, Alessandra

    2014-10-01

    Phytochemical investigation of Sideritis pullulans aerial part and root extracts allowed to isolate six ent-kaurane diterpenes, two phenylpropanoids, and one coumarin, identified as 1α,3α,7β,18-tetrahydroxy-ent-kaur-16-ene (sideripullol A) (1), 3α,11α,18-trihydroxy-ent-kaur-16-ene (sideripullol B) (2), 3α,7β,18-trihydroxy-17-nor-ent-kauran-16-one (sideritone A) (3), 3α,7β-dihydroxy-18-acetyloxy-17-nor-ent-kauran-16-one (sideritone B) (4), 3α,7β,16α,17-tetrahydroxy-18-acetyloxy-ent-kaurane (sideripullol C) (5), 7β,16α,17,18-tetrahydroxy-ent-kaurane (sideripullol D) (6), β-(3-methoxy-4-hydroxyphenyl)-ethyl-O-α-l-arabinopiranosyl-(1→2)-α-l-rhamnopyranosyl-(1→3)-6-O-t-feruloyl-β-d-glucopyranoside (sideritiside A) (7), β-(3,4-dihydroxyphenyl)-ethyl-O-α-l-arabinopiranosyl-(1→2)-α-l-rhamnopyranosyl-(1→3)-6-O-t-feruloyl-β-d-glucopyranoside (sideritiside B) (8), and 7-demethyl-8-methoxycoumarsabin (9), respectively. Twenty known compounds, including phenolics, flavonol glycosides, iridoids, diterpenes, sesquiterpenes, lignans, coumarins, and phenylpropanoids were also isolated and characterized. All diterpenes were evaluated for their cytotoxic activity.

  13. RNA Protection is Effectively Achieved by Pullulan Film Formation.

    PubMed

    Hsieh, Ping-Yao; Monsur Ali, M; Tram, Kha; Jahanshahi-Anbuhi, Sana; Brown, Christine L; Brennan, John D; Filipe, Carlos D M; Li, Yingfu

    2017-03-16

    RNA is a functionally versatile polymer but suffers from susceptibility to spontaneous and RNase-catalyzed degradation. This vulnerability makes it difficult to preserve RNA for extended periods of time, thus limiting its use in various contexts, including practical applications as functional nucleic acids. Here we present a simple method to preserve RNA by pullulan (a complex sugar produced by Aureobasidium pullulans fungus) film formation. This strategy can markedly suppress both spontaneous and RNase degradation. Importantly, the pullulan film readily dissolves in aqueous solution, thus allowing retrieval of fully functional RNA species. In order to illustrate the advantage of this protective method in a practical application, we engineered a simple paper sensor containing a bacteria-detecting RNA-cleaving DNAzyme. This detection capability of the device was unchanged after storage at room temperature for six months.

  14. Pasting investigation, SEM observation and the possible interaction study on rice starch-pullulan combination.

    PubMed

    Chen, Long; Ren, Fei; Yu, Xueping; Zhang, Zipei; Xu, Dejun; Tong, Qunyi

    2015-02-01

    The pasting properties of rice starch (RS) with high concentration (10%, w/w) were investigated in the presence or absence of pullulan (PUL) using a rapid visco-analyzer (RVA). Addition of pullulan resulted in the reduction of peak viscosity, trough viscosity, final viscosity, and setback value of RS. Furthermore, an interesting phenomenon, i.e. a small viscosity peak appeared in the RVA curves of RS-PUL mixtures, was observed. It indicated that addition of pullulan might suppress the gelatinization of starch granules by maintaining the integration of some granules. The scanning electron microscope (SEM) observation of samples suggested that starch granules could be wrapped by a thin membrane composed of pullulan and/or pullulan-amylose associations. The coating ability of pullulan and/or the possible molecular interactions between pullulan and amylose could be responsible for these results.

  15. Enhanced production of Ca²⁺-polymalate (PMA) with high molecular mass by Aureobasidium pullulans var. pullulans MCW.

    PubMed

    Wang, Yu-Kuang; Chi, Zhe; Zhou, Hai-Xiang; Liu, Guang-Lei; Chi, Zhen-Ming

    2015-08-07

    Polymalic acid (PMA) has many applications in food and medical industries. However, so far it has not been commercially produced by fermentation. Therefore, it is very important how to develop an economical process for a large scale production of PMA by one step fermentation. After over 200 strains of Aureobasidium spp. isolated from the mangrove systems in the South of China were screened for their ability to produce Ca(2+)-polymalate (PMA), it was found that Aureobasidium pullulans var. pullulans MCW strain among them could produce high level of Ca(2+)-PMA. The medium containing only 140.0 g/L glucose, 65.0 g/L CaCO3 and 7.5 g/L corn steep liquor was found to be the most suitable for Ca(2+)-PMA production. Then, 121.3 g/L of Ca(2+)-PMA was produced by A. pullulans var. pullulans MCW strain within 120 h at flask level. During 10-L batch fermentation, 152.52 g/L of Ca(2+)-PMA in the culture and 8.6 g/L of cell dry weight were obtained within 96 h, leaving 4.5 g/L of reducing sugar in the fermented medium. After purification of the Ca(2+)-PMA from the culture and acid hydrolysis of the purified Ca(2+)-PMA, HPLC analysis showed that A. pullulans var. pullulans MCW strain produced only one main component of Ca(2+)-PMA and the hydrolysate of the purified Ca(2+)-PMA was mainly composed of L-malic acid. Mw (the apparent molecular weight) of the purified PMA was 2.054 × 10(5) (g/moL) and the purified PMA was estimated to be composed of 1784 L-malic acids. It was found that A. pullulans var. pullulans MCW strain obtained in this study could yield 152.52 g/L of Ca(2+)-PMA within the short time, the produced PMA had the highest molecular weight and the medium for production of Ca(2+)- PMA by this yeast was very simple.

  16. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 45 Public Welfare 1 2010-10-01 2010-10-01 false Definition. 17.1 Section 17.1 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION RELEASE OF ADVERSE INFORMATION TO NEWS MEDIA... Department or any principal operating component made to the news media inviting public attention to an action...

  17. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 45 Public Welfare 1 2013-10-01 2013-10-01 false Definition. 17.1 Section 17.1 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION RELEASE OF ADVERSE INFORMATION TO NEWS MEDIA... Department or any principal operating component made to the news media inviting public attention to an action...

  18. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 45 Public Welfare 1 2014-10-01 2014-10-01 false Definition. 17.1 Section 17.1 Public Welfare Department of Health and Human Services GENERAL ADMINISTRATION RELEASE OF ADVERSE INFORMATION TO NEWS MEDIA... Department or any principal operating component made to the news media inviting public attention to an action...

  19. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 45 Public Welfare 1 2011-10-01 2011-10-01 false Definition. 17.1 Section 17.1 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION RELEASE OF ADVERSE INFORMATION TO NEWS MEDIA... Department or any principal operating component made to the news media inviting public attention to an action...

  20. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 45 Public Welfare 1 2012-10-01 2012-10-01 false Definition. 17.1 Section 17.1 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION RELEASE OF ADVERSE INFORMATION TO NEWS MEDIA... Department or any principal operating component made to the news media inviting public attention to an action...

  1. Bioproducts from diverse phylogenetic clades of Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    More than 90 isolates of the fungus A. pullulans from tropical and temperate climates were classified into 13 phylogenetic clades using multilocus sequence analyses (ITS, IGS, BT2, RPB2, and EF-1a). Tropical isolates appeared to exhibit the greatest genetic diversity. Representatives of each clade...

  2. Laccase production by diverse phylogenetic clades of Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    Laccases (EC 1.10.3.2) have numerous potential industrial applications including the degradation of dyes and toxic materials. Novel sources of this enzyme would be desirable to improve activity yields and substrate specificities. In this study we tested 51 strains of A. pullulans representing 13 d...

  3. Production of novel antibacterial liamocins by strains of Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    Certain strains of Aureobasidium pullulans produce liamocins, heavier-than-water “oils” that accumulate in liquid cultures. Liamocins are surface active, and inhibit mammalian cancer cell lines. Recently, we discovered that liamocins have antibacterial activity with specificity against Streptococcus...

  4. Fabrication of pullulan and pectin submicron fibers by electrospinning

    USDA-ARS?s Scientific Manuscript database

    Pullulan (PUL), a food grade polysaccharide, was fabricated into fibrous mats from fibers of submicron size by electrospinning. The effects of inorganic salts and polyanions present in the electrospinning solution on the properties of the resultant fibers was investigated. The inclusion of exogenous...

  5. Isolation and Identification of Trehalase from Pullularia pullulans

    PubMed Central

    Merdinger, Emanuel; Lange, Charles F.; Booker, Ben F.

    1971-01-01

    Trehalase has been isolated from Pullularia pullulans. The enzyme, which is specific for trehalose, was purified approximately 800-fold. The optimal pH was found to be 4.0 and the Michaelis dissociation constant, Km, was determined to be 3.2 × 10−3m. PMID:5104982

  6. Characterization of Pullulan/Chitosan Oligosaccharide/Montmorillonite Nanofibers Prepared by Electrospinning Technique.

    PubMed

    Rabbani, Mohammad Mahbub; Yang, Seong Baek; Park, Soo-Jin; Oh, Weontae; Yeum, Jeong Hyun

    2016-06-01

    Pullulan/Chitosan oligosaccharide (COS)/Montmorillonite (MMT) hybrid nanofibers were electrospun from their aqueous solution using different Pullulan/COS mass ratios and variable amounts of MMT. The effects of Pullulan/COS mass ratios and MMT contents on the morphologies and properties of PulluIan/COS/MMT hybrid nanofibers were investigated. The obtained nanofibers were characterized with field emission-scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), thermo gravimetric analysis (TGA), and tensile strength measurement. The Pullulan/COS mass ratio and MMT contents significantly influence the morphologies and properties of the Pullulan/COS/MMT hybrid nanofibers. Higher Pullulan contents than COS contents forms uniform and bead free nanofibers. The addition of COS to Pullulan improves the thermal stability of Pullulan/COS blend nanofibers. The incorporation of MMT to the Pullulan/COS/MMT hybrid nanofibers increase their fiber diameter, improves their thermal stability and tensile strength. These morphological changes and property enhancement depend on the amount of MMT added. The XRD and TEM results suggest the coexistence of Pullulan, COS and MMT within polymer matrix through intercalation of polymer chain between silicate layers forming well-ordered multiplayer morphology with alternating polymeric and silicate layers.

  7. Cholesterol-Modified Amino-Pullulan Nanoparticles as a Drug Carrier: Comparative Study of Cholesterol-Modified Carboxyethyl Pullulan and Pullulan Nanoparticles

    PubMed Central

    Tao, Xiaojun; Xie, Yongchao; Zhang, Qiufang; Qiu, Ximin; Yuan, Liming; Wen, Yi; Li, Min; Yang, Xiaoping; Tao, Ting; Xie, Minghui; Lv, Yanwei; Wang, Qinyi; Feng, Xing

    2016-01-01

    To search for nano-drug preparations with high efficiency in tumor treatment, we evaluated the drug-loading capacity and cell-uptake toxicity of three kinds of nanoparticles (NPs). Pullulan was grafted with ethylenediamine and hydrophobic groups to form hydrophobic cholesterol-modified amino-pullulan (CHAP) conjugates. Fourier transform infrared spectroscopy and nuclear magnetic resonance were used to identify the CHAP structure and calculate the degree of substitution of the cholesterol group. We compared three types of NPs with close cholesterol hydrophobic properties: CHAP, cholesterol-modified pullulan (CHP), and cholesterol-modified carboxylethylpullulan (CHCP), with the degree of substitution of cholesterol of 2.92%, 3.11%, and 3.46%, respectively. As compared with the two other NPs, CHAP NPs were larger, 263.9 nm, and had a positive surface charge of 7.22 mV by dynamic light-scattering measurement. CHAP NPs showed low drug-loading capacity, 12.3%, and encapsulation efficiency of 70.8%, which depended on NP hydrophobicity and was affected by surface charge. The drug release amounts of all NPs increased in the acid media, with CHAP NPs showing drug-release sensitivity with acid change. Cytotoxicity of HeLa cells was highest with mitoxantrone-loaded CHAP NPs on MTT assay. CHAP NPs may have potential as a high-efficiency drug carrier for tumor treatment. PMID:28335293

  8. Low-melanin containing pullulan production from sugarcane bagasse hydrolysate by Aureobasidium pullulans in fermentations assisted by light-emitting diode.

    PubMed

    Terán Hilares, Ruly; Orsi, Camila Ayres; Ahmed, Muhammad Ajaz; Marcelino, Paulo Franco; Menegatti, Carlos Renato; da Silva, Silvio Silvério; Dos Santos, Júlio César

    2017-04-01

    Pullulan is a polymer produced by Aureobasidium pullulans and the main bottleneck for its industrial production is the presence of melanin pigment. In this study, light-emitting diodes (LEDs) of different wavelengths were used to assist the fermentation process aiming to produce low-melanin containing pullulan by wild strain of A. pullulans LB83 with different carbon sources. Under white light using glucose-based medium, 11.75g.L(-1) of pullulan with high melanin content (45.70UA540nm.g(-1)) was obtained, this production improved in process assisted by blue LED light, that resulted in 15.77g.L(-1) of pullulan with reduced content of melanin (4.46UA540nm.g(-1)). By using sugarcane bagasse (SCB) hydrolysate as carbon source, similar concentration of pullulan (about 20g.L(-1)) was achieved using white and blue LED lights, with lower melanin contents in last. Use of LED light was found as a promising approach to assist biotechnological process for low-melanin containing pullulan production. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. 43 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... DEPARTMENT OF THE INTERIOR Nondiscrimination on the Basis of Race, Color, or National Origin § 17.1 Purpose... to the end that no person in the United States shall, on the grounds of race, color, or national...

  10. 43 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... DEPARTMENT OF THE INTERIOR Nondiscrimination on the Basis of Race, Color, or National Origin § 17.1 Purpose... to the end that no person in the United States shall, on the grounds of race, color, or national...

  11. 43 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... DEPARTMENT OF THE INTERIOR Nondiscrimination on the Basis of Race, Color, or National Origin § 17.1 Purpose... to the end that no person in the United States shall, on the grounds of race, color, or national...

  12. 43 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... DEPARTMENT OF THE INTERIOR Nondiscrimination on the Basis of Race, Color, or National Origin § 17.1 Purpose... to the end that no person in the United States shall, on the grounds of race, color, or national...

  13. 43 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... DEPARTMENT OF THE INTERIOR Nondiscrimination on the Basis of Race, Color, or National Origin § 17.1 Purpose... to the end that no person in the United States shall, on the grounds of race, color, or national...

  14. Deoiledjatropha seed cake is a useful nutrient for pullulan production.

    PubMed

    Choudhury, Anirban Roy; Sharma, Nishat; Prasad, G S

    2012-03-30

    Ever increasing demand for fossil fuels is a major factor for rapid depletion of these non-renewable energy resources, which has enhanced the interest of finding out alternative sources of energy. In recent years jatropha seed oil has been used extensively for production of bio-diesel and has shown significant potential to replace petroleum fuels at least partially. De-oiled jatropha seed cake (DOJSC) which comprises of approximately 55 to 65% of the biomass is a byproduct of bio-diesel industry. DOJSC contains toxic components like phorbol esters which restricts its utilization as animal feed. Thus along with the enhancement of biodiesel production from jatropha, there is an associated problem of handling this toxic byproduct. Utilization of DOJSC as a feed stock for production of biochemicals may be an attractive solution to the problem.Pullulan is an industrially important polysaccharide with several potential applications in food, pharmaceuticals and cosmetic industries. However, the major bottleneck for commercial utilization of pullulan is its high cost. A cost effective process for pullulan production may be developed using DOJSC as sole nutrient source which will in turn also help in utilization of the byproduct of bio-diesel industry. In the present study, DOJSC has been used as a nutrient for production of pullulan, in place of conventional nutrients like yeast extract and peptone. Process optimization was done in shake flasks, and under optimized conditions (8% DOJSC, 15% dextrose, 28°C temperature, 200 rpm, 5% inoculum, 6.0 pH) 83.98 g/L pullulan was obtained. The process was further validated in a 5 L laboratory scale fermenter. This is the first report of using DOJSC as nutrient for production of an exopolysaccharide. Successful use of DOJSC as nutrient will help in finding significant application of this toxic byproduct of biodiesel industry. This in turn also have a significant impact on cost reduction and may lead to development of a cost

  15. Media optimization for elevated molecular weight and mass production of pigment-free pullulan.

    PubMed

    Yu, Xiaoliu; Wang, Yulei; Wei, Gongyuan; Dong, Yingying

    2012-07-01

    In this study, an Aureobasidium pullulans SZU 1001 mutant, deficient in pigment production, was screened by complex UV and γ-ray mutagenesis. Medium composition optimization for increased pullulan molecular weight and production was conducted using this mutant. Six nutrients: yeast extract, (NH4)2SO4, K2HPO4, NaCl, MgSO4·7H2O and CaCl2 were detected within pullulan production in flasks. It is shown that NaCl and K2HPO4 have significant influences on molecular weight of pullulan, while yeast extract and (NH4)2SO4 significantly affect pullulan yield. To achieve a higher molecular weight and more efficient pullulan production, a response surface methodology approach was introduced to predict an optimal nutrient combination. A molecular weight of 5.74 × 10(6) and pullulan yield on glucose of 51.30% were obtained under batch pullulan fermentation with the optimized media, which increased molecular weight and pullulan production by 97.25% and 11.04%, respectively compared with the control media. Copyright © 2012 Elsevier Ltd. All rights reserved.

  16. Pasting and rheological properties of rice starch as affected by pullulan.

    PubMed

    Chen, Long; Tong, Qunyi; Ren, Fei; Zhu, Guilan

    2014-05-01

    Effect of pullulan (PUL) on the pasting, rheological properties of rice starch (RS) was investigated. The swelling power, amylose leaching, and confocal laser scanning microscopy (CLSM) observation of the samples were also conducted to explore the possible interaction between starch and pullulan. Rapid visco-analysis (RVA) showed that PUL significantly changed viscosity parameters of rice starch-pullulan (RS-PUL) mixtures. Dynamic rheological measurements revealed that the modulus (G', G″) of the mixtures increased with the increase of pullulan concentration from 0.01% to 0.07%, but then decreased with the increase of pullulan concentration from 0.07% to 0.50%. The pasting and rheological properties of samples indicated that pullulan could blend well with rice starch and promote the gelatinization of starch granules at low concentration of pullulan, but suppress the gelatinization of starch granules at high concentration of pullulan. The results of swelling power, leached amylose and CLSM observation of samples further suggest that the interaction between starch and pullulan occurred in the RS-PUL system and the interaction was hypothesized to be responsible for these results.

  17. Production of novel types of antibacterial liamocins by diverse strains of Aureobasidium pullulans grown on different culture media

    USDA-ARS?s Scientific Manuscript database

    Objective: The objective was to compare production of antibacterial liamocins by diverse strains of A. pullulans grown on different culture media. Results: Liamocins produced by strains of A. pullulans have potential agricultural and pharmaceutical applications as antibacterials with specificity aga...

  18. Adsorption of phospholipid bilayers onto pullulan-modified cellulose surfaces

    NASA Astrophysics Data System (ADS)

    Choi, Heejun; Liu, Zelin; Esker, Alan

    2009-03-01

    1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC) vesicle adsorption onto regenerated cellulose and pullulan 4-bromocinnamate (P4BC) modified cellulose surfaces was investigated via surface plasmon resonance (SPR) spectroscopy and quartz crystal microbalance with dissipation monitoring (QCM-D). P4BC with a degree of substitution (DS) of 0.061 ± 0.002 from UV measurements and 0.058 from ^1H NMR was synthesized from pullulan and 4-bromocinnamic acid to yield P4BC. The deduced thicknesses from SPR for DMPC layers were ˜3.7 nm (bilayer) on regenerated cellulose surfaces and ˜2.1 nm (monolayer) on P4BC modified cellulose surfaces. Qualitative analysis of the QCM-D data also indicated that the DMPC layers on P4BC modified cellulose surfaces were thinner than on regenerated cellulose surfaces.

  19. Surface plasmon resonance studies of pullulan and pullulan cinnamate adsorption onto cellulose.

    PubMed

    Kaya, Abdulaziz; Du, Xiaosong; Liu, Zelin; Lu, Jessica W; Morris, John R; Glasser, Wolfgang G; Heinze, Thomas; Esker, Alan R

    2009-09-14

    Surface plasmon resonance studies showed pullulan cinnamates (PCs) with varying degrees of substitution (DS) adsorbed onto regenerated cellulose surfaces from aqueous solutions below their critical aggregation concentrations. Results on cellulose were compared to PC adsorption onto hydrophilic and hydrophobic self-assembled thiol monolayers (SAMs) on gold to probe how different interactions affected PC adsorption. PC adsorbed onto methyl-terminated SAMs (SAM-CH(3)) > cellulose > hydroxyl-terminated SAMs (SAM-OH) for high DS and increased with DS for each surface. Data for PC adsorption onto cellulose and SAM-OH surfaces were effectively fit by Langmuir isotherms; however, Freundlich isotherms were required to fit PC adsorption isotherms for SAM-CH(3) surfaces. Atomic force microscopy images from the solid/liquid interfaces revealed PC coatings were uniform with surface roughnesses <2 nm for all surfaces. This study revealed hydrogen bonding alone could not explain PC adsorption onto cellulose and hydrophobic modification of water-soluble polysaccharides was a facile strategy for their conversion into surface modifying agents.

  20. 15 CFR 17.1 - Licensing rules.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... THE CUSTODY OF THE DEPARTMENT OF COMMERCE Licensing of Rights in Domestic Patents and Patent Applications § 17.1 Licensing rules. (a) The Government-wide rules for the licensing of rights in domestic... applicable to all such licensing activities of the Department of Commerce, subject to the following...

  1. Phylogenetic classification of Aureobasidium pullulans strains for production of feruloyl esterase

    USDA-ARS?s Scientific Manuscript database

    The objective was to phylogenetically classify diverse strains of A. pullulans and determine their production of feruloyl esterase. Seventeen strains from the A. pullulans literature were phylogenetically classified. Phenotypic traits of color variation and endo-ß-1,4-xylanase overproduction were as...

  2. Effect of rice wax on water vapor permeability and sorption properties of edible pullulan films

    USDA-ARS?s Scientific Manuscript database

    Edible pullulan films were prepared by substituting the pullulan with various ratios of rice wax. Freestanding composite films were obtained with up to 50% rice wax. Water vapor barrier properties of the film were improved with increased addition of the rice wax. Moisture sorption isotherms were ...

  3. Aureobasidium pullulans as a biocontrol agent of postharvest pathogens of apples in Uruguay

    USDA-ARS?s Scientific Manuscript database

    The yeast, Aureobasidium pullulans, was the microorganism most frequently recovered from the surface of apple fruit (cv. Red Delicious) stored in commercial cold chambers for six months. In the present work, ten isolates of Aureobasidium pullulans were assayed to determine if they could control blu...

  4. Aureobasidium pullulans as a source of liamocins (heavy oils) with anticancer activity

    USDA-ARS?s Scientific Manuscript database

    Liamocins are structurally unique, heavier-than-water “oils” produced by certain strains of Aureobasidium pullulans. Nine strains of A. pullulans from phylogenetic clades 8, 9, and 11 were examined for the first time for production of liamocins. Strains in these clades have only been isolated from t...

  5. Poly (beta-L-malic acid) production by diverse phylogenetic clades of Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    Poly (beta-L-malic acid) (PMA) is a natural biopolyester that has pharmaceutical applications and other potential uses. Here we examine PMA production by genetically diverse phylogenetic clades of the fungus A. pullulans. Thirty-six strains of A. pullulans were isolated for this study from various...

  6. Production of anti-streptococcal liamocins from agricultural biomass by Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    Liamocins are unique heavier-than-water “oils” produced by certain strains of the fungus Aureobasidium pullulans. Liamocins have antibacterial activity with specificity for Streptococcus sp. Previous studies reported that liamocin yields were highest from strains of A. pullulans belonging to phylog...

  7. A new pullulan-producing yeast and medium optimization for its exopolysaccharide production

    NASA Astrophysics Data System (ADS)

    Shuangzhi, Zhao; Zhenming, Chi

    2003-04-01

    Yeast strain Y68 producing high level of pullulan was isolated from the phyton collected in Toulouse, France. This strain was identified to be Rhodotorula bacarum by BIOLOG analysis. This is the first report that pullulan was produced by Rhodotorula bacarum. The optimal medium (g L-1) for pullulan production by this strain was 80 glucose, 20 soybean cake hydrolysate, 5 K2HPO4, 1 NaCl, 0.2 MgSO4·7H2O, 0.6 (NH4)2SO4, pH 7.0. Under this condition, 54 gL-1 pullulan was produced within 60 h at 30°C. Pullulan is a better starting material for producing marine prodrugs.

  8. Effect of oligosaccharides derived from Laminaria japonica-incorporated pullulan coatings on preservation of cherry tomatoes.

    PubMed

    Wu, Shengjun; Lu, Mingsheng; Wang, Shujun

    2016-05-15

    Laminaria japonica-derived oligosaccharides (LJOs) exhibit antibacterial and antioxidant activities, and pullulan is a food thickener that can form impermeable films. The ability of pullulan coatings with various LJO concentrations (1% pullulan+0.1%, 0.2% or 0.3% LJOs) to preserve cherry tomatoes during storage at room temperature was investigated. The LJO-incorporated pullulan coatings were found to effectively reduce respiratory intensity, vitamin C loss, weight loss and softening, as well as to increase the amount of titratable acid and the overall likeness of fruit compared with the control. These effects were observed to be dose-dependent. Therefore, using LJO-incorporated pullulan coatings can extend the shelf life of cherry tomatoes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Single-Leaf Resolution of the Temporal Population Dynamics of Aureobasidium pullulans on Apple Leaves

    PubMed Central

    Woody, Scott T.; Spear, Russell N.; Nordheim, Erik V.; Ives, Anthony R.; Andrews, John H.

    2003-01-01

    The abundance of phylloplane microorganisms typically varies over several orders of magnitude among leaves sampled concurrently. Because the methods traditionally used to sample leaves are destructive, it has remained unclear whether this high variability is due to fixed differences in habitat quality among leaves or to asynchronous temporal variation in the microbial population density on individual leaves. We developed a novel semidestructive assay to repeatedly sample the same apple leaves from orchard trees over time by removing progressively more proximal ∼1-cm-wide transverse segments. Aureobasidium pullulans densities were determined by standard leaf homogenization and plating procedures and were expressed as CFU per square centimeter of segment. The A. pullulans population densities among leaves were lognormally distributed. The variability in A. pullulans population densities among subsections of a given leaf was one-third to one-ninth the variability among whole leaves harvested concurrently. Sequential harvesting of leaf segments did not result in detectable changes in A. pullulans density on residual leaf surfaces. These findings implied that we could infer whole-leaf A. pullulans densities over time by using partial leaves. When this successive sampling regimen was applied over the course of multiple 7- to 8-day experiments, the among-leaf effects were virtually always the predominant source of variance in A. pullulans density estimates. Changes in A. pullulans density tended to be synchronous among leaves, such that the rank order of leaves arrayed with respect to A. pullulans density was largely maintained through time. Occasional periods of asynchrony were observed, but idiosyncratic changes in A. pullulans density did not contribute appreciably to variation in the distribution of populations among leaves. This suggests that persistent differences in habitat (leaf) quality are primarily responsible for the variation in A. pullulans density among

  10. Comparative proteomic analysis of Aureobasidium pullulans in the presence of high and low levels of nitrogen source.

    PubMed

    Sheng, Long; Zhu, Guilan; Tong, Qunyi

    2014-10-29

    Pullulan, produced by Aureobasidium pullulans strain, has been broadly used in the food and medical industries. However, relatively little is known concerning the molecular basis of pullulan biosynthesis of this strain. In this paper, the effect of different concentrations of (NH4)2SO4 on pullulan fermentation was studied. Proteomics containing two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS) were used to analyze the protein with different expressions of A. pullulans cells between the nitrogen limitation and nitrogen repletion. Maximum pullulan production reached 37.72 g/L when 0.6 g/L of initial (NH4)2SO4 was added. Excess nitrogen source would impel carbon flux flow toward biomass production, but decreased the pullulan production. Nitrogen limitation in A. pullulans seemed to influence the flux change of carbon flux flow toward exopolysaccharide accumulation. The findings indicated that 12 identified protein spots were involved in energy-generating enzymes, antioxidant-related enzymes, amino acid biosynthesis, glycogen biosynthesis, glycolysis, protein transport, and transcriptional regulation. These results presented more evidence of pullulan biosynthesis under nitrogen-limited environment, which would provide a molecular understanding of the physiological response of A. pullulans for optimizing the performance of industrial pullulan fermentation.

  11. Preservation of Brussels Sprouts by Pullulan Coating Containing Oregano Essential Oil.

    PubMed

    Kraśniewska, Karolina; Gniewosz, Małgorzata; Kosakowska, Olga; Cis, Aneta

    2016-03-01

    In this study, the effectiveness of pullulan (a fungal polysaccharide) film containing oregano essential oil (OEO) at 1.0 to 10.0% was evaluated against bacteria, yeasts, and molds. The quality of the sprouts, as determined by weight loss, color, and appearance, was monitored during storage at 2 and 16°C. An organoleptic evaluation of odor preference and odor acceptability of OEO on the Brussels sprouts was also conducted. The antimicrobial activity of pullulan films with OEO increased significantly with the increase in OEO concentration (1 to 10%). Pullulan films with OEO were more effective for inhibiting the growth of yeasts and molds than for inhibiting gram-positive and gram-negative bacteria. Pullulan with 1.0% OEO was an effective combination and was used subsequently as the base coating for maintaining the safety and quality of fresh Brussels sprouts stored at 16°C for 14 days. The pullulan coating containing 1.0% OEO reduced Aspergillus niger populations by 2 log CFU/g. This coating also reduced weight loss in the sprouts. Compared with uncoated Brussels sprouts, the percent weight loss after 14 days was reduced in samples coated with pullulan and with pullulan plus 1% OEO by 3.81 and 6.06%, respectively, after storage at 2°C and by 8.04 and 9.30%, respectively, after storage 16°C. The coating also significantly reduced changes in general appearance and color during storage. Evaluation of the organoleptic properties indicated that pullulan containing OEO had only a slight detrimental effect on odor properties. Incorporating OEO into a delivery system for antimicrobial compounds in pullulan coatings extended the microbiological shelf life of Brussels sprouts.

  12. Production, Purification, and Properties of a Thermostable beta-Glucosidase from a Color Variant Strain of Aureobasidium pullulans.

    PubMed

    Saha, B C; Freer, S N; Bothast, R J

    1994-10-01

    A color variant strain of Aureobasidium pullulans (NRRL Y-12974) produced beta-glucosidase activity when grown in liquid culture on a variety of carbon sources, such as cellobiose, xylose, arabinose, lactose, sucrose, maltose, glucose, xylitol, xylan, cellulose, starch, and pullulan. An extracellular beta-glucosidase was purified 129-fold to homogeneity from the cell-free culture broth of the organism grown on corn bran. The purification protocol included ammonium sulfate treatment, CM Bio-Gel A agarose column chromatography, and gel filtrations on Bio-Gel A-0.5m and Sephacryl S-200. The beta-glucosidase was a glycoprotein with native molecular weight of 340,000 and was composed of two subunits with molecular weights of about 165,000. The enzyme displayed optimal activity at 75 degrees C and pH 4.5 and had a specific activity of 315 mumol . min . mg of protein under these conditions. The purified beta-glucosidase was active against p-nitrophenyl-beta-d-glucoside, cellobiose, cellotriose, cellotetraose, cellopentaose, cellohexaose, and celloheptaose, with K(m) values of 1.17, 1.00, 0.34, 0.36, 0.64, 0.68, and 1.65 mM, respectively. The enzyme activity was competitively inhibited by glucose (K(i) = 5.65 mM), while fructose, arabinose, galactose, mannose, and xylose (each at 56 mM) and sucrose and lactose (each at 29 mM) were not inhibitory. The enzyme did not require a metal ion for activity, and its activity was not affected by p-chloromercuribenzoate (0.2 mM), EDTA (10 mM), or dithiothreitol (10 mM). Ethanol (7.5%, vol/vol) stimulated the initial enzyme activity by 15%. Glucose production was enhanced by 7.9% when microcrystalline cellulose (2%, wt/vol) was treated for 48 h with a commercial cellulase preparation (5 U/ml) that was supplemented with the purified beta-glucosidase (0.21 U/ml) from A. pullulans.

  13. Economic co-production of poly(malic acid) and pullulan from Jerusalem artichoke tuber by Aureobasidium pullulans HA-4D.

    PubMed

    Xia, Jun; Xu, Jiaxing; Liu, Xiaoyan; Xu, Jiming; Wang, Xingfeng; Li, Xiangqian

    2017-02-23

    poly(L-malic acid) (PMA) is a water-soluble polyester with many attractive properties in medicine and food industries, but the high cost of PMA fermentation has restricted its further application for large-scale production. To overcome this problem, PMA production from Jerusalem artichoke tubers was successfully performed. Additionally, a valuable exopolysaccharide, pullulan, was co-produced with PMA by Aureobasidum pullulans HA-4D. The Jerusalem artichoke medium for PMA and pullulan co-production contained only 100 g/L hydrolysate sugar, 30 g/L CaCO3 and 1 g/L NaNO3. Compared with the glucose medium, the Jerusalem artichoke medium resulted in a higher PMA concentration (114.4 g/L) and a lower pullulan concentration (14.3 g/L) in a 5 L bioreactor. Meanwhile, the activity of pyruvate carboxylase and malate dehydrogenas was significantly increased, while the activity of α-phosphoglucose mutase, UDP-glucose pyrophosphorylase and glucosyltransferase was not affected. To assay the economic-feasibility, large-scale production in a 1 t fermentor was performed, yielding 117.5 g/L PMA and 15.2 g/L pullulan. In this study, an economical co-production system for PMA and pullulan from Jerusalem artichoke was developed. The medium for PMA and pullulan co-production was significantly simplified when Jerusalem artichoke tubers were used. With the simplified medium, PMA production was obviously stimulated, which would be associated with the improved activity of pyruvate carboxylase and malate dehydrogenas.

  14. Optimization of fast disintegration tablets using pullulan as diluent by central composite experimental design.

    PubMed

    Patel, Dipil; Chauhan, Musharraf; Patel, Ravi; Patel, Jayvadan

    2012-03-01

    The objective of this work was to apply central composite experimental design to investigate main and interaction effect of formulation parameters in optimizing novel fast disintegration tablets formulation using pullulan as diluents. Face centered central composite experimental design was employed to optimize fast disintegration tablet formulation. The variables studied were concentration of diluents (pullulan, X(1)), superdisintigrant (sodium starch glycolate, X(2)), and direct compression aid (spray dried lactose, X(3)). Tablets were characterized for weight variation, thickness, disintegration time (Y(1)) and hardness (Y(2)). Good correlation between the predicted values and experimental data of the optimized formulation methodology in optimizing fast disintegrating tablets using pullulan as a diluent.

  15. New improved method for fructooligosaccharides production by Aureobasidium pullulans.

    PubMed

    Dominguez, Ana; Nobre, Clarisse; Rodrigues, Lígia R; Peres, António M; Torres, Duarte; Rocha, Isabel; Lima, Nelson; Teixeira, José

    2012-08-01

    Fructooligosaccharides are prebiotics with numerous health benefits within which the improvement of gut microbiota balance can be highlighted, playing a key role in individual health. In this study, an integrated one-stage method for FOS production via sucrose fermentation by Aureobasidium pullulans was developed and optimized using experimental design tools. Optimization of temperature and agitation speed for maximizing the FOS production was performed using response surface methodology. Temperature was found to be the most significant parameter. The optimum fermentation conditions were found to be 32 °C and 385 rpm. Under these conditions, the model predicted a total FOS production yield of 64.7 gFOS/gsucrose. The model was validated at optimal conditions in order to check its adequacy and accuracy and an experimental yield of 64.1 (±0.0) gFOS/gsucrose was obtained. A significant improvement of the total FOS production yields by A. pullulans using a one-stage process was obtained.

  16. Effects of plastic composite support and pH profiles on pullulan production in a biofilm reactor.

    PubMed

    Cheng, Kuan-Chen; Demirci, Ali; Catchmark, Jeffrey M

    2010-04-01

    Pullulan is a linear homopolysaccharide which is composed of glucose units and often described as alpha-1, 6-linked maltotriose. The applications of pullulan range from usage as blood plasma substitutes to environmental pollution control agents. In this study, a biofilm reactor with plastic composite support (PCS) was evaluated for pullulan production using Aureobasidium pullulans. In test tube fermentations, PCS with soybean hulls, defatted soy bean flour, yeast extract, dried bovine red blood cells, and mineral salts was selected for biofilm reactor fermentation (due to its high nitrogen content, moderate nitrogen leaching rate, and high biomass attachment). Three pH profiles were later applied to evaluate their effects on pullulan production in a PCS biofilm reactor. The results demonstrated that when a constant pH at 5.0 was applied, the time course of pullulan production was advanced and the concentration of pullulan reached 32.9 g/L after 7-day cultivation, which is 1.8-fold higher than its respective suspension culture. The quality analysis demonstrated that the purity of produced pullulan was 95.8% and its viscosity was 2.4 centipoise. Fourier transform infrared spectroscopy spectra also supported the supposition that the produced exopolysaccharide was mostly pullulan. Overall, this study demonstrated that a biofilm reactor can be successfully implemented to enhance pullulan production and maintain its high purity.

  17. POLYSACCHARIDES FROM CELL WALLS OF AUREOBASIDIUM (PULLULARIA) PULLULANS. PART I. GLUCANS,

    DTIC Science & Technology

    The cell wall of Aureobasidium (Pullularia) pullulans contains three types of beta - glucan . One, extracted with dilute alkali, has a linear backbone...insoluble in dilute alkali contains a highly crystalline, essentially linear linked glucan and an amorphous glucan . (Author)

  18. Isolation of new aureobasidium strains that produce high-molecular-weight pullulan with reduced pigmentation.

    PubMed

    Pollock, T J; Thorne, L; Armentrout, R W

    1992-03-01

    New isolates of Aureobasidium pullulans were obtained from plant leaf surfaces gathered in San Diego County. The new fungal isolates were identified as A. pullulans on the basis of the appearance of polymorphic colonies formed on agar plates, the electrophoretic profiles of repeated genomic DNA sequences, and the production of pullulan in shake flask cultures. The isolates showed different degrees of pigmentation. One of the natural isolates was nonpigmented under mock production conditions in liquid culture, but was still able to synthesize a reduced amount of pigment on agar plates at late times. A mutagenic treatment with ethidium bromide produced derivatives of normally pigmented natural isolates that exhibited an increased tendency toward yeastlike growth and reduced pigmentation. Additionally, some of the new isolates and mutant derivatives accumulated pullulan of relatively high molecular weight in the culture broths.

  19. Isolation of New Aureobasidium Strains That Produce High-Molecular-Weight Pullulan with Reduced Pigmentation

    PubMed Central

    Pollock, Thomas J.; Thorne, Linda; Armentrout, Richard W.

    1992-01-01

    New isolates of Aureobasidium pullulans were obtained from plant leaf surfaces gathered in San Diego County. The new fungal isolates were identified as A. pullulans on the basis of the appearance of polymorphic colonies formed on agar plates, the electrophoretic profiles of repeated genomic DNA sequences, and the production of pullulan in shake flask cultures. The isolates showed different degrees of pigmentation. One of the natural isolates was nonpigmented under mock production conditions in liquid culture, but was still able to synthesize a reduced amount of pigment on agar plates at late times. A mutagenic treatment with ethidium bromide produced derivatives of normally pigmented natural isolates that exhibited an increased tendency toward yeastlike growth and reduced pigmentation. Additionally, some of the new isolates and mutant derivatives accumulated pullulan of relatively high molecular weight in the culture broths. Images PMID:16348676

  20. Using pullulan-based edible coatings to extend shelf-life of fresh-cut 'Fuji' apples.

    PubMed

    Wu, Shengjun; Chen, Jinhua

    2013-04-01

    Pullulan is a thickener that can form semipermeable films, and glutathione is an effective reducing agent, while chitooligosaccharide has antibacterial activity. In this study, effect of pullulan-based coatings in combination with antibrowning and antibacterial agents (1% pullulan; 0.8% glutathione+1% chitooligosaccharides; and 0.8% glutathione+1% chitooligosaccharides+1% pullulan) on apple slices was investigated during hypothermia storage. Pullulan-coating treatments effectively retarded enzymatic browning, maintained firmness, decreased weight loss, and inhibited microbial growth and respiration rate of apple slices during hypothermia storage compared with that of the control (p<0.05). Results indicate that using pullulan-based coatings in combination with glutathione and chitooligosaccharides is a promising way to extend the shelf-life of apple slices. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Protection of Lactobacillus acidophilus NRRL-B 4495 under in vitro gastrointestinal conditions with whey protein/pullulan microcapsules.

    PubMed

    Çabuk, Burcu; Tellioğlu Harsa, Şebnem

    2015-12-01

    In this research, whey protein/pullulan (WP/pullulan) microcapsules were developed in order to assess its protective effect on the viability of Lactobacillus acidophilus NRRL-B 4495 under in vitro gastrointestinal conditions. Results demonstrated that WP/pullulan microencapsulated cells exhibited significantly (p ≤ 0.05) higher resistance to simulated gastric acid and bile salt. Pullulan incorporation into protein wall matrix resulted in improved survival as compared to free cells after 3 h incubation in simulated gastric solution. Moreover WP/pullulan microcapsules were found to release over 70% of encapsulated L. acidophilus NRRL-B 4495 cells within 1 h. The effect of encapsulation during refrigerated storage was also studied. Free bacteria exhibited 3.96 log reduction while, WP/pullulan encapsulated bacteria showed 1.64 log reduction after 4 weeks of storage. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  2. A biocleavable pullulan-based vector via ATRP for liver cell-targeting gene delivery.

    PubMed

    Yang, Xin-Chao; Niu, Yan-Lan; Zhao, Na-Na; Mao, Chun; Xu, Fu-Jian

    2014-04-01

    Pullulan due to its specificity for liver has been widely exploited for biomedical applications. In this work, a tailor-made biocleavable pullulan-based gene vector (PuPGEA) with good hemocompatibility was successfully proposed via atom transfer radical polymerization (ATRP) for efficient liver cell-targeting gene delivery. A two-step method involving the reaction of hydroxyl groups of pullulan with cystamine was developed to introduce reduction-sensitive disulfide-linked initiation sites of ATRP onto pullulan. The poly(glycidyl methacrylate) (PGMA) side chains prepared subsequently via ATRP were functionalized with ethanolamine (EA) to produce the resultant biocleavable comb-shaped PuPGEA vectors consisting of nonionic pullulan backbones and disulfide-linked cationic EA-functionalized PGMA (PGEA) side chains with plentiful secondary amine and nonionic hydroxyl units. The cationic PGEA side chains can be readily cleavable from the pullulan backbones of PuPGEA under reducible conditions. Due to the liver targeting performance of pullulan backbones, such PuPGEA vectors exhibited much higher gene transfection efficiency and cellular uptake rates in HepG2 cell lines than in Hella cell lines. In addition, in vitro transfection efficiency and uptake mechanism of polyplex in HepG2 cells were evaluated in the presence of different endocytosis inhibitors, indicating that the asialoglycoprotein receptor was involved in transfection process of hepatocytes. More importantly, in comparison with gold standard polyethylenimine (PEI, ∼25 kDa), PuPGEA vectors possessed excellent hemocompatibility without causing undesirable hemolysis. Properly grafting short bioreducible PGEA polycation side chains from a liver cell-targeting pullulan backbone is an effective means to produce new hemocompatible polysaccharide-based gene delivery vectors. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Utilization of corn steep liquor for biosynthesis of pullulan, an important exopolysaccharide.

    PubMed

    Sharma, Nishat; Prasad, G S; Choudhury, Anirban Roy

    2013-03-01

    Five different agricultural wastes viz. rice bran oil cake, soya bean oil cake, cotton seed oil cake, mustard seed oil cake and corn steep liquor (CSL) were evaluated for their use as nutrient along with 15% (w/v) glucose as carbon source for biosynthesis of pullulan using Aureobasidium pullulans RBF 4A3. Among the selected agricultural wastes, CSL was found to be the best and supported production of 77.92gL(-1) pullulan under un-optimized conditions. Single point optimization technique resulted in increase in 18% pullulan (88.59gL(-1)) production. The process was successfully validated in a 7-L fermenter and a process economic analysis has suggested that use of CSL as nutrient may result in 3-fold reduction of cost of raw materials for pullulan production as compared to a process where conventional nitrogen sources were used. These observations may be helpful in development of a cost effective process for pullulan production. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Statistical studies on high molecular weight pullulan production in solid state fermentation using jack fruit seed.

    PubMed

    Sugumaran, K R; Sindhu, R V; Sukanya, S; Aiswarya, N; Ponnusami, V

    2013-10-15

    The purpose of the work was to optimize the medium variables for maximizing pullulan production using jack fruit seed as a low cost substrate by Aureobasidium pullulans in solid state fermentation. Effects of K2HPO4, KH2PO4, ZnSO4·5H2O, MgSO4·7H2O, NaCl, (NH4)2SO4·5H2O, yeast extract, moisture content (%, w/w) in the production medium on pullulan production were studied using Plackett-Burman design. Production of pullulan was significantly affected by the medium variables namely KH2PO4, ZnSO4·5H2O, NaCl and moisture content (%, w/w). Then screened variables were optimized by Box Behnken experiment design. The pullulan obtained was characterized and confirmed by FTIR, (1)H NMR and (13)C NMR. Molecular weight of pullulan was found to be 1.733×10(6) g/mol by gel permeation chromatography (GPC). Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Deterioration of expanded polystyrene caused by Aureobasidium pullulans var. melanogenum.

    PubMed

    Castiglia, Valeria C; Kuhar, Francisco

    2015-01-01

    An expanded-polystyrene factory located in northern Buenos Aires reported unusual dark spots causing esthetic damage in their production. A fungal strain forming black-olive colonies on extract malt agar medium was isolated from the damaged material and identified as Aureobasidium pullullans var. melanogenum. This fungus is particularly known for its capacity to produce hydrolytic enzymes and a biodegradable extracellular polysaccharide known as pullulan, which is used in the manufacture of packaging material for food and medicine. Laboratory tests were conducted to characterize its growth parameters. It was found that the organism was resistant to a wide range of pHs but did not survive at temperatures over 65°C. The proposed action plan includes drying of the material prior to packaging and disinfection of the machinery used in the manufacturing process and of the silos used for raw material storage.

  6. Cellularized Bilayer Pullulan-Gelatin Hydrogel for Skin Regeneration.

    PubMed

    Nicholas, Mathew N; Jeschke, Marc G; Amini-Nik, Saeid

    2016-05-01

    Skin substitutes significantly reduce the morbidity and mortality of patients with burn injuries and chronic wounds. However, current skin substitutes have disadvantages related to high costs and inadequate skin regeneration due to highly inflammatory wounds. Thus, new skin substitutes are needed. By combining two polymers, pullulan, an inexpensive polysaccharide with antioxidant properties, and gelatin, a derivative of collagen with high water absorbency, we created a novel inexpensive hydrogel-named PG-1 for "pullulan-gelatin first generation hydrogel"-suitable for skin substitutes. After incorporating human fibroblasts and keratinocytes onto PG-1 using centrifugation over 5 days, we created a cellularized bilayer skin substitute. Cellularized PG-1 was compared to acellular PG-1 and no hydrogel (control) in vivo in a mouse excisional skin biopsy model using newly developed dome inserts to house the skin substitutes and prevent mouse skin contraction during wound healing. PG-1 had an average pore size of 61.69 μm with an ideal elastic modulus, swelling behavior, and biodegradability for use as a hydrogel for skin substitutes. Excellent skin cell viability, proliferation, differentiation, and morphology were visualized through live/dead assays, 5-bromo-2'-deoxyuridine proliferation assays, and confocal microscopy. Trichrome and immunohistochemical staining of excisional wounds treated with the cellularized skin substitute revealed thicker newly formed skin with a higher proportion of actively proliferating cells and incorporation of human cells compared to acellular PG-1 or control. Excisional wounds treated with acellular or cellularized hydrogels showed significantly less macrophage infiltration and increased angiogenesis 14 days post skin biopsy compared to control. These results show that PG-1 has ideal mechanical characteristics and allows ideal cellular characteristics. In vivo evidence suggests that cellularized PG-1 promotes skin regeneration and may

  7. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 1 General Provisions 1 2014-01-01 2012-01-01 true Receipt and processing. 17.1 Section 17.1... PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing § 17.1 Receipt and processing. Unless special arrangements are made with the Director of the Federal Register...

  8. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 1 General Provisions 1 2012-01-01 2012-01-01 false Receipt and processing. 17.1 Section 17.1... PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing § 17.1 Receipt and processing. Unless special arrangements are made with the Director of the Federal Register...

  9. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 1 General Provisions 1 2011-01-01 2011-01-01 false Receipt and processing. 17.1 Section 17.1... PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing § 17.1 Receipt and processing. Unless special arrangements are made with the Director of the Federal Register...

  10. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 1 General Provisions 1 2010-01-01 2010-01-01 false Receipt and processing. 17.1 Section 17.1... PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing § 17.1 Receipt and processing. Unless special arrangements are made with the Director of the Federal Register...

  11. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 1 General Provisions 1 2013-01-01 2012-01-01 true Receipt and processing. 17.1 Section 17.1... PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing § 17.1 Receipt and processing. Unless special arrangements are made with the Director of the Federal Register...

  12. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 1 2010-10-01 2010-10-01 false Application-T/ALL. 34.17-1 Section 34.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY TANK VESSELS FIREFIGHTING EQUIPMENT Fixed Foam Extinguishing Systems, Details § 34.17-1 Application—T/ALL. (a) Where a fixed foam extinguishing system...

  13. Genome sequencing of four Aureobasidium pullulans varieties: biotechnological potential, stress tolerance, and description of new species

    DOE PAGES

    Gostinčar, Cene; Ohm, Robin A.; Kogej, Tina; ...

    2014-07-01

    Aureobasidium pullulans is a black-yeast-like fungus used for production of the polysaccharide pullulan and the antimycotic aureobasidin A, and as a biocontrol agent in agriculture. It can cause opportunistic human infections, and it inhabits various extreme environments. To promote the understanding of these traits, we performed de-novo genome sequencing of the four varieties of A. pullulans. The 25.43-29.62 Mb genomes of these four varieties of A. pullulans encode between 10266 and 11866 predicted proteins. Their genomes encode most of the enzyme families involved in degradation of plant material and many sugar transporters, and they have genes possibly associated with degradationmore » of plastic and aromatic compounds. Proteins believed to be involved in the synthesis of pullulan and siderophores, but not of aureobasidin A, are predicted. Putative stress-tolerance genes include several aquaporins and aquaglyceroporins, large numbers of alkali-metal cation transporters, genes for the synthesis of compatible solutes and melanin, all of the components of the high-osmolarity glycerol pathway, and bacteriorhodopsin-like proteins. All of these genomes contain a homothallic mating-type locus. The differences between these four varieties of A. pullulans are large enough to justify their redefinition as separate species: A. pullulans, A. melanogenum, A. subglaciale and A. namibiae. We observed redundancy in several gene families that can be linked to the nutritional versatility of these species and their particular stress tolerance. In conclusions, the availability of the genome sequences of the four Aureobasidium species should improve their biotechnological exploitation and promote our understanding of their stress-tolerance mechanisms, diverse lifestyles, and pathogenic potential.« less

  14. Genome sequencing of four Aureobasidium pullulans varieties: biotechnological potential, stress tolerance, and description of new species

    SciTech Connect

    Gostinčar, Cene; Ohm, Robin A.; Kogej, Tina; Sonjak, Silva; Turk, Martina; Zajc, Janja; Zalar, Polona; Grube, Martin; Sun, Hui; Han, James; Sharma, Aditi; Chiniquy, Jennifer; Ngan, Chew; Lipzen, Anna; Barry, Kerrie; Grigoriev, Igor V.; Gunde-Cimerman, Nina

    2014-07-01

    Aureobasidium pullulans is a black-yeast-like fungus used for production of the polysaccharide pullulan and the antimycotic aureobasidin A, and as a biocontrol agent in agriculture. It can cause opportunistic human infections, and it inhabits various extreme environments. To promote the understanding of these traits, we performed de-novo genome sequencing of the four varieties of A. pullulans. The 25.43-29.62 Mb genomes of these four varieties of A. pullulans encode between 10266 and 11866 predicted proteins. Their genomes encode most of the enzyme families involved in degradation of plant material and many sugar transporters, and they have genes possibly associated with degradation of plastic and aromatic compounds. Proteins believed to be involved in the synthesis of pullulan and siderophores, but not of aureobasidin A, are predicted. Putative stress-tolerance genes include several aquaporins and aquaglyceroporins, large numbers of alkali-metal cation transporters, genes for the synthesis of compatible solutes and melanin, all of the components of the high-osmolarity glycerol pathway, and bacteriorhodopsin-like proteins. All of these genomes contain a homothallic mating-type locus. The differences between these four varieties of A. pullulans are large enough to justify their redefinition as separate species: A. pullulans, A. melanogenum, A. subglaciale and A. namibiae. We observed redundancy in several gene families that can be linked to the nutritional versatility of these species and their particular stress tolerance. In conclusions, the availability of the genome sequences of the four Aureobasidium species should improve their biotechnological exploitation and promote our understanding of their stress-tolerance mechanisms, diverse lifestyles, and pathogenic potential.

  15. Properties of edible films based on pullulan-chitosan blended film-forming solutions at different pH

    USDA-ARS?s Scientific Manuscript database

    Influences of solution pH on the properties of pullulan-chitosan blended (Pul-Chi) films and the rheological properties of film-forming solutions were investigated. The extended conformation of chitosan in pH 4.0 solution increased intermolecular interactions with pullulan compared to the more compa...

  16. Polyols, not sugars, determine the structural diversity of anti-streptococcal liamocins produced by Aureobasidium pullulans strain NRRL 50380

    USDA-ARS?s Scientific Manuscript database

    Liamocins are polyol-lipids produced by the fungus Aureobasidium pullulans, and have selective antibacterial activity against Streptococcus species. Liamocins produced by A. pullulans strain NRRL 50380 on sucrose medium have a D-mannitol head-group ester linked to 3,5-dihydroxydecanoate acyl chains,...

  17. Modification of the mannitol biosynthetic pathway in Aureobasidium pullulans to alter the structure of the polyol lipid liamocin

    USDA-ARS?s Scientific Manuscript database

    Aureobasidium pullulans is an important industrial organism that is utilized for the production of numerous bioproducts, such as pullulan, ß-glucan, aureobasidin, and polymalic acid. Our laboratory is also interested in production of the extracellular polyol lipids, called liamocins, produced by cer...

  18. Improvement on Physical Properties of Pullulan Films by Novel Cross-Linking Strategy.

    PubMed

    Chen, Chieh-Ting; Chen, Kuan-I; Chiang, Hsin-Han; Chen, Yu-Kuo; Cheng, Kuan-Chen

    2017-01-01

    Pullulan based films possess several advantages, including high transparency, low toxicity, good biodegradability, good mechanical properties, and low oxygen permeability, are preferable for food packaging. The application of pullulan films on food packaging, however, has inherent disadvantage of high water solubility. In this study, glutaraldehyde and glycerol were used as the cross-linking reagent and the plasticizer respectively to improve water resistance and physical properties of the pullulan films. Effects of cross-linking degree on physical properties, including water absorptions, swelling behaviors, water vapor permeability and tensile strengths of films were evaluated. FTIR results demonstrated that the pullulan films were successfully cross-linked by glutaraldehyde. The tensile strength of pullulan films could be enhanced significantly (P < 0.05) when glutaraldehyde was between 1% and 5% (w/w); nevertheless, the amount of glutaraldehyde above 20% (w/w) led to films brittleness. With the addition of glycerol as a plasticizer enhanced the extensibility of films as well as the hydrophilicity, resulting in higher water vapor permeability.

  19. Ketoprofen-loaded pomegranate seed oil nanoemulsion stabilized by pullulan: Selective antiglioma formulation for intravenous administration.

    PubMed

    Ferreira, Luana M; Cervi, Verônica F; Gehrcke, Mailine; da Silveira, Elita F; Azambuja, Juliana H; Braganhol, Elizandra; Sari, Marcel H M; Zborowski, Vanessa A; Nogueira, Cristina W; Cruz, Letícia

    2015-06-01

    This study aimed to prepare pomegranate seed oil nanoemulsions containing ketoprofen using pullulan as a polymeric stabilizer, and to evaluate antitumor activity against in vitro glioma cells. Formulations were prepared by the spontaneous emulsification method and different concentrations of pullulan were tested. Nanoemulsions presented adequate droplet size, polydispersity index, zeta potential, pH, ketoprofen content and encapsulation efficiency. Nanoemulsions were able to delay the photodegradation profile of ketoprofen under UVC radiation, regardless of the concentration of pullulan. In vitro release study indicates that nanoemulsions were able to release approximately 95.0% of ketoprofen in 5h. Free ketoprofen and formulations were considered hemocompatible at 1 μg/mL, in a hemolysis study, for intravenous administration. In addition, a formulation containing the highest concentration of pullulan was tested against C6 cell line and demonstrated significant activity, and did not reduce fibroblasts viability. Thus, pullulan can be considered an interesting excipient to prepare nanostructured systems and nanoemulsion formulations can be considered promising alternatives for the treatment of glioma.

  20. Rheological properties of pullulan-sodium alginate based solutions during film formation.

    PubMed

    Xiao, Qian; Tong, Qunyi; Zhou, Yujia; Deng, Fangming

    2015-10-05

    During film formation, the rheological properties of pullulan, sodium alginate, and blends, dried at 50°C were studied using an oscillatory rheometer. According to the drying curves, the drying process of pullulan, alginate, and blend films was divided into three stages. At the first drying stage, four samples exhibited typical liquid-like viscoelastic behavior. As the drying proceeded (polysaccharide concentration up to 75%), pure pullulan chains formed an entangled network, whereas coupling of alginate molecules gave a weak gel. At this drying stage, complex viscosity data for 75% alginate and blends were fitted with the power law equation. The effects of drying on the mechanical properties of pullulan-sodium alginate based samples were analyzed using the generalized Maxwell model, and their relaxation spectra were determined. The rheological properties during drying obtained from this study is essential for understanding film-forming mechanism and predicting the properties of pullulan-sodium alginate based edible films. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Development of chitosan-pullulan composite nanoparticles for nasal delivery of vaccines: in vivo studies.

    PubMed

    Cevher, Erdal; Salomon, Stefan K; Somavarapu, Satyanarayana; Brocchini, Steve; Alpar, H Oya

    2015-01-01

    Here, we aimed at developing chitosan/pullulan composite nanoparticles and testing their potential as novel systems for the nasal delivery of diphtheria toxoid (DT). All the chitosan derivatives [N-trimethyl (TMC), chloride and glutamate] and carboxymethyl pullulan (CMP) were synthesised and antigen-loaded composites were prepared by polyion complexation of chitosan and pullulan derivatives (particle size: 239-405 nm; surface charge: +18 and +27 mV). Their immunological effects after intranasal administration to mice were compared to intramuscular route. Composite nanoparticles induced higher levels of IgG responses than particles formed with chitosan derivative and antigen. Nasally administered TMC-pullulan composites showed higher DT serum IgG titre when compared with the other composites. Co-encapsulation of CpG ODN within TMC-CMP-DT nanoparticles resulted in a balanced Th1/Th2 response. TMC/pullulan composite nanoparticles also induced highest cytokine levels compared to those of chitosan salts. These findings demonstrated that TMC-CMP-DT composite nanoparticles are promising delivery system for nasal vaccination.

  2. Synthesis of some new antianemics I. Iron pullulan complexes of pharmaceutical interest.

    PubMed

    Nikolic, G; Cakic, M; Ilic, Lj; Ristic, S; Cakic, Z

    2002-03-01

    First experiments were performed in order to obtain a polynuclear iron(III) complex with pullulan, which could be applied in therapy of sideropenic anemia. Complete synthesis of the complex can be realized with oligomers of depolymerized pullulan at an average molar mass 8000-10,000 g.mol-1. Maximum iron contents of 51.4 mg.cm-3 is effected by pullulan depolymerizates (Mw approximately 9000 g.mol-1, [eta] = 0.085 dl.g-1) at 130 degrees C for 240 min, at pH = 10.5 and at an initial mass ratio Fe/ligand of 1:3. The effected concentrations and complex stability respond to requirements of pharmacological application of parenteral antianemic preparations.

  3. Pullulan-based coatings for preservation of razor clam Sinonovacula constricta.

    PubMed

    Jiang, Longfa

    2016-11-01

    Glutathione has antioxidant activity, and citric acid exhibits acidity. Potassium sorbate displays antibacterial activity. Effects of pullulan-based coatings with 0.05% glutathione, 0.5% citric acid and 0.2% potassium sorbate on the preservation of razor clam Sinonovacula constricta meats (SCMs) during refrigerated storage (∼2°C) were investigated. The glutathione-, citric acid- and potassium sorbate-incorporated pullulan-based coatings inhibited bacterial growth. Moreover, results showed reduced increase in pH, decreased total volatile basic nitrogen and enhanced overall acceptability score of SCMs during refrigerated storage (∼2°C). These results indicated that treatment with pullulan-based coatings with glutathione, citric acid and potassium sorbate could prolong the shelf life of SCM for up to 10 days.

  4. Exceptional oxygen barrier performance of pullulan nanocomposites with ultra-low loading of graphene oxide

    NASA Astrophysics Data System (ADS)

    Uysal Unalan, Ilke; Wan, Chaoying; Figiel, Łukasz; Olsson, Richard T.; Trabattoni, Silvia; Farris, Stefano

    2015-07-01

    Polymer nanocomposites are increasingly important in food packaging sectors. Biopolymer pullulan is promising in manufacturing packaging films or coatings due to its excellent optical clarity, mechanical strength, and high water-solubility as compared to other biopolymers. This work aims to enhance its oxygen barrier properties and overcome its intrinsic brittleness by utilizing two-dimensional planar graphene oxide (GO) nanoplatelets. It has been found that the addition of only 0.2 wt% of GO enhanced the tensile strength, Young’s modulus, and elongation at break of pullulan films by about 40, 44 and 52%, respectively. The light transmittance at 550 nm of the pullulan/GO films was 92.3% and haze values were within 3.0% threshold, which meets the general requirement for food packaging materials. In particular, the oxygen permeability coefficient of pullulan was reduced from 6337 to 2614 mL μm m-2 (24 h-1) atm-1 with as low as 0.05 wt% of GO loading and further to 1357 mL μm m-2 (24 h-1) atm-1 when GO concentration reached 0.3 wt%. The simultaneous improvement of the mechanical and oxygen barrier properties of pullulan was ascribed to the homogeneous distribution and prevalent unidirectional alignment of GO nanosheets, as determined from the characterization and theoretical modelling results. The exceptional oxygen barrier properties of pullulan/GO nanocomposites with enhanced mechanical flexibility and good optical clarity will add new values to high performance food packaging materials.

  5. 38 CFR 17.1 - Incorporation by reference.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... reference. 17.1 Section 17.1 Pensions, Bonuses, and Veterans' Relief DEPARTMENT OF VETERANS AFFAIRS MEDICAL....archives.gov/federal_register/code_of_federal_regulations/ibr_locations.html. Copies may be obtained from... Reference (IBR) approved for §§ 17.63 and 17.81. (2) NFPA 101, Life Safety Code (2009 edition), IBR...

  6. Effect of pullulan on the water distribution, microstructure and textural properties of rice starch gels during cold storage.

    PubMed

    Chen, Long; Tian, Yaoqi; Tong, Qunyi; Zhang, Zipei; Jin, Zhengyu

    2017-01-01

    The effects of pullulan on the water distribution, microstructure and textural properties of rice starch gels during cold storage were investigated by low field-nuclear magnetic resonance (LF-NMR), scanning electron microscope (SEM), and texture profile analysis (TPA). The addition of pullulan reduced the transversal relaxation time of rice starch gels during cold storage. The microstructure of rice starch gel with 0.5% pullulan was denser and more uniform compared with that of rice starch without pullulan in each period of storage time. With regard to textural properties, 0.01% pullulan addition did not significantly change the texture of rice starch gels, while 0.5% pullulan addition appeared to reduce the hardness and retain the springiness of rice starch gels (P⩽0.05). The restriction effects of pullulan on water mobility and starch retrogradation were hypothesized to be mainly responsible for the water retention, gel structure maintenance, and modification of the textural attributes of rice starch gels. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Liamocin oil from Aureobasidium pullulans has antibacterial activity with specificity for species of Streptococcus

    USDA-ARS?s Scientific Manuscript database

    Liamocin oil from Aureobasidium pullulans NRRL 50380 was tested for antibacterial activity. Liamocins inhibited growth of Streptococcus agalactiae, S. uberis, S. mitis, S. infantarius, and S. mutans, with minimum inhibitory concentrations from 20 'g/ml to 78 'g/ml. Enterococcus faecalis was less sus...

  8. Antibacterial activity of liamocins oil from Aureobasidium pullulans is specific for species of Streptococcus

    USDA-ARS?s Scientific Manuscript database

    Liamocins are a heterogeneous mixture of denser-than-water oils produced by the fungus Aureobasidium pullulans. Liamocins have unique chemical structures with a mannitol head group linked to long chain polyester tails consisting of multiple 3,5-dihydroxydecanoic acid ester groups, some of which are ...

  9. Liamocin oil from Aureobasidium pullulans has antibacterial activity with specificity for species of Streptococcus

    USDA-ARS?s Scientific Manuscript database

    The fungus Aureobasidium pullulans produces denser-than-water oils called liamocins. Liamocins have unique chemical structures with a mannitol head group linked to long chain polyester tails consisting of three, four or five 3,5-dihydroxydecanoic acid esters, some of which are O-acetylated. Broth di...

  10. Water vapor barrier and sorption properties of edible films from pullulan and rice wax.

    USDA-ARS?s Scientific Manuscript database

    Edible films were prepared by using various ratios of pullulan and rice wax. Freestanding composite films were obtained with up to 46.4% rice wax. Water vapor barrier properties of the film were improved with increased addition of rice wax. Moisture sorption isotherms were also studied to examine...

  11. Bioproducts and morphological features of diverse isolates of the fungus Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    Aureobasidium pullulans is a fungus included among the “black yeasts.” Although many strains are predominantly yeast-like, the species is actually polymorphic, exhibiting a variety of complex forms. The fungus is ubiquitous, routinely found on the surface of leaves, wood, painted walls, etc. We rece...

  12. Production of poly(beta-L-malic acid) (PMA) from agricultural biomass substrates by Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    We report here for the first time the production of poly(beta-L-malic acid) (PMA) from agricultural biomass substrates by the yeastlike fungus Aureobasidium pullulans. Strains NRRL Y 2311-1, NRRL 50382, NRRL 50383, and NRRL 50384, representing diverse isolation sources and phylogenetic clades, prod...

  13. Influence of polysaccharide composition on the biocompatibility of pullulan/dextran-based hydrogels.

    PubMed

    Abed, Aicha; Assoul, Nabila; Ba, Maguette; Derkaoui, Sidi Mohamed; Portes, Patrick; Louedec, Liliane; Flaud, Patrice; Bataille, Isabelle; Letourneur, Didier; Meddahi-Pellé, Anne

    2011-03-01

    The implantation of a biomaterial for tissue engineering requires the presence of a suitable scaffold on which the tissue repair and regeneration will take place. Polymers have been frequently used for that purpose because they show similar properties to that of the natural extracellular matrix. Scaffold properties and biocompatibility are modulated by the composition of the polymers used. In this work four polysaccharide-based hydrogels (PSH) made of dextran and pullulan were synthesized. Their in vitro properties were determined and then tested in vivo in a rat model. As pullulan concentration increased in dextran hydrogels, the glass transition temperature and the maximum modulus decreased. In vitro degradation studies for 30 days demonstrated no significant degradation of PSH except for 100% pullulan hydrogel. In vivo tissue response evaluated 30 days after PSH subcutaneous implantation in rats indicated that all PSH were surrounded by a fibrous capsule. Adding pullulan to dextran induced an increased inflammatory reaction compared to PSH-D(100% dextran) or PSH-D(75)P(25)(75% dextran). This in vitro and in vivo data can be used in the design of hydrogels appropriate for tissue engineering applications.

  14. Genome sequence of Aureobasidium pullulans AY4, an emerging opportunistic fungal pathogen with diverse biotechnological potential.

    PubMed

    Chan, Giek Far; Bamadhaj, Hasima Mustafa; Gan, Han Ming; Rashid, Noor Aini Abdul

    2012-11-01

    Aureobasidium pullulans AY4 is an opportunistic pathogen that was isolated from the skin of an immunocompromised patient. We present here the draft genome of strain AY4, which reveals an abundance of genes relevant to bioindustrial applications, including biocontrol and biodegradation. Putative genes responsible for the pathogenicity of strain AY4 were also identified.

  15. Poly(beta-L-malic acid) from agricultural substrates by Aureobasidium pullulans

    USDA-ARS?s Scientific Manuscript database

    We report here for the first time the production of poly(beta-L-malic acid) (PMA) from agricultural substrates by the yeastlike fungus Aureobasidium pullulans. PMA is a natural biopolyester that has primarily been studied for biomedical uses as a drug carrier. However, PMA also has potential as a ...

  16. FTIR spectroscopic characterization of Cu(II) coordination compounds with exopolysaccharide pullulan and its derivatives

    NASA Astrophysics Data System (ADS)

    Mitić, Ž.; Nikolić, G. S.; Cakić, M.; Premović, P.; Ilić, Lj.

    2009-04-01

    Pullulan is a water-soluble, extracellular neutral polysaccharide with a linear flexible chain of α-(1 → 6)-linked maltotriose units, the structure of which is intermediate between pullulan and amylose structures because of the co-existence of both α-(1 → 6) and α-(1 → 4)-glycosidic linkages in single compounds. In alkali solutions Cu(II) ion forms complexes with reduced low-molar pullulan (RLMP). The metal content and the solution composition depends on pH. The complexing process begins in a weak alkali solution (pH > 7), and involves OH groups in C(2) and C(3) or C(6) pullulan monomer units (α- D-glucopyranose). Complexes of Cu(II) ion with reduced low-molar pullulan were synthesized in the water solutions, at the boiling temperature and at different pH values (7.512). Fourier-Transform Infrared (FTIR) spectroscopic data of synthesized complexes are rare in literature. FTIR spectroscopic characterization (FTIR, LNT-FTIR, ATR-FTIR, and FTIR microspectroscopy) of Cu(II) ion complexes with RLMP ( M w 6000 g mol -1) was carried out in this work. The similarities of the γ(C sbnd H) range in a part of FTIR spectra indicate that there is no difference in the conformation of the C 1 glucopyranose (Glc) unit in the RLMP and synthesized Cu(II) complexes. The complexing Cu(II) ion with RLMP in the dependence from the pH form different types of complex (pH 7-8: Cu(II)(Glc) 2(H 2O) 2, pH 8-10: Cu(II)(Glc) 2(H 2O)(OH), pH 10-12: Cu(II)(Glc) 2(OH) 2).

  17. Aureobasidium pullulans as a source of liamocins (heavy oils) with anticancer activity.

    PubMed

    Manitchotpisit, Pennapa; Watanapokasin, Ramida; Watanapoksin, Ramida; Price, Neil P J; Bischoff, Kenneth M; Tayeh, Malatee; Teeraworawit, Sudarat; Kriwong, Saranya; Leathers, Timothy D

    2014-08-01

    Liamocins are structurally unique, heavier-than-water “oils” produced by certain strains of Aureobasidium pullulans. The aim of the current study is to identify new sources of liamocins and evaluate their potential as anticancer agents. Nine strains of A. pullulans from phylogenetic clades 8, 9, and 11 were examined for the first time for production of liamocins. Strains in these clades have only been isolated from tropical environments, and all strains tested here were from various locations in Thailand. Strains RSU 9, RSU 21, and RSU 29, all from clade 11, produced from 7.0 to 8.6 g liamocins/l from medium containing 5 % sucrose. These are the highest yields of liamocins that we have found thus far. These strains also produced from 9.4 to 17 g pullulan/l. The structural identity of liamocins was confirmed by matrix-assisted laser desorption/ionization mass spectrometry; differential spectra were obtained in which the dominant ion was either at about m/z 805.5 or m/z 949.6, consistent with the structure of liamocins. Liamocins from A. pullulans strains RSU 9 and RSU 21 inhibited two human breast cancer cell lines and a human cervical cancer cell line (IC50 values of 32.2 ± 1.4 to 63.1 ± 2.4 μg liamocins/ml) but were not toxic to a normal cell line. Liamocins weakly inhibited a strain of Enterococcus faecalis, but did not inhibit strains of Lactobacillus fermentum, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Thus, A. pullulans phylogenetic clade 11 is a promising source of liamocins, and these compounds merit further examination as potential anticancer agents.

  18. Successful treatment of Aureobasidium pullulans central catheter-related fungemia and septic pulmonary emboli.

    PubMed

    Mehta, Sanjay R; Johns, Scott; Stark, Paul; Fierer, Joshua

    2017-01-01

    Aureobasidium pullulans is a saprophytic fungus that is widely distributed in the environment, and in the right host can be an opportunistic human pathogen. A 66-year-old man with Crohn's disease with a single kidney, and requiring total parenteral nutrition via a Hickman catheter, was admitted with a 10-week history of progressive shortness of breath, fevers and weight loss. Chest imaging demonstrated new multifocal lung parenchymal opacities compatible with septic pulmonary emboli. Blood culture grew a yeast-like organism that transformed into a black mold on subculture, eventually identified as A. pullulans. Due to triazole resistance, the patient was treated with liposomal amphotericin and micafungin. Serum (1,3)-β-d-glucan level was used to monitor therapy, initially measured at >500 pg/mL and decreasing to 66 pg/mL after one year of therapy. We describe the successful treatment of a case of catheter related fungemia and septic pulmonary emboli due A. pullulans. While initially appearing as an oval yeast on blood culture, subsequent growth as a black mold led to identification of the fungus as A. pullulans. The infection was cured with a combination of antifungal agents, even though the foreign body could not be safely removed. Nephrotoxicity required dosing adjustment of the amphotericin to biweekly during the maintenance phase of treatment. The serum (1,3)-β-d-glucan level proved to be useful in monitoring response to therapy. We report here successful treatment of a disseminated A. pullulans infection with an induction and maintenance approach to liposomal amphotericin dosing, and monitoring response to therapy with serum (1,3)-β-d-glucan levels.

  19. Characteristics, protein engineering and applications of microbial thermostable pullulanases and pullulan hydrolases.

    PubMed

    Nisha, M; Satyanarayana, T

    2016-07-01

    Pullulan hydrolyzing enzymes are endoacting, classified based on the substrate specificity and hydrolysis products as pullulanases (type I and II) and pullulan hydrolases (type I, II and III). Pullulanases and pullulan hydrolase type I are produced by bacteria and archaea. Among bacteria, many mesophilic, thermophilic and hyperthermophilic bacteria produce pullulanases and neopullulanases. While pullulan hydrolase type II and type III are produced by fungi and archaea, respectively. These are multi-domain proteins with three conserved catalytic acidic residues of the glycosyl hydrolases. The recent advances in molecular biology and protein engineering via mutagenesis and truncation led to improvement in thermostability, catalytic activity and substrate specificity. Pullulanases are debranching enzymes, which are widely employed in starch saccharification that minimizes the use of glucoamylase (approx. 50 %) and reduces the total reaction time of the industrial starch conversion process. The thermostable amylopullulanases are useful in one-step starch liquefaction and saccharification, which replaces amylolytic enzymes like α-amylase and glucoamylase, thus resulting in the reduction in the cost of sugar production. The enzymes also find application in making resistant starches and as an antistale in bread making. Panose and isopanose containing syrups are useful as prebiotics, while panose has also been reported to display anticarcinogenic activity. This review focuses on the distinguishing features of these enzymes based on the analysis of amino acid sequences and domain structure, besides highlighting recent advances in the molecular biology and protein engineering for enhancing their thermostability, catalytic activity and substrate specificity. This review also briefly summarizes the potential applications of pullulanases and pullulan hydrolases.

  20. Effect of Pullulan Coating on Postharvest Quality and Shelf-Life of Highbush Blueberry (Vaccinium corymbosum L.)

    PubMed Central

    Kraśniewska, Karolina; Ścibisz, Iwona; Mitek, Marta; Pobiega, Katarzyna; Cendrowski, Andrzej

    2017-01-01

    Fruits form an important part of a healthy human diet as they contain many ingredients with proven pro-health effects such as vitamins, phenolic compounds, organic acids, fiber, and minerals. The purpose of this work was to evaluate the effect of pullulan coating on the quality and shelf life of highbush blueberry during storage. General appearance, weight loss, dry matter, soluble solid content, reducing sugars, content of L-ascorbic acid, phenolic compounds (total phenolics, phenolics acids and anthocyanins) were determined in uncoated and coated blueberries fruits. The microbiological efficiency of pullulan coating was also evaluated. All parameters were monitored during storage at 4 °C and 16 °C by 28 and 14 days, respectively. The study showed that pullulan coating protects perishable food products especially susceptible to mechanical injury including fruits such as blueberries. Pullulan acts as a barrier that minimizes respiration rate, delaying deterioration and controlling microbial growth. PMID:28820473

  1. Improved viability of Lactobacillus acidophilus NRRL-B 4495 during freeze-drying in whey protein-pullulan microcapsules.

    PubMed

    Çabuk, Burcu; Harsa, Şebnem Tellioglu

    2015-01-01

    In this research, pullulan was incorporated in protein-based encapsulation matrix in order to assess its cryoprotective effect on the viability of freeze-dried (FD) probiotic Lactobacillus acidophilus NRRL-B 4495. This study demonstrated that pullulan in encapsulation matrix resulted in a 90.4% survival rate as compared to 88.1% for whey protein (WPI) encapsulated cells. The protective effects of pullulan on the survival of FD-encapsulated cells in gastrointestinal conditions were compared. FD WPI-pullulan capsules retained higher survived cell numbers (7.10 log CFU/g) than those of FD WPI capsules (6.03 log CFU/g) after simulated gastric juice exposure. Additionally, use of pullulan resulted in an increased viability after bile exposure. FD-free bacteria exhibited 2.18 log CFU/g reduction, while FD WPI and FD WPI-pullulan encapsulated bacteria showed 0.95 and 0.49 log CFU/g reduction after 24 h exposure to bile solution, respectively. Morphology of the FD microcapsules was visualized by scanning electron microscopy.

  2. Efficacy of antimicrobial pullulan-based coating to improve internal quality and shelf-life of chicken eggs during storage.

    PubMed

    Morsy, Mohamed K; Sharoba, Ashraf M; Khalaf, Hassan H; El-Tanahy, Hassan H; Cutter, Catherine N

    2015-05-01

    There has been a growing interest in the use of natural materials as a delivery mechanism for antimicrobials and coatings in foods. The aim of the present study was to evaluate the effectiveness of pullulan coatings to improve internal quality and shelf-life of fresh eggs during 10 wk of storage at 25 and 4 °C. Three treatments of eggs were evaluated as follows; non-coated (control; C), coated with pullulan (P), and coated with pullulan containing nisin (N). The effects of the pullulan coatings on microbiological qualities, physical properties, and freshness parameters were investigated and compared with non-coated eggs. For non-coated eggs, as storage time increased, yolk index, albumen index, and Haugh unit value decreased and weight loss increased. However, pullulan coatings (P or N) minimized weight loss (<1.5%) and preserved the albumen and yolk quality of eggs (with a final B grade) 3 wk longer than non-coated eggs at 25 °C. At 4 °C, both P- and N-coated eggs went from AA to A grade after 9 wk and maintained the grade for 10 wk (4 wk longer than that of non-coated eggs). This study is the first to demonstrate that pullulan coatings can preserve the internal quality, prolong the shelf-life, and minimize weight loss of fresh eggs.

  3. Charged pullulan derivatives for the development of nanocarriers by polyelectrolyte complexation.

    PubMed

    Dionísio, M; Braz, L; Corvo, M; Lourenço, J P; Grenha, A; Rosa da Costa, A M

    2016-05-01

    Pullulan, a neutral polysaccharide, was chemically modified in order to obtain two charged derivatives: reaction with SO3(.)DMF complex afforded a sulfate derivative (SP), while reaction with glycidyltrimethylammonium chloride gave a quaternary ammonium salt (AP). The presence of the charged groups was confirmed by FTIR. Assessment of the positions where the reaction took place was based on (1)H- and (13)C NMR (COSY, HSQC-TOCSY, HSQC-DEPT, and HMBC) experiments. Estimation of the degree of substitution (DS) was made from elemental analysis data, and further confirmed by NMR peak areas in the case of AP. These new derivatives showed the capability to condense with each other, forming nanoparticles with the ability to associate a model protein (BSA) and displaying adequate size for drug delivery applications, therefore making them good candidates for the production of pullulan-based nanocarriers by polyelectrolyte complexation.

  4. Green Fluorescent Protein as a Novel Indicator of Antimicrobial Susceptibility in Aureobasidium pullulans

    PubMed Central

    Webb, Jeremy S.; Barratt, Sarah R.; Sabev, Hristo; Nixon, Marianne; Eastwood, Ian M.; Greenhalgh, Malcolm; Handley, Pauline S.; Robson, Geoffrey D.

    2001-01-01

    Presently there is no method available that allows noninvasive and real-time monitoring of fungal susceptibility to antimicrobial compounds. The green fluorescent protein (GFP) of the jellyfish Aequoria victoria was tested as a potential reporter molecule for this purpose. Aureobasidium pullulans was transformed to express cytosolic GFP using the vector pTEFEGFP (A. J. Vanden Wymelenberg, D. Cullen, R. N. Spear, B. Schoenike, and J. H. Andrews, BioTechniques 23:686–690, 1997). The transformed strain Ap1 gfp showed bright fluorescence that was amenable to quantification using fluorescence spectrophotometry. Fluorescence levels in Ap1 gfp blastospore suspensions were directly proportional to the number of viable cells determined by CFU plate counts (r2 > 0.99). The relationship between cell viability and GFP fluorescence was investigated by adding a range of concentrations of each of the biocides sodium hypochlorite and 2-n-octylisothiozolin-3-one (OIT) to suspensions of Ap1 gfp blastospores (pH 5 buffer). These biocides each caused a rapid (<25-min) loss of fluorescence of greater than 90% when used at concentrations of 150 μg of available chlorine ml−1 and 500 μg ml−1, respectively. Further, loss of GFP fluorescence from A. pullulans cells was highly correlated with a decrease in the number of viable cells (r2 > 0.92). Losses of GFP fluorescence and cell viability were highly dependent on external pH; maximum losses of fluorescence and viability occurred at pH 4, while reduction of GFP fluorescence was absent at pH 8.0 and was associated with a lower reduction in viability. When A. pullulans was attached to the surface of plasticized poly(vinylchloride) containing 500 ppm of OIT, fluorescence decreased more slowly than in cell suspensions, with >95% loss of fluorescence after 27 h. This technique should have broad applications in testing the susceptibility of A. pullulans and other fungal species to antimicrobial compounds. PMID:11722914

  5. Transparent Pullulan/Mica Nanocomposite Coatings with Outstanding Oxygen Barrier Properties

    PubMed Central

    Boyacı, Derya; Trabattoni, Silvia; Tavazzi, Silvia

    2017-01-01

    This study presents a new bionanocomposite coating on poly(ethylene terephthalate) (PET) made of pullulan and synthetic mica. Mica nanolayers have a very high aspect ratio (α), at levels much greater than that of conventional exfoliated clay layers (e.g., montmorillonite). A very small amount of mica (0.02 wt %, which is ϕ ≈ 0.00008) in pullulan coatings dramatically improved the oxygen barrier performance of the nanocomposite films under dry conditions, however, this performance was partly lost as the environmental relative humidity (RH) increased. This outcome was explained in terms of the perturbation of the spatial ordering of mica sheets within the main pullulan phase, because of RH fluctuations. This was confirmed by modelling of the experimental oxygen transmission rate (OTR) data according to Cussler’s model. The presence of the synthetic nanobuilding block (NBB) led to a decrease in both static and kinetic coefficients of friction, compared with neat PET (≈12% and 23%, respectively) and PET coated with unloaded pullulan (≈26% reduction in both coefficients). In spite of the presence of the filler, all of the coating formulations did not significantly impair the overall optical properties of the final material, which exhibited haze values below 3% and transmittance above 85%. The only exception to this was represented by the formulation with the highest loading of mica (1.5 wt %, which is ϕ ≈ 0.01). These findings revealed, for the first time, the potential of the NBB mica to produce nanocomposite coatings in combination with biopolymers for the generation of new functional features, such as transparent high oxygen barrier materials. PMID:28925951

  6. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party may...

  7. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party may...

  8. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party may...

  9. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party may...

  10. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party may...

  11. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean or coastwise service must be fitted with a magnetic compass. (b) All... gyrocompass in addition to the magnetic compass. (c) Each vessel must have an illuminated repeater for...

  12. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean or coastwise service must be fitted with a magnetic compass. (b) All... gyrocompass in addition to the magnetic compass. (c) Each vessel must have an illuminated repeater for...

  13. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean or coastwise service must be fitted with a magnetic compass. (b) All... gyrocompass in addition to the magnetic compass. (c) Each vessel must have an illuminated repeater for...

  14. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean or coastwise service must be fitted with a magnetic compass. (b) All... gyrocompass in addition to the magnetic compass. (c) Each vessel must have an illuminated repeater for...

  15. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean or coastwise service must be fitted with a magnetic compass. (b) All... gyrocompass in addition to the magnetic compass. (c) Each vessel must have an illuminated repeater for...

  16. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge. ...

  17. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge. ...

  18. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge. ...

  19. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge. ...

  20. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge. ...

  1. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17.1 Purpose of regulations. (a) The regulations in this part implement the Endangered Species Act of... Convention on International Trade in Endangered Species of Wild Fauna and Flora, for which regulations...

  2. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17.1 Purpose of regulations. (a) The regulations in this part implement the Endangered Species Act of... Convention on International Trade in Endangered Species of Wild Fauna and Flora, for which regulations...

  3. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17.1 Purpose of regulations. (a) The regulations in this part implement the Endangered Species Act of... Convention on International Trade in Endangered Species of Wild Fauna and Flora, for which regulations...

  4. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17.1 Purpose of regulations. (a) The regulations in this part implement the Endangered Species Act of... Convention on International Trade in Endangered Species of Wild Fauna and Flora, for which regulations...

  5. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17.1 Purpose of regulations. (a) The regulations in this part implement the Endangered Species Act of... Convention on International Trade in Endangered Species of Wild Fauna and Flora, for which regulations...

  6. Printed paper sensors for serum lactate dehydrogenase using pullulan-based inks to immobilize reagents.

    PubMed

    Kannan, Balamurali; Jahanshahi-Anbuhi, Sana; Pelton, Robert H; Li, Yingfu; Filipe, Carlos D M; Brennan, John D

    2015-09-15

    In this study, a paper-based point-of-care (POC) colorimetric biosensor was developed for the detection of lactate dehydrogenase in serum using a nonporous, oxygen impermeable reversibly gelling polysaccharide material based on pullulan. The pullulan could be printed onto paper surfaces along with all required assay reagents, providing a means for high-stability immobilization of all reagents on paper. Serum containing lactate dehydrogenase (LDH) was directly spotted on to the pullulan-coated bioactive paper and provided quantitative colorimetric data that was comparable to that obtained with a conventional plate-reader method. The paper strip was found to be highly stable and could be stored at 4 °C for at least 10 weeks with no loss in performance, as compared to a complete loss in performance within 1 day when the reagents were printed without the stabilizing polysaccharide. The ease of fabrication coupled with the high stability of the printed reagents provides a facile platform for easily manufactured POC sensors.

  7. PAMAM-pullulan conjugates as targeted gene carriers for liver cell.

    PubMed

    Askarian, Saeedeh; Abnous, Khalil; Ayatollahi, Sara; Farzad, Sara Amel; Oskuee, Reza Kazemi; Ramezani, Mohammad

    2017-02-10

    Targeted nano-carriers are highly needed to promote nucleic acid delivery into the specific cell for therapeutic approaches. Pullulan as a linear carbohydrate has an intrinsic liver targeting property interacting with asialoglycoprotein receptor (ASGPR) found on liver cells. In the present study, we developed polyamidoamine (PAMAM)-pullulan conjugates and investigated their targeting activity in delivering gene into liver cells. The particle size, zeta potential, buffering capacity and ethidium bromide exclusion assays of the conjugates were evaluated. The cytotoxicity and transfection efficiency of new derivatives were assessed following in vitro transfection of HepG2 (receptor positive) and N2A (receptor negative) cell lines. Size of conjugated polymers ranged between 118 and 184 nanometers and their cytotoxicity were similar to PAMAM. Among six produced nanocarriers, G4PU4 and G5PU4 enhanced transfection efficiency in HepG2 cells compared to unmodified PAMAM. Therefore, the PAMAM-pullulan derivatives seem to improve delivery of nucleic acids into the liver cells expressing asialoglycoprotein receptor with minimal transfection in non-targeted cells.

  8. Synthesis and characterization of pullulan-polycaprolactone core-shell nanospheres encapsulated with ciprofloxacin.

    PubMed

    Shady, Sally Fouad; Gaines, Peter; Garhwal, Rahul; Leahy, Charles; Ellis, Edward; Crawford, Kathryn; Schmidt, Daniel F; McCarthy, Stephen P

    2013-09-01

    Nanosphere-encapsulated drugs offer a means to overcome many drug delivery limitations by localizing the site of delivery and providing controlled release. This research details the synthesis and encapsulation of ciprofloxacin in pullulan-polycaprolactone (PCL) core shell nanospheres and the characterization of these materials by 1H-NMR, UV spectroscopy, dynamic light scattering (DLS) and scanning electron microscopy (SEM).1H-NMR results confirm that the pullulan-PCL grafted copolymer was successfully synthesized. UV spectroscopy showed that the ciprofloxacin loaded nanospheres contain 35-40% ciprofloxacin by weight. DLS and SEM results indicate that the loaded nanospheres are spherical in shape and approximately 142+/-12 nm in size. Under in vitro test conditions, approximately 20% of the ciprofloxacin is released in the first 4 hours, with additional release over 10 days. The nanoparticles demonstrate bioactivity against Escheria coli and do not affect the proliferation of two human cell lines. These results demonstrate the potential of pullulan-PCL core-shell nanospheres as delivery vehicles of hydrophobic drugs, including antibiotics for localized treatments applicable to a wide-range of human bacterial infections.

  9. Functional Biomaterials: Solution Electrospinning and Gelation of Whey Protein and Pullulan

    NASA Astrophysics Data System (ADS)

    Sullivan, Stephanie Tolstedt

    Utilizing biomaterials that are biodegradable, biocompatible and edible serve well for food products as well as biomedical applications. Biomaterials whey protein and pullulan both have these characteristics. Whey proteins (WP) have been used in food products for many years and more recently in pharmaceutical products. They have the ability to form both gels and stable foams. Pullulan (PULL) has also been used in both food and pharmaceutical products, and is a highly water soluble, non-gelling polysaccharide and has been used primarily as a film former. Herein, we investigate the ability of whey protein and pullulan to form nanofibers and gels. Combining their distinct properties allows the ability to uniquely manipulate nanofiber and gel characteristics and behavior for a variety of applications, from food to even tissue scaffolding. First, we determined the electrospinnability of aqueous whey protein solutions. Both whey protein isolate (WPI) and one of its major components beta--lactoglobulin (BLG), either in native or denatured form, yielded interesting micro and nanostructures when electrosprayed; while nanofiber production required blending with a spinnable polymer, poly(ethylene oxide) (PEO). WP:PEO solutions were also successfully electrospun at acidic pH (2≤pH≤3), which could improve shelf life. Fourier Transform Infrared Reflectance (FTIR) analysis of WP:PEO fiber mat indicated some variation in WP secondary structure with varying WPI concentration (as WPI increased, % alpha-helix increased and beta-turn decreased) and pH (as pH decreased from neutral (7.5) to acidic (2), % beta-sheet decreased and alpha-helix increased). X-ray Photoelectron Spectroscopy (XPS) also confirmed the presence of WP on the surface of the blend fibers, augmenting the FTIR analysis. Interestingly, WP:PEO composite nanofibers maintained its fibrous morphology at temperatures as high as 100 °C, above the 60 °C PEO melting point. Further, we show that the blend mats retained a

  10. Genetic Modification of the Marine-Isolated Yeast Aureobasidium melanogenum P16 for Efficient Pullulan Production from Inulin.

    PubMed

    Ma, Zai-Chao; Liu, Nan-Nan; Chi, Zhe; Liu, Guang-Lei; Chi, Zhen-Ming

    2015-08-01

    In this study, in order to directly and efficiently convert inulin into pullulan, the INU1 gene from Kluyveromyces maximum KM was integrated into the genomic DNA and actively expressed in the high pullulan producer Aureobasidium melanogenum P16 isolated from the mangrove ecosystem. After the ability to produce pullulan from inulin by different transformants was examined, it was found that the recombinant strain EI36, one of the transformants, produced 40.92 U/ml of inulinase activity while its wild-type strain P16 only yielded 7.57 U/ml of inulinase activity. Most (99.27 %) of the inulinase produced by the recombinant strain EI36 was secreted into the culture. During the 10-l fermentation, 70.57 ± 1.3 g/l of pullulan in the fermented medium was attained from inulin (138.0 g/l) within 108 h, high inulinase activity (42.03 U/ml) was produced within 60 h, the added inulin was actively hydrolyzed by the secreted inulinase, and most of the reducing sugars were used by the recombinant strain EI36. This confirmed that the genetically engineered yeast of A. melanogenum strain P16 was suitable for direct pullulan production from inulin.

  11. LX-17-1 Stockpile Returned Material Lot Comparison

    SciTech Connect

    Gagliardi, F.; Pease, S.; Willey, T.

    2015-02-18

    Many different lots of LX-17 have been produced over the years. Two varieties of LX-17, LX-17-0 and LX-17-1, have at one point or another been a part of the Livermore stockpile systems. LX-17-0 was made with dry-aminated TATB whereas LX-17-1 was made with wet-aminated TATB. Both versions have the same TATB to Kel-F 800 mass ratio of 92.5%/7.5%. Both kinds of LX-17 were formulated at Holston during the late 1970s or early to mid-1980s and were certified to have met the necessary specifications that cover the purity, particle size range, explosive to binder ratio, etc. In recent years, Trevor Willy and others have performed a detailed evaluation of solid parts made from each of the LX-17 lots manufactured at Holston. Using the Advanced Light Source at LBNL, Willey and his colleagues radiographed many samples from isostatic pressings using the same scanning conditions. In their investigation they identified that even though the bulk composition can be the same, there may exist a large spread in how smoothly the TATB and binder were distributed within the radiographed volume of different lots of material.1 Overall, the dry-aminated TATB-based material, LX-17-0, had a smooth TATB and binder distribution, whereas the wet-aminated TATB-based LX-17-1 showed a wide range of binder distributions. The results for five different LX-17-1 lots are shown in Figure 1. The wide variation in material distribution has raised the question about whether or not this sort variability will cause significant differences in mechanical behavior.

  12. Preparation and characterization of fast dissolving pullulan films containing BCS class II drug nanoparticles for bioavailability enhancement.

    PubMed

    Krull, Scott M; Ma, Zhelun; Li, Meng; Davé, Rajesh N; Bilgili, Ecevit

    2016-01-01

    The aim of this study is to assess pullulan as a novel steric stabilizer during the wet-stirred media milling (WSMM) of griseofulvin, a model poorly water-soluble drug, and as a film-former in the preparation of strip films via casting-drying the wet-milled drug suspensions for dissolution and bioavailability enhancement. To this end, pullulan films, with xanthan gum (XG) as thickening agent and glycerin as plasticizer, were loaded with griseofulvin nanoparticles prepared by WSMM using pullulan in combination with sodium dodecyl sulfate (SDS) as an ionic stabilizer. The effects of drug loading and milling time on the particle size and suspension stability were investigated, as well as XG concentration and casting thickness on film properties and dissolution rate. The nanosuspensions prepared with pullulan-SDS combination were relatively stable over 7 days; hence, this combination was used for the film preparation. All pullulan-based strip films exhibited excellent content uniformity (most <3% RSD) despite containing only 0.3-1.3 mg drug, which was ensured by the use of precursor suspensions with >5000 cP viscosity. USP IV dissolution tests revealed fast/immediate drug release (t80 < 30 min) from films <120 μm thick. Thinner films, films with lower XG loading, or smaller drug particles led to faster drug dissolution, while drug loading had no discernible effect. Overall, these results suggest that pullulan may serve as an acceptable stabilizer for media milling in combination with surfactant as well as a fast-dissolving film former for the fast release of poorly water-soluble drug nanoparticles.

  13. Plasticizers Increase Adhesion of the Deteriogenic Fungus Aureobasidium pullulans to Polyvinyl Chloride

    PubMed Central

    Webb, Jeremy S.; Van der Mei, Henny C.; Nixon, Marianne; Eastwood, Ian M.; Greenhalgh, Malcolm; Read, Simon J.; Robson, Geoffrey D.; Handley, Pauline S.

    1999-01-01

    Initial adhesion of fungi to plasticized polyvinyl chloride (pPVC) may determine subsequent colonization and biodeterioration processes. The deteriogenic fungus Aureobasidium pullulans was used to investigate the physicochemical nature of adhesion to both unplasticized PVC (uPVC) and pPVC containing the plasticizers dioctyl phthalate (DOP) and dioctyl adipate (DOA). A quantitative adhesion assay using image analysis identified fundamental differences in the mechanism of adhesion of A. pullulans blastospores to these substrata. Adhesion to pPVC was greater than that to uPVC by a maximum of 280% after a 4-h incubation with 108 blastospores ml−1. That plasticizers enhance adhesion to PVC was confirmed by incorporating a dispersion of both DOA and DOP into the blastospore suspension. Adhesion to uPVC was increased by up to 308% in the presence of the dispersed plasticizers. Hydrophobic interactions were found to dominate adhesion to uPVC because (i) a strong positive correlation was observed between substratum hydrophobicity (measured by using a dynamic contact angle analyzer) and adhesion to a range of unplasticized polymers including uPVC, and (ii) neither the pH nor the electrolyte concentration of the suspension buffer, both of which influence electrostatic interactions, affected adhesion to uPVC. In contrast, adhesion to pPVC is principally controlled by electrostatic interactions. Enhanced adhesion to pPVC occurred despite a relative reduction of 13° in the water contact angle of pPVC compared to that of uPVC. Furthermore, adhesion to pPVC was strongly dependent on both the pH and electrolyte concentration of the suspension medium, reaching maximum levels at pH 8 and with an electrolyte concentration of 10 mM NaCl. Plasticization with DOP and DOA therefore increases adhesion of A. pullulans blastospores to pPVC through an interaction mediated by electrostatic forces. PMID:10427051

  14. Growth inhibition of Candida species by Wickerhamomyces anomalus mycocin and a lactone compound of Aureobasidium pullulans.

    PubMed

    Tay, Sun-Tee; Lim, Su-Lin; Tan, Hui-Wee

    2014-11-08

    The increasing resistance of Candida yeasts towards antifungal compounds and the limited choice of therapeutic drugs have spurred great interest amongst the scientific community to search for alternative anti-Candida compounds. Mycocins and fungal metabolites have been reported to have the potential for treatment of fungal infections. In this study, the growth inhibition of Candida species by a mycocin produced by Wickerhamomyces anomalus and a lactone compound from Aureobasidium pullulans were investigated. Mycocin was purified from the culture supernatant of an environmental isolate of W. anomalus using Sephadex G-75 gel filtration column chromatography. The mycocin preparation was subjected to SDS-PAGE analysis followed by MALDI TOF/TOF mass spectrometry analysis. The thermal and temperature stability of the mycocin were determined. The glucanase activity of the mycocin was investigated by substrate staining of the mycocin with 4-methyl-umbelliferyl-ß-D-glucoside (MUG). Gas chromatography mass spectrometry (GCMS) analysis was used to identify anti-Candida metabolite in the culture supernatant of an environmental isolate of Aureobasidium pullulans. The inhibitory effects of the anti-Candida compound against planktonic and biofilm cultures of various Candida species were determined using broth microdilution and biofilm quantitation methods. A mycocin active against Candida mesorugosa but not C. albicans, C. parapsilosis and C. krusei was isolated from the culture supernatant of W. anomalus in this study. The mycocin, identified as exo-ß-1,3 glucanase by MALDI TOF/TOF mass spectrometry, was stable at pH 3-6 and temperature ranging from 4-37°C. The glucanase activity of the mycocin was confirmed by substrate staining with MUG. 5-hydroxy-2-decenoic acid lactone (HDCL) was identified from the culture supernatant of A. pullulans. Using a commercial source of HDCL, the planktonic and biofilm MICs of HDCL against various Candida species were determined in this study

  15. Composite pullulan-dextran polysaccharide scaffold with interfacial polyelectrolyte complexation fibers: a platform with enhanced cell interaction and spatial distribution.

    PubMed

    Cutiongco, Marie Francene Arnobit; Tan, Ming Hao; Ng, Martin Yoke Kuang; Le Visage, Catherine; Yim, Evelyn King Fai

    2014-10-01

    Hydrogels are highly preferred in soft tissue engineering because they recapitulate the hydrated extracellular matrix. Naturally derived polysaccharides, like pullulan and dextran, are attractive materials with which to form hydrophilic polymeric networks due to their non-immunogenic and non-antigenic properties. However, their inherent hydrophilicity prevents adherent cell growth. In this study, we modified pullulan-dextran scaffolds with interfacial polyelectrolyte complexation (IPC) fibers to improve their ability to support adherent cell growth. We showed that the pullulan-dextran-IPC fiber composite scaffold laden with extracellular matrix protein has improved cell adhesion and proliferation compared to the plain polysaccharide scaffold. We also demonstrated the zero-order release kinetics of the biologics bovine serum albumin and vascular endothelial growth factor (VEGF) incorporated in the composite scaffold. Lastly, we showed that the VEGF released from the composite scaffold retained its capacity to stimulate endothelial cell growth. The incorporation of IPC fibers in the pullulan-dextran hydrogel scaffold improved its functionality and biological activity, thus enhancing its potential in tissue engineering applications.

  16. Facile and rapid thermo-regulated biomineralization of gold by pullulan and study of its thermodynamic parameters.

    PubMed

    Choudhury, Anirban Roy; Malhotra, Ankit; Bhattacharjee, Paramita; Prasad, G S

    2014-06-15

    A novel method for the production of gold nanoparticles (AuNPs) using pullulan as reducing and stabilizing agent has been developed. Quasi-spherical shaped AuNPs in the range of 50-100 nm were produced at three different temperature regimes 80°C, 90°C and 100°C as characterized using UV-vis spectrophotometer, TEM and DLS. Study of reaction kinetics and thermodynamic parameters indicated that the reaction between pullulan and chloroauric acid for AuNPs formation followed first order reaction kinetics and higher temperature was favorable for the synthesis of smaller sized AuNPs. FT-IR data analyses, provided an insight towards the mechanism of gold nanoparticle formation which suggested that, the free CH2OH groups of pullulan molecule were oxidized to carboxylate ions resulted in formation of AuNPs whereas the basic skeletal structure of pullulan remained unaltered. This study may open up new avenues for synthesis of tailor made biogenic AuNPs with possible application in biomedical field.

  17. Production of novel types of antibacterial liamocins by diverse strains of Aureobasidium pullulans grown on different culture media.

    PubMed

    Leathers, Timothy D; Price, Neil P J; Bischoff, Kenneth M; Manitchotpisit, Pennapa; Skory, Christopher D

    2015-10-01

    To compare production of antibacterial liamocins (polyol lipids) by diverse strains of Aureobasidium pullulans grown on different culture media. Liamocins produced by strains of A. pullulans have potential agricultural and pharmaceutical applications as antibacterials with specificity against Streptococcus spp. Six strains of A. pullulans were characterized for liamocin production on four different culture media. The choice of strain and culture medium affected growth, liamocin yields, and production of contaminating pigments. Best growth and highest liamocin yields were obtained using A. pullulans strain NRRL 50384 grown on a sucrose basal medium. Unexpectedly, the choice of strain and culture medium also affected the structure of liamocins produced, providing novel types of liamocins. Liamocins varied not only in the ratios of trimer and tetramer polyester tail groups, but also in the nature of the polyol headgroup, which could include mannitol, arabitol, or glycerol. The ability to conveniently produce novel types of liamocins in good yields will provide novel antibacterials for applied uses, and facilitate structure-function studies on the mechanism of antibacterial activity.

  18. Ascochyta blight of chickpea reduced 38% by application of Aureobasidium pullulans (anamorphic Dothioraceae, Dothideales) to post harvest debris.

    USDA-ARS?s Scientific Manuscript database

    In 2004-2005, early winter application of suspensions of Aureobasidium pullulans (AuP) conidia to post-harvest chickpea debris resulted in significantly fewer spring-time Ascochyta blight lesions on chickpea test plants relative to controls. Survival of plants adjacent to treated debris was higher t...

  19. Improving Erythritol Production of Aureobasidium pullulans from Xylose by Mutagenesis and Medium Optimization.

    PubMed

    Guo, Jian; Li, Junxia; Chen, Yefu; Guo, Xuewu; Xiao, Dongguang

    2016-10-01

    The biotransformation of xylose, the second most abundant sugar, has been a hot topic in recent years. In this work, Aureobasidium pullulans CGMCC3.0837 was subjected to UV mutagenesis to improve its erythritol production from xylose. The erythritol production of the obtained mutant ER35 was 50.92 % (17.28 g/L) higher than that of the parent strain. Response surface methodology was applied to optimize the medium composition. Yeast extract, KH2PO4, and citric acid were the key factors influencing erythritol synthesis, and the optimal concentrations were 17.82, 0.76, and 6.60 g/L, respectively. Under the optimized conditions, 31.75 and 31.42 g/L erythritol were obtained in shake flasks and in a 5-L fermentor, respectively. ER35 also showed a good consuming ability on xylose mother liquor with a final erythritol production of 26.35 g/L. This report provided insights into the potential of A. pullulans for the production of erythritol using xylose as a carbon source.

  20. Production of anti-streptococcal liamocins from agricultural biomass by Aureobasidium pullulans.

    PubMed

    Leathers, Timothy D; Price, Neil P J; Manitchotpisit, Pennapa; Bischoff, Kenneth M

    2016-12-01

    Liamocins are unique heavier-than-water "oils" produced by certain strains of the fungus Aureobasidium pullulans. Liamocins have antibacterial activity with specificity for Streptococcus sp. Previous studies reported that liamocin yields were highest from strains of A. pullulans belonging to phylogenetic clades 8, 9, and 11, cultured on medium containing sucrose. In this study, 27 strains from these clades were examined for the first time for production of liamocins from agricultural biomass substrates. Liamocin yields were highest from strains in phylogenetic clade 11, and yields were higher from cultures grown on sucrose than from those grown on pretreated wheat straw. However, when supplementary enzymes (cellulase, β-glucosidase, and xylanase) were added, liamocin production on pretreated wheat straw was equivalent to that on sucrose. Liamocins produced from wheat straw were free of the melanin contamination common in sucrose-grown cultures. Furthermore, MALDI-TOF MS analysis showed that liamocins produced from wheat straw were under-acetylated, resulting in higher proportions of the mannitol A1 and B1 species of liamocin, the latter of which has the highest biological activity against Streptococcus sp.

  1. Microwave-assisted rapid synthesis, characterization and application of poly (D,L-lactide)-graft-pullulan.

    PubMed

    Tang, Xiao-Jiao; Huang, Jun; Xu, Liang-Yu; Li, Yang; Song, Juan; Ma, Yue; Yang, Li; Yuan, Dan; Wu, Hai-Yang

    2014-07-17

    A novel microwave-assisted method was developed to synthetize amphiphilic copolymer poly (d,l-lactide)-graft-pullulan (PL) in a monomode microwave reactor. The effects of microwave power, ratio of catalyst/lactide, ratio of lactide/hydroxyl group of pullulan (lactide/OH-P) and solvent on the synthesis were further investigated. Three samples (designated as PL 8, 9, and 6), characterized by FT-IR and NMR, were applied to form nanoparticles and microparticles investigated by dynamic light scattering, fluorescence spectroscopy and transmission electron microscopy. PL9 and PL6 were used for loading model drug curcumin. The results indicated that microwave-assisted synthesis shortened the copolymerization of PL, with higher yield and lactide conversion, from 24h to 5 min and showed some specific microwave effects compared with conventional oil heating. PL with a relative higher substitution degree gave nanoparticles with smaller sizes and critical aggregation concentrations. The solubility of curcumin was increased to 1.97 mg mL(-1) as the forms of nanoparticles. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. An Injectable Enzymatically Crosslinked Carboxymethylated Pullulan/Chondroitin Sulfate Hydrogel for Cartilage Tissue Engineering.

    PubMed

    Chen, Feng; Yu, Songrui; Liu, Bing; Ni, Yunzhou; Yu, Chunyang; Su, Yue; Zhu, Xinyuan; Yu, Xiaowei; Zhou, Yongfeng; Yan, Deyue

    2016-01-28

    In this study, an enzymatically cross-linked injectable and biodegradable hydrogel system comprising carboxymethyl pullulan-tyramine (CMP-TA) and chondroitin sulfate-tyramine (CS-TA) conjugates was successfully developed under physiological conditions in the presence of both horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) for cartilage tissue engineering (CTTE). The HRP crosslinking method makes this injectable system feasible, minimally invasive and easily translatable for regenerative medicine applications. The physicochemical properties of the mechanically stable hydrogel system can be modulated by varying the weight ratio and concentration of polymer as well as the concentrations of crosslinking reagents. Additionally, the cellular behaviour of porcine auricular chondrocytes encapsulated into CMP-TA/CS-TA hydrogels demonstrates that the hydrogel system has a good cyto-compatibility. Specifically, compared to the CMP-TA hydrogel, these CMP-TA/CS-TA composite hydrogels have enhanced cell proliferation and increased cartilaginous ECM deposition, which significantly facilitate chondrogenesis. Furthermore, histological analysis indicates that the hydrogel system exhibits acceptable tissue compatibility by using a mouse subcutaneous implantation model. Overall, the novel injectable pullulan/chondroitin sulfate composite hydrogels presented here are expected to be useful biomaterial scaffold for regenerating cartilage tissue.

  3. Preparation of maltotriose from fermentation broth by hydrolysis of pullulan using pullulanase.

    PubMed

    Wu, Sheng-Jun; Chen, Jing

    2014-07-17

    In this study, we prepared maltotriose from the fermentation broth of Auerobasidium Pullulans CJ001 isolated from the sea mud by hydrolysis of pullulan with pullulanase. The fermentation broth was centrifuged to remove the microorganisms and then hydrolysed by pullulanase. The optimal hydrolysis conditions were obtained as follows: time, 9.40 h; pH, 4.92; temperature, 47.88°C; pullulanase, 10ASPU/g. Under these optimum hydrolysis conditions, the maximum dextrose equivalent value reached 31.86. The hydrolysates were filtrated through a filter membrane to separate any particle with molecular weight higher than 1,000 Da, concentrated to ∼20%, and precipitated with 8 volumes of absolute ethanol. The precipitate was dried at 80°C for 2h to yield the maltotriose product. The maltotriose content in the product and the yield of maltotriose were 92.13% and 90.23%, respectively. The results indicate that this was a promising way of maltotriose production. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. An Injectable Enzymatically Crosslinked Carboxymethylated Pullulan/Chondroitin Sulfate Hydrogel for Cartilage Tissue Engineering

    NASA Astrophysics Data System (ADS)

    Chen, Feng; Yu, Songrui; Liu, Bing; Ni, Yunzhou; Yu, Chunyang; Su, Yue; Zhu, Xinyuan; Yu, Xiaowei; Zhou, Yongfeng; Yan, Deyue

    2016-01-01

    In this study, an enzymatically cross-linked injectable and biodegradable hydrogel system comprising carboxymethyl pullulan-tyramine (CMP-TA) and chondroitin sulfate-tyramine (CS-TA) conjugates was successfully developed under physiological conditions in the presence of both horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) for cartilage tissue engineering (CTTE). The HRP crosslinking method makes this injectable system feasible, minimally invasive and easily translatable for regenerative medicine applications. The physicochemical properties of the mechanically stable hydrogel system can be modulated by varying the weight ratio and concentration of polymer as well as the concentrations of crosslinking reagents. Additionally, the cellular behaviour of porcine auricular chondrocytes encapsulated into CMP-TA/CS-TA hydrogels demonstrates that the hydrogel system has a good cyto-compatibility. Specifically, compared to the CMP-TA hydrogel, these CMP-TA/CS-TA composite hydrogels have enhanced cell proliferation and increased cartilaginous ECM deposition, which significantly facilitate chondrogenesis. Furthermore, histological analysis indicates that the hydrogel system exhibits acceptable tissue compatibility by using a mouse subcutaneous implantation model. Overall, the novel injectable pullulan/chondroitin sulfate composite hydrogels presented here are expected to be useful biomaterial scaffold for regenerating cartilage tissue.

  5. Cloning, sequencing, and regulation of a xylanase gene from the fungus Aureobasidium pullulans Y-2311-1

    SciTech Connect

    Li, Xin-Liang; Ljungdahl, L.G.

    1994-09-01

    Aureobasidium pullulans Y-2311-1 growing on xylan secretes four major xylanases with different masses and isoelectric points. Two of these enzymes, named APX-I and APX-II, have been purified previously. Their N-terminal amino acid sequences are identical except that APX-I has Asp and APX-II has Asn at position 7. An 83-bp DNA region was amplified by PCR and used as a probe for the xylanase gene cloning. The longest cDNA (xynA) obtained by cDNA cloning and PCR amplification consisted of 895 bp. A. pullulans xynA had an open reading frame encoding a polypeptide of 221 amino acids with a calculated mass of 23,531 Da and contained a putative 34-amino-acid signal peptide in front of the amino terminus of the mature enzyme. Strong homology was found between the deduced amino acid sequence of XynA and some xylanases from bacterial and fungal sources. It is suggested that A. pullulans XynA belongs to the family G glycanases. Northern (RNA blot) analysis revealed that only one transcript of 900 bases was present in cultures grown in medium containing D-xylose or oat spelt xylan. Transcription was completely repressed in the presence of glucose in the medium. Southern blot analysis indicated that A. pullulans xynA was present as a single copy in the genome. Comparison between the genomic and cDNA sequences revealed that one intron of 59 bp was present in the coding region. The data presented suggest that the highly active xylanases, APX-I and APX-II, secreted by A. pullulans are encoded by the same gene. 36 refs., 7 figs., 3 tabs.

  6. Synthesis, characterization, and cytotoxicity of the plasmid EGFP-p53 loaded on pullulan-spermine magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Eslaminejad, Touba; Nematollahi-Mahani, Seyed Noureddin; Ansari, Mehdi

    2016-03-01

    Magnetic nanoparticles have been used as effective vehicles for the targeted delivery of therapeutic agents that can be controlled in their concentration and distribution to a desired part of the body by using externally driven magnets. This study focuses on the synthesis, characterization, and functionalization of pullulan-spermine (PS) magnetic nanoparticles for medical applications. Magnetite nanopowder was produced by thermal decomposition of goethite (FeOOH) in oleic acid and 1-octadecene; pullulan-spermine was deposited on the magnetite nanoparticles in the form of pullulan-spermine clusters. EGFP-p53 plasmid was loaded on functionalized iron oleate to transfer into cells. Synthesized nanoparticles were characterized by Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), vibrating sample magnetometry (VSM), and transmission electron microscopy (TEM). The encapsulation efficiency and drug loading efficiency of the nanocomplexes were tested. FTIR studies showed the presence of oleic acid and 1-octadecene in the iron oleate nanopowder and verified the interaction between spermine and pullulan. The characteristic bands of PS in the spectrum of the pullulan-spermine-coated iron oleate (PSCFO) confirmed that PS covered the surface of the iron oleate particles. TEM studies showed the average size of the iron oleate nanopowder, the PSCFO, and the plasmid-carrying PSCFO (PSCFO/pEGFP-p53) to be 34±12 nm, 100±50 nm and 172±3 nm, respectively. Magnetic measurements revealed that magnetic saturation of the PSCFO was lower in comparison with the iron oleate nanopowder due to the presence of organic compounds in the former. In cytotoxicity tests performed using U87 cells as glioblastoma cells, a 92% survival rate was observed at 50 μg/μl of the plasmid-carrying PSCFO, with an IC50 value of 189 μg/μl.

  7. In situ quantification of biocide efficacy using GFP transformed Aureobasidium pullulans.

    PubMed

    Sabev, H A; Handley, P S; Robson, G D

    2004-01-01

    To develop a real-time in situ method to quantify loss of viability of Aureobasidium pullulans PRAFS8 cells attached to plasticized polyvinyl chloride (pPVC) with incorporated biocides, and to use the method to compare biocide efficacy in situ. A. pullulans PRAFS8, transformed with green fluorescent protein (GFP), was used to quantify the efficacy of a range of biocides incorporated into pPVC. Experimentally, it was found that a density of 1.53 x 10(6) yeast cells per cm(2) of pPVC was optimal as increasing the density of the yeast cells to 6.12 x 10(6) cm(-2) attached to pPVC containing the biocide 2-n-octyl-4-isothiazolin-3-one (OIT) decreased the rate of fluorescence loss. A strong positive correlation between fluorescence and viable yeast cell number was observed and fluorescence was used as a direct indicator of cell viability. The effectiveness of five commercial biocides, commonly incorporated into pPVC at their in-use concentrations, was tested against yeast cells attached to the pPVC surface. The loss of fluorescence and hence viability in situ was quantified using image analysis. The biocides N-(trichloromethylthio) phthalimide (NCMP), 10,10'-oxybisphenoxarsine (OBPA), OIT and 2,3,5,6-tetrachloro-4-(methylsulphonyl) pyridine (TCMP) caused complete loss of fluorescence within 30-50 h. In contrast the biocide dichloro-octyl-isothiazoline caused only 55 +/- 15% fluorescence loss after 50 h. Starvation of the yeast cells in suspension for 24 h prior to attachment reduced their initial sensitivity to OBPA, NCMP, OIT and TCMP by 15-20%, but eventually the fluorescence was also completely lost. The use of A. pullulans expressing cytosolic GFP enables the in situ quantification of loss of viability when cells are attached to pPVC with incorporated biocides. GFP fluorescence was used as a real-time indicator of cell viability and thus can be applied for direct quantification of the effectiveness of a broad range of biocides, incorporated into the polymer mass and

  8. [Effect of conditions od Trichosporon pullulans culturing on the synthesis of immunomodulating glycoprotein].

    PubMed

    Eroshin, V K; Dediukhina, E G; Vagabov, V M; Kulaev, I S

    1999-01-01

    An extracellular glycoprotein (GP) exhibiting immunomodulating activity produced by the yeast Trichosporon pullulans grown in a defined ethanol-containing medium differed substantially in its composition from that of the yeast cell walls: therefore, it cannot be considered a structural component of the cell walls. In batch culture, the greatest GP production (40 mg/l) occurred in the exponential phase of the yeast growth. Under continuous cultivation, in both chemostat and pH-auxostat regimes, the specific rate of GP synthesis (qGP) increased with the increasing specific growth rate (mu) and reached 1.55 mg/(g h) at mumax. Under limitation of the yeast growth by zinc qGP was three times lower than under nitrogen or iron limitation. The rate of GP production depended inversely on the oxygen concentration.

  9. Synthesis and physicochemical characterization of a novel ampholytic pullulan derivative with amphiphilic behavior in alkaline media.

    PubMed

    Souguir, Z; Roudesli, S; About-Jaudet, E; Le Cerf, D; Picton, L

    2007-09-01

    Pullulan derivative was synthesized by coupling carboxymethylpullulan (degree of substitution DS(0)(in)=0.7) and dimethylaminopropylamine (DMAPA), activated by a hydrosoluble carbodiimide N(')-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDAC). FTIR and (13)C and (1)H NMR spectroscopic analyses have evidenced that the polysaccharide has been successfully modified. (1)H NMR, nitrogen analysis, and conductimetric measurements allow determination of the degree of substitution in both carboxylic acid and amine functions. We have found that both functions present a similar DS of 0.35, which is characteristic of an ampholytic polymer with possible zwitterionic-type properties. Solution properties have been studied by flow field flow fractionation (F4) coupled on-line with multiangle laser light scattering (MALLS) and quasi elastic light scattering (QELS), surface tension, and viscosity measurements. The behavior has been found largely pH dependent and an amphiphilic behavior has been evidenced in alkaline media.

  10. Coating with spermine-pullulan polymer enhances adenoviral transduction of mesenchymal stem cells

    PubMed Central

    Wan, Li; Yao, Xinglei; Faiola, Francesco; Liu, Bojun; Zhang, Tianyuan; Tabata, Yasuhiko; Mizuguchi, Hiroyuki; Nakagawa, Shinsaku; Gao, Jian-Qing; Zhao, Robert Chunhua

    2016-01-01

    Mesenchymal stem cells (MSCs) are adult stem cells with multilineage potential, which makes them attractive tools for regenerative medicine applications. Efficient gene transfer into MSCs is essential not only for basic research in developmental biology but also for therapeutic applications involving gene-modification in regenerative medicine. Adenovirus vectors (Advs) can efficiently and transiently introduce an exogenous gene into many cell types via their primary receptors, the coxsackievirus and adenovirus receptors, but not into MSCs, which are deficient in coxsackievirus and adenovirus receptors expression. To overcome this problem, we developed an Adv coated with a spermine-pullulan (SP) cationic polymer and investigated its physicochemical properties and internalization mechanisms. We demonstrated that the SP coating could enhance adenoviral transduction of MSCs without detectable cytotoxicity or effects on differentiation. Our results argue in favor of the potentiality of the SP-coated Adv as a prototype vector for efficient and safe transduction of MSCs. PMID:28008251

  11. A modified model for non-newtonian viscosity behavior of Aureobasidium pullulans culture fluid.

    PubMed

    Furuse, Hisamoto; Yabe, Isamu; Asakura, Tomoko; Miyawaki, Osato; Toda, Kiyoshi

    2003-01-01

    The culture fluid of the fungus Aureobasidium pullulans and the exopolysaccharide solution obtained by removal of the microbial cells exhibit a marked shear dependence of viscosity. The viscosity in a high shear rate region was a little higher than that predicted by a non-Newtonian viscosity equation derived previously on the basis of the concept of traveling force. In a sample exhibiting such high shear rate dependence, a hydrodynamic effect based on the fluid structure of the binding of contacting polymers and suspended microbial cells on viscosity becomes comparatively significant. A model for the shear rate dependence of the viscosity is needed to elucidate the mechanism of the viscosity behavior. A term concerning the increase in viscosity caused by the binding of polymers and the microbial cells suspended in a medium was added to the previous viscosity equation. The experimental shear dependence of the viscosity was well simulated by the modified viscosity equation.

  12. Nontoxic Genetic Engineering of Mesenchymal Stem Cells Using Serum-Compatible Pullulan-Spermine/DNA Anioplexes

    PubMed Central

    Thakor, Devang K.; Obata, Hideaki; Nagane, Kentaro; Saito, Shigeru

    2011-01-01

    Genetic modification of stem cells could be applied to initiate/enhance their secretion of therapeutic molecules, alter their biological properties, or label them for in vivo tracking. We recently developed a negatively charged gene carrier (“anioplex”) based on pullulan-spermine, a conjugate prepared from a natural polysaccharide and polyamine. In rat mesenchymal stem cells (MSCs), anioplex-derived reporter gene activity was comparable to or exceeded that obtained using a commercial cationic lipid reagent. Transfection in the growth medium with 15% serum and antibiotics was approximately sevenfold more effective than in serum-free conditions. Cytotoxicity was essentially indiscernible after 24 h of anioplex transfection with 20 μg/mL DNA, in contrast to cationic lipid transfection that resulted in 40%–60% death of target MSCs. Anioplex-derived reporter gene activity persisted throughout the entire 3-week study, with post-transfection MSCs appearing to maintain osteogenic, adipogenic, and chondrogenic multipotency. In particular, chondrogenic pellet formation of differentiating human MSCs was significantly inhibited after lipofection but not after aniofection, which further indicates the biological inertness of pullulan-spermine/DNA anioplexes. Collectively, these data introduce a straightforward technology for genetic engineering of adult stem/progenitor cells under physiological niche-like conditions. Moreover, reporter gene activity was observed in rat spinal cords after minimally invasive intrathecal implantation, suggesting effective engraftment of donor MSCs. It is therefore plausible that anioplex-transfected MSCs or other stem/progenitor cells with autologous potential could be applied to disorders such as neurotrauma or neuropathic pain that involve the spinal cord and brain. PMID:20698746

  13. Biocontrol of Monilinia laxa by Aureobasidium pullulans strains: Insights on competition for nutrients and space.

    PubMed

    Di Francesco, Alessandra; Ugolini, Luisa; D'Aquino, Salvatore; Pagnotta, Eleonora; Mari, Marta

    2017-02-17

    Two Aureobasidium pullulans strains (L1 and L8), able to prevent postharvest fruit decay, were evaluated in order to elucidate how the competition for nutrients and space was involved in their activity against Monilinia laxa, the causal agent of peach brown rot. The competition for nutrients was studied by co-culturing pathogen conidia and antagonists in different conditions of nutrient availability and avoiding contact between them. Both antagonists prevented the germination of conidia of M. laxa in water, reducing germination rate by >35%. However, L1 and L8 showed the lowest inhibition of conidial germination in peach juice at 5%, with a reduction of 12.6% and 13.9% respectively. HPLC amino acid analysis of peach juice revealed that the addition of the yeast suspension greatly modified their composition: asparagine was completely depleted soon after 12h of incubation and was probably hydrolyzed to aspartic acid by the yeasts, as aspartic acid content markedly increased. Pure asparagine and aspartic acid were tested by in vitro trials at the concentrations found in peach juice: both influenced M. laxa growth, but in opposite ways. Asparagine stimulated pathogen growth; conversely, amended medium with aspartic acid significantly inhibited the conidia germination and mycelial development of M. laxa. Scanning Electron Microscopy revealed that both strains showed a great capability to compete with M. laxa for space (starting 8h after treatment), colonizing the wound surface and inhibiting pathogen growth. This study clearly showed that A. pullulans L1 and L8 strains could compete with M. laxa for nutrients and space; this mode of action may play an important role in the antagonistic activity, especially in the first hours of tritrophic host-pathogen-antagonist interaction, although several other mechanisms can interact each other.

  14. 29 CFR 17.1 - What is the purpose of these regulations?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 1 2014-07-01 2013-07-01 true What is the purpose of these regulations? 17.1 Section 17.1... ACTIVITIES § 17.1 What is the purpose of these regulations? (a) The regulations in this part implement E.O.... These regulations also implement applicable provisions of section 401 of the Intergovernmental...

  15. Incorporation of essential oils and nanoparticles in pullulan films to control foodborne pathogens on meat and poultry products.

    PubMed

    Morsy, Mohamed K; Khalaf, Hassan H; Sharoba, Ashraf M; El-Tanahi, Hassan H; Cutter, Catherine N

    2014-04-01

    The incorporation of essential oils and nanotechnology into edible films has the potential to improve the microbiological safety of foods. The aim of this study was to evaluate the effectiveness of pullulan films containing essential oils and nanoparticles against 4 foodborne pathogens. Initial experiments using plate overlay assays demonstrated that 2% oregano essential oil was active against Staphylococcus aureus and Salmonella Typhimurium, whereas Listeria monocytogenes and Escherichia coli O157:H7 were not inhibited. Two percent rosemary essential oil was active against S. aureus, L. monocytogenes, E. coli O157:H7, and S. Typhimurium, when compared with 1%. Zinc oxide nanoparticles at 110 nm were active against S. aureus, L. monocytogenes, E. coli O157:H7, and S. Typhimurium, when compared with 100 or 130 nm. Conversely, 100 nm silver (Ag) nanoparticles were more active against S. aureus than L. monocytogenes. Using the results from these experiments, the compounds exhibiting the greatest activity were incorporated into pullulan films and found to inhibit all or some of the 4 pathogens in plate overlay assays. In challenge studies, pullulan films containing the compounds effectively inhibited the pathogens associated with vacuum packaged meat and poultry products stored at 4 °C for up to 3 wk, as compared to control films. Additionally, the structure and cross-section of the films were evaluated using electron microscopy. The results from this study demonstrate that edible films made from pullulan and incorporated with essential oils or nanoparticles may improve the safety of refrigerated, fresh or further processed meat and poultry products. © 2014 Institute of Food Technologists®

  16. Similarity of permeabilities for Ficoll, pullulan, charge-modified albumin and native albumin across the rat peritoneal membrane.

    PubMed

    Asgeirsson, D; Axelsson, J; Rippe, C; Rippe, B

    2009-08-01

    Compared to neutral globular proteins, neutral polysaccharides, such as dextran, pullulan and Ficoll, appear hyperpermeable across the glomerular filtration barrier. This has been attributed to an increased flexibility and/or asymmetry of polysaccharides. The present study investigates whether polysaccharides are hyperpermeable also across the continuous capillaries in the rat peritoneum. In anaesthetized Wistar rats, FITC-Ficoll or FITC-pullulan together with (125)I-human serum albumin (RISA) or neutralized (125)I-bovine serum albumin (nBSA) were given intravenously, after which peritoneal dialysis (PD) using conventional PD fluid (Gambrosol 1.5%) was performed for 120 min. Concentrations of FITC-polysaccharides and radioactive albumin species in plasma and dialysis fluid were analysed with high-performance size exclusion chromatography and a gamma counter respectively. Transperitoneal clearance values were calculated for polysaccharides in the molecular radius range 36-150 A, and for RISA and nBSA. Ficoll and pullulan showed more or less identical permeabilities, compared to RISA and nBSA, across the peritoneal membrane. Although RISA-clearance, 5.50 +/- 0.28 (microL min(-1); +/-SEM), tended to be lower than the clearances of Ficoll(36A) (6.55 +/- 0.25), pullulan(36A) (6.08 +/- 0.22) and nBSA (6.56 +/- 0.23), the difference was not statistically significant. This is in contrast to the hyperpermeability exhibited by polysaccharides across the glomerular filtration barrier and also contrasts with the charge selectivity of the latter. The phenomenon of molecular flexibility is more important for a macromolecule's permeability through the glomerular filter than across the continuous peritoneal capillary endothelium. Furthermore, it seems that charge plays a subordinate role in the steady-state transport across the combined peritoneal capillary-interstitial barrier.

  17. NMR spectroscopic structural characterization of a water-soluble β-(1→3, 1→6)-glucan from Aureobasidium pullulans.

    PubMed

    Kono, Hiroyuki; Kondo, Nobuhiro; Hirabayashi, Katsuki; Ogata, Makoto; Totani, Kazuhide; Ikematsu, Shinya; Osada, Mitsumasa

    2017-10-15

    An unambiguous structural characterization of the water-soluble Aureobasidium pullulans β-(1→3, 1→6)-glucan is yet to be achieved, although this β-(1→3, 1→6)-glucan is expected to exhibit excellent biofunctional properties. Thus, we herein report the elucidation of the primary structure of the A. pullulans β-(1→3, 1→6)-glucan using nuclear magnetic resonance spectroscopy, followed by comparison of the obtained structure with that of schizophyllan (SPG). Structural characterization of the A. pullulans β-(1→3, 1→6)-glucan revealed that the structural units are a β-(1→3)-d-glucan backbone with four β-(1→6)-d-glucosyl side branching units every six residues. In addition, circular dichroism spectroscopic analysis revealed that the β-(1→3, 1→6)-glucan interacted with polyadenylic acid (poly(A)) chains in DMSO solution to form a complex similar to that obtained in the complexation of SPG/poly(A). This finding indicates that β-(1→3, 1→6)-glucan forms a triple-helical conformation in aqueous solution but exhibits a random coil structure in DMSO solution, which is similar to the behavior of SPG. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. [Agrobacterium tumefaciens-mediated transformation of Aureobasidium pullulans and high-efficient screening for polymalic acid producing strain].

    PubMed

    Tu, Guangwei; Wang, Yongkang; Feng, Jun; Li, Xiaorong; Guo, Meijin; Zou, Xiang

    2015-07-01

    To develop a genetic transformation method of Aureobasidium pullulans and T-DNA insertion for high-efficient screening of polymalic acid (PMA) producing strain. Agrobacterium tumefaciens-AGL1, containing the selection genes encoding hygromycin B phosphotase or phosphinothricin acetyltranferase, was used to transform Aureobasidium pullulans CCTCC M2012223 and transformants were confirmed by colony PCR method. Transferred DNA (T-DNA) insertional mutants were cultured in microwell plate, and screened for high-titer PMA producing strain according to the pH response model. DNA walking was used to detect the insertion sites in the mutant. Results show that the selection markers could stably generated in the transformants, and 80 to 120 transformants could be found per 10(7) single cells. A high-titer PMA mutant H27 was obtained, giving a good PMA production caused by the disruption of phosphoglycerate mutase, that increased by 24.5% compared with the control. Agrobacterium tumefaciens-mediated transformation and high-efficient screening method were successfully developed, which will be helpful for genetic transformation of Aureobasidium pullulans and its functional genes discovery.

  19. Occurrence and diversity of marine yeasts in Antarctica environments

    NASA Astrophysics Data System (ADS)

    Zhang, Xue; Hua, Mingxia; Song, Chunli; Chi, Zhenming

    2012-03-01

    A total of 28 yeast strains were obtained from the sea sediment of Antarctica. According to the results of routine identification and molecular characterization, the strains belonged to species of Yarrowia lipolytica, Debaryomyces hansenii, Rhodotorula slooffiae, Rhodotorula mucilaginosa, Sporidiobolus salmonicolor, Aureobasidium pullulans, Mrakia frigida and Guehomyces pullulans, respectively. The Antarctica yeasts have wide potential applications in biotechnology, for some of them can produce β-galactosidase and killer toxins.

  20. Effect of meadowsweet flower extract-pullulan coatings on rhizopus rot development and postharvest quality of cold-stored red peppers.

    PubMed

    Synowiec, Alicja; Gniewosz, Małgorzata; Kraśniewska, Karolina; Chlebowska-Śmigiel, Anna; Przybył, Jarosław L; Bączek, Katarzyna; Węglarz, Zenon

    2014-08-25

    The study involved an examination of the antifungal activity on red peppers of pullulan coating (P) and pullulan coating containing either water-ethanol (P + eEMF) or ethanol extract of meadowsweet flowers (P + eEMF). Pullulan was obtained from a culture of Aureobasidium pullulans B-1 mutant. Both non-inoculated peppers and those artificially inoculated with Rhizopus arrhizus were coated and incubated at 24 °C for 5 days. The intensity of the decay caused by Rhizopus arrhizus in the peppers with P and P + eEMF coatings was nearly 3-fold lower, and in the case of P + weEMF 5-fold lower, than that observed in the control peppers. Additionally, the P + weEMF coating decreased, almost two-fold the severity of pepper decay compared to other samples. The influence of coating of pepper postharvest quality was examined after 30 days of storage at 6 °C and 70%-75% RH. All coatings formed a thin and well-attached additional layer of an intensified gloss. During storage, color, total soluble solid content and weight loss of coated peppers were subject to lower changes in comparison with uncoated ones. The results indicate the possibility of the application of pullulan coatings containing MFEs as an alternative to the chemical fungicides used to combat pepper postharvest diseases.

  1. Green synthesis of pullulan stabilized gold nanoparticles for cancer targeted drug delivery.

    PubMed

    Ganeshkumar, Moorthy; Ponrasu, Thangavel; Raja, Modhugoor Devendiran; Subamekala, Muthaiya Kannappan; Suguna, Lonchin

    2014-09-15

    The aim of this study was to synthesize green chemistry based gold nanoparticles using liver specific biopolymer and to develop a liver cancer targeted drug delivery system with enhanced efficacy and minimal side effects. Pullulan stabilized gold nanoparticles (PAuNPs) were coupled with 5-Fluorouracil (5-Fu) and folic acid (Fa) which could be used as a tool for targeted drug delivery and imaging of cancer. The toxicity of 5-Fu, 5-Fu adsorbed gold nanoparticles (5-Fu@AuNPs), Fa-coupled 5-Fu adsorbed gold nanoparticles (5-Fu@AuNPs-Fa), was studied using zebrafish embryo as an in vivo model. The in vitro cytotoxicity of free 5-Fu, 5-Fu@AuNPs, 5-Fu@AuNPs-Fa against HepG2 cells was studied and found that the amount of 5-Fu required to achieve 50% of growth of inhibition (Ic50) was much lower in 5-Fu@AuNP-Fa than in free 5-Fu, 5-Fu@AuNPs. The in vivo biodistribution of PAuNPs showed that higher amount of gold had been accumulated in liver (54.42±5.96 μg) than in other organs.

  2. Cholesteryl Pullulan Encapsulated TNF-α Nanoparticles Are an Effective Mucosal Vaccine Adjuvant against Influenza Virus

    PubMed Central

    Nagatomo, Daiki; Taniai, Madoka; Ariyasu, Harumi; Taniguchi, Mutsuko; Aga, Miho; Ariyasu, Toshio; Ohta, Tsunetaka; Fukuda, Shigeharu

    2015-01-01

    We encapsulated tumor necrosis factor-α (TNF-α), a major proinflammatory cytokine, into cholesteryl pullulan (CHP) to prepare TNF/CHP nanoparticles. In this report, we describe the immune-enhancing capability of the nanoparticles to act as a vaccine adjuvant. TNF/CHP nanoparticles showed excellent storage stability and enhanced host immune responses to external immunogens. The nanoparticles were effective via the nasal route of administration for inducing systemic IgG1 as well as mucosal IgA. We applied the nanoparticles in a model experimental influenza virus infection to investigate their adjuvant ability. TNF/CHP nanoparticles combined with a conventional split vaccine protected mice via nasal administration against a lethal challenge of A/PR/8/34 (H1N1) influenza virus. Mechanistic studies showed that the nanoparticles enhanced antigen uptake by dendritic cells (DCs) and moderately induced the expression of inflammation-related genes in nasopharynx lymphoid tissue (NALT), leading to the activation of both B and T cells. Preliminary safety study revealed no severe toxicity to TNF/CHP nanoparticles. Slight-to-moderate influences in nasal mucosa were observed only in the repeated administration and they seemed to be reversible. Our data show that TNF/CHP nanoparticles effectively enhance both humoral and cellular immunity and could be a potential adjuvant for vaccines against infectious diseases, especially in the mucosa. PMID:26421290

  3. Green synthesis of pullulan stabilized gold nanoparticles for cancer targeted drug delivery

    NASA Astrophysics Data System (ADS)

    Ganeshkumar, Moorthy; Ponrasu, Thangavel; Raja, Modhugoor Devendiran; Subamekala, Muthaiya Kannappan; Suguna, Lonchin

    2014-09-01

    The aim of this study was to synthesize green chemistry based gold nanoparticles using liver specific biopolymer and to develop a liver cancer targeted drug delivery system with enhanced efficacy and minimal side effects. Pullulan stabilized gold nanoparticles (PAuNPs) were coupled with 5-Fluorouracil (5-Fu) and folic acid (Fa) which could be used as a tool for targeted drug delivery and imaging of cancer. The toxicity of 5-Fu, 5-Fu adsorbed gold nanoparticles (5-Fu@AuNPs), Fa-coupled 5-Fu adsorbed gold nanoparticles (5-Fu@AuNPs-Fa), was studied using zebrafish embryo as an in vivo model. The in vitro cytotoxicity of free 5-Fu, 5-Fu@AuNPs, 5-Fu@AuNPs-Fa against HepG2 cells was studied and found that the amount of 5-Fu required to achieve 50% of growth of inhibition (Ic50) was much lower in 5-Fu@AuNP-Fa than in free 5-Fu, 5-Fu@AuNPs. The in vivo biodistribution of PAuNPs showed that higher amount of gold had been accumulated in liver (54.42 ± 5.96 μg) than in other organs.

  4. Production of polymalic acid and malic acid by Aureobasidium pullulans fermentation and acid hydrolysis.

    PubMed

    Zou, Xiang; Zhou, Yipin; Yang, Shang-Tian

    2013-08-01

    Malic acid is a dicarboxylic acid widely used in the food industry and also a potential C4 platform chemical that can be produced from biomass. However, microbial fermentation for direct malic acid production is limited by low product yield, titer, and productivity due to end-product inhibition. In this work, a novel process for malic acid production from polymalic acid (PMA) fermentation followed by acid hydrolysis was developed. First, a PMA-producing Aureobasidium pullulans strain ZX-10 was screened and isolated. This microbe produced PMA as the major fermentation product at a high-titer equivalent to 87.6 g/L of malic acid and high-productivity of 0.61 g/L h in free-cell fermentation in a stirred-tank bioreactor. Fed-batch fermentations with cells immobilized in a fibrous-bed bioreactor (FBB) achieved the highest product titer of 144.2 g/L and productivity of 0.74 g/L h. The fermentation produced PMA was purified by adsorption with IRA-900 anion-exchange resins, achieving a ∼100% purity and a high recovery rate of 84%. Pure malic acid was then produced from PMA by hydrolysis with 2 M sulfuric acid at 85°C, which followed the first-order reaction kinetics. This process provides an efficient and economical way for PMA and malic acid production, and is promising for industrial application.

  5. Development of ecofriendly bionanocomposite: Whey protein isolate/pullulan films with nano-SiO2.

    PubMed

    Hassannia-Kolaee, Mahbobeh; Khodaiyan, Faramarz; Pourahmad, Rezvan; Shahabi-Ghahfarrokhi, Iman

    2016-05-01

    During the past decade, the limitation of petroleum based polymers, the high price of oil, and the environmental concern were attracted the attention of researchers to develop biobased polymers. The composition of different biopolymers and the reinforcement with nano filler are common methods to improve the drawbacks of biopolymers. In this study whey protein isolate/pullulan (WPI/PUL) films contain 1%, 3%, and 5% (w/w) nano-SiO2 (NS) were prepared by a casting method. Tensile strength of nanocomposite films increased after increasing NS content, but elongation at break decreased, simultaneously. Water absorption, moisture content, solubility in water improved in the wake of increasing NS content because NS increase the cohesiveness of the polymer matrix and improved the barrier and water resistance properties of the films. water vapor permeability of film specimens decreased by increasing NS content. Uniform distribution of NS into polymer matrix was confirmed by scanning electron microscopy (SEM). XRD pattern and thermal analysis revealed increasing crystallinity and increasing Tg of film specimens with increasing NS content, respectively. According to our result WPI/PUL/NS films possess potential to be used as environment friendly packaging films to improve shelf life of food and can be used as promising alternative to petroleum based packaging films.

  6. Hydrophobicity, thermal and micro-structural properties of whey protein concentrate-pullulan-beeswax films.

    PubMed

    Jafari, Seid Mahdi; Khanzadi, Mehrdad; Mirzaei, Habibollah; Dehnad, Danial; Chegini, Faramarz Khodaian; Maghsoudlou, Yayha

    2015-09-01

    In this research, effects of beeswax (BW) on functional properties of whey protein concentrates (WPC):pullulan (PUL) films were investigated. For this purpose, 0, 10, 20 and 30w/w(glycerol)% BW rates and 30:70, 50:50 and 70:30w/w% WPC:PUL ratios were applied. Films containing 70% WPC:30% PUL (WPC70) and 30% BW (BW30) justified the highest contact angle (92.4°) among all films; SEM micrographs indicated that BW could come toward the surface of films during drying stage and resulted in a higher hydrophobic behavior of bilayer films compared with blend films. WPC70 supplied the lowest T(g) values (36-48 °C) among different proportions of WPC-PUL; the highest melting points were just assured in the absence of BW regardless of combination ratio for WPI:PUL. BW30 films deserved lower roughness rates than BW20 (and even BW10) films, indicating more advantageous microstructure and higher hydrogen connections in BW30 films and justifying similar melting points attained for BW30 films to BW20 or 10 ones. Overall, application of WPC70 and BW30 was recommended to obtain optimum combination of final properties for WPC-PUL-BW bilayer films as SEM exhibited flexible and elastic structures of such films.

  7. Activities of Aureobasidium pullulans cell filtrates against Monilinia laxa of peaches.

    PubMed

    Di Francesco, Alessandra; Roberti, Roberta; Martini, Camilla; Baraldi, Elena; Mari, Marta

    2015-12-01

    The Aureobasidium pullulans L1 and L8 strains are known as efficient biocontrol agents against several postharvest fungal pathogens. In order to better understand the mechanism of action underneath the antifungal activity of L1 and L8 strains, yeast cell filtrates grown at different times were evaluated in vivo against Monilinia laxa on peach. Lesion diameters on peach fruit were reduced by L1 and L8 culture filtrates of 42.5% and 67% respectively. The ability of these filtrates to inhibit M. laxa conidia germination and germ tube elongation was studied by in vitro assays. The results showed a 70% reduction of conidia germination for both strains while for germ tube elongation, it was 52% and 41% for L1 and L8 culture filtrates respectively. Finally, the activity of cell wall hydrolytic enzymes such as chitinase and glucanase in cell filtrates was analysed and the expression of genes encoding these activities was quantified during yeast growth. From 24h onward, both culture filtrates contained β,1-3,glucanase and. chitinase activities, the most pronounced of which was N-β-acetylglucosaminidase. Gene expression level encoding for these enzymes in L1 and L8 varied according to the strain. These results indicate that L1 and L8 strains culture filtrates retain the yeast antagonistic activity and suggest that the production of hydrolytic enzymes plays an important role in this activity.

  8. Antimicrobial activity of a pullulan-caraway essential oil coating on reduction of food microorganisms and quality in fresh baby carrot.

    PubMed

    Gniewosz, Małgorzata; Kraśniewska, Karolina; Woreta, Marcin; Kosakowska, Olga

    2013-08-01

    This research evaluated the antimicrobial efficacy of pullulan films containing caraway essential oil (CEO). The films were prepared from a 10% of pullulan, containing from 0.12% to 10.0% of CEO. The composition of the CEO was analyzed with the use of gas chromatography. The antimicrobial activity of the CEO was evaluated with the method of serial microdilutions, and the films containing CEO-with the agar diffusion method against selected Gram-negative, Gram-positive bacteria, and fungi. The structure of the film surface and its cross-section were analyzed using a scanning electron microscope (SEM). Analyses were also carried out to determine the efficacy of a pullulan coating with 10% CEO on baby carrots experimentally inoculated with Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae, or Aspergillus niger and stored at a room temperature for 7 d. At a concentration of 0.12%, CEO inhibited the growth of all the tested microorganisms. Pullulan films containing 8% to 10% of CEO were active against all tested microorganisms. Populations of S. aureus on carrot samples were reduced by approximately 3 log CFU/g, while those of A. niger and S. cerevisiae by, respectively, 5 and 4 log CFU/g, after 7 d of storage. S. enteritidis was the most resistant among the tested species, since it was not significantly reduced after 7 d of storage. At the end of storage, samples treated with pullulan-caraway oil coating maintained better visual acceptability than control samples. Results of this study suggest the feasibility of applying a pullulan film with incorporated CEO to extend the microbiological stability of minimally processed foods.

  9. 26 CFR 31.3121(b)(17)-1 - Services in employ of Communist organization.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Services in employ of Communist organization. 31.3121(b)(17)-1 Section 31.3121(b)(17)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... COLLECTION OF INCOME TAX AT SOURCE Federal Insurance Contributions Act (Chapter 21, Internal Revenue Code...

  10. Influence of starvation, surface attachment and biofilm growth on the biocide susceptibility of the biodeteriogenic yeast Aureobasidium pullulans.

    PubMed

    Sabev, H A; Robson, G D; Handley, P S

    2006-08-01

    To investigate the effect of starvation, surface attachment and growth in a biofilm on the susceptibility of Aureobasidium pullulans to the biocides 2-n-octyl-4-isothiazolin-3-one (OIT) and sodium hypochlorite (NaOCl). Fluorescence loss from a green fluorescent protein (GFP)-transformed strain was used to monitor real-time loss in viability as previously described in situ in 96-well plates. Exponential phase, yeast-like (YL) cells were settled in the bottom of the wells as a low-density monolayer (LDM) and were susceptible to all biocide concentrations (25-100 mug ml(-1)). The exponential phase YL cells were either starved for 48 h in suspension or starved for 48 h as LDMs in the wells. Starvation in both cases led to a small reduction in susceptibility to the biocides. In contrast, 48-h biofilms grown in malt extract broth showed an apparent lack of susceptibility to 25 and 50 mug ml(-1) OIT and to 25-100 mug ml(-1) NaOCl. However, when the OIT concentration was increased to compensate for the higher cell density in the biofilm, the biofilms were found to be equally susceptible to the LDM. Starvation of A. pullulans YL cells either in suspension or as attached LDM resulted in a decrease in susceptibility to low concentrations of both OIT and NaOCl while the apparent reduced susceptibility of mature biofilms was due to the increase in biofilm cell density rather than true biofilm resistance per se. Monitoring fluorescence loss from the GFP-transformed strain of A. pullulans can be used as a fast and reliable method for monitoring cell death in real time as a response to biocide and antimicrobial challenge.

  11. Effect of Pullulan Nanoparticle Surface Charges on HSA Complexation and Drug Release Behavior of HSA-Bound Nanoparticles

    PubMed Central

    Tao, Xiaojun; Zhang, Qiufang; Ling, Kai; Chen, Yicun; Yang, Wenzhi; Gao, Fenfei; Shi, Ganggang

    2012-01-01

    Nanoparticle (NP) compositions such as hydrophobicity and surface charge are vital to determine the presence and amount of human serum albumin (HSA) binding. The HSA binding influences drug release, biocompatibility, biodistribution, and intercellular trafficking of nanoparticles (NPs). Here, we prepared 2 kinds of nanomaterials to investigate HSA binding and evaluated drug release of HSA-bound NPs. Polysaccharides (pullulan) carboxyethylated to provide ionic derivatives were then conjugated to cholesterol groups to obtain cholesterol-modified carboxyethyl pullulan (CHCP). Cholesterol-modified pullulan (CHP) conjugate was synthesized with a similar degree of substitution of cholesterol moiety to CHCP. CHCP formed self-aggregated NPs in aqueous solution with a spherical structure and zeta potential of −19.9±0.23 mV, in contrast to −1.21±0.12 mV of CHP NPs. NPs could quench albumin fluorescence intensity with maximum emission intensity gradually decreasing up to a plateau at 9 to 12 h. Binding constants were 1.12×105 M−1 and 0.70×105 M−1 to CHP and CHCP, respectively, as determined by Stern-Volmer analysis. The complexation between HSA and NPs was a gradual process driven by hydrophobic force and inhibited by NP surface charge and shell-core structure. HSA conformation was altered by NPs with reduction of α-helical content, depending on interaction time and particle surface charges. These NPs could represent a sustained release carrier for mitoxantrone in vitro, and the bound HSA assisted in enhancing sustained drug release. PMID:23166632

  12. A carboxymethyl cellulase from a marine yeast ( Aureobasidium pullulans 98): Its purification, characterization, gene cloning and carboxymethyl cellulose digestion

    NASA Astrophysics Data System (ADS)

    Rong, Yanjun; Zhang, Liang; Chi, Zhenming; Wang, Xianghong

    2015-10-01

    We have reported that A. pullulans 98 produces a high yield of cellulase. In this study, a carboxymethyl cellulase (CMCase) in the supernatant of the culture of A. pullulans 98 was purified to homogeneity, and the maximum production of CMCase was 4.51 U (mg protein)-1. The SDS-PAGE analysis showed that the molecular mass of the purified CMCase was 67.0 kDa. The optimal temperature of the purified enzyme with considerable thermosensitivity was 40°C, much lower than that of the CMCases from other fungi. The optimal pH of the enzyme was 5.6, and the activity profile was stable in a range of acidity (pH 5.0-6.0). The enzyme was activated by Na+, Mg2+, Ca2+, K+, Fe2+ and Cu2+, however, it was inhibited by Fe3+, Ba2+, Zn2+, Mn2+ and Ag+. K m and V max values of the purified enzyme were 4.7 mg mL-1 and 0.57 µmol L-1 min-1 (mg protein)-1, respectively. Only oligosaccharides with different sizes were released from carboxymethylcellulose (CMC) after hydrolysis with the purified CMCase. The putative gene encoding CMCase was cloned from A. pullulans 98, which contained an open reading frame of 954 bp (EU978473). The protein deduced contained the conserved domain of cellulase superfamily (glucosyl hydrolase family 5). The N-terminal amino acid sequence of the purified CMCase was M-A-P-H-A-E-P-Q-S-Q-T-T-E-Q-T-S-S-G-Q-F, which was consistent with that deduced from the cloned gene. This suggested that the purified CMCase was indeed encoded by the cloned CMCase gene in this yeast.

  13. Bone engineering by phosphorylated-pullulan and β-TCP composite.

    PubMed

    Takahata, Tomohiro; Okihara, Takumi; Yoshida, Yasuhiro; Yoshihara, Kumiko; Shiozaki, Yasuyuki; Yoshida, Aki; Yamane, Kentaro; Watanabe, Noriyuki; Yoshimura, Masahide; Nakamura, Mariko; Irie, Masao; Van Meerbeek, Bart; Tanaka, Masato; Ozaki, Toshifumi; Matsukawa, Akihiro

    2015-11-20

    A multifunctional biomaterial with the capacity bond to hard tissues, such as bones and teeth, is a real need for medical and dental applications in tissue engineering and regenerative medicine. Recently, we created phosphorylated-pullulan (PPL), capable of binding to hydroxyapatite in bones and teeth. In the present study, we employed PPL as a novel biocompatible material for bone engineering. First, an in vitro evaluation of the mechanical properties of PPL demonstrated both PPL and PPL/β-TCP composites have higher shear bond strength than materials in current clinical use, including polymethylmethacrylate (PMMA) cement and α-tricalcium phosphate (TCP) cement, Biopex-R. Further, the compressive strength of PPL/β-TCP composite was significantly higher than Biopex-R. Next, in vivo osteoconductivity of PPL/β-TCP composite was investigated in a murine intramedular injection model. Bone formation was observed 5 weeks after injection of PPL/β-TCP composite, which was even more evident at 8 weeks; whereas, no bone formation was detected after injection of PPL alone. We then applied PPL/β-TCP composite to a rabbit ulnar bone defect model and observed bone formation comparable to that induced by Biopex-R. Implantation of PPL/β-TCP composite induced new bone formation at 4 weeks, which was remarkably evident at 8 weeks. In contrast, Biopex-R remained isolated from the surrounding bone at 8 weeks. In a pig vertebral bone defect model, defects treated with PPL/β-TCP composite were almost completely replaced by new bone; whereas, PPL alone failed to induce bone formation. Collectively, our results suggest PPL/β-TCP composite may be useful for bone engineering.

  14. Ultrasound-assisted pullulan/montmorillonite bionanocomposite coating with high oxygen barrier properties.

    PubMed

    Introzzi, Laura; Blomfeldt, Thomas O J; Trabattoni, Silvia; Tavazzi, Silvia; Santo, Nadia; Schiraldi, Alberto; Piergiovanni, Luciano; Farris, Stefano

    2012-07-31

    In this paper, the preparation and characterization of oxygen barrier pullulan sodium montmorillonite (Na(+)-MMT) nanocomposite coatings are presented for the first time. Full exfoliation of platelets during preparation of the coating water dispersions was mediated by ultrasonic treatment, which turned out to be a pivotal factor in the oxygen barrier performance of the final material even at high relative humidity (RH) conditions [oxygen permeability coefficients ~1.43 ± 0.39 and 258.05 ± 13.78 mL·μm·m(-2)·(24 h)(-1)·atm(-1) at 23 °C and 0% RH and 70% RH, respectively]. At the micro- and nanoscale, the reasons are discussed. The final morphology of the coatings revealed that clay lamellae were stacked on top of one another, probably due to the forced confinement of the platelets within the coating thickness after solvent evaporation. This was also confirmed by modeling the experimental oxygen permeability data with the well-known Nielsen and Cussler permeation theoretical models, which suggested a reasonable aspect ratio (α) of ~100. Electron microscopic analyses also disclosed a peculiar cell-like arrangement of the platelets. The stacking of the clay lamellae and the cell-like arrangement create the excellent oxygen barrier properties. Finally, we demonstrated that the slight haze increase in the bionanocomposite coating materials arising from the addition of the clays depends on the clay concentration but not so much on the sonication time, due to the balance of opposite effects after sonication (an increase in the number of scattering centers but a reduction in their size).

  15. Crosslinked pullulan/cellulose acetate fibrous scaffolds for bone tissue engineering.

    PubMed

    Atila, Deniz; Keskin, Dilek; Tezcaner, Ayşen

    2016-12-01

    Natural polymer based fibrous scaffolds have been explored for bone tissue engineering applications; however, their inadequate 3-dimensionality and poor mechanical properties are among the concerns for their use as bone substitutes. In this study, pullulan (P) and cellulose acetate (CA), two polysaccharides, were electrospun at various P/CA ratios (P80/CA20, P50/CA50, and P20/CA80%) to develop 3D fibrous network. The scaffolds were then crosslinked with trisodium trimetaphosphate (STMP) to improve the mechanical properties and to delay fast weight loss. The lowest weight loss was observed for the groups that were crosslinked with P/STMP 2/1 for 10min. Fiber morphologies of P50/CA50 were more uniform without phase separation and this group was crosslinked most efficiently among groups. It was found that mechanical properties of P20/CA80 and P50/CA50 were higher than that of P80/CA20. After crosslinking strain values of P50/CA50 scaffolds were improved and these scaffolds became more stable. Unlike P80/CA20, uncrosslinked P50/CA50 and P20/CA80 were not lost in PBS. Among all groups, crosslinked P50/CA50 scaffolds had more uniform pores; therefore this group was used for bioactivity and cell culture studies. Apatite-like structures were observed on fibers after SBF incubation. Human Osteogenic Sarcoma Cell Line (Saos-2) seeded onto crosslinked P50/CA50 scaffolds adhered and proliferated. The functionality of cells was tested by measuring ALP activity of the cells and the results indicated their osteoblastic differentiation. In vitro tests showed that scaffolds were cytocompatible. To sum up, crosslinked P50/CA50 scaffolds were proposed as candidate cell carriers for bone tissue engineering applications.

  16. CAM 17.1--a new diagnostic marker in pancreatic cancer.

    PubMed Central

    Gansauge, F.; Gansauge, S.; Parker, N.; Beger, M. I.; Poch, B.; Link, K. H.; Safi, F.; Beger, H. G.

    1996-01-01

    CAM 17.1-Ab is a recently described monoclonal antibody that detects a mucus glycoprotein with high specificity for intestinal mucus, particularly in the colon, small intestine, biliary tract and pancreas. We investigated the expression and release of CAM 17.1 in pancreatic carcinoma cell lines and tissue specimens of normal pancreas, chronic pancreatitis and pancreatic cancer. CAM 17.1 was weakly expressed on normal ductal cells and chronic pancreatitis, whereas it was overexpressed in pancreatic cancer. Serum analysis using a new enzyme-linked antibody sandwich assay (CAM 17.1/WGA) of patients with chronic pancreatitis, pancreatic cancer or other gastrointestinal cancer and of healthy blood donors revealed a high sensitivity (67%) and excellent specificity (90%) of CAM 17.1/WGA assay in pancreatic cancer. In comparison with the tumour marker CA19-9, the sensitivity of the CAM 17.1/WGA assay was similar to the sensitivity of CA 19-9 (67% and 76%, P = 0.22), whereas the specificity of CAM 17.1/WGA assay was higher than in CA 19-9 (90% compared with 78% in chronic pancreatitis, P > 0.05). Images Figure 2 PMID:8980403

  17. Synthesis, physicochemical and spectroscopic characterization of copper(II)-polysaccharide pullulan complexes by UV-vis, ATR-FTIR, and EPR.

    PubMed

    Mitić, Zarko; Cakić, Milorad; Nikolić, Goran M; Nikolić, Ružica; Nikolić, Goran S; Pavlović, Radmila; Santaniello, Enzo

    2011-02-15

    Bioactive copper(II) complexes with polysaccharides, like pullulan and dextran, are important in both veterinary and human medicine for the treatment of hypochromic microcitary anemia and hypocupremia. In aqueous alkaline solutions, Cu(II) ion forms complexes with the exopolysaccharide pullulan and its reduced low-molecular derivative. The metal content and the solution composition depend on pH, temperature, and time of the reaction. The complexing process begins in a weak alkali solution (pH >7) and involves OH groups of pullulan monomer (glucopyranose) units. Complexes of Cu(II) ion with reduced low-molecular pullulan (RLMP, M(w) 6000 g mol(-1)) were synthesized in water solutions, at the boiling temperature and at different pH values ranging from 7.5 to 12. The Cu(II) complex formation with RLMP was analyzed by UV-vis spectrophotometry and other physicochemical methods. Spectroscopic characterizations (ATR-FTIR, FT-IRIS, and EPR) and spectra-structure correlation of Cu(II)-RLMP complexes were also carried out.

  18. Edible Active Coatings Based on Pectin, Pullulan, and Chitosan Increase Quality and Shelf Life of Strawberries (Fragaria ananassa).

    PubMed

    Treviño-Garza, Mayra Z; García, Santos; del Socorro Flores-González, Ma; Arévalo-Niño, Katiushka

    2015-08-01

    Edible active coatings (EACs) based on pectin, pullulan, and chitosan incorporated with sodium benzoate and potassium sorbate were employed to improve the quality and shelf life of strawberries. Fruits were washed, disinfected, coated by dipping, packed, and stored at 4 °C for 15 d. Application of EACs reduced (P < 0.05) weight loss and fruit softening and delayed alteration of color (redness) and total soluble solids content. In contrast, pH and titratable acidity were not affected (P > 0.05) throughout storage, and ascorbic acid content was maintained in pectin-EAC coated strawberries. Microbiological analyses showed that application of EACs reduced (P < 0.05) microbial growth (total aerobic counts, molds, and yeasts) on strawberries. Chitosan-EAC coated strawberries presented the best results in microbial growth assays. Sensory quality (color, flavor, texture, and acceptance) improved and decay rate decreased (P < 0.05) in pectin-EAC, pullulan-EAC, and chitosan-EAC coated strawberries. In conclusion, EACs based on polysaccharides improved the physicochemical, microbiological, and sensory characteristics, increasing the shelf life of strawberries from 6 (control) to 15 d (coated fruits). © 2015 Institute of Food Technologists®

  19. Microfibrillated cellulose and borax as mechanical, O₂-barrier, and surface-modulating agents of pullulan biocomposite coatings on BOPP.

    PubMed

    Cozzolino, Carlo A; Campanella, Gaetano; Türe, Hasan; Olsson, Richard T; Farris, Stefano

    2016-06-05

    Multifunctional composite coatings on bi-oriented polypropylene (BOPP) films were obtained using borax and microfibrillated cellulose (MFC) added to the main pullulan coating polymer. Spectroscopy analyses suggested that a first type of interaction occurred via hydrogen bonding between the C6OH group of pullulan and the hydroxyl groups of boric acid, while monodiol and didiol complexation represented a second mechanism. The deposition of the coatings yielded an increase in the elastic modulus of the entire plastic substrate (from ∼2GPa of the neat BOPP to ∼3.1GPa of the P/B+/MFC-coated BOPP). The addition of MFC yielded a decrease of both static and kinetic coefficients of friction of approximately 22% and 25%, respectively, as compared to the neat BOPP. All composite coatings dramatically increased the oxygen barrier performance of BOPP, especially under dry conditions. The deposition of the high hydrophilic coatings allowed to obtain highly wettable surfaces (water contact angle of ∼18°). Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Polyols, not sugars, determine the structural diversity of anti-streptococcal liamocins produced by Aureobasidium pullulans strain NRRL 50380.

    PubMed

    Price, Neil Pj; Bischoff, Kenneth M; Leathers, Timothy D; Cossé, Allard A; Manitchotpisit, Pennapa

    2017-02-01

    Liamocins are polyol lipids produced by the fungus Aureobasidium pullulans, and have selective antibacterial activity against Streptococcus species. Liamocins produced by A. pullulans strain NRRL 50380 on sucrose medium have a d-mannitol head group ester-linked to 3,5-dihydroxydecanoate acyl chains, three or four of which are joined together by 1,5-polyester bonds (liamocins Man-A1 and Man-B1), and similar 3'-O-acetylated analogs (Man-A2 and Man-B2). However, other types of liamocins are produced depending on the choice of strain and growth conditions. In the current study, growth on different polyols, but not sugars, resulted in considerable structural variation, including liamocins with d-galactitol (dulcitol), d-sorbitol (glucitol), d- and l-arabitol, d-xylitol, l-threitol and glycerol head groups. The head groups of liamocins produced on arabitol were shown to be entirely composed of d-arabitol. These liamocin variants were structurally characterized by NMR and MS, and tested for antibacterial activity. The new liamocin variants also had selective activity against Streptococcus. Liamocin structural variants are novel antibacterials against Streptococcus sp. that merit further investigation.

  1. Blending of soluble corn fiber with pullulan, sorbitol, or fructose attenuates glycemic and insulinemic responses in the dog and affects hydrolytic digestion in vitro.

    PubMed

    de Godoy, M R C; Knapp, B K; Bauer, L L; Swanson, K S; Fahey, G C

    2013-08-01

    The objective of these experiments was to measure in vitro hydrolytic digestion and glycemic and insulinemic responses of select carbohydrate blends, all containing the novel carbohydrate soluble corn fiber (SCF). Two SCF that varied in their method of production were used to formulate the carbohydrate blends. One set of blends contained a SCF that was spray dried (SCFsd) and then blended with different amounts of either pullulan, sorbitol, or fructose. The other set of blends contained a SCF produced using longer evaporation time (SCF) and then blended with different ratios of pullulan, sorbitol, and fructose. Free sugar concentrations found in the individual SCFsd and SCF substrates were low but varied. Spray-dried soluble corn fiber had a reduced free sugar concentration compared with SCF (2.8 vs. 14.2%). Glucose was the main free sugar found in both SCFsd and SCF but at different concentrations (2.7 vs. 12.7%, respectively). The majority of the SCFsd blends were completely hydrolyzed to their monosaccharide components. Glucose accounted for most of the hydrolyzed monosaccharides for SCFsd and all the SCFsd blends. Hydrolyzed monosaccharide concentrations for the SCF:pullulan:sorbitol:fructose blends followed similar trends to the SCFsd blends where greater percentages of fructose and sorbitol resulted in decreased (P < 0.05) hydrolyzed monosaccharide concentrations. The SCFsd blends had intermediate to high amounts of monosaccharides released as a result of in vitro hydrolytic digestion. The SCFsd:pullulan blends were more digestible in vitro (approximately 91%; P < 0.05) than SCFsd:fructose or SCFsd:sorbitol. Total released monosaccharides were high in SCFsd blends containing either 50% fructose or sorbitol, but the combination resulted in reduced concentrations of glucose released (P < 0.05). The SCF:pullulan:sorbitol:fructose blends also had intermediate to high released monosaccharides as a result of in vitro hydrolytic digestion. All SCF blends resulted in

  2. Effects of Particle Hydrophobicity, Surface Charge, Media pH Value and Complexation with Human Serum Albumin on Drug Release Behavior of Mitoxantrone-Loaded Pullulan Nanoparticles

    PubMed Central

    Tao, Xiaojun; Jin, Shu; Wu, Dehong; Ling, Kai; Yuan, Liming; Lin, Pingfa; Xie, Yongchao; Yang, Xiaoping

    2015-01-01

    We prepared two types of cholesterol hydrophobically modified pullulan nanoparticles (CHP) and carboxyethyl hydrophobically modified pullulan nanoparticles (CHCP) substituted with various degrees of cholesterol, including 3.11, 6.03, 6.91 and 3.46 per polymer, and named CHP−3.11, CHP−6.03, CHP−6.91 and CHCP−3.46. Dynamic laser light scattering (DLS) showed that the pullulan nanoparticles were 80–120 nm depending on the degree of cholesterol substitution. The mean size of CHCP nanoparticles was about 160 nm, with zeta potential −19.9 mV, larger than CHP because of the carboxyethyl group. A greater degree of cholesterol substitution conferred greater nanoparticle hydrophobicity. Drug-loading efficiency depended on nanoparticle hydrophobicity, that is, nanoparticles with the greatest degree of cholesterol substitution (6.91) showed the most drug encapsulation efficiency (90.2%). The amount of drug loading increased and that of drug release decreased with enhanced nanoparticle hydrophobicity. Nanoparticle surface-negative charge disturbed the amount of drug loading and drug release, for an opposite effect relative to nanoparticle hydrophobicity. The drug release in pullulan nanoparticles was higher pH 4.0 than pH 6.8 media. However, the changed drug release amount was not larger for negative-surface nanoparticles than CHP nanoparticles in the acid release media. Drug release of pullulan nanoparticles was further slowed with human serum albumin complexation and was little affected by nanoparticle hydrophobicity and surface negative charge. PMID:28344259

  3. Effects of nitrogen availability on polymalic acid biosynthesis in the yeast-like fungus Aureobasidium pullulans.

    PubMed

    Wang, Yongkang; Song, Xiaodan; Zhang, Yongjun; Wang, Bochu; Zou, Xiang

    2016-08-22

    Polymalic acid (PMA) is a novel polyester polymer that has been broadly used in the medical and food industries. Its monomer, L-malic acid, is also a potential C4 platform chemical. However, little is known about the mechanism of PMA biosynthesis in the yeast-like fungus, Aureobasidium pullulans. In this study, the effects of different nitrogen concentration on cell growth and PMA biosynthesis were investigated via comparative transcriptomics and proteomics analyses, and a related signaling pathway was also evaluated. A high final PMA titer of 44.00 ± 3.65 g/L (49.9 ± 4.14 g/L of malic acid after hydrolysis) was achieved in a 5-L fermentor under low nitrogen concentration (2 g/L of NH4NO3), which was 18.3 % higher yield than that obtained under high nitrogen concentration (10 g/L of NH4NO3). Comparative transcriptomics profiling revealed that a set of genes, related to the ribosome, ribosome biogenesis, proteasome, and nitrogen metabolism, were significantly up- or down-regulated under nitrogen sufficient conditions, which could be regulated by the TOR signaling pathway. Fourteen protein spots were identified via proteomics analysis, and were found to be associated with cell division and growth, energy metabolism, and the glycolytic pathway. qRT-PCR further confirmed that the expression levels of key genes involved in the PMA biosynthetic pathway (GLK, CS, FUM, DAT, and MCL) and the TOR signaling pathway (GS, TOR1, Tap42, and Gat1) were upregulated due to nitrogen limitation. Under rapamycin stress, PMA biosynthesis was obviously inhibited in a dose-dependent manner, and the transcription levels of TOR1, MCL, and DAT were also downregulated. The level of nitrogen could regulate cell growth and PMA biosynthesis. Low concentration of nitrogen was beneficial for PMA biosynthesis, which could upregulate the expression of key genes involved in the PMA biosynthesis pathway. Cell growth and PMA biosynthesis might be mediated by the TOR signaling pathway in

  4. Short communication: Electrospinning of casein/pullulan blends for food-grade applications.

    PubMed

    Tomasula, P M; Sousa, A M M; Liou, S-C; Li, R; Bonnaillie, L M; Liu, L S

    2016-03-01

    Electrospinning is a complex process that produces fibers with diameters on the micrometer or nano-scale from an electrified jet of a polymer solution. The objective of this study was to create electrospun fibers for food use from aqueous solutions of calcium (CaCAS) or sodium caseinate (NaCAS). Fibers were not formed from electrospinning of solutions of either caseinate (CAS) at 50 °C, but were formed from blends of either CAS solution with aqueous solutions of the food-grade polysaccharide, pullulan (PUL), when using mass ratios from 2:1 to 1:4 of PUL/CAS. The CAS in the spinning solutions ranged from 3 to 15% (wt/wt) and the PUL ranged from 5 to 15% (wt/wt). The PUL/CaCAS 1:2 fibers showed the lowest fiber diameter sizes (FDS) of 172 ± 43 nm, as determined by scanning electron microscopy, and were smaller in size than fibers electrospun from 15% (wt/wt) PUL solution. The PUL/NaCAS solutions were more viscous and formed fibers with occasional branching and less uniform FDS at higher NaCAS contents. Reductions in NaCAS in these solutions reduced viscosity and improved jet stabilities with consequent improvement in fiber morphology leading to more uniform FDS. Fibers with less defects and more homogeneous FDS were formed from PUL/CaCAS blends with more CaCAS, showing that each CAS interacted differently with PUL and formed the best fibers at different solution conditions. Calcium bridging may also underlie the anomalous behavior of the PUL/CaCAS blends by forming crosslinks with the phosphoserine residues, further enabling chain entanglements for fiber formation. The PUL/NaCAS fibers tended to be larger than the PUL/CaCAS fibers, which may also be due to other factors such as solution surface tension and conductivity, which also affect fiber quality and size. The shear viscosities at 100 s(-1) of the solutions producing fibers were within the range of 0.07 to 0.16 Pa/s, with the smallest standard deviations in FDS noted for solutions with viscosities within about

  5. CTE and Ratchet Growth Measurements on LX17-1 and Constituents

    SciTech Connect

    Cunningham, B; Weese, R; Lewis, P; Harwood, P; Tran, T

    2002-02-22

    Dimensional changes in PBX materials resulting from temperature change are of interest to engineers, designers and modelers. In this paper we present data from recent measurements made on LX17-1, as well as on the material's binder and its energetic constituent. LX17-1 is made from 7.5% KEL-F 800 binder combined with 92.5% wet aminated TATB energetic crystals. Due apparently to the anisotropic expansionary behavior of the TATB, the material exhibits irreversible growth, in addition to the usual reversible expansions and contractions associated with temperature change. In an effort understand reversible and irreversible growth behavior and to verify consistency between our measurements and those made historically, measurements were performed on billet pressed LX17-1, on die pressed TATB, and on KEL-F alone. It is important to realize that, for materials involving TATB, expansionary behavior results from the combined effects of reversible and irreversible (ratchet growth) phenomena.

  6. Development of tannic acid/chitosan/pullulan composite nanofibers from aqueous solution for potential applications as wound dressing.

    PubMed

    Xu, Fenghua; Weng, Baicheng; Gilkerson, Robert; Materon, Luis Alberto; Lozano, Karen

    2015-01-22

    This study presents the successful development of biocompatible tannic acid (TA)/chitosan (CS)/pullulan (PL) composite nanofibers (NFs) with synergistic antibacterial activity against the Gram-negative bacteria Escherichia coli. The NFs were developed utilizing the forcespinning(®) (FS) technique from CS-CA aqueous solutions to avoid the usage of toxic organic solvents. The ternary nanofibrous membranes were crosslinked to become water stable for potential applications as wound dressing. The morphology, structure, water solubility, water absorption capability and thermal properties of the NFs were characterized. The ternary composite membrane exhibits good water absorption ability with rapid uptake rate. This novel membrane favors fibroblast cell attachment and growth by providing a 3D environment which mimics the extracellular matrix (ECM) in skin and allows cells to move through the fibrous structure resulting in interlayer growth throughout the membrane, thus favoring potential for deep and intricate wound healing.

  7. Assessment of the physical, mechanical, and moisture-retention properties of pullulan-based ternary co-blended films.

    PubMed

    Pan, Hongyang; Jiang, Bo; Chen, Jie; Jin, Zhengyu

    2014-11-04

    Multi-component substances made through direct blending or blending with co-drying can form films on the surfaces of intermediate moisture foods (IMFs), which help retain moisture and protect food texture and flavor. An IMF film system based on pullulan, with glycerol serving as the plasticizer, was studied using alginate and four different types of polysaccharides (propyleneglycol alginate, pectin, carrageenan, and aloe polysaccharide) as the blend-modified substances. The physical, mechanical, color, transparency, and moisture-retention properties of the co-blended films with the polysaccharides were assessed. A new formula was established for the average moisture retention property, water barrier, tensile strength, elongation at break, and oxygen barrier property of the ternary co-blended films using the Design Expert software. The new model established for moisture content measurement used an indirect method of film formation on food surfaces by humectants, which should expedite model validation and allow a better comprehension of moisture transfer through edible films.

  8. Cyanoethylation of the glucans dextran and pullulan: Substitution pattern and formation of nanostructures and entrapment of magnetic nanoparticles

    PubMed Central

    Fiege, Kathrin; Lünsdorf, Heinrich; Atarijabarzadeh, Sevil

    2012-01-01

    Summary Cyanoethylglucans with a degree of substitution in the range of 0.74 to 2.40 for dextran and 0.84 to 2.42 for pullulan were obtained by Michael addition of acrylonitrile to the glucans under various conditions. Products were thoroughly characterized, comprising elementary analysis, NMR and ATR–IR spectroscopy, and analysis of the substituent distribution in the glucosyl units by GC–FID and GC–MS of the constituting monosaccharide derivatives. Nanostructuring of the highly substituted cyanoethylpolysaccharides was performed by dialysis against a non-solvent. In the presence of ferromagnetic iron-oxide nanoparticles, multicore cyanoethylglucan-coated ferromagnetic nanoparticles were formed by selective entrapment. The specific interaction between cyano groups and iron could be proven. The size distribution and morphology of the nanoparticles were analyzed by dynamic light scattering (DLS), scanning electron microscopy (SEM) and energy-filtered transmission electron microscopy (EF–TEM) with parallel electron energy loss spectroscopy (PEELS). PMID:22563354

  9. Adaptation and transcriptome analysis of Aureobasidium pullulans in corncob hydrolysate for increased inhibitor tolerance to malic acid production.

    PubMed

    Zou, Xiang; Wang, Yongkang; Tu, Guangwei; Zan, Zhanquan; Wu, Xiaoyan

    2015-01-01

    Malic acid is a dicarboxylic acid widely used in the food industry, and is also a potential C4 platform chemical. Corncob is a low-cost renewable feedstock from agricultural industry. However, side-reaction products (furfural, 5-hydroxymethylfurfural (HMF), formic acid, and acetic acid) that severely hinder fermentation are formed during corncob pretreatment. The process for producing malic acid from a hydrolysate of corncob was investigated with a polymalic acid (PMA)-producing Aureobasidium pullulans strain. Under the optimal hydrolysate sugar concentration 110 g/L, A. pullulans was further adapted in an aerobic fibrous bed bioreactor (AFBB) by gradually increasing the sugar concentration of hydrolysate. After nine batches of fermentation, the production and productivity of malic acid reached 38.6 g/L and 0.4 g/L h, respectively, which was higher than that in the first batch (27.6 g/L and 0.29 g/L h, respectively). The adapted strain could grow under the stress of 0.5 g/L furfural, 3 g/L HMF, 2g/L acetic acid, and 0.5 g/L formic acid, whereas the wild type did not. Transcriptome analysis revealed that the differentially expressed genes were related to carbohydrate transport and metabolism, lipid transport and metabolism, signal transduction mechanism, redox metabolism, and energy production and conversion under 0.5 g/L furfural and 3 g/L HMF stress conditions. In total, 42 genes in the adapted strain were upregulated by 15-fold or more, and qRT-PCR also confirmed that the expression levels of key genes (i.e. SIR, GSS, CYS, and GSR) involved in sulfur assimilation pathway were upregulated by over 10-fold in adapted strain for cellular protection against oxidative stress.

  10. Adaptation and Transcriptome Analysis of Aureobasidium pullulans in Corncob Hydrolysate for Increased Inhibitor Tolerance to Malic Acid Production

    PubMed Central

    Zou, Xiang; Wang, Yongkang; Tu, Guangwei; Zan, Zhanquan; Wu, Xiaoyan

    2015-01-01

    Malic acid is a dicarboxylic acid widely used in the food industry, and is also a potential C4 platform chemical. Corncob is a low-cost renewable feedstock from agricultural industry. However, side-reaction products (furfural, 5-hydroxymethylfurfural (HMF), formic acid, and acetic acid) that severely hinder fermentation are formed during corncob pretreatment. The process for producing malic acid from a hydrolysate of corncob was investigated with a polymalic acid (PMA)-producing Aureobasidium pullulans strain. Under the optimal hydrolysate sugar concentration 110 g/L, A. pullulans was further adapted in an aerobic fibrous bed bioreactor (AFBB) by gradually increasing the sugar concentration of hydrolysate. After nine batches of fermentation, the production and productivity of malic acid reached 38.6 g/L and 0.4 g/L h, respectively, which was higher than that in the first batch (27.6 g/L and 0.29 g/L h, respectively). The adapted strain could grow under the stress of 0.5 g/L furfural, 3 g/L HMF, 2g/L acetic acid, and 0.5 g/L formic acid, whereas the wild type did not. Transcriptome analysis revealed that the differentially expressed genes were related to carbohydrate transport and metabolism, lipid transport and metabolism, signal transduction mechanism, redox metabolism, and energy production and conversion under 0.5 g/L furfural and 3 g/L HMF stress conditions. In total, 42 genes in the adapted strain were upregulated by 15-fold or more, and qRT-PCR also confirmed that the expression levels of key genes (i.e. SIR, GSS, CYS, and GSR) involved in sulfur assimilation pathway were upregulated by over 10-fold in adapted strain for cellular protection against oxidative stress. PMID:25793624

  11. 26 CFR 17.1 - Industrial development bonds used to provide solid waste disposal facilities; temporary rules.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 14 2010-04-01 2010-04-01 false Industrial development bonds used to provide solid waste disposal facilities; temporary rules. 17.1 Section 17.1 Internal Revenue INTERNAL REVENUE... UNDER 26 U.S.C. 103(c) § 17.1 Industrial development bonds used to provide solid waste...

  12. 26 CFR 17.1 - Industrial development bonds used to provide solid waste disposal facilities; temporary rules.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 14 2011-04-01 2010-04-01 true Industrial development bonds used to provide solid waste disposal facilities; temporary rules. 17.1 Section 17.1 Internal Revenue INTERNAL REVENUE... UNDER 26 U.S.C. 103(c) § 17.1 Industrial development bonds used to provide solid waste...

  13. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    .... § 1.401(a)(17)-1 Limitation on annual compensation. (a) Compensation limit requirement—(1) In general... an annual compensation limit for each employee under a qualified plan. This limit applies to a... annual compensation limit. Second, the amount of an employee's annual compensation that may be taken into...

  14. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    .... § 1.401(a)(17)-1 Limitation on annual compensation. (a) Compensation limit requirement—(1) In general... an annual compensation limit for each employee under a qualified plan. This limit applies to a... annual compensation limit. Second, the amount of an employee's annual compensation that may be taken into...

  15. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...(a)(17)-1 Limitation on annual compensation. (a) Compensation limit requirement—(1) In general. In... annual compensation limit for each employee under a qualified plan. This limit applies to a qualified... compensation limit. Second, the amount of an employee's annual compensation that may be taken into account in...

  16. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 15 2012-04-01 2012-04-01 false Fishing services. 31.3306(c)(17)-1 Section 31... Fishing services. (a) In general. Subject to the limitations prescribed in paragraphs (b) and (c) of this...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or...

  17. Expression of fatty-acid-modifying enzymes in the halotolerant black yeast Aureobasidium pullulans (de Bary) G. Arnaud under salt stress.

    PubMed

    Gostincar, C; Turk, M; Trbuha, T; Vaupotic, T; Plemenitas, A; Gunde-Cimerman, N

    2008-01-01

    Multiple tolerance to stressful environmental conditions of the black, yeast-like fungus Aureobasidium pullulans is achieved through different adaptations, among which there is the restructuring of the lipid composition of their membranes. Here, we describe three novel genes encoding fatty-acid-modifying enzymes in A. pullulans, along with the levels of their mRNAs under different salinity conditions. High levels of Delta(9)-desaturase and Delta(12)-desaturase mRNAs were seen at high salinities, which were consistent with an increased desaturation of the fatty acids in the cell membranes. Elevated levels of elongase mRNA were also detected. Surprisingly, increases in the levels of these mRNAs were also seen following hypo-osmotic shock, while hyperosmotic shock had exactly the opposite effect, demonstrating that data that are obtained from up-shift and down-shift salinity studies should be interpreted with caution.

  18. Expression of fatty-acid-modifying enzymes in the halotolerant black yeast Aureobasidium pullulans (de Bary) G. Arnaud under salt stress

    PubMed Central

    Gostinčar, C.; Turk, M.; Trbuha, T.; Vaupotič, T.; Plemenitaš, A.; Gunde-Cimerman, N.

    2008-01-01

    Multiple tolerance to stressful environmental conditions of the black, yeast-like fungus Aureobasidium pullulans is achieved through different adaptations, among which there is the restructuring of the lipid composition of their membranes. Here, we describe three novel genes encoding fatty-acid-modifying enzymes in A. pullulans, along with the levels of their mRNAs under different salinity conditions. High levels of Δ9-desaturase and Δ12-desaturase mRNAs were seen at high salinities, which were consistent with an increased desaturation of the fatty acids in the cell membranes. Elevated levels of elongase mRNA were also detected. Surprisingly, increases in the levels of these mRNAs were also seen following hypo-osmotic shock, while hyperosmotic shock had exactly the opposite effect, demonstrating that data that are obtained from up-shift and down-shift salinity studies should be interpreted with caution. PMID:19287526

  19. A new pullulan and a branched (1-->3)-, (1-->6)-linked beta-glucan from the lichenised ascomycete Teloschistes flavicans.

    PubMed

    Reis, Rodrigo A; Tischer, Cesar A; Gorin, Philip A J; Iacomini, Marcello

    2002-04-23

    The polysaccharides formed on hot alkaline extraction of the ascomycetous lichen Teloschistes flavicans were fractionated to give two glucans, which were characterised by methylation analysis and 1D and 2D NMR spectroscopy. One was a branched beta-glucan containing (1-->3) and (1-->6) linkages, a structure which is more typical of basidiomycetes rather than ascomycetes, which have linear glucans. The other was an alpha-glucan with alternating (1-->4) and (1-->6) linkages, found for the first time in Nature. This structure can be classified as a pullulan, which has been isolated from the fungi Aureobasidium pullulans, Tremella mesenterica, and Cyttaria harioti, but has different ratios of the component glycosidic linkages. The significance of the presence of the isolated alpha- and beta-glucans is discussed.

  20. Variations in grain lipophilic phytochemicals, proteins and resistance to Fusarium spp. growth during grain storage as affected by biological plant protection with Aureobasidium pullulans (de Bary).

    PubMed

    Wachowska, Urszula; Tańska, Małgorzata; Konopka, Iwona

    2016-06-16

    Modern agriculture relies on an integrated approach, where chemical treatment is reduced to a minimum and replaced by biological control that involves the use of active microorganisms. The effect of the antagonistic yeast-like fungus Aureobasidium pullulans on proteins and bioactive compounds (alkylresorcinols, sterols, tocols and carotenoids) in winter wheat grain and on the colonization of wheat kernels by fungal microbiota, mainly Fusarium spp. pathogens, was investigated. Biological treatment contributed to a slight increase contents of tocols, alkylresorcinols and sterols in grain. At the same time, the variation of wheat grain proteins was low and not significant. Application of A. pullulans enhanced the natural yeast colonization after six months of grain storage and inhibited growth of F. culmorum pathogens penetrating wheat kernel. This study demonstrated that an integrated approach of wheat grain protection with the use of the yeast-like fungus A. pullulans reduced kernel colonization by Fusarium spp. pathogens and increased the content of nutritionally beneficial phytochemicals in wheat grain without a loss of gluten proteins responsible for baking value.

  1. Novel Pullulan-Eudragit® S100 blend microparticles for oral delivery of risedronate: formulation, in vitro evaluation and tableting of blend microparticles.

    PubMed

    de Arce Velasquez, Aline; Ferreira, Luana Mota; Stangarlin, Mônica Fabiele Lorensi; da Silva, Cristiane de Bona; Rolim, Clarice Madalena Bueno; Cruz, Letícia

    2014-05-01

    Polymer blends have been considered a promising strategy to tailor drug release. In order to achieve gastroresistance and controlled release, Pullulan, a polysaccharide, and Eudragit® S100, an enteric polymer were selected to prepare microparticles for oral delivery of risedronate, an antiresorptive drug associated with GI tract injuries. Blend microparticles were prepared by spray-drying technique at 3 Pullulan and Eudragit® S100 ratios (MP2:1, MP1:1 and MP1:2) and were characterized in terms of yield, particle size, encapsulation efficiency, morphology, moisture content, flowability and in vitro drug release profiles. Microparticles presented yields between 31 and 42%, encapsulation efficiencies close to 100%, moisture contents lower than 11%, particle size ranging from 2.9 to 4.8 μm and narrow distribution. In the gastric medium, MP1:2 showed the best gastroresistance profile. In the intestinal fluid, all samples were able to prolong drug release. MP1:2 was compressed into tablets with or without polyvinylpyrrolidone. Both tableted microparticles could be obtained with acceptable average weights, drug content close to 100%, sufficient hardness and low friability. In vitro studies showed that tablets maintained the gastroresistance observed for microparticles and were also able to prolong risedronate release. In conclusion, Pullulan/Eudragit® S100 microparticles are promising alternatives for the oral delivery of risedronate in the future. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Induction of stilbene phytoalexins in grapevine (Vitis vinifera) and transgenic stilbene synthase-apple plants (Malus domestica) by a culture filtrate of Aureobasidium pullulans.

    PubMed

    Rühmann, Susanne; Pfeiffer, Judith; Brunner, Philipp; Szankowski, Iris; Fischer, Thilo C; Forkmann, Gert; Treutter, Dieter

    2013-11-01

    Products containing the epiphytic yeast Aureobasidium pullulans are commercially available and applied by fruit growers to prevent several fungal and bacterial diseases of fruit trees. The proposed beneficial mechanisms relate to limitations of space and nutrients for the pathogens in presence of the rapidly proliferating yeast cells. These explanations ignore the potential of yeasts to elicit the plant's defense. Our experiments aim at clarifying if an autoclaved and centrifuged suspension of A. pullulans may induce defense mechanisms. As a model system, the biosynthesis and accumulation of stilbene phytoalexins in callus and shoots of grapevine Vitis vinifera grown in vitro was used. Yeast application to the plant tissue stimulated stilbene biosynthesis, sometimes at the cost of flavonoids. The expression of the gene encoding stilbene synthase was enhanced and the enzyme showed higher activity while chalcone synthase activity and expression was reduced in some cases. An accumulation of stilbenes was also found in transgenic apple trees (Malus domestica cv. Holsteiner Cox) harboring the stilbene synthase-gene under control of its own promoter. These results clearly show that the application of A. pullulans may induce defense mechanisms of the treated plants. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  3. Biocontrol activity of an alkaline serine protease from Aureobasidium pullulans expressed in Pichia pastoris against four postharvest pathogens on apple.

    PubMed

    Banani, Houda; Spadaro, Davide; Zhang, Dianpeng; Matic, Slavica; Garibaldi, Angelo; Gullino, Maria Lodovica

    2014-07-16

    The yeast-like fungus Aureobasidium pullulans PL5 is a microbial antagonist against postharvest pathogens of fruits. The strain is able to produce hydrolases, including glucanases, chitinases and proteases. The alkaline serine protease gene ALP5 from A. pullulans was cloned, inserted into the vector pPIC9 to construct pPIC9/ALP5, and then expressed in Pichia pastoris strain KM71. ALP5 had a molecular mass of 42.9kDa after 5days growth with 1% methanol induction at 28°C. The recombinant protease expressed in P. pastoris showed its highest activity under alkaline conditions (at pH10) and a temperature of 50°C. The antifungal activity of the recombinant protease was investigated against Penicillium expansum, Botrytis cinerea, Monilinia fructicola and Alternaria alternata in vitro and on apple. The recombinant protease reduced significantly the spore germination and the germ tube length of the tested pathogens in PDB medium. The highest level of protease efficacy was observed against M. fructicola and B. cinerea, whereas a lower efficacy was observed against P. expansum and A. alternata indicating a possible effect of the pathogen cell wall composition on the proteolytic activity of the recombinant protease. The presence of protease was able to cause the swelling of the hyphae of B. cinerea, under an optical microscope. The recombinant protease expressed in P. pastoris was more active against the pathogens in vitro than the same enzyme expressed in E. coli in previous studies. The efficacy of ALP5 was also evaluated against the pathogens in vivo on cv Golden Delicious apples. The protease was more efficient in controlling M. fructicola, B. cinerea and P. expansum than A. alternata. However, the extent of the activity was dependent on the enzyme concentration and the length of fruit storage. This study demonstrated the capacity of the alkaline serine protease to keep its enzymatic activity for some days in the unfavorable environment of the fruit wounds. The alkaline

  4. Self-assembled nanocomplexes of anionic pullulan and polyallylamine for DNA and pH-sensitive intracellular drug delivery

    NASA Astrophysics Data System (ADS)

    Vora, Lalit; Tyagi, Monica; Patel, Ketan; Gupta, Sanjay; Vavia, Pradeep

    2014-12-01

    The amalgamation of chemotherapy and gene therapy is promising treatment option for cancer. In this study, novel biocompatible self-assembled nanocomplexes (NCs) between carboxylmethylated pullulan t335 (CMP) with polyallylamine (CMP-PAA NCs) were developed for plasmid DNA (pDNA) and pH-sensitive doxorubicin (DOX) delivery. DOX was conjugated to CMP (DOX-CMP) via hydrazone and confirmed by FTIR and 1H-NMR. In vitro release studies of pH-sensitive DOX-CMP conjugate showed 23 and 85 % release after 48 h at pH 7.4 (physiological pH) and pH 5 (intracellular/tumoral pH), respectively. The CMP-PAA NCs or DOX-CMP-PAA NCs self-assembled into a nanosized (<250 nm) spherical shape as confirmed by DLS and TEM. The hemolysis and cytotoxicity study indicated that the CMP-PAA NCs did not show cytotoxicity in comparison with plain polyallylamine. Gel retardation assay showed complete binding of pDNA with CMP-PAA NCs at 1:2 weight ratio. CMP-PAA NCs/pDNA showed significantly higher transfection in HEK293 cells compared to PAA/pDNA complexes. Confocal imaging demonstrated successful cellular uptake of DOX-CMP-PAA NCs in HEK293 cells. Thus, NCs hold great potential for targeted pDNA and pH-sensitive intratumoral drug delivery.

  5. A Novel Biosurfactant Produced by Aureobasidium pullulans L3-GPY from a Tiger Lily Wild Flower, Lilium lancifolium Thunb.

    PubMed Central

    Kim, Jong Shik; Lee, In Kyoung; Yun, Bong Sik

    2015-01-01

    Yeast biosurfactants are important biotechnological products in the food industry, and they have medical and cosmeceutical applications owing to their specific modes of action, low toxicity, and applicability. Thus, we have isolated and examined biosurfactant-producing yeast for various industrial and medical applications. A rapid and simple method was developed to screen biosurfactant-producing yeasts for high production of eco-friendly biosurfactants. Using this method, several potential niches of biosurfactant-producing yeasts, such as wild flowers, were investigated. We successfully selected a yeast strain, L3-GPY, with potent surfactant activity from a tiger lily, Lilium lancifolium Thunb. Here, we report the first identification of strain L3-GPY as the black yeast Aureobasidium pullulans. In addition, we isolated a new low-surface-tension chemical, designated glycerol-liamocin, from the culture supernatant of strain L3-GPY through consecutive chromatography steps, involving an ODS column, solvent partition, silica gel, Sephadex LH-20, and an ODS Sep-Pak cartridge column. The chemical structure of glycerol-liamocin, determined by mass spectrometry and nuclear magnetic resonance spectroscopy, indicates that it is a novel compound with the molecular formula C33H62O12. Furthermore, glycerol-liamocin exhibited potent biosurfactant activity (31 mN/m). These results suggest that glycerol-liamocin is a potential novel biosurfactantfor use in various industrial applications. PMID:25849549

  6. Effects of cholesterol-bearing pullulan (CHP)-nanogels in combination with prostaglandin E1 on wound healing.

    PubMed

    Kobayashi, Hiroshi; Katakura, Osamu; Morimoto, Nobuyuki; Akiyoshi, Kazunari; Kasugai, Shohei

    2009-10-01

    The cholesterol-bearing pullulan (CHP)-nanogels are able to trap hydrophobic drugs or proteins inside the nanogels, which is potential in application to drug delivery system and tissue engineering. On the other hand, prostaglandin E1 (PGE1) plays important roles in wound healing and PGE1 ointment has been clinically used to treat chronic skin ulcers and wounds. The purpose of this study is to evaluate effects of CHP nanogels in combination with prostaglandin E1 on wound healing in full thickness skin defect model. A square skin defect (1 x 1 cm(2)) of full thickness was created on the dorsal of Wistar rats. The wound was treated with CHP nanogels without PGE1 (CHP group) or CHP nanogels containing with PGE1 (CHP/PGE1 group) or PGE1 ointment (PGE1 ointment group). In both CHP/PGE1 and PGE1 ointment groups, approximately 6 microg of PGE1 was applied to each wound. In the control group, the wound was untreated. The wound was evaluated in measuring wound area and histologically. In CHP/PGE1 group, the rate of wound size reduction was significantly higher than the ones of other groups. Histologically, CHP/PGE1 promoted neoepithelialization, neovascularization, and wound closure compared to the other treatments. These results suggest that CHP in combination with PGE1 can promote wound healing, which confirms the efficiency of CHP nanogels-based drug delivery system. (c) 2009 Wiley Periodicals, Inc.

  7. Production of poly(malic acid) from sugarcane juice in fermentation by Aureobasidium pullulans: Kinetics and process economics.

    PubMed

    Wei, Peilian; Cheng, Chi; Lin, Meng; Zhou, Yipin; Yang, Shang-Tian

    2017-01-01

    Poly(β-l-malic acid) (PMA) is a biodegradable polymer with many potential biomedical applications. PMA can be readily hydrolyzed to malic acid (MA), which is widely used as an acidulant in foods and pharmaceuticals. PMA production from sucrose and sugarcane juice by Aureobasidium pullulans ZX-10 was studied in shake-flasks and bioreactors, confirming that sugarcane juice can be used as an economical substrate without any pretreatment or nutrients supplementation. A high PMA titer of 116.3g/L and yield of 0.41g/g were achieved in fed-batch fermentation. A high productivity of 0.66g/L·h was achieved in repeated-batch fermentation with cell recycle. These results compared favorably with those obtained from glucose and other biomass feedstocks. A process economic analysis showed that PMA could be produced from sugarcane juice at a cost of $1.33/kg, offering a cost-competitive bio-based PMA for industrial applications. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Physical and mechanical properties in biodegradable films of whey protein concentrate-pullulan by application of beeswax.

    PubMed

    Khanzadi, Mehrdad; Jafari, Seid Mahdi; Mirzaei, Habibollah; Chegini, Faramarz Khodaian; Maghsoudlou, Yayha; Dehnad, Danial

    2015-03-15

    Different ratios of whey protein concentrate (WPC):pullulan (PUL) (70:30, 50:50, 30:70%w/w) and various rates of beeswax (BW) (0, 10, 20, and 30%w/wglycerol) were applied to prepare biodegradable WPC-PUL films containing glycerol as a plasticizer, for the first time. Thickness, moisture content, water solubility, water vapour permeability, colour, and mechanical properties of prepared films were measured. Higher ratios of WPC:PUL led to more desirable physical and mechanical properties; in other words, lower rates of thickness, moisture content, water solubility and water vapour permeability, and higher elongations were achieved. Application of BW (especially in higher contents) could successfully improve colour indices, diminish water solubility (nearly 12%) and water vapour permeability (approximately 3×10(-11)gm(-1)s(-1)Pa(-1)), and increase tensile strength (by about 7MPa) of WPC-PUL blend films. Our edible films enjoyed great whiteness and ignorable yellowness indices, making it a suitable alternative for application in food products. Overall, WPC70-PUL30 containing 30% BW resulted in the best performance of physical and mechanical aspects as an optimum film.

  9. Intensification of β-poly(L: -malic acid) production by Aureobasidium pullulans ipe-1 in the late exponential growth phase.

    PubMed

    Cao, Weifeng; Luo, Jianquan; Zhao, Juan; Qiao, Changsheng; Ding, Luhui; Qi, Benkun; Su, Yi; Wan, Yinhua

    2012-07-01

    β-Poly(malic acid) (PMLA) has attracted industrial interest because this polyester can be used as a prodrug or for drug delivery systems. In PMLA production by Aureobasidium pullulans ipe-1, it was found that PLMA production was associated with cell growth in the early exponential growth phase and dissociated from cell growth in the late exponential growth phase. To enhance PMLA production in the late phase, different fermentation modes and strategies for controlling culture redox potential (CRP) were studied. The results showed that high concentrations of produced PMLA (above 40 g/l) not only inhibited PMLA production, but also was detrimental to cell growth. Moreover, when CRP increased from 57 to 100 mV in the late exponential growth phase, the lack of reducing power in the broth also decreased PMLA productivity. PMLA productivity could be enhanced by repeated-batch culture to maintain cell growth in the exponential growth phase, or by cell-recycle culture with membrane to remove the produced PMLA, or by maintaining CRP below 70 mV no matter which kind of fermentation mode was adopted. Repeated-batch culture afforded a high PMLA concentration (up to 63.2 g/l) with a productivity of 1.15 g l(-1) h(-1). Cell-recycle culture also confirmed that PMLA production by the strain ipe-1 was associated with cell growth.

  10. Polycan, a β-glucan from Aureobasidium pullulans SM-2001, mitigates ovariectomy-induced osteoporosis in rats

    PubMed Central

    Jung, Mi Young; Kim, Joo Wan; Kim, Ki Young; Choi, Seong Hun; Ku, Sae Kwang

    2016-01-01

    The present study aimed to investigate the protective effects of Polycan, a β-glucan from Aureobasidium pullulans SM-2001, in a rat model of ovariectomy-induced osteoporosis. Ovariectomized (OVX) rats were orally administered 31.25, 62.5 or 125 mg/kg/day Polycan for 126 days, and alterations in body weight, bone mineral content, bone mineral density, failure load, histological profiles and histomorphometric indices were analyzed. In particular, serum levels of osteocalcin, bone-specific alkaline phosphatase (bALP), calcium and phosphorus, and the urine deoxypyridinoline/creatinine ratio, were measured. Furthermore, the femur, tibia and lumbar vertebrae were harvested from all rats, and histomorphometrical analyses were conducted in order to assess the mass and structure of the bones, and the rates of bone resorption and formation. One group of rats was treated with alendronate, which served as the reference drug. The results of the present study suggested that Polycan treatment was able to inhibit ovariectomy-induced alterations in bone resorption and turnover in a dose-dependent manner. In addition, the serum expression levels of bALP and all histomorphometrical indices for bone formation were markedly increased in the Polycan-treated groups. These results indicated that Polycan was able to preserve bone mass and strength, and increase the rate of bone formation in OVX rats; thus suggesting that Polycan may be considered a potential effective anti-osteoporosis agent. PMID:27588046

  11. High-level production of poly (β-L: -malic acid) with a new isolated Aureobasidium pullulans strain.

    PubMed

    Zhang, Huili; Cai, Jin; Dong, Jiaqi; Zhang, Danping; Huang, Lei; Xu, Zhinan; Cen, Peilin

    2011-10-01

    Poly (β-L: -malic acid) (PMLA) is a water-soluble polyester with many attractive properties in chemical industry and medicine development. However, the low titer of PMLA in the available producer strains limits further industrialization efforts and restricts its many potential applications. In order to solve this problem, a new strain with the distinguished high productivity of PMLA was isolated from fresh plants samples. It was characterized as the candidate of Aureobasidium pullulans based on the morphology and phylogenetic analyses of the internal transcribed spacer sequences. After the optimization of culture conditions, the highest PMLA concentration (62.27 g l(-1)) could be achieved in the shake flask scale. In addition, the contribution of the carbon flux to exopolysaccharide (EPS) and PMLA could be regulated by the addition of CaCO₃ in the medium. This high-level fermentation process was further scaled up in the 10 l benchtop fermentor with a high PMLA concentration (57.2 g l(-1)) and productivity (0.35 g l(-1) h(-1)), which are the highest level in all the literature. Finally, the suitable acid hydrolysis conditions of PMLA were also investigated with regard to the production of L: -malic acid, and the kinetics of PMLA acid hydrolysis was modeled to simulate the whole degradation process. The present work paved the road to produce this multifunctional biomaterial (PMLA) at industrial scale and promised one alternative method to produce L: -malic acid in the future.

  12. Resistant starch and pullulan reduce postprandial glucose, insulin, and GLP-1, but have no effect on satiety in healthy humans.

    PubMed

    Klosterbuer, Abby S; Thomas, William; Slavin, Joanne L

    2012-12-05

    The aim of this study was to determine the effects of three novel fibers on satiety and serum parameters. In a randomized, double-blind, crossover design, fasted subjects (n=20) consumed a low-fiber control breakfast or one of four breakfasts containing 25 g of fiber from soluble corn fiber (SCF) or resistant starch (RS), alone or in combination with pullulan (SCF+P and RS+P). Visual analog scales assessed appetite, and blood samples were collected to measure glucose, insulin, ghrelin, and glucagon-like peptide-1 (GLP-1). The fiber treatments did not influence satiety or energy intake compared to control. RS+P significantly reduced glucose, insulin, and GLP-1, but neither SCF treatment differed from control. To conclude, these fibers have little impact on satiety when provided as a mixed meal matched for calories and macronutrients. Additional research regarding the physiological effects of these novel fibers is needed to guide their use as functional ingredients in food products.

  13. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... engaged in catching fish. 31.3401(a)(17)-1 Section 31.3401(a)(17)-1 Internal Revenue INTERNAL REVENUE... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms...

  14. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... engaged in catching fish. 31.3401(a)(17)-1 Section 31.3401(a)(17)-1 Internal Revenue INTERNAL REVENUE... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms...

  15. Cloning, characterization, expression and antifungal activity of an alkaline serine protease of Aureobasidium pullulans PL5 involved in the biological control of postharvest pathogens.

    PubMed

    Zhang, Dianpeng; Spadaro, Davide; Valente, Silvia; Garibaldi, Angelo; Gullino, Maria Lodovica

    2012-02-15

    An alkaline protease gene was amplified from genomic DNA and cDNA of the antagonistic yeast-like fungus Aureobasidium pullulans PL5, a biocontrol agent effective against Monilinia laxa on stone fruit and Botrytis cinerea and Penicillium expansum on pome fruits. An open reading frame of 1248 bp encoding a 415-amino acid (aa) protein with a calculated molecular weight (M(r)) of 42.9 kDa and an isoelectric point (pI) of 4.5 was characterized. The cDNAALP5 gene had an 18-amino acid signal peptide, one N-gylcosylation, one histidine active site, and one serine active site. The ALP5 gene with a M(r) of 1351 bp contained two introns. One intron was of 54 bp, while the other was of 50 bp. Protein BLAST and phylogenetic tree analysis of the deduced amino sequences from the cDNAALP5 gene showed that the encoded protein had 100% homology to a protease enzyme (ALP2) of a sea strain of A. pullulans, suggesting that the protein ALP5 was an alkaline serine protease. Expression of ALP5 in Escherichia coli BL21 (DE3), followed by identification with Western-blotting, purification with Ni-NTA and analysis of enzymatic activity, yielded an homogeneous recombinant ALP5 which hydrolysed the substrate casein and inhibited the mycelial growth of the pathogens. At its optimal pH of 10.0 and reaction temperature of 50°C, the recombinant protease exhibited the highest activity towards the substrate casein, though the highest stability was at lower temperatures and pH between 7.0 and 9.0. This study provided the direct evidence that extracellular proteases secreted by the antagonist A. pullulans PL5 played a role in the biocontrol activities against some postharvest pathogens of apple and peach.

  16. Experimental high gradient testing of a 17.1 GHz photonic band-gap accelerator structure

    NASA Astrophysics Data System (ADS)

    Munroe, Brian J.; Zhang, JieXi; Xu, Haoran; Shapiro, Michael A.; Temkin, Richard J.

    2016-03-01

    We report the design, fabrication, and high gradient testing of a 17.1 GHz photonic band-gap (PBG) accelerator structure. Photonic band-gap (PBG) structures are promising candidates for electron accelerators capable of high-gradient operation because they have the inherent damping of high order modes required to avoid beam breakup instabilities. The 17.1 GHz PBG structure tested was a single cell structure composed of a triangular array of round copper rods of radius 1.45 mm spaced by 8.05 mm. The test assembly consisted of the test PBG cell located between conventional (pillbox) input and output cells, with input power of up to 4 MW from a klystron supplied via a TM01 mode launcher. Breakdown at high gradient was observed by diagnostics including reflected power, downstream and upstream current monitors and visible light emission. The testing procedure was first benchmarked with a conventional disc-loaded waveguide structure, which reached a gradient of 87 MV /m at a breakdown probability of 1.19 ×10-1 per pulse per meter. The PBG structure was tested with 100 ns pulses at gradient levels of less than 90 MV /m in order to limit the surface temperature rise to 120 K. The PBG structure reached up to 89 MV /m at a breakdown probability of 1.09 ×10-1 per pulse per meter. These test results show that a PBG structure can simultaneously operate at high gradients and low breakdown probability, while also providing wakefield damping.

  17. Inhibition of UVB-induced skin damage by exopolymers from Aureobasidium pullulans SM-2001 in hairless mice.

    PubMed

    Kim, Kyung Hu; Park, Soo Jin; Lee, Young Joon; Lee, Ji Eun; Song, Chang Hyun; Choi, Seong Hun; Ku, Sae Kwang; Kang, Su Jin

    2015-02-01

    Because antioxidants from natural sources may be an effective approach to the treatment and prevention of UV radiation-induced skin damage, the effects of purified exopolymers from Aureobasidium pullulans SM-2001 ('E-AP-SM2001') were evaluated in UVB-induced hairless mice. E-AP-SM2001 consists of 1.7% β-1,3/1,6-glucan, fibrous polysaccharides and other organic materials, such as amino acids, and mono- and di-unsaturated fatty acids (linoleic and linolenic acids) and shows anti-osteoporotic and immunomodulatory effects, through antioxidant and anti-inflammatory mechanisms. Hairless mice were treated topically with vehicle, E-AP-SM2001 stock and two and four times diluted solutions once per day for 15 weeks against UVB irradiation (three times per week at 0.18 J/cm(2) ). The following parameters were evaluated in skin samples: myeloperoxidase (MPO) activity, cytokine levels [interleukin (IL)-1β and IL-10], endogenous antioxidant content (glutathione, GSH), malondialdehyde (MDA) levels, superoxide anion production; matrix metalloproteases (MMP-1, -9 and -13), GSH reductase and Nox2 (gp91phox) mRNA levels, and immunoreactivity for nitrotyrosine (NT), 4-hydroxynonenal (HNE), caspase-3, and cleaved poly(ADP-ribose) polymerase (PARP). Photoageing was induced by UVB irradiation through ROS-mediated inflammation, which was related to the depletion of endogenous antioxidants, activation of MMPs and keratinocyte apoptosis. Topical treatment with all three doses of E-AP-SM2001 and 5 nm myricetin attenuated the UV-induced depletion of GSH, activation of MMPs, production of IL-1β, the decrease in IL-10 and keratinocyte apoptosis. In this study, E-AP-SM2001 showed potent inhibitory effects against UVB-induced skin photoageing. Thus, E-AP-SM2001 may be useful as a functional ingredient in cosmetics, especially as a protective agent against UVB-induced skin photoageing.

  18. Purification and Characterization of a Novel Thermostable α-l-Arabinofuranosidase from a Color-Variant Strain of Aureobasidium pullulans

    PubMed Central

    Saha, Badal C.; Bothast, Rodney J.

    1998-01-01

    A color-variant strain of Aureobasidium pullulans (NRRL Y-12974) produced α-l-arabinofuranosidase (α-l-AFase) when grown in liquid culture on oat spelt xylan. An extracellular α-l-AFase was purified 215-fold to homogeneity from the culture supernatant by ammonium sulfate treatment, DEAE Bio-Gel A agarose column chromatography, gel filtration on a Bio-Gel A-0.5m column, arabinan-Sepharose 6B affinity chromatography, and SP-Sephadex C-50 column chromatography. The purified enzyme had a native molecular weight of 210,000 and was composed of two equal subunits. It had a half-life of 8 h at 75°C, displayed optimal activity at 75°C and pH 4.0 to 4.5, and had a specific activity of 21.48 μmol · min−1 · mg−1 of protein against p-nitrophenyl-α-l-arabinofuranoside (pNPαAF). The purified α-l-AFase readily hydrolyzed arabinan and debranched arabinan and released arabinose from arabinoxylans but was inactive against arabinogalactan. The Km values of the enzyme for the hydrolysis of pNPαAF, arabinan, and debranched arabinan at 75°C and pH 4.5 were 0.26 mM, 2.14 mg/ml, and 3.25 mg/ml, respectively. The α-l-AFase activity was not inhibited at all by l-arabinose (1.2 M). The enzyme did not require a metal ion for activity, and its activity was not affected by p-chloromercuribenzoate (0.2 mM), EDTA (10 mM), or dithiothreitol (10 mM). PMID:9435077

  19. The concentration of polysaccharides and proteins in EPS of Pseudomonas putida and Aureobasidum pullulans as revealed by 13C CPMAS NMR spectroscopy.

    PubMed

    Metzger, Ulrich; Lankes, Ulrich; Fischpera, Kai; Frimmel, Fritz H

    2009-11-01

    Extracellular polymeric substances were extracted from the bacterial strain Pseudomonas putida and the fungal species Aureobasidium pullulans using three different methods (formaldehyde-NaOH, ethylenediaminetetraacetic acid (EDTA) and cation-exchange-resin). The composition of the extracellular polymeric substances (EPS) was analysed by biochemical and high-resolution solid state 13C nuclear magnetic resonance (NMR) spectroscopic methods. The EPS yield was strongly dependent on the extraction method, with the formaldehyde-NaOH method showing the best extraction efficiency. The NMR method revealed that when using the EDTA extraction method, about 40% of the EDTA accumulated in the EPS and that was responsible for the apparent high extraction yields. EPS protein content determined by the NMR method was up to 30% higher than the protein content determined using the biochemical (Lowry) method for P. putida and for A. pullulans. The average protein carbon content determined by the NMR method was approximately 70% of the total carbon content. NMR results could be supported by elemental analysis, which showed a high nitrogen content (approximately 10%) in the EPS. The carbohydrate carbon content detected with both methods in the cell aggregates and the EPS was approximately 20% in each. In this study, quantitative 13C cross-polarisation magic angle spinning NMR spectroscopy was conducted on unlabeled cell strains, and EPS and could be used to quantify protein and carbohydrate of different samples.

  20. Incorporation of nisin Z and lauric arginate into pullulan films to inhibit foodborne pathogens associated with fresh and ready-to-eat muscle foods.

    PubMed

    Pattanayaiying, Rinrada; H-Kittikun, Aran; Cutter, Catherine N

    2015-08-17

    A combination of food grade compounds with edible films, used to inhibit foodborne pathogens associated with fresh or further processed muscle foods, is receiving considerable attention. In this study, pullulan films containing lauric arginate (LAE) and nisin Z (produced by Lactococcus lactis subsp. lactis I8-7-3 and isolated from catfish gut), alone or in combination, were investigated for controlling foodborne pathogens on fresh and further processed muscle foods after long-term refrigerated storage. Salmonella Typhimurium and Salmonella Enteritidis on raw turkey breast slices wrapped with a film containing LAE or the combination of LAE with nisin Z were reduced throughout the experiment, 2.5 to 4.5 log10 CFU/cm(2) and 3.5 to 5.1 log10 CFU/cm(2), respectively. Film containing a combination of LAE with nisin Z reduced Staphylococcus aureus and Listeria monocytogenes Scott A inoculated onto ham surfaces by approximately 5.53 and 5.62 log10 CFU/cm(2), respectively during refrigerated storage. Escherichia coli O157:H7, O111, and O26 also were reduced by >4 log 10CFU/cm(2) on raw beef slices after treatment with the combination film and refrigerated storage. The results obtained from this study indicate the LAE- and LAE-nisin Z-containing pullulan films displayed excellent inhibition against foodborne pathogens on fresh and further processed muscle foods.

  1. Idiotypic cascades in cancer patients treated with monoclonal antibody CO17-1A.

    PubMed Central

    Wettendorff, M; Iliopoulos, D; Tempero, M; Kay, D; DeFreitas, E; Koprowski, H; Herlyn, D

    1989-01-01

    We have previously shown that gastrointestinal cancer patients treated with monoclonal antibody CO17-1A (Ab1) developed anti-idiotypic antibodies (Ab2) to the Ab1. We now demonstrate that patients produce anti-anti-idiotypic antibodies (Ab3) to their autologous Ab2. Ab3 were demonstrated in culture supernatants of peripheral blood mononuclear cells from five Ab1-treated patients after stimulation of the cells with heterologous Ab2 that functionally mimicked the tumor antigen (Ag) defined by Ab1 and immunologically cross reacted with the patients' Ab2. Ab3 shared idiotopes with Ab1 and were Ab1-like in their binding specificities to tumor cells, Ag, and Ab2. Such antibodies were also elicited by stimulating cells with Ag. However, they were not produced by stimulating posttreatment mononuclear cells with control proteins or by stimulating pretreatment cells with either Ag or Ab2. Our results demonstrate idiotypic cascades in cancer patients treated with monoclonal antibody. Ag-specific Ab3 responses may underlie delayed clinical responses often observed in cancer patients treated with monoclonal antibodies of various specificities. Images PMID:2786203

  2. 17.1%-Efficient Multi-Scale-Textured Black Silicon Solar Cells without Dielectric Antireflection Coating

    SciTech Connect

    Toor, F.; Page, M. R.; Branz, H. M.; Yuan, H. C.

    2011-01-01

    In this work we present 17.1%-efficient p-type single crystal Si solar cells with a multi-scale-textured surface and no dielectric antireflection coating. Multi-scale texturing is achieved by a gold-nanoparticle-assisted nanoporous etch after conventional micron scale KOH-based pyramid texturing (pyramid black etching). By incorporating geometric enhancement of antireflection, this multi-scale texturing reduces the nanoporosity depth required to make silicon `black' compared to nanoporous planar surfaces. As a result, it improves short-wavelength spectral response (blue response), previously one of the major limiting factors in `black-Si' solar cells. With multi-scale texturing, the spectrum-weighted average reflectance from 350- to 1000-nm wavelength is below 2% with a 100-nm deep nanoporous layer. In comparison, roughly 250-nm deep nanopores are needed to achieve similar reflectance on planar surface. Here, we characterize surface morphology, reflectivity and solar cell performance of the multi-scale textured solar cells.

  3. A novel multicompartimental system based on aminated poly(vinyl alcohol) microspheres/succinoylated pullulan microspheres for oral delivery of anionic drugs.

    PubMed

    Constantin, M; Fundueanu, G; Bortolotti, F; Cortesi, R; Ascenzi, P; Menegatti, E

    2007-02-07

    Poly(vinyl alcohol) (PVA) microspheres were prepared by dispersion reticulation with glutaraldehyde and further aminated. These microspheres were firstly loaded with diclofenac (DF) and then entrapped in cellulose acetate butyrate (CAB) microcapsules by an o/w solvent evaporation technique for intestinal delivery of drug. The encapsulated PVA microspheres due to their low swelling degree in intestinal fluids, do not have enough force to produce the disruption of CAB shell, therefore different amounts of succinoylated pullulan microspheres (SP-Ms) (exchange capacity up to 5.2 meq/g) were co-encapsulated. The SP-Ms do not swell in acidic pH, but swell up to 20-times in intestinal fluids causing the rupture of CAB shell and facilitating the escape of loaded PVA microspheres.

  4. 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

  5. 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.

  6. Efficient Delivery of DOX to Nuclei of Hepatic Carcinoma Cells in the Subcutaneous Tumor Model Using pH-Sensitive Pullulan-DOX Conjugates.

    PubMed

    Li, Huanan; Cui, Yani; Sui, Junhui; Bian, Shaoquan; Sun, Yong; Liang, Jie; Fan, Yujiang; Zhang, Xingdong

    2015-07-29

    A series of pullulan-doxorubicin conjugates (Pu-DOXs) were investigated for effectively delivering DOX to nuclei of hepatic carcinoma cells in subcutaneous tumor model. These Pu-DOXs were prepared by conjugating DOX onto pullulan molecule via pH-responsive hydrazone bond using spacers with different alkane chain length. The highest drug loading content of Pu-DOXs went up to nearly 50%, and the diameter of Pu-DOX nanoparticles ranged from 50 to 170 nm, as measured by DLS and TEM. These Pu-DOX nanoparticles could rapidly release DOX in the acidic environment at pH = 5.0 while being kept relatively stable in neural conditions. The in vitro cell coculture experiments revealed that these Pu-DOX nanoparticles were selectively internalized by hepatic carcinoma cells through receptor-mediated endocytosis via asialoglycoprotein receptor on the hepatic carcinoma cell surface. DOX was rapidly released from Pu-DOX nanoparticles in acidic endosome/lysosome, diffused into cell nuclei due to its strong affinity to nucleic acid, inhibited the cell proliferation, and accelerated the cell apoptosis. In the nude mice subcutaneous hepatic carcinoma model, Pu-DOX nanoparticles efficiently accumulated in the tumor site through the enhanced permeation and retention effect. Then DOX was specifically internalized by hepatic carcinoma cells and rapidly diffused into the nuclei of cells. Compared with the control group in in vivo experiments, these Pu-DOX nanoparticles effectively inhibited solid tumor growth, prolonging the lifetime of the experimental animal. These pH sensitive nanoparticles might provide an important clinical implication for targeted hepatic carcinoma therapy with high efficiency and low systematic toxicity.

  7. Crystal structure of Thermoactinomyces vulgaris R-47 alpha-amylase II (TVAII) hydrolyzing cyclodextrins and pullulan at 2.6 A resolution.

    PubMed

    Kamitori, S; Kondo, S; Okuyama, K; Yokota, T; Shimura, Y; Tonozuka, T; Sakano, Y

    1999-04-16

    The crystal structure of Thermoactinomyces vulgaris R-47 alpha-Amylase II (TVAII) has been determined by multiple isomorphous replacement at 2.6 A resolution. TVAII was crystallized in an orthorhombic system with the space group P212121 and the cell dimensions a=118.5 A, b=119.5 A, c=114.5 A. There are two molecules in an asymmetric unit, related by the non-crystallographic 2-fold symmetry. Diffraction data were collected at 113 K and the cell dimensions reduced to a=114.6 A, b=117.9 A, c=114.2 A, and the model was refined against 7.0-2.6 A resolution data giving an R-factor of 0.204 (Rfree=0.272). The final model consists of 1170 amino acid residues (two molecules) and 478 water molecules with good chemical geometry. TVAII has three domains, A, B, and C, like other alpha-amylases. Domain A with a (beta/alpha)8 barrel structure and domain C with a beta-sandwich structure are very similar to those found in other alpha-amylases. Additionally, TVAII has an extra domain N composed of 121 amino acid residues at the N-terminal site, which has a beta-barrel-like structure consisting of seven antiparallel beta-strands. Domain N is one of the driving forces in the formation of the dimer structure of TVAII, but its role in the enzyme activity is still not clear. TVAII does not have the Ca2+ binding site that connects domains A and B in other alpha-amylases, rather the NZ atom of Lys299 of TVAII serves as the connector between these domains. TVAII can hydrolyze cyclodextrins and pullulan as well as starch. Based on a structural comparison with the complex between a mutant cyclodextrin glucanotransferase and a beta-cyclodextrin derivative, Phe286 located at domain B is considered the residue most likely to recognize the hydrophobic cavity of cyclodextrins. The active-site cleft of TVAII is wider and shallower than that of other alpha-amylases, and seems to be suitable for the binding of pullulan which is expected not to adopt the helical structure of amylose.

  8. Biocompatible cationic pullulan-g-desoxycholic acid-g-PEI micelles used to co-deliver drug and gene for cancer therapy.

    PubMed

    Chen, Lili; Ji, Fangling; Bao, Yongming; Xia, Jing; Guo, Lianying; Wang, Jingyun; Li, Yachen

    2017-01-01

    The greatest crux in the combination of chemotherapy and gene therapy is the construction of a feasible and biocompatible carrier for loading the therapeutic drug and gene simultaneously. Here, a new amphiphilic bifunctional pullulan derivative (named as PDP) synthesized by grafting lipophilic desoxycholic acid and low-molecular weight (1kDa) branched polyethylenimine onto the backbone of pullulan was evaluated as a nano-carrier for the co-delivery of drug and gene for potential cancer therapy. PDP exhibited good blood compatibility and low cytotoxicity in the hemolysis and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, respectively. By self-assembly process, the amphiphilic PDP polymer formed cationic core-shell nanomicelles in aqueous solution with an average diameter of 160.8nm and a zeta potential of approximate 28mV. The PDP micelles had relative higher drug encapsulation efficiency (84.05%) and loading capacity (7.64%) for doxorubicin (DOX), an effective anti-tumor drug, demonstrating sustained drug release profile and good DNA-binding ability. The flow cytometry and confocal laser scanning microscopy showed that PDP/DOX micelles could be successfully internalized by MCF-7 cells, and presenting higher cytotoxicity against MCF-7 cells than that of free DOX. Furthermore, PDP micelles could efficiently transport tumor suppressor gene p53 into MCF-7 cells, and the expressed exogenous p53 protein induced MCF-7 cells to die. More excitedly, in comparison with single DOX or p53 delivery, the co-delivery of DOX and gene p53 using PDP micelles displayed higher cytotoxicity, induced higher apoptosis rate of tumor cells and blocked more effectively the migration of cancer cells in vitro. In tumor-bearing mice, co-delivery of DOX and p53 also exhibited enhanced antitumor efficacy as compared with single delivery of DOX or p53 alone. In summary, these results demonstrated that it is highly promising to use cationic PDP micelles for effectively

  9. Polymalic acid fermentation by Aureobasidium pullulans for malic acid production from soybean hull and soy molasses: Fermentation kinetics and economic analysis.

    PubMed

    Cheng, Chi; Zhou, Yipin; Lin, Meng; Wei, Peilian; Yang, Shang-Tian

    2017-01-01

    Polymalic acid (PMA) production by Aureobasidium pullulans ZX-10 from soybean hull hydrolysate supplemented with corn steep liquor (CSL) gave a malic acid yield of ∼0.4g/g at a productivity of ∼0.5g/L·h. ZX-10 can also ferment soy molasses, converting all carbohydrates including the raffinose family oligosaccharides to PMA, giving a high titer (71.9g/L) and yield (0.69g/g) at a productivity of 0.29g/L·h in fed-batch fermentation under nitrogen limitation. A higher productivity of 0.64g/L·h was obtained in repeated batch fermentation with cell recycle and CSL supplementation. Cost analysis for a 5000 MT plant shows that malic acid can be produced at $1.10/kg from soy molasses, $1.37/kg from corn, and $1.74/kg from soybean hull. At the market price of $1.75/kg, malic acid production from soy molasses via PMA fermentation offers an economically competitive process for industrial production of bio-based malic acid. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Pullulan microbeads/Si-HPMC hydrogel injectable system for the sustained delivery of GDF-5 and TGF-β1: new insight into intervertebral disc regenerative medicine.

    PubMed

    Henry, Nina; Clouet, Johann; Fragale, Audrey; Griveau, Louise; Chédeville, Claire; Véziers, Joëlle; Weiss, Pierre; Le Bideau, Jean; Guicheux, Jérôme; Le Visage, Catherine

    2017-11-01

    Discogenic low back pain is considered a major health concern and no etiological treatments are today available to tackle this disease. To clinically address this issue at early stages, there is a rising interest in the stimulation of local cells by in situ injection of growth factors targeting intervertebral disc (IVD) degenerative process. Despite encouraging safety and tolerability results in clinic, growth factors efficacy may be further improved. To this end, the use of a delivery system allowing a sustained release, while protecting growth factors from degradation appears of particular interest. We propose herein the design of a new injectable biphasic system, based on the association of pullulan microbeads (PMBs) into a cellulose-based hydrogel (Si-HPMC), for the TGF-β1 and GDF-5 growth factors sustained delivery. We present for the first time the design and mechanical characterization of both the PMBs and the called biphasic system (PMBs/Si-HPMC). Their loading and release capacities were also studied and we were able to demonstrate a sustained release of both growth factors, for up to 28 days. Noteworthy, the growth factors biological activity on human cells was maintained. Altogether, these data suggest that this PMBs/Si-HPMC biphasic system may be a promising candidate for the development of an innovative bioactive delivery system for IVD regenerative medicine.

  11. Oral administration of the Aureobasidium pullulans-derived β-glucan effectively prevents the development of high fat diet-induced fatty liver in mice

    PubMed Central

    Aoki, Shiho; Iwai, Atsushi; Kawata, Koji; Muramatsu, Daisuke; Uchiyama, Hirofumi; Okabe, Mitsuyasu; Ikesue, Masahiro; Maeda, Naoyoshi; Uede, Toshimitsu

    2015-01-01

    Aureobasidium pullulans-derived β-glucan (AP-PG) consisting of a β-(1,3)-linked glucose main chain and β-(1,6)-linked glucose branches is taken as a supplement to improve health. This study demonstrates that oral administration of AP-PG is effective to prevent the development of high-fat diet (HFD)-induced fatty liver in mice. Here, C57BL/6N mice were fed with a normal diet or HFD, and AP-PG diluted in drinking water was administered orally. After 16 weeks, the serological analysis showed that HFD-induced high blood cholesterol and triglyceride levels were reduced by the oral administration of AP-PG. Further, HFD induced-fatty liver was significantly reduced by the oral administration of AP-PG. The triglyceride accumulation in the liver was also significantly reduced in mice administered AP-PG. Liver injury as indicated by an increase in serum alanine aminotransferase (ALT) in the HFD-fed mice was significantly reduced in the mice administered AP-PG orally, and the gene expression of cholesterol 7 alpha-hydroxylase (CYP7A1) which is known to be involved in cholesterol degradation in the liver was significantly increased in the AP-PG administered mice. These results suggest the possibility that the oral administration of AP-PG is effective to prevent the development of non-alcoholic fatty liver disease (NAFLD). PMID:26179949

  12. β-(1 → 3)-Glucanolytic yeasts from Brazilian grape microbiota: production and characterization of β-glucanolytic enzymes by Aureobasidium pullulans 1WA1 cultivated on fungal Mycelium.

    PubMed

    Bauermeister, Anelize; Amador, Ismael R; Pretti, Carla P; Giese, Ellen C; Oliveira, André L M

    2015-01-14

    A total of 95 yeast strains were isolated from the microbiota of different grapes collected at vineyards in southern Brazil. The yeasts were screened for β-(1 → 3)-glucanases using a newly developed zymogram method that relies upon the appearance of clearance zones around growing colonies cultured on agar–botryosphaeran medium and also by submerged fermentation on nutrient medium containing botryosphaeran, a (1 → 3),(1 → 6)-β-d-glucan. Among 14 β-(1 → 3)-glucanase-positive yeasts identified, four strains produced the highest β-glucanolytic activities and were evaluated for enzyme production on cellobiose, botryosphaeran, and mycelial biomass from Botryosphaeria rhodina (MAMB-05). Yeast strain 1WA1 produced the highest β-(1 → 3)-glucanase and β-glucosidase activities and was identified by molecular characterization as Aureobasidium pullulans. The physicochemical properties of the crude β-glucanolytic enzyme preparation were characterized, and the preparation was used to hydrolyze several β-d-glucans (laminarin, botryosphaeran, lasiodiplodan, pustulan, and curdlan). The production and physicochemical properties of the β-glucanolytic preparation enable its potential applications in wine enology and production of prebiotics through hydrolysis of β-d-glucans.

  13. T cell immunomonitoring and tumor responses in patients immunized with a complex of cholesterol-bearing hydrophobized pullulan (CHP) and NY-ESO-1 protein.

    PubMed

    Uenaka, Akiko; Wada, Hisashi; Isobe, Midori; Saika, Takashi; Tsuji, Kazuhide; Sato, Eiichi; Sato, Shuichiro; Noguchi, Yuji; Kawabata, Ryohei; Yasuda, Takushi; Doki, Yuichiro; Kumon, Hiromi; Iwatsuki, Keiji; Shiku, Hiroshi; Monden, Morito; Jungbluth, Achim A; Ritter, Gerd; Murphy, Roger; Hoffman, Eric; Old, Lloyd J; Nakayama, Eiichi

    2007-04-19

    We recently showed that vaccination with a complex of cholesterol-bearing hydrophobized pullulan and NY-ESO-1 protein (CHP-NY-ESO-1) elicited antibody responses in 9 of 9 patients vaccinated in a clinical trial. In this study, we performed T cell immunomonitoring and analyzed tumor responses in these patients. To evaluate CD4 and CD8 T cell responses, an IFN-gamma secretion assay was used. The assay showed low background and was sensitive for detecting antigen-specific T cells. An increase in the CD4 T cell response was observed in 2 of 2 initially sero-positive and 5 of 7 initially sero-negative patients after vaccination. An increase in the CD8 T cell response was also observed in 2 of 2 sero-positive and 5 of 7 sero-negative patients after vaccination. Analysis of peptides recognized by CD4 and CD8 T cells revealed two dominant NY-ESO-1 regions, 73-114 and 121-144. Tumor responses were observed in 3 esophageal cancer patients and a malignant melanoma patient. In 3 of 4 prostate cancer patients, prostate-specific antigen (PSA) values stabilized during the course of vaccination. The use of whole protein, containing multiple CD4 and CD8 epitopes, may be beneficial for cancer vaccines to prevent tumors from evading the immune response.

  14. Novel hydrogels based on carboxyl pullulan and collagen crosslinking with 1, 4-butanediol diglycidylether for use as a dermal filler: initial in vitro and in vivo investigations.

    PubMed

    Li, Xian; Xue, Wenjiao; Zhu, Chenhui; Fan, Daidi; Liu, Yannan; XiaoxuanMa

    2015-12-01

    Novel hydrogels based on carboxyl pullulan (PC) and human-like collagen (HLC) crosslinking with 1,4-butanediol diglycidyl ether (BDDE) are promising soft fillers for tissue engineering due to their highly tunable properties. Recent studies, however, have shown that incorporating hyaluronic acid and BDDE results in hydrogels with a microporous structure, a large pore size and high porosity, which reduce cell adhesion and enhance degradation in vivo. To improve biocompatibility and prevent biodegradation, the use of PC to replace hyaluronic acid in the fabrication of PC/BDDE (PCB) and PC/BDDE/HLC (PCBH) hydrogels was investigated. Preparation of gels with PC is a promising strategy due to the high reactivity, superb selectivity, and mild reaction conditions of PC. In particular, the Schiff base reaction of HLC and PC produces the novel functional group -RCONHR' in PCBH hydrogels. Twenty-four weeks after subcutaneous injection of either PCB or PCBH hydrogel in mice, the surrounding tissue inflammation, enzymatic response and cell attachment were better compared to hyaluronic acid-based hydrogels. However, the biocompatibility, cytocompatibility and non-biodegradability of PCBH were milder than those of the PCB hydrogels both in vivo and in vitro. These results show that the proposed use of PC and HLC for the fabrication of hydrogels is a promising strategy for generating soft filler for tissue engineering.

  15. Effect of whey protein isolate-pullulan edible coatings on the quality and shelf life of freshly roasted and freeze-dried Chinese chestnut.

    PubMed

    Gounga, M E; Xu, S-Y; Wang, Z; Yang, W G

    2008-05-01

    Harvested chestnut is characterized by a short shelf life, exposing many Chinese producers to a storage problem as product losses are very high. The objective of this study was to develop a suitable technology to extend the shelf life of harvested chestnut fruits for commercial use. The effect of whey protein isolate-pullulan (WPI-Pul) coating on fresh-roasted chestnuts (FRC) and roasted freeze-dried chestnut (RFDC) quality and shelf life was studied under 2 different storage temperature (4 and 20 degrees C) conditions. Coatings were formed directly onto the surface of the fruits by dipping them into a film solution. SEM micrographs showed homogeneous WPI-Pul to cover the whole surface of chestnut with good adherence and perfect integrity. Moisture loss or gain, fruit quality, and shelf life were evaluated by weight loss or gain, surface color development, and visible decay during the storage period of 15 to 120 d at 4 and 20 degrees C, respectively. WPI-Pul coating had a low, yet significant effect on reducing moisture loss and decay incidence of FRC, hence delaying changes in their external color. The results were satisfactory when the coating was done with freeze-drying at low temperature storage, thus improving the quality and increasing the shelf life. This provides an alternative strategy to minimize the significant losses in harvested chestnut.

  16. Antitumor and antimetastatic activity of a novel water-soluble low molecular weight beta-1, 3-D-glucan (branch beta-1,6) isolated from Aureobasidium pullulans 1A1 strain black yeast.

    PubMed

    Kimura, Yoshuiyuki; Sumiyoshi, Maho; Suzuki, Toshio; Sakanaka, Masahiro

    2006-01-01

    Though it has been reported that beta-glucans or protein-binding hetero-glucans isolated from mushrooms have antitumor activity, the antitumor and antimetastatic actions of purified, structurally defined polysaccharides (such as beta-glucans) have not been proven yet. A new low molecular weight (approximately 100 kDa) beta-glucan was isolated from Aureobasidium pullulans black yeast, and was found to have low viscosity and to be water-soluble. The industrial production of this P-glucan was achieved from the culture media of A. pullulans. The effects of water-soluble low-molecular-weight (LMW) beta-(1-->3) and 50-80% branched beta-(1-->6) glucan isolated from A. pullulans on tumor growth and metastasis to the liver were examined in mice intrasplenically with implanted colon 26 tumor cells. In addition, to clarify the antitumor and antimetastatic actions of LMW beta-(1,3-1,6) glucan, the effects on immune functions in the small intestine were also examined. The intraperitoneal (5 and 15 mg/kg) and oral (50 mg/kg) administration of LMW P-(1.3-1.6) glucan inhibited the tumor growth and liver metastasis in mice intrasplenically implanted with colon 26 cells. The numbers of natural killer (NK)- and interferon (IFN)-gamma-positive cells in the small intestine of colon 26-bearing mice were lower than those in normal mice. The intraperitoneally and orally administered LMW beta-(1,3-1,6) glucan prevented the reduction of the number of NK- and IFN-gamma-positive cells induced by the tumor growth after implantation of colon 26 cells. These findings suggest that the antitumor and antimetastatic actions of LMW beta-(1,3-1,6) glucan may involve the enhancement of intestinal immune functions through increases in NK- and IFN-gamma-positive cell numbers.

  17. Experimental high gradient testing of a 17.1 GHz photonic band-gap accelerator structure

    DOE PAGES

    Munroe, Brian J.; Zhang, JieXi; Xu, Haoran; ...

    2016-03-29

    In this paper, we report the design, fabrication, and high gradient testing of a 17.1 GHz photonic band-gap (PBG) accelerator structure. Photonic band-gap (PBG) structures are promising candidates for electron accelerators capable of high-gradient operation because they have the inherent damping of high order modes required to avoid beam breakup instabilities. The 17.1 GHz PBG structure tested was a single cell structure composed of a triangular array of round copper rods of radius 1.45 mm spaced by 8.05 mm. The test assembly consisted of the test PBG cell located between conventional (pillbox) input and output cells, with input power ofmore » up to 4 MW from a klystron supplied via a TM01 mode launcher. Breakdown at high gradient was observed by diagnostics including reflected power, downstream and upstream current monitors and visible light emission. The testing procedure was first benchmarked with a conventional disc-loaded waveguide structure, which reached a gradient of 87 MV=m at a breakdown probability of 1.19 × 10–1 per pulse per meter. The PBG structure was tested with 100 ns pulses at gradient levels of less than 90 MV=m in order to limit the surface temperature rise to 120 K. The PBG structure reached up to 89 MV=m at a breakdown probability of 1.09 × 10–1 per pulse per meter. These test results show that a PBG structure can simultaneously operate at high gradients and low breakdown probability, while also providing wakefield damping.« less

  18. Effects of co-fermented Pleurotus eryngii stalk residues and soybean hulls by Aureobasidium pullulans on performance and intestinal morphology in broiler chickens.

    PubMed

    Lai, L P; Lee, M T; Chen, C S; Yu, B; Lee, T T

    2015-12-01

    Soybean hulls are a by-product of soybean processing for oil and meal production; Pleurotus eryngii stalk residues (PESR) are by-products of the edible portion of the fruiting body enriched in bioactive metabolites. This study evaluated the effects of co-fermented PESR and soybean hulls with Aureobasidium pullulans on performance and intestinal morphology in broiler chickens. The in vitro experimental results showed that xylananse and mannanase activity of solid-state fermented soybean hulls (100% SBH) and soybean hulls partially replaced with PESR (75:25, SHP) reached peak at day 12; solid-state fermentation (SSF) enhanced the total phenolic content and trolox equivalency in both products as well. Additionally, FSHP had higher xylotriose and mannobiose levels than fermented FSBH did. A total of 400 broilers (Ross 308) were assigned randomly into four groups receiving the basal diet (control) or the basal diet supplemented with 0.5% fermented SBH (0.5% FSBH), 0.5% fermented SBHP (0.5% FSHP) and 1.0% fermented SBHP (1.0% FSHP) until 35 d of age, respectively. Results demonstrated that 0.5% FSHP addition increased body weight gain as compared with corresponding normal diet fed control in birds during entire experimental period. Compared with the control group, 0.5% FSHP group significantly increased the ratio of lactic acid bacteria to Clostridium perfringens in ceca as well as ileum villus height and jejunum villus height/crypt depth ratio of 35 d old birds. In conclusion, 0.5% FSHP supplementation in the diet could obtain not only improved body weight gain, but optimal intestinal morphology by exerting its bioactive metabolite properties when fed to broilers. © 2015 Poultry Science Association Inc.

  19. Stimulation of macrophages with the β-glucan produced by aureobasidium pullulans promotes the secretion of tumor necrosis factor-related apoptosis inducing ligand (TRAIL).

    PubMed

    Kawata, Koji; Iwai, Atsushi; Muramatsu, Daisuke; Aoki, Shiho; Uchiyama, Hirofumi; Okabe, Mitsuyasu; Hayakawa, Sumio; Takaoka, Akinori; Miyazaki, Tadaaki

    2015-01-01

    A β-glucan produced by Aureobasidium pullulans (AP-PG) is consisting of a β-(1,3)-linked main chain with β-(1,6)-linked glucose side residues. Various β-glucans consisting of β-(1,3)-linked main chain including AP-PG are believed to exhibit anti-tumor activities, and actually, anti-tumor activities of AP-PG in mice have been demonstrated. In this study, we demonstrate that stimulation with AP-PG induces TRAIL expression in mouse and human macrophage-like cell lines. TRAIL is known to be a cytokine which specifically induces apoptosis in transformed cells, but not in untransformed cells. The expression of TRAIL mRNA after stimulation with AP-PG was increased in RAW264.7 cells, Mono Mac 6 cells, and macrophage-differentiated THP-1 cells. The mRNA expression of TNF-α and FasL is only weakly increased after stimulation with AP-PG. The induction activity of TRAIL by curdlan, a bacterial β-glucan, was very similar to that by AP-PG in RAW264.7 cells, but weaker in macrophage-differentiated THP-1 cells. Activation of caspases was found in HeLa cells after treatment with the supernatant of cultured medium from AP-PG-stimulated Mono Mac 6 cells, and was inhibited by the anti-TRAIL neutralizing antibody. These findings suggest that the stimulation with AP-PG effectively induces TRAIL in macrophages, and that it may be related to apoptosis induction of tumor cells.

  20. Stimulation of Macrophages with the β-Glucan Produced by Aureobasidium pullulans Promotes the Secretion of Tumor Necrosis Factor-Related Apoptosis Inducing Ligand (TRAIL)

    PubMed Central

    Kawata, Koji; Iwai, Atsushi; Muramatsu, Daisuke; Aoki, Shiho; Uchiyama, Hirofumi; Okabe, Mitsuyasu; Hayakawa, Sumio; Takaoka, Akinori; Miyazaki, Tadaaki

    2015-01-01

    A β-glucan produced by Aureobasidium pullulans (AP-PG) is consisting of a β-(1,3)-linked main chain with β-(1,6)-linked glucose side residues. Various β-glucans consisting of β-(1,3)-linked main chain including AP-PG are believed to exhibit anti-tumor activities, and actually, anti-tumor activities of AP-PG in mice have been demonstrated. In this study, we demonstrate that stimulation with AP-PG induces TRAIL expression in mouse and human macrophage-like cell lines. TRAIL is known to be a cytokine which specifically induces apoptosis in transformed cells, but not in untransformed cells. The expression of TRAIL mRNA after stimulation with AP-PG was increased in RAW264.7 cells, Mono Mac 6 cells, and macrophage-differentiated THP-1 cells. The mRNA expression of TNF-α and FasL is only weakly increased after stimulation with AP-PG. The induction activity of TRAIL by curdlan, a bacterial β-glucan, was very similar to that by AP-PG in RAW264.7 cells, but weaker in macrophage-differentiated THP-1 cells. Activation of caspases was found in HeLa cells after treatment with the supernatant of cultured medium from AP-PG-stimulated Mono Mac 6 cells, and was inhibited by the anti-TRAIL neutralizing antibody. These findings suggest that the stimulation with AP-PG effectively induces TRAIL in macrophages, and that it may be related to apoptosis induction of tumor cells. PMID:25875639

  1. The mitochondrial protein import component, TRANSLOCASE OF THE INNER MEMBRANE17-1, plays a role in defining the timing of germination in Arabidopsis.

    PubMed

    Wang, Yan; Law, Simon R; Ivanova, Aneta; van Aken, Olivier; Kubiszewski-Jakubiak, Szymon; Uggalla, Vindya; van der Merwe, Margaretha; Duncan, Owen; Narsai, Reena; Whelan, James; Murcha, Monika W

    2014-11-01

    In Arabidopsis (Arabidopsis thaliana), small gene families encode multiple isoforms for many of the components of the mitochondrial protein import apparatus. There are three isoforms of the TRANSLOCASE OF THE INNER MEMBRANE17 (Tim17). Transcriptome analysis indicates that AtTim17-1 is only detectable in dry seed. In this study, two independent transfer DNA insertional mutant lines of tim17-1 exhibited a germination-specific phenotype, showing a significant increase in the rate of germination. Microarray analyses revealed that Attim17-1 displayed alterations in the temporal sequence of transcriptomic events during germination, peaking earlier compared with the wild type. Promoter analysis of AtTim17-1 further identified an abscisic acid (ABA)-responsive element, which binds ABA-responsive transcription factors, acting to repress the expression of AtTim17-1. Attim17-1 dry seeds contained significantly increased levels of ABA and gibberellin, 2- and 5-fold, respectively. These results support the model that mitochondrial biogenesis is regulated in a tight temporal sequence of events during germination and that altering mitochondrial biogenesis feeds back to alter the germination rate, as evidenced by the altered levels of the master regulatory hormones that define germination.

  2. Characterization of the molecular mass distribution of pullulans by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using 2,5-dihydroxybenzoic acid butylamine (DHBB) as liquid matrix.

    PubMed

    Schnöll-Bitai, Irene; Ullmer, Roman; Hrebicek, Thomas; Rizzi, Andreas; Lacik, Igor

    2008-10-01

    The performances of several matrices were investigated for the accurate determination of the molecular mass distributions of pullulans by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The ionic liquid matrix (ILM) 2,5-dihydroxybenzoic acid butylamine (DHBB) gave better and more reliable results than the crystalline matrices 2,5-dihydroxybenzoic acid (DHB) and 2,4,6-trihydroxyacetophenone (THAP). With the ILM it was possible to obtain spectra of pullulans up to more than 100 kDa, the highest molar mass reported thus far. Owing to the known advantages of liquid matrices providing better spot-to-spot reproducibility, an almost noise-free spectrum and constant baselines were obtained when working under optimized conditions. In particular, the extent of in-source fragmentation occurring with this group of fragile polymers was considerably and decisively reduced. Thus, a more reliable representation of the true oligomer and polymer distributions is experimentally attainable, especially for distributions with small polydispersity values. The maximum error in the measured distribution associated with fragmentation was estimated by model calculations describing the changes in the polymer distribution upon different probabilities of fragmentation events. These simulation results indicated that the data obtained by MALDI-TOFMS using the liquid DHBB matrix were of high reliability. In particular, the average value of the distributions, M(w), and the polydispersity were obtained with predicted uncertainties of between 3 and 15% depending on the width of the distribution and the mass of the polymers.

  3. 17.1%-Efficient Multi-Scale-Textured Black Silicon Solar Cells without Dielectric Antireflection Coating: Preprint

    SciTech Connect

    Toor, F.; Page, M. R.; Branz, H. M.; Yuan, H. C.

    2011-07-01

    In this work we present 17.1%-efficient p-type single crystal Si solar cells with a multi-scale-textured surface and no dielectric antireflection coating. Multi-scale texturing is achieved by a gold-nanoparticle-assisted nanoporous etch after conventional micron scale KOH-based pyramid texturing (pyramid black etching). By incorporating geometric enhancement of antireflection, this multi-scale texturing reduces the nanoporosity depth required to make silicon 'black' compared to nanoporous planar surfaces. As a result, it improves short-wavelength spectral response (blue response), previously one of the major limiting factors in 'black-Si' solar cells. With multi-scale texturing, the spectrum-weighted average reflectance from 350- to 1000-nm wavelength is below 2% with a 100-nm deep nanoporous layer. In comparison, roughly 250-nm deep nanopores are needed to achieve similar reflectance on planar surface. Here, we characterize surface morphology, reflectivity and solar cell performance of the multi-scale textured solar cells.

  4. Time-dependent transition of the immunoglobulin G subclass and immunoglobulin E response in cancer patients vaccinated with cholesteryl pullulan-melanoma antigen gene-A4 nanogel

    PubMed Central

    Kyogoku, Noriaki; Ikeda, Hiroaki; Tsuchikawa, Takahiro; Abiko, Takehiro; Fujiwara, Aki; Maki, Takehiro; Yamamura, Yoshiyuki; Ichinokawa, Masaomi; Tanaka, Kimitaka; Imai, Naoko; Miyahara, Yoshihiro; Kageyama, Shinichi; Shiku, Hiroshi; Hirano, Satoshi

    2016-01-01

    A phase I+II clinical trial of vaccination with MAGE-A4 protein complexed with cholesteryl pullulan melanoma antigen gene-A4 nanogel (CHP-MAGE-A4) is currently underway in patients with MAGE-A4-expressing cancer. In the present study, the primary phase I endpoint was to test the safety of the administration of 300 µg CHP-MAGE-A4 with and without OK-432. Another aim of the study was to clarify the details of the specific humoral immune response to vaccination. The 9 patients enrolled for phase I were vaccinated 6 times, once every 2 weeks: 3 patients with 100 µg and 3 patients with 300 µg CHP-MAGE-A4, and 3 patients with 300 µg CHP-MAGE-A4 plus 0.5 clinical units of OK-432. Toxicities were assessed using Common Terminology Criteria for Adverse Events v3.0. Clinical response was evaluated by modified Response Evaluation Criteria in Solid Tumours. Immunological monitoring of anti-MAGE-A4-specific antibodies was performed by ELISA of pre- and post-vaccination patient sera. The 6 vaccinations produced no severe adverse events. Stable disease was assessed in 4/9 patients. Anti-MAGE-A4 total immunoglobulin (Ig)G titers increased in 7/9 patients. Efficacious anti-MAGE-A4 IgG1, 2 and 3 antibody responses were observed in 7/9 patients. Among them, positive conversions to T helper 2 (Th2)-type antibody responses (IgG4 and IgE) were observed after frequent vaccination in 4/7 patients. The Th2 conversion was possibly associated with undesirable clinical observations, including progressive disease and the appearance of a new relapse lesion. The present study suggested that frequent vaccinations activated a Th2-dominant status in the cancer patients. The identification of a time-dependent IgG subclass and IgE antibody production during vaccination protocols may be a useful surrogate marker indicating a potentially undesirable change of the immunological environment for an effective antitumor immune response in cancer patients. PMID:28105158

  5. Characterization of a glycoside hydrolase family-51 α-l-arabinofuranosidase gene from Aureobasidium pullulans ATCC 20524 and its encoded product.

    PubMed

    Ohta, Kazuyoshi; Fujii, Shinya; Higashida, Chihiro

    2013-09-01

    The genomic DNA and cDNA encoding α-l-arabinofuranosidase were cloned from the dimorphic fungus Aureobasidium pullulans ATCC 20524 and sequenced. The open reading frame (2097 bp) of the α-l-arabinofuranosidase gene abfB was interrupted by five introns of 49, 49, 50, 65, and 49 bp. The gene encoded a presumed signal peptide of 17 residues and a mature protein of 682 residues with a calculated Mr of 74,230 Da and a theoretical isoelectric point of 4.95. Glu-362 and Glu-440 residues are likely involved in catalytic reactions as an acid/base and a nucleophile, respectively. The protein possessed 15 potential N-glycosylation sites. The deduced amino acid sequence of the abfB gene product was 58% identical to the Penicillium purpurogenum ABF 2, which belongs to the glycoside hydrolase family-51 α-l-arabinofuranosidase. The abfB cDNA was functionally expressed in the yeast Pichia pastoris. The recombinant enzyme, AbfB, was purified from the culture filtrate, and it appeared as a single band on SDS-PAGE with an apparent Mr of 110 kDa. AbfB showed α-l-arabinofuranosidase activity of 56.6 U/mg of protein toward p-nitrophenyl (pNP) α-l-arabinofuranoside at optimal pH 4.5 and 75°C. The enzyme exhibited apparent Km and Vmax values of 6.27 mM and 78.1 μmol/mg/min, respectively, for pNP α-l-arabinofuranoside. The enzyme was highly active on rye arabinoxylan as well as pNP α-l-arabinofuranoside, but it showed weak activity toward α-(1→5)-l-arabinobiose, α-(1→5)-l-arabinotriose, branched l-arabinan, linear α-(1→5)-l-arabinan, and arabinogalactan.

  6. Characterization of N-glycan structures and biofunction of anti-colorectal cancer monoclonal antibody CO17-1A produced in baculovirus-insect cell expression system.

    PubMed

    Song, Mira; Park, Da-Young; Kim, Youngkwan; Lee, Kyung-Jin; Lu, Zhe; Ko, Kinarm; Choo, Young Kug; Han, Yeon Soo; Ahn, Mi-Hyun; Oh, Doo-Byoung; Ko, Kisung

    2010-08-01

    Advantages of the baculovirus insect cell expression system for production of recombinant proteins include high capacity, flexibility, and glycosylation capability. In this study, this expression system was exploited to produce anti-cancer monoclonal antibody (mAb) CO17-1A, which recognizes the antigen GA733. The heavy chain (HC) and light chain (LC) genes of mAb CO17-1A were cloned under the control of P(10) and Polyhedrin promoters in the pFastBac dual vector, respectively. Gene expression cassettes carrying the HC and LC genes were transposed into a bacmid in Escherichia coli (DH10Bac). The transposed bacmid was transfected to Sf9 insect cells to generate baculovirus expressing mAb CO17-1A. Confocal immunofluorescence and Western blot analyses confirmed expression of mAb CO17-1A in baculovirus-infected insect cells. The optimum conditions for mAb expression were evaluated at 24, 48, and 72 h after the virus infection at an optimum virus multiplicity of infection of 1. Expression of mAb CO17-1A in insect cells significantly increased at 72 h after infection. HPLC analysis of glycosylation status revealed that the insect-derived mAb (mAb(I)) CO17-1A had insect specific glycan structures. ELISA showed that the purified mAb(I) from cell culture supernatant specifically bound to SW948 human colorectal cancer cells. Fluorescence-activated cell sorting analysis showed that, although mAb(I) had insect specific glycan structures that differed from their mammalian counterparts, mAb(I) similarly interacted with CD64 (FcgammaRI) and Fc of IgG, compared to the interactions of mammalian-derived mAb. These results suggest that the baculovirus insect cell expression system is able to express, assemble, and secrete biofunctional full size mAb.

  7. A small scale study on the effects of oral administration of the β-glucan produced by Aureobasidium pullulans on milk quality and cytokine expressions of Holstein cows, and on bacterial flora in the intestines of Japanese black calves

    PubMed Central

    2012-01-01

    Background The β–(1 → 3),(1 → 6)-D-glucan extracellularly produced by Aureobasidium pullulans exhibits immunomodulatory activity, and is used for health supplements. To examine the effects of oral administration of the β–(1 → 3),(1 → 6)-D-glucan to domestic animals, a small scale study was conducted using Holstein cows and newborn Japanese Black calves. Findings Holstein cows of which somatic cell count was less than 3 x 105/ml were orally administered with or without the β-(1 → 3),(1 → 6)-D-glucan-enriched A. pullulans cultured fluid (AP-CF) for 3 months, and the properties of milk and serum cytokine expression were monitored. Somatic cell counts were not significantly changed by oral administration of AP-CF, whereas the concentration of solid non fat in the milk tended to increase in the AP-CF administered cows. The results of cytokine expression analysis in the serum using ELISA indicate that the expressions of tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 in all cows which were orally administered with AP-CF became slightly lower than that of control cows after the two-month treatment. On the other hand, IL-8 expression tended to indicate a moderately higher level in all treated cows after the three-month administration of AP-CF in comparison with that of the control cows. Peripartum Japanese Black beef cows and their newborn calves were orally administered with AP-CF, and bacterial flora in the intestines of the calves were analyzed by T-RFLP (terminal restriction fragment length polymorphism). The results suggest that bacterial flora are tendentiously changed by oral administration of AP-CF. Conclusions Our data indicated the possibility that oral administration of the β–(1 → 3),(1 → 6)-D- glucan produced by A. pullulans affects cytokine expressions in the serum of Holstein cows, and influences bacterial flora in the intestines of Japanese Black calves. The findings may be helpful for

  8. Antibody responses against NY-ESO-1 and HER2 antigens in patients vaccinated with combinations of cholesteryl pullulan (CHP)-NY-ESO-1 and CHP-HER2 with OK-432.

    PubMed

    Aoki, Masatoshi; Ueda, Shugo; Nishikawa, Hiroyoshi; Kitano, Shigehisa; Hirayama, Michiko; Ikeda, Hiroaki; Toyoda, Hideki; Tanaka, Kyosuke; Kanai, Michiyuki; Takabayashi, Arimichi; Imai, Hiroshi; Shiraishi, Taizo; Sato, Eiichi; Wada, Hisashi; Nakayama, Eiichi; Takei, Yoshiyuki; Katayama, Naoyuki; Shiku, Hiroshi; Kageyama, Shinichi

    2009-11-16

    Combination vaccines of the NY-ESO-1 protein complexed with cholesteryl pullulan (CHP), CHP-NY-ESO-1, and the truncated 146HER2 protein with CHP, CHP-HER2, were subcutaneously administered with the immuno-adjuvant OK-432 to eight esophageal cancer patients. Vaccination was well-tolerated. NY-ESO-1- and HER2-specific antibody responses were analyzed using the patients' sera and samples from previous single CHP-NY-ESO-1 or CHP-HER2 vaccine trial. The responses to NY-ESO-1 in the combination vaccine study were comparable to the single vaccine. For responses to HER2, there were fewer antibody responses in the combination vaccines. Although there were marked individual variations in the antibody responses to the NY-ESO-1 and HER2 antigens, the reaction patterns to these antigens were comparable within each patient. Antibodies to OK-432 were not augmented. Protein cancer vaccines targeting multiple antigens are feasible.

  9. Genome sequence of "Candidatus Microthrix parvicella" Bio17-1, a long-chain-fatty-acid-accumulating filamentous actinobacterium from a biological wastewater treatment plant.

    PubMed

    Muller, Emilie E L; Pinel, Nicolás; Gillece, John D; Schupp, James M; Price, Lance B; Engelthaler, David M; Levantesi, Caterina; Tandoi, Valter; Luong, Khai; Baliga, Nitin S; Korlach, Jonas; Keim, Paul S; Wilmes, Paul

    2012-12-01

    "Candidatus Microthrix" bacteria are deeply branching filamentous actinobacteria which occur at the water-air interface of biological wastewater treatment plants, where they are often responsible for foaming and bulking. Here, we report the first draft genome sequence of a strain from this genus: "Candidatus Microthrix parvicella" strain Bio17-1.

  10. Inhibitory effects of water-soluble low-molecular-weight beta-(1,3-1,6) d-glucan purified from Aureobasidium pullulans GM-NH-1A1 strain on food allergic reactions in mice.

    PubMed

    Kimura, Yoshiyuki; Sumiyoshi, Maho; Suzuki, Takahiro; Suzuki, Toshio; Sakanaka, Masahiro

    2007-07-01

    There have been a number of reports showing that the crude beta-glucan fraction prepared from various kinds of Basidiomycetes mushrooms acts as anti-cancer and anti-allergic reagent through stimulation of IFN-gamma production. It has been reported, however, that the exposure of the airway to beta-(1,3) d-glucan, contained in house dust, indoor moulds and some bacteria, potentiates the airway allergic response. It seems likely that the discrepant effects on immune function may be related to such factors as differences of the administration route, average molecular weight and water solubility. We isolated a new low-molecular-weight (about 100 kDa) beta-glucan from Aureobasidium pullulans 1A1 strain of black yeast, and found that it had low viscosity and was water-soluble. In this study, we examined the effects of water-soluble low-molecular-weight beta-(1-->3) and 50-80% branched beta-(1-->6) glucan (LMW-beta-Glucan) isolated from A. pullulans on the ova-albumin (OVA)-treated allergic reaction in mice. Feeding standard laboratory diets containing 0.5 and 1% LMW-beta-Glucan significantly inhibited the OVA-specific IgE elevation compared to that in OVA-sensitized mice fed standard laboratory diet alone (control). Furthermore, feeding standard laboratory diets containing 0.5 and 1% LMW-beta-Glucan inhibited the reduction of IL-12 and IFN-gamma production from splenocytes and the reduction of CD8- and IFN-gamma-positive cell number in the small intestine of the OVA-sensitized mice. These findings suggest that anti-food allergic action of LMW-beta-Glucan may be due to the inducing IFN-gamma production in the small intestine and splenocytes.

  11. Correlation of maple sap composition with bacterial and fungal communities determined by multiplex automated ribosomal intergenic spacer analysis (MARISA).

    PubMed

    Filteau, Marie; Lagacé, Luc; LaPointe, Gisèle; Roy, Denis

    2011-08-01

    During collection, maple sap is contaminated by bacteria and fungi that subsequently colonize the tubing system. The bacterial microbiota has been more characterized than the fungal microbiota, but the impact of both components on maple sap quality remains unclear. This study focused on identifying bacterial and fungal members of maple sap and correlating microbiota composition with maple sap properties. A multiplex automated ribosomal intergenic spacer analysis (MARISA) method was developed to presumptively identify bacterial and fungal members of maple sap samples collected from 19 production sites during the tapping period. Results indicate that the fungal community of maple sap is mainly composed of yeast related to Mrakia sp., Mrakiella sp., Guehomyces pullulans, Cryptococcus victoriae and Williopsis saturnus. Mrakia, Mrakiella and Guehomyces peaks were identified in samples of all production sites and can be considered dominant and stable members of the fungal microbiota of maple sap. A multivariate analysis based on MARISA profiles and maple sap chemical composition data showed correlations between Candida sake, Janthinobacterium lividum, Williopsis sp., Leuconostoc mesenteroides, Mrakia sp., Rhodococcus sp., Pseudomonas tolaasii, G. pullulans and maple sap composition at different flow periods. This study provides new insights on the relationship between microbial community and maple sap quality. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. Micrometastatic cancer cells in bone marrow: in vitro detection with anti-cytokeratin and in vivo labeling with anti-17-1A monoclonal antibodies

    SciTech Connect

    Schlimok, G.; Funke, I.; Holzmann, B.; Goettlinger, G.; Schmidt, G.; Haeser, H.; Swierkot, S.; Warnecke, H.H.; Schneider, B.; Koprowski, H.; Riethmueller, G.

    1987-12-01

    The detection of early micrometastasis or disseminated single tumor cells poses a problem for conventional diagnosis procedures. Using a panel of monoclonal antibodies against cytokeratin and the 17-1A epithelial antigen the authors identified immunocytochemically tumor cells in bone marrow of patients with breast cancer and colorectal cancer at the time of surgery of the primary tumor. Monoclonal antibody CK2, recognizing the human cytokeratin component 18 in simple epithelia, appeared to be the most suitable reagent because of its negative reaction with bone marrow samples of the noncarcinoma patients. Its specificity was further demonstrated in a double-marker staining procedure using an anti-leukocyte common antigen monoclonal antibody (T200) as counterstain. A comparative analysis showed that immunocytology was clearly superior to conventional cytology and histology. In 9.5-20.5% of patients without distant metastasis, tumor cells could be detected in bone marrow. They found a significant correlation between tumor cells in bone marrow and conventional risk factors, such as distant metastasis or lymph node involvement. In a first approach toward immunotherapy they demonstrated in 3 patients that infused monoclonal antibody 17-1A can label single tumor cells in bone marrow in vivo. They then used this single approach to follow up on 7 patients undergoing 17-1A therapy in an adjuvant clinical trial.

  13. Selective chromosomal damage and cytotoxicity of sup 125 I-labeled monoclonal antibody 17-1a in human cancer cells

    SciTech Connect

    Woo, D.V.; Li, D.; Mattis, J.A.; Steplewski, Z. )

    1989-06-01

    A monoclonal antibody, 17-1a, which reacts with antigen expressed in human colon cancers was radiolabeled in high specific activity with {sup 125}I. The combination of the antibody and this radionuclide was observed to elicit specific cellular damage after being internalized into cells of the SW1116 human colon cancer cell line. The degree of internalization was quantitatively measured and found to increase over time to 49% after a 48-h incubation period. During this period, significant chromosome aberrations were observed in the SW1116 cell line due to the Auger electrons of {sup 125}I. This damage was not observed using Na{sup 125}I, a nonimmunoreactive radiolabeled antibody, or cells which did not contain the requisite antigen. The number of chromosomal aberrations increased with increasing radioactive concentration of {sup 125}I-17-1a. The nuclear damage resulted in specific cellular cytotoxicity and decreased cell survival of SW1116 cells exposed to various concentrations of {sup 125}I-17-1a.

  14. Effect of monoclonal antibody 17-1A and GM-CSF in patients with advanced colorectal carcinoma--long-lasting, complete remissions can be induced.

    PubMed

    Ragnhammar, P; Fagerberg, J; Frödin, J E; Hjelm, A L; Lindemalm, C; Magnusson, I; Masucci, G; Mellstedt, H

    1993-03-12

    Antibody-dependent cellular cytotoxicity (ADCC) is considered to be one of the effector functions of unconjugated monoclonal antibodies (MAbs) in tumor therapy. The antitumor activity of MAbs might therefore be augmented if the cytotoxic capability of the effector cells could be increased. In an in vitro system, the killing capacity of MAb was significantly enhanced by pre-treatment of the effector cells with granulocyte-macrophage colony-stimulating factor (GM-CSF). Based on these findings, the therapeutic effect of the combination of mouse MAb 17-1A (IgG2a) and GM-CSF was evaluated in 20 patients with metastatic colorectal carcinoma (CRC). The patients received GM-CSF for 10 days and a single i.v. infusion of MAb 17-1A on day 3 of the cycle. Four cycles were given at 1-monthly intervals. There was a continuous increase in blood monocytes and lymphocytes during all 4 GM-CSF cycles. Neutrophils and eosinophils were also significantly augmented but in a biphasic manner and the cell counts on day 10 of cycle IV were significantly lower than in cycles I and II. GM-CSF-related side-effects were of no major clinical importance. During the third cycle, an immediate-type allergic reaction (ITAR) against MAb 17-1A occurred in most patients, necessitating reduction of the MAb dose as well as of the infusion rate. Two patients achieved complete remission. One patient had a minor response, and 3 other patients were considered to have stable disease > 3 months.

  15. Single-step purification and characterization of an extreme halophilic, ethanol tolerant and acidophilic xylanase from Aureobasidium pullulans NRRL Y-2311-1 with application potential in the food industry.

    PubMed

    Yegin, Sirma

    2017-04-15

    An extracellular xylanase from Aureobasidium pullulans NRRL Y-2311-1 produced on wheat bran was purified by a single-step chromatographic procedure. The enzyme had a molecular weight of 21.6kDa. The optimum pH and temperature for xylanase activity were 4.0 and 30-50°C, respectively. The enzyme was stable in the pH range of 3.0-8.0. The inactivation energy of the enzyme was calculated as 218kJmol(-1). The xylanase was ethanol tolerant and kept complete activity in the presence of 10% ethanol. Likewise, it retained almost complete activity at a concentration range of 0-20% NaCl. In general, the enzyme was resistant to several metal ions and reagents. Mg(2+), Zn(2+), Cu(2+), K(1+), EDTA and β-mercaptoethanol resulted in enhanced xylanase activity. The Km and Vmax values on beechwood xylan were determined to be 19.43mgml(-1) and 848.4Uml(-1), respectively. The enzyme exhibits excellent characteristics and could, therefore, be a promising candidate for application in food and bio-industries. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Growth suppression of colorectal cancer by plant-derived multiple mAb CO17-1A × BR55 via inhibition of ERK1/2 phosphorylation.

    PubMed

    Kwak, Dong Hoon; Moussavou, Ghislain; Lee, Ju Hyoung; Heo, Sung Youn; Ko, Kisung; Hwang, Kyung-A; Jekal, Seung-Joo; Choo, Young-Kug

    2014-11-14

    We have generated the transgenic Tabaco plants expressing multiple monoclonal antibody (mAb) CO7-1A × BR55 by cross-pollinating with mAb CO17-1A and mAb BR55. We have demonstrated the anti-cancer effect of plant-derived multiple mAb CO17-1A × BR55. We find that co-treatment of colorectal mAbs (anti-epithelial cellular adhesion molecule (EpCAM), plant-derived monoclonal antibody (mAb(P)) CO17-1A and mAb(P) CO17-1A × BR55) with RAW264.7 cells significantly inhibited the cell growth in SW620 cancer cells. In particular, multi mAb(P) CO17-1A × BR55 significantly and efficiently suppressed the growth of SW620 cancer cells compared to another mAbs. Apoptotic death-positive cells were significantly increased in the mAb(P) CO17-1A × BR55-treated. The mAb(P) CO17-1A × BR55 treatment significantly decreased the expression of B-Cell lymphoma-2 (BCl-2), but the expression of Bcl-2-associated X protein (Bax), and cleaved caspase-3 were markedly increased. In vivo, the mAb(P) CO17-1A × BR55 significantly and efficiently inhibited the growth of colon tumors compared to another mAbs. The apoptotic cell death and inhibition of pro-apoptotic proteins expression were highest by treatment with mAb(P) CO17-1A × BR55. In addition, the mAb(P) CO17-1A × BR55 significantly inhibited the extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation in cancer cells and tumors. Therefore, this study results suggest that multiple mAb(P) CO17-1A × BR55 has a significant effect on apoptosis-mediated anticancer by suppression of ERK1/2 phosphorylation in colon cancer compared to another mAbs. In light of these results, further clinical investigation should be conducted on mAb(P) CO17-1A × BR55 to determine its possible chemopreventive and/or therapeutic efficacy against human colon cancer.

  17. Growth Suppression of Colorectal Cancer by Plant-Derived Multiple mAb CO17-1A × BR55 via Inhibition of ERK1/2 Phosphorylation

    PubMed Central

    Kwak, Dong Hoon; Moussavou, Ghislain; Lee, Ju Hyoung; Heo, Sung Youn; Ko, Kisung; Hwang, Kyung-A; Jekal, Seung-Joo; Choo, Young-Kug

    2014-01-01

    We have generated the transgenic Tabaco plants expressing multiple monoclonal antibody (mAb) CO7-1A × BR55 by cross-pollinating with mAb CO17-1A and mAb BR55. We have demonstrated the anti-cancer effect of plant-derived multiple mAb CO17-1A × BR55. We find that co-treatment of colorectal mAbs (anti-epithelial cellular adhesion molecule (EpCAM), plant-derived monoclonal antibody (mAbP) CO17-1A and mAbP CO17-1A × BR55) with RAW264.7 cells significantly inhibited the cell growth in SW620 cancer cells. In particular, multi mAbP CO17-1A × BR55 significantly and efficiently suppressed the growth of SW620 cancer cells compared to another mAbs. Apoptotic death-positive cells were significantly increased in the mAbP CO17-1A × BR55-treated. The mAbP CO17-1A × BR55 treatment significantly decreased the expression of B-Cell lymphoma-2 (BCl-2), but the expression of Bcl-2-associated X protein (Bax), and cleaved caspase-3 were markedly increased. In vivo, the mAbP CO17-1A × BR55 significantly and efficiently inhibited the growth of colon tumors compared to another mAbs. The apoptotic cell death and inhibition of pro-apoptotic proteins expression were highest by treatment with mAbP CO17-1A × BR55. In addition, the mAbP CO17-1A × BR55 significantly inhibited the extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation in cancer cells and tumors. Therefore, this study results suggest that multiple mAbP CO17-1A × BR55 has a significant effect on apoptosis-mediated anticancer by suppression of ERK1/2 phosphorylation in colon cancer compared to another mAbs. In light of these results, further clinical investigation should be conducted on mAbP CO17-1A × BR55 to determine its possible chemopreventive and/or therapeutic efficacy against human colon cancer. PMID:25405740

  18. Dose-dependent effects of NY-ESO-1 protein vaccine complexed with cholesteryl pullulan (CHP-NY-ESO-1) on immune responses and survival benefits of esophageal cancer patients.

    PubMed

    Kageyama, Shinichi; Wada, Hisashi; Muro, Kei; Niwa, Yasumasa; Ueda, Shugo; Miyata, Hiroshi; Takiguchi, Shuji; Sugino, Sahoko H; Miyahara, Yoshihiro; Ikeda, Hiroaki; Imai, Naoko; Sato, Eiichi; Yamada, Tomomi; Osako, Masaharu; Ohnishi, Mami; Harada, Naozumi; Hishida, Tadashi; Doki, Yuichiro; Shiku, Hiroshi

    2013-10-05

    Cholesteryl pullulan (CHP) is a novel antigen delivery system for cancer vaccines. This study evaluated the safety, immune responses and clinical outcomes of patients who received the CHP-NY-ESO-1 complex vaccine, Drug code: IMF-001. Patients with advanced/metastatic esophageal cancer were enrolled and subcutaneously vaccinated with either 100 μg or 200 μg of NY-ESO-1 protein complexed with CHP. The primary endpoints were safety and humoral immune responses, and the secondary endpoint was clinical efficacy. A total of 25 patients were enrolled. Thirteen and twelve patients were repeatedly vaccinated with 100 μg or 200 μg of CHP-NY-ESO-1 with a median of 8 or 9.5 doses, respectively. No serious adverse events related to the vaccine were observed. Three out of 13 patients in the 100-μg cohort and 7 out of 12 patients in the 200-μg cohort were positive for anti-NY-ESO-1 antibodies at baseline. In the 100-μg cohort, an antibody response was observed in 5 out of 10 pre-antibody-negatives patients, and the antibody levels were augmented in 2 pre-antibody-positive patients after vaccination. In the 200-μg cohort, all 5 pre-antibody-negative patients became seropositive, and the antibody level was amplified in all 7 pre-antibody-positive patients. No tumor shrinkage was observed. The patients who received 200 μg of CHP-NY-ESO-1 survived longer than patients receiving 100 μg of CHP-NY-ESO-1, even those who exhibited unresponsiveness to previous therapies or had higher tumor burdens. The safety and immunogenicity of CHP-NY-ESO-1 vaccine were confirmed. The 200 μg dose more efficiently induced immune responses and suggested better survival benefits. (Clinical trial registration number NCT01003808).

  19. Comparison of monoclonal antibodies 17-1A and 323/A3: the influence of the affinity on tumour uptake and efficacy of radioimmunotherapy in human ovarian cancer xenografts.

    PubMed Central

    Kievit, E.; Pinedo, H. M.; Schlüper, H. M.; Haisma, H. J.; Boven, E.

    1996-01-01

    The low-affinity monoclonal antibody (MAb) chimeric 17-1A(c-17-1A) and the high-affinity MAb mouse 323/A3 (m-323/A3) were used to study the effect of the MAb affinity on the tumour uptake and efficacy of radioimmunotherapy in nude mice bearing subcutaneously the human ovarian cancer xenografts FMa, OVCAR-3 and Ov.Pe. Both MAbs are directed against the same pancarcinoma glycoprotein. In vitro, the number of binding sites on tumour cells at 4 degrees C was similar for both MAbs, but m-323/A3 had an approximately 5-fold higher affinity (1.3-3.0x10(9) M-1) than c-17-1A (3.0-5.4x10(8) M-1). This difference in affinity was more extreme at 37 degrees C, when no binding of c-17-1A could be observed. MAb m-323/A3 completely blocked binding of c-17-1A to tumour cells, whereas the reverse was not observed. Immunohistochemistry showed a similar but more intense staining pattern of m-323/A3 in human ovarian cancer xenografts than of c-17-1A. In vivo, the blood clearance in non-tumour-bearing nude mice was similar for both MAbs with terminal half-lives of 71.4 h for m-323/A3 and 62.7 h for c-17-1A. MAb m-323/A3 targeted better to tumour tissue, but was more heterogeneously distributed than c-17-1A. The cumulative absorbed radiation dose delivered by m-323/A3 to tumour tissue was 2.5- to 4.7-fold higher than that delivered by c-17-1A. When mice were treated with equivalent radiation doses of 131(I)m-323/A3 and 131(I)c-17-1A, based on a correction for the immunoreactivity of the radiolabelled MAbs, m-323/A3 induced a better growth inhibition in two of the three xenografts. When the radiation doses were adjusted to obtain a similar amount of radiation in the tumour c-17-1A was more effective in tumour growth inhibition in all three xenografts. Images Figure 3 Figure 4 PMID:8595159

  20. Dose-dependent effects of NY-ESO-1 protein vaccine complexed with cholesteryl pullulan (CHP-NY-ESO-1) on immune responses and survival benefits of esophageal cancer patients

    PubMed Central

    2013-01-01

    Background Cholesteryl pullulan (CHP) is a novel antigen delivery system for cancer vaccines. This study evaluated the safety, immune responses and clinical outcomes of patients who received the CHP-NY-ESO-1 complex vaccine, Drug code: IMF-001. Methods Patients with advanced/metastatic esophageal cancer were enrolled and subcutaneously vaccinated with either 100 μg or 200 μg of NY-ESO-1 protein complexed with CHP. The primary endpoints were safety and humoral immune responses, and the secondary endpoint was clinical efficacy. Results A total of 25 patients were enrolled. Thirteen and twelve patients were repeatedly vaccinated with 100 μg or 200 μg of CHP-NY-ESO-1 with a median of 8 or 9.5 doses, respectively. No serious adverse events related to the vaccine were observed. Three out of 13 patients in the 100-μg cohort and 7 out of 12 patients in the 200-μg cohort were positive for anti-NY-ESO-1 antibodies at baseline. In the 100-μg cohort, an antibody response was observed in 5 out of 10 pre-antibody-negatives patients, and the antibody levels were augmented in 2 pre-antibody-positive patients after vaccination. In the 200-μg cohort, all 5 pre-antibody-negative patients became seropositive, and the antibody level was amplified in all 7 pre-antibody-positive patients. No tumor shrinkage was observed. The patients who received 200 μg of CHP-NY-ESO-1 survived longer than patients receiving 100 μg of CHP-NY-ESO-1, even those who exhibited unresponsiveness to previous therapies or had higher tumor burdens. Conclusions The safety and immunogenicity of CHP-NY-ESO-1 vaccine were confirmed. The 200 μg dose more efficiently induced immune responses and suggested better survival benefits. (Clinical trial registration number NCT01003808). PMID:24093426

  1. The influence of Aster x salignus Willd. Invasion on the diversity of soil yeast communities

    NASA Astrophysics Data System (ADS)

    Glushakova, A. M.; Kachalkin, A. V.; Chernov, I. Yu.

    2016-07-01

    The annual dynamics of yeast communities were studied in the soddy-podzolic soil under the thickets of Aster x salignus Willd., one of the widespread invasive plant species in central Russia. Yeast groups in the soils under continuous aster thickets were found to differ greatly from the yeast communities in the soils under the adjacent indigenous meadow vegetation. In both biotopes the same species ( Candida vartiovaarae, Candida sake, and Cryptococcus terreus) are dominants. However, in the soils under indigenous grasses, eurybiontic yeasts Rhodotorula mucilaginosa, which almost never occur in the soil under aster, are widespread. In the soil under aster, the shares of other typical epiphytic and pedobiontic yeast fungi (ascomycetic species Wickerhamomyces aniomalus, Barnettozyma californica and basidiomycetic species Cystofilobasidium macerans, Guehomyces pullulans) significantly increase. Thus, the invasion of Aster x salignus has a clear effect on soil yeast complexes reducing their taxonomic and ecological diversity.

  2. Anti-EpCAM monoclonal antibody (MAb17-1A) based treatment combined with alpha-interferon, 5-fluorouracil and granulocyte-macrophage colony-stimulating factor in patients with metastatic colorectal carcinoma.

    PubMed

    Liljefors, Maria; Ragnhammar, Peter; Nilsson, Bo; Ullenhag, Gustav; Mellstedt, Håkan; Frödin, Jan-Erik

    2004-09-01

    Monoclonal antibodies (MAbs) have different modes of action and toxicity profile compared to chemotherapeutics, which makes it interesting to combine these drugs. Addition of cytokines to MAb therapy may also augment immune effector functions utilized by MAb. In an effort to improve the therapeutic effect of a MAb-based regimen in colorectal carcinoma (CRC) patients, the effects of a combination of alpha-interferon (alpha-IFN), 5-fluorouracil (5-FU), granulocyte-macrophage colony-stimulating factor (GM-CSF) and mouse MAb17-1A was evaluated in 27 patients with metastatic disease. alpha-IFN was given s.c. once daily for 5 consecutive days and at days 4 and 5, 5-FU was administered as a daily i.v. bolus injection. After 2 days rest, GM-CSF was given s.c. once daily, days 8-14 and on day 10, MAb17-1A was given i.v. The treatment cycle was repeated every 4th week. One patient achieved a partial remission and 13 patients showed a minor response or stable disease >3 months, inducing an overall response rate of 54%. Responding patients survived significantly longer than non-responding patients (p=0.021). Median overall survival time for all patients was 75 weeks and progression-free survival time 15 weeks. Adverse events related to alpha-IFN, GM-CSF and 5-FU were as expected. The frequency of patients with an immediate-type allergic reaction (ITAR) against MAb17-1A at the 1st, 2nd, 3rd and 4th treatment cycles was 11%, 52%, 62% and 64% respectively. The planned MAb17-1A dose had to be reduced by repeated infusions. No patient received full dose of MAb17-1A from the 3rd cycle and onward. Compared to historical control patients treated with MAb17-1A alone, the present combination regimen seemed to improve the response rate (54% vs 15%) as well as progression-free survival (15 vs 7 weeks; p<0.05).

  3. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... act that relate to prescription drug marketing practices. (b) Section 303(f)(1) of the act authorizing... mitigation strategies for drugs. (d) Section 303(g)(1) of the act authorizing civil money penalties...

  4. 7 CFR 17.1 - General.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ..., as amended (hereinafter called “the Act”). (b) Agricultural commodities agreements. (1) Under the Act, the Government of the United States enters into Agricultural Commodities Agreements with...

  5. 32 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... conduct of trials by a military commission appointed pursuant to 32 CFR part 9 and Military Order of November 13, 2001, “Detention, Treatment, and Trial of Certain Non-Citizens in the War Against Terrorism...

  6. 32 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... conduct of trials by a military commission appointed pursuant to 32 CFR part 9 and Military Order of November 13, 2001, “Detention, Treatment, and Trial of Certain Non-Citizens in the War Against Terrorism...

  7. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation and mitigation strategies for drugs. (d) Section 303(g)(1) of the act authorizing civil money penalties for...

  8. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation and mitigation strategies for drugs. (d) Section 303(g)(1) of the act authorizing civil money penalties for...

  9. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation and mitigation strategies for drugs. (d) Section 303(g)(1) of the act authorizing civil money penalties for...

  10. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation and mitigation strategies for drugs. (d) Section 303(g)(1) of the act authorizing civil money penalties for...

  11. 36 CFR 17.1 - Authority.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... acquired by the Secretary from non-Federal sources within any unit of the National Park System except national parks and those national monuments of scientific significance. This legislation is referred to as...

  12. 36 CFR 17.1 - Authority.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... acquired by the Secretary from non-Federal sources within any unit of the National Park System except national parks and those national monuments of scientific significance. This legislation is referred to as...

  13. Maple sap predominant microbial contaminants are correlated with the physicochemical and sensorial properties of maple syrup.

    PubMed

    Filteau, Marie; Lagacé, Luc; Lapointe, Gisèle; Roy, Denis

    2012-03-01

    Maple sap processing and microbial contamination are significant aspects that affect maple syrup quality. In this study, two sample sets from 2005 and 2008 were used to assess the maple syrup quality variation and its relationship to microbial populations, with respect to processing, production site and harvesting period. The abundance of maple sap predominant bacteria (Pseudomonas fluorescens group and two subgroups, Rahnella spp., Janthinobacterium spp., Leuconostoc mesenteroides) and yeast (Mrakia spp., Mrakiella spp.,Guehomyces pullulans) was assessed by quantitative PCR. Maple syrup properties were analyzed by physicochemical and sensorial methods. Results indicate that P. fluorescens, Mrakia spp., Mrakiella spp. G. pullulans and Rahnella spp. are stable contaminants of maple sap, as they were found for every production site throughout the flow period. Multiple factor analysis reports a link between the relative abundance of P. fluorescens group and Mrakia spp. in maple sap with maple and vanilla odor as well as flavor of maple syrup. This evidence supports the contribution of these microorganisms or a consortium of predominant microbial contaminants to the characteristic properties of maple syrup.

  14. Cyclostratigraphy and eccentricity tuning of the early Oligocene through early Miocene (30.1-17.1 Ma): Cibicides mundulus stable oxygen and carbon isotope records from Walvis Ridge Site 1264

    NASA Astrophysics Data System (ADS)

    Liebrand, Diederik; Beddow, Helen M.; Lourens, Lucas J.; Pälike, Heiko; Raffi, Isabella; Bohaty, Steven M.; Hilgen, Frederik J.; Saes, Mischa J. M.; Wilson, Paul A.; van Dijk, Arnold E.; Hodell, David A.; Kroon, Dick; Huck, Claire E.; Batenburg, Sietske J.

    2016-09-01

    Few astronomically calibrated high-resolution (≤5 kyr) climate records exist that span the Oligocene-Miocene time interval. Notably, available proxy records show responses varying in amplitude at frequencies related to astronomical forcing, and the main pacemakers of global change on astronomical time-scales remain debated. Here we present newly generated X-ray fluorescence core scanning and benthic foraminiferal stable oxygen and carbon isotope records from Ocean Drilling Program Site 1264 (Walvis Ridge, southeastern Atlantic Ocean). Complemented by data from nearby Site 1265, the Site 1264 benthic stable isotope records span a continuous ∼13-Myr interval of the Oligo-Miocene (30.1-17.1 Ma) at high resolution (∼3.0 kyr). Spectral analyses in the stratigraphic depth domain indicate that the largest amplitude variability of all proxy records is associated with periods of ∼ 3.4 m and ∼ 0.9 m, which correspond to 405- and ∼110-kyr eccentricity, using a magnetobiostratigraphic age model. Maxima in CaCO3 content, δ18O and δ13C are interpreted to coincide with ∼110 kyr eccentricity minima. The strong expression of these cycles in combination with the weakness of the precession- and obliquity-related signals allow construction of an astronomical age model that is solely based on tuning the CaCO3 content to the nominal (La2011_ecc3L) eccentricity solution. Very long-period eccentricity maxima (∼2.4-Myr) are marked by recurrent episodes of high-amplitude ∼110-kyr δ18O cycles at Walvis Ridge, indicating greater sensitivity of the climate/cryosphere system to short eccentricity modulation of climatic precession. In contrast, the responses of the global (high-latitude) climate system, cryosphere, and carbon cycle to the 405-kyr cycle, as expressed in benthic δ18O and especially δ13C signals, are more pronounced during ∼2.4-Myr minima. The relationship between the recurrent episodes of high-amplitude ∼110-kyr δ18O cycles and the ∼1.2-Myr amplitude

  15. Yeast and yeast-like diversity in the southernmost glacier of Europe (Calderone Glacier, Apennines, Italy).

    PubMed

    Branda, Eva; Turchetti, Benedetta; Diolaiuti, Guglielmina; Pecci, Massimo; Smiraglia, Claudio; Buzzini, Pietro

    2010-06-01

    The present study reports the characterization of psychrophilic yeast and yeast-like diversity in cold habitats (superficial and deep sediments, ice cores and meltwaters) of the Calderone Glacier (Italy), which is the southernmost glacier in Europe. After incubation at 4 and 20 degrees C, sediments contained about 10(2)-10(3) CFU of yeasts g(-1). The number of viable yeast cells in ice and meltwaters was several orders of magnitude lower. The concomitant presence of viable bacteria and filamentous fungi has also been observed. In all, 257 yeast strains were isolated and identified by 26S rRNA gene D1/D2 and internal transcribed spacers (1 and 2) sequencing as belonging to 28 ascomycetous and basidiomycetous species of 11 genera (Candida, Cystofilobasidium, Cryptococcus, Dioszegia, Erythrobasidium, Guehomyces, Mastigobasidium, Mrakia, Mrakiella, Rhodotorula and Sporobolomyces). Among them, the species Cryptococcus gastricus accounted for almost 40% of the total isolates. In addition, 12 strains were identified as belonging to the yeast-like species Aureobasidium pullulans and Exophiala dermatitidis, whereas 15 strains, presumably belonging to new species, yet to be described, were also isolated. Results herein reported indicate that the Calderone Glacier, although currently considered a vanishing ice body due to the ongoing global-warming phenomenon, still harbors viable psychrophilic yeast populations. Differences of yeast and yeast-like diversity between the glacier under study and other worldwide cold habitats are also discussed.

  16. Bioprospection of cold-adapted yeasts with biotechnological potential from Antarctica.

    PubMed

    Martorell, María Martha; Ruberto, Lucas Adolfo Mauro; Fernández, Pablo Marcelo; Castellanos de Figueroa, Lucía Inés; Mac Cormack, Walter Patricio

    2017-06-01

    The aim of this study was to investigate the ability to produce extracellular hydrolytic enzymes at low temperature of yeasts isolated from 25 de Mayo island, Antarctica, and to identify those exhibiting one or more of the evaluated enzymatic activities. A total of 105 yeast isolates were obtained from different samples and 66 were identified. They belonged to 12 basidiomycetous and four ascomycetous genera. Most of the isolates were ascribed to the genera Cryptococcus, Mrakia, Cystobasidium, Rhodotorula, Gueomyces, Phenoliferia, Leucosporidium, and Pichia. Results from enzymes production at low temperatures revealed that the Antarctic environment contains metabolically diverse cultivable yeasts, which represent potential tools for biotechnological applications. While most the isolates proved to produce 2-4 of the investigated exoenzymes, two of them evidenced the six evaluated enzymatic activities: Pichia caribbica and Guehomyces pullulans, which were characterized as psycrotolerant and psycrophilic, respectively. In addition, P. caribbica could assimilate several n-alkanes and diesel fuel. The enzyme production profile and hydrocarbons assimilation capacity, combined with its high level of biomass production and the extended exponential growth phase make P. caribbica a promising tool for cold environments biotechnological purposes in the field of cold-enzymes production and oil spills bioremediation as well. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. 38 CFR 17.1 - Incorporation by reference.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... this part. (1) NFPA 10, Standard for Portable Fire Extinguishers (2010 edition), Incorporation by..., Standard for the Installation of Sprinkler Systems (2010 edition), IBR approved for § 17.74. (5) NFPA 13D... (2010 edition), IBR approved for § 17.74. (6) NFPA 13R, Standard for the Installation of...

  18. China Report, Red Flag, No. 17, 1 September 1983.

    DTIC Science & Technology

    2007-11-02

    heart disease, nyctalopia, beriberi, and scurvy . Today it has been proven that many of these cures correspond to modern theories of nutrition...on Chinese boats many of the sailors used to catch scurvy . "First it makes the legs swell, then it continues every day 51 until it reaches the...Although today everyone is obscurely aware that scurvy is linked to one’s diet, people do not know that the real reason is a lack of vitamin Bl

  19. 38 CFR 17.1 - Incorporation by reference.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Code (2008 edition), IBR approved for § 17.74. (9) NFPA 72, National Fire Alarm and Signaling Code... the National Fire Protection Association, 1 Batterymarch Park, Quincy, MA 02269. (For ordering... this part. (1) NFPA 10, Standard for Portable Fire Extinguishers (2010 edition), Incorporation by...

  20. 38 CFR 17.1 - Incorporation by reference.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Code (2008 edition), IBR approved for § 17.74. (9) NFPA 72, National Fire Alarm and Signaling Code... the National Fire Protection Association, 1 Batterymarch Park, Quincy, MA 02269. (For ordering... this part. (1) NFPA 10, Standard for Portable Fire Extinguishers (2010 edition), Incorporation by...

  1. 26 CFR 1.381(c)(17)-1 - Deficiency dividend of personal holding company.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 4 2010-04-01 2010-04-01 false Deficiency dividend of personal holding company... Deficiency dividend of personal holding company. (a) Carryover requirement. If a determination (as defined in... 381(a) applies is liable for personal holding company tax imposed by section 541 (or by...

  2. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or... taking of salmon or halibut, for commercial purposes, nor the services of any other individual...

  3. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or... taking of salmon or halibut, for commercial purposes, nor the services of any other individual...

  4. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or... taking of salmon or halibut, for commercial purposes, nor the services of any other individual...

  5. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or... taking of salmon or halibut, for commercial purposes, nor the services of any other individual...

  6. Millwright Apprenticeship. Related Training Modules. 17.1-17.13 Hydraulics.

    ERIC Educational Resources Information Center

    Lane Community Coll., Eugene, OR.

    This packet of 13 learning modules on hydraulics is 1 of 6 such packets developed for apprenticeship training for millwrights. Introductory materials are a complete listing of all available modules and a supplementary reference list. Each module contains some or all of these components: goal, performance indicators, study guide (a check list of…

  7. Stationary Engineers Apprenticeship. Related Training Modules. 17.1-17.3 Feedwater.

    ERIC Educational Resources Information Center

    Lane Community Coll., Eugene, OR.

    This learning module, one in a series of 20 related training modules for apprentice stationary engineers, deals with feedwater. Addressed in the individual instructional packages included in the module are the following topics: types of feedwater, equipment for use in working with feedwater, water treatments, and procedures for testing feedwater.…

  8. Laboratory for Computer Science Progress Report 17, 1 July 1979-30 June 1980

    DTIC Science & Technology

    1980-12-11

    use of interconnected computers for strategic planning; as well as the sociological impact of computers on individuals and the ethical problems of...Ludwig J. Thomas J. Marrgraff S. Toner A. Mondori R. Vieraitis P. O’Donnell E. Wylen H. Peterson 2 Support Staff R. Bisbee E. Poithier D. Fagin M. Webber ...class. Each node may or may not have restriction predicates attached to the underlying class. The interpretation is that each restricted non-root node

  9. 26 CFR 1.381(c)(17)-1 - Deficiency dividend of personal holding company.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... (or by a corresponding provision of prior income tax law) for any taxable year ending on or before the... section 547(d)(1) if paid by the distributor or transferor corporation to its own shareholders except that it shall be paid by the acquiring corporation to its own shareholders and shall be paid after the...

  10. 26 CFR 1.381(c)(17)-1 - Deficiency dividend of personal holding company.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... corresponding provision of prior income tax law) for any taxable year ending on or before the date of...)(1) if paid by the distributor or transferor corporation to its own shareholders except that it shall be paid by the acquiring corporation to its own shareholders and shall be paid after the date of...

  11. Ecology and Oil Spills, 17-1. Military Curriculum Materials for Vocational and Technical Education.

    ERIC Educational Resources Information Center

    Army Quartermaster School, Ft. Lee, VA.

    This single-lesson course in ecology and oil spills for the secondary/postsecondary level comprises one of a number of military-developed curriculum packages selected for adaptation to vocational instruction and curriculum development in a civilian setting. The purpose stated for the 2-hour course is to provide students with an understanding of…

  12. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 7 2014-04-01 2013-04-01 true Supplemental unemployment benefit trusts. 1.501(c... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  13. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Supplemental unemployment benefit trusts. 1.501(c... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  14. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 7 2011-04-01 2009-04-01 true Supplemental unemployment benefit trusts. 1.501(c... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  15. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 7 2012-04-01 2012-04-01 false Supplemental unemployment benefit trusts. 1.501... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  16. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 7 2013-04-01 2013-04-01 false Supplemental unemployment benefit trusts. 1.501... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  17. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... authority to amend the plan means the legislature, board, commission, council, or other governing body with... effective date, annual compensation limit means $200,000, adjusted as provided by the Commissioner. The... after the OBRA '93 effective date, annual compensation limit means $150,000, adjusted as provided by the...

  18. METRO-APEX Volume 17.1: Industrialist's Manual No. 7, Shick Cannery. Revised.

    ERIC Educational Resources Information Center

    University of Southern California, Los Angeles. COMEX Research Project.

    The Industrialist's Manual No. 7 (Shick Cannery) is one of a set of twenty-one manuals used in METRO-APEX 1974, a computerized college and professional level, computer-supported, role-play, simulation exercise of a community with "normal" problems. Stress is placed on environmental quality considerations. APEX 1974 is an expansion of…

  19. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... other business or is a common-law employee in any business. For the 1994 calendar year, Employee C has... beginning on or after January 1, 1996, of the governing body with authority to amend the plan, if that body does not meet continuously. For purposes of this paragraph (d)(4), the term governing body...

  20. Enhanced poly(L-malic acid) production from pretreated cane molasses by Aureobasidium pullulans in fed-batch fermentation.

    PubMed

    Xia, Jun; Xu, Jiaxing; Hu, Lei; Liu, Xiaoyan

    2016-11-16

    Poly(L-malic acid) (PMA) is a natural polyester with many attractive properties for biomedical application. However, the cost of PMA production is high when glucose is used as a carbon source. To solve this problem, cane molasses as a low-cost feedstock was applied for the production of PMA. Six pretreatment methods were applied to cane molasses before fermentation. Pretreatment with combined tricalcium phosphate, potassium ferrocyanide, and sulfuric acid (TPFSA) removed significant amounts of metal ions from cane molasses. The PMA concentration increased from 5.4 g/L (untreated molasses) to 36.9 g/L (TPFSA-pretreated molasses) after fermentation in shake flasks. A fed-batch fermentation strategy was then developed. In this method, TPFSA-pretreated cane molasses solution was continuously fed into the fermentor to maintain the total sugar concentration at 20 g/L. This technique generated approximately 95.4 g/L PMA with a productivity of 0.57 g/L/hr. The present study indicated that fed-batch fermentation using pretreated cane molasses is a feasible technique for producing high amounts of PMA.

  1. Itzï (version 17.1): an open-source, distributed GIS model for dynamic flood simulation

    NASA Astrophysics Data System (ADS)

    Guillaume Courty, Laurent; Pedrozo-Acuña, Adrián; Bates, Paul David

    2017-05-01

    Worldwide, floods are acknowledged as one of the most destructive hazards. In human-dominated environments, their negative impacts are ascribed not only to the increase in frequency and intensity of floods but also to a strong feedback between the hydrological cycle and anthropogenic development. In order to advance a more comprehensive understanding of this complex interaction, this paper presents the development of a new open-source tool named Itzï that enables the 2-D numerical modelling of rainfall-runoff processes and surface flows integrated with the open-source geographic information system (GIS) software known as GRASS. Therefore, it takes advantage of the ability given by GIS environments to handle datasets with variations in both temporal and spatial resolutions. Furthermore, the presented numerical tool can handle datasets from different sources with varied spatial resolutions, facilitating the preparation and management of input and forcing data. This ability reduces the preprocessing time usually required by other models. Itzï uses a simplified form of the shallow water equations, the damped partial inertia equation, for the resolution of surface flows, and the Green-Ampt model for the infiltration. The source code is now publicly available online, along with complete documentation. The numerical model is verified against three different tests cases: firstly, a comparison with an analytic solution of the shallow water equations is introduced; secondly, a hypothetical flooding event in an urban area is implemented, where results are compared to those from an established model using a similar approach; and lastly, the reproduction of a real inundation event that occurred in the city of Kingston upon Hull, UK, in June 2007, is presented. The numerical approach proved its ability at reproducing the analytic and synthetic test cases. Moreover, simulation results of the real flood event showed its suitability at identifying areas affected by flooding, which were verified against those recorded after the event by local authorities.

  2. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 15 2012-04-01 2012-04-01 false Remuneration for services performed on a boat... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms of...

  3. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 15 2014-04-01 2014-04-01 false Remuneration for services performed on a boat... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms of...

  4. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 15 2013-04-01 2013-04-01 false Remuneration for services performed on a boat... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms of...

  5. Covalent Immobilization and Characterization of a Novel Pullulanase from Fontibacillus sp. Strain DSHK 107 onto Florisil® and Nano-silica for Pullulan Hydrolysis.

    PubMed

    Alagöz, Dilek; Yildirim, Deniz; Güvenmez, Hatice Korkmaz; Sihay, Damla; Tükel, S Seyhan

    2016-08-01

    A novel pullulanase partially purified from Fontibacillus sp. was covalently immobilized on Florisil® and nano-silica through both glutaraldehyde and (3-glycidyloxypropyl)trimethoxysilane spacer arms. The pullulanase immobilized on Florisil® and nano-silica through glutaraldehyde spacer arm showed 85 and 190 % activity of its free form, respectively, whereas no activity was observed when it was immobilized on the same supports through (3-glycidyloxypropyl)trimethoxysilane spacer arm. The maximum working pHs of both the immobilized pullulanases on Florisil® and nano-silica through glutaraldehyde spacer arm were determined as 5.0; however, the maximum working pH of the free pullulanase was pH 6.0. The maximum temperatures of all the pullulanase preparations were determined as 35 °C. The apparent K m values were 1.49, 1.54, and 0.59 mg/mL pullunan, respectively, for the free and immobilized pullulanases on Florisil® and nano-silica. The corresponding apparent V max values were 0.59, 1.53, and 1.57 U mg prot.(-1) min.(-1). Thermal stability of pullulanases immobilized on Florisil® and nano-silica was enhanced 6.5- and 15.6-folds, respectively at 35 °C and 6.6- and 16.0-folds, respectively, at 50 °C. The pullulanases immobilized on Florisil® and nano-silica protected 71 and 90 % of their initial activities after 10 reuses.

  6. High molecular weight β-poly(L-malic acid) produced by A. pullulans with Ca²⁺ added repeated batch culture.

    PubMed

    Cao, Weifeng; Chen, Xiangrong; Luo, Jianquan; Yin, Junxiang; Qiao, Changsheng; Wan, Yinhua

    2016-04-01

    β-Poly(malic acid) (PMLA) has attracted increasing attentions because of its potential application in medicine and other industries. In this study, the variation of PMLA molecular weight (Mw) in the batch culture and the strategies to enhance PMLA Mw were studied. Adding exogenous Ca(2+) (0.1g/L CaCl2) to the medium caused a significant increase in both PMLA concentration and Mw (11.38% and 26.3%, respectively) when Na2CO3 was used as the neutralizer. The Mw of PMLA during the process of batch culture, which associated with the specific PMLA production per unit cell mass (Yp/x) before glucose was depleted, increased from 12.522 KDa to its maximum 18.693 KDa and then kept decreasing until the end of the culture. Compared with the results in batch culture, Mw increased by 84.4% (up to 19.51 kDa) with a productivity of 1.1 gh(-1)L(-1) when the cells were maintained in exponential growth phase during Ca(2+) added repeated batch culture. The present work provides an efficient approach to obtain superior quality PMLA product with high Mw. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Role of Community and Technical Colleges in Producing Nursing Graduates: Rethinking the Pipeline for Guided Pathways. Research Report 17-1

    ERIC Educational Resources Information Center

    Washington State Board for Community and Technical Colleges, 2017

    2017-01-01

    Though it is still possible to enter the nursing profession with a Licensed Practical Nursing Certificate (LPN) or an Associate's Degree in Nursing (ADN), it is becoming increasingly necessary to get a Bachelor of Science in Nursing (BSN) in order to be assured of continued employment. The Associate in Applied Science-T Nursing Degree prepares…

  8. Quarterly Management Document – FY17, 1st Quarter, Physics-based Creep Simulations of Thick Section Welds in High Temperature and Pressure Applications

    SciTech Connect

    Lillo, Thomas Martin

    2017-01-01

    During the first quarter of FY17, research has focused on: 1. Generation of synthetic microstructures of welds 2. Aging of gamma prime 3. Short term creep tests 4. Preparation for stress drop tests to determine the threshold stress.

  9. SY 17-1 DYNAMIC REGULATION OF REDOX REGULATING FACTOR APE1/REF-1 ON THE OXIDATIVE STRESS AND VASCULAR INFLAMMATION.

    PubMed

    Jeon, Byeong Hwa

    2016-09-01

    Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein that plays a central role in the cellular response to DNA damage and redox regulation against oxidative stress. APE1/Ref-1 is essential for cellular survival and embryonic lethal in knockout mouse models. Heterozygous APE1/Ref-1 mice showed impaired endothelium-dependent vasorelaxation, reduced vascular NO levels, and are hypertensive. APE1/Ref-1 reduces intracellular reactive oxygen species production by negatively regulating the activity of the NADPH oxidase. APE1/Ref-1 is predominantly localized in the nucleus; however, its subcellular localization is dynamically regulated. Recently, it was shown that APE1/Ref-1 is secreted in response to hyperacetylation at specific lysine residues. We investigated the functions of extracellular APE1/Ref-1 with respect to leading anti-inflammatory signaling in TNF-α-stimulated endothelial cells in response to acetylation. Trichostatin A (TSA), an inhibitor of histone deacetylase, considerably suppressed vascular cell adhesion molecule-1 (VCAM-1) in TNF-α-stimulated endothelial cells. During TSA-mediated acetylation in culture, a time-dependent increase in secreted APE1/Ref-1 was confirmed. Recombinant human APE1/Ref-1 with reducing activity induced a conformational change in TNFR1 by thiol-disulfide exchange. Following treatment with the neutralizing anti-APE1/Ref-1 antibody, inflammatory signals via the binding of TNF-α to TNFR1 were remarkably recovered. Furthermore, rhAPE1/Ref-1 inhibited IL-1β-induced VCAM-1 expression in endothelial cells, and it inhibited iNOS or COX-2 expression in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. These results strongly indicate that anti-inflammatory effects of secreted APE1/Ref-1 and its property of secreted APE1/Ref-1 may be useful as a therapeutic biomolecule in cardiovascular disease.

  10. Biopolymer production from wastewater sludges for encapsulation and other applications. Final report

    SciTech Connect

    Baker, F.R.

    1988-01-01

    The main purpose of the research is to determine the feasibility of producing biopolymers from waste sludges by growing suitable microorganisms on the sludge hydrolysate. Primary and secondary municipal sludges and a bakery waste sludge were used in the study. Enzymatic hydrolysis was found to be ineffective in producing a substrate suitable for the growth of Aureobasidium pullulans (A. pullulans). High pressure acid hydrolysis of municipal sludge tended to yield a better substrate in terms of pullulan yield than low pressure acid hydrolysis. Acid hydrolysis of the bakery waste sludge resulted in a fermentation medium that was highly conductive to the growth of A. pullulans and pullulan elaboration. The feasibility of employing pullulan as a carrier material in the controlled-release of pesticides on stored grains was examined by determining the effect of pullulan-chlorpyrifos-methyl encapsulates on the mortality rate of flour beetles (Tribolilum castaneum).

  11. Design Rules for Fluorocarbon-Free Omniphobic Solvent Barriers in Paper-Based Devices.

    PubMed

    Jahanshahi-Anbuhi, Sana; Pennings, Kevin; Leung, Vincent; Kannan, Balamurali; Brennan, John D; Filipe, Carlos D M; Pelton, Robert H

    2015-11-18

    The utility of hydrophobic wax barriers in paper-based lateral flow and multiwell devices for containment of aqueous solvents was extended to organic solvents and challenging aqueous surfactant solutions by preparation of a three layer barrier, consisting of internal pullulan impregnated paper barriers surrounded by external wax barriers. When paper impregnated with pullulan solution dries, the polymer forms solvent blocking lenses in the paper structure. Lens formation was illustrated by forming pullulan lenses in glass capillaries. The lens shapes were less curved compared to the predictions of a model based upon minimizing surface area. For barriers on Whatman # 1 filter paper, the pullulan molecular weight must be greater than ∼70 kDa, the mass fraction of pullulan in the barrier zone must be at least 32%, and there are restrictions on the minimum width of the pullulan impregnated zone.

  12. Taxonomy of Aureobasidium spp. and biosynthesis and regulation of their extracellular polymers.

    PubMed

    Li, Yang; Chi, Zhe; Wang, Guang-Yuan; Wang, Zhi-Peng; Liu, Guang-Lei; Lee, Ching-Fu; Ma, Zhai-Chao; Chi, Zhen-Ming

    2015-06-01

    The genus Aureobasidium spp. have been divided into three species, A. pullulans. A. leucospermi and A. proteae, and A. pullulans has been known to have five varieties. However, after analysis of many strains of this yeast isolated from different environments, they do not belong to any of the three species or the five varieties. Although pullulan produced by A. pullulans has been widely used in different fields in industry and different strains of this yeast has been known to produce poly(β-L-malic acid) (PMA), heavy oils and β-1,3-glucan, it is still unknown how the black yeast synthesizes and secretes the extracellular polymers at molecular level. In this review article, new biosynthetic pathways of pullulan, PMA and heavy oils, the enzymes and their genes related to their biosynthesis and regulation are proposed. Furthermore, some enzymes and their genes related to pullulan biosynthesis in A. pullulans have been characterized. But it is completely unknown how pullulan is secreted and how PMA, heavy oils and β-1,3-glucan are synthesized and secreted. Therefore, there is much work to be done about taxonomy and biosynthesis, secretion and regulation of pullulan, PMA, heavy oils and β-1,3-glucan at molecular levels in Aureobasidium spp.

  13. Production of novel antistreptococcal liamocins by fermentation of agricultural biomass

    USDA-ARS?s Scientific Manuscript database

    Liamocins are unique heavier-than-water “oils” produced by certain strains of the fungus Aureobasidium pullulans. Liamocins have antibacterial activity with specificity for Streptococcus sp. Previous studies reported that liamocin yields were highest from strains of A. pullulans belonging to phyloge...

  14. Picky, hungry eaters in the cold: persistent substrate selectivity among polar pelagic microbial communities

    PubMed Central

    Steen, Andrew D.; Arnosti, Carol

    2014-01-01

    Polar pelagic microbial communities access a narrower range of polysaccharide substrates than communities at lower latitudes. For example, the glucose-containing polysaccharide pullulan is typically not hydrolyzed in fjord waters of Svalbard, even though pullulan is rapidly hydrolyzed in sediments from Svalbard fjords, other polysaccharides are hydrolyzed rapidly in Svalbard waters, and pullulan is hydrolyzed rapidly in temperate waters. The purpose of this study was to investigate potential factors preventing hydrolysis of pullulan in Svalbard fjord waters. To this end, in two separate years, water from Isfjorden, Svalbard, was amended with different carbon sources and/or additional nutrients in order to determine whether increasing the concentration of these potentially-limiting factors would lead to measurable enzymatic activity. Addition of nitrate, phosphate, glucose, or amino acids did not yield detectable pullulan hydrolysis. The only treatment that led to detectable pullulan hydrolysis was extended incubation after the addition of maltotriose (a subunit of pullulan, and potential inducer of pullulanase). In these fjords, the ability to enzymatically access pullulan is likely confined to numerically minor members of the pelagic microbial community. These results are consistent with the hypothesis that pelagic microbial communities at high latitudes exhibit streamlined functionality, focused on a narrower range of substrates, than their temperate counterparts. PMID:25339946

  15. Efficient access to novel androsteno-17-(1',3',4')-oxadiazoles and 17β-(1',3',4')-thiadiazoles via N-substituted hydrazone and N,N'-disubstituted hydrazine intermediates, and their pharmacological evaluation in vitro.

    PubMed

    Kovács, Dóra; Wölfling, János; Szabó, Nikoletta; Szécsi, Mihály; Minorics, Renáta; Zupkó, István; Frank, Éva

    2015-06-15

    A series of novel 17-exo-oxadiazoles and -thiadiazoles in the Δ(5) androstene series were efficiently synthesized from pregnenolone acetate and pregnadienolone acetate via multistep pathways. 17β-(1',3',4')-Oxadiazoles were obtained in high yields by the phenyliodonium diacetate-induced oxidative ring closure of semicarbazone and N-acylhydrazones derived from 3β-acetoxy- and 3β-hydroxyandrost-5-ene-17β-carbaldehydes. For the synthesis of analogous Δ(16)-17-oxadiazolyl derivatives, N,N'-disubstituted hydrazine intermediates were prepared from 3β-acetoxyandrosta-5,16-diene-17-carboxylic acid, which then underwent cyclodehydration in the presence of POCl3. The cyclization of steroidal N,N'-diacylhydrazines containing a saturated ring D with the Lawesson reagent afforded 17β-(1',3',4')-thiadiazoles in good yields. Most of the products were subjected to deacetylation in basic media in order to enlarge the compound library available for pharmacological studies. All of these derivatives were screened in vitro for their antiproliferative effects against four malignant human adherent cell lines (HeLa, A2780, MCF7 and A431) by means of the MTT assay. The 3β-hydroxy derivatives of the newly-synthesized 17-exo-heterocycles were tested in vitro to investigate their inhibitory effects on rat testicular C17,20-lyase. One of the 1,3,4-oxadiazolyl derivatives proved to exert noteworthy enzyme-inhibitory action, with an IC50 (0.065 μM) of the same order of magnitude as that of abiraterone.

  16. Paul's gospel and the rhetoric of apostolic rejection: A study of Galatians 1:15--17, 1 Corinthians 15:8, F. C. Baur, and the origins of Paul's Gentile mission

    NASA Astrophysics Data System (ADS)

    Mitchell, Matthew Wesley

    This dissertation proposes a new understanding of Paul's Gentile mission and its relationship to his so-called "conversion." This dissertation examines the origins of Paul's mission to the Gentiles, and locates it in his claims to have been personally commissioned to undertake such a mission by Jesus. Specifically, I argue that it is the rejection of Paul's claim to be an apostle, a claim founded upon his "conversion" experience, that precipitates his mission to the Gentiles. In arguing this view, I draw upon Ferdinand Christian Baur's nineteenth century theories concerning both the unreliability of Acts as a historical source, and his proposal of a clear division between Paul and the other apostles. In establishing the methodological and theoretical framework of the dissertation, I discuss the "New Perspective on Paul" that has dominated New Testament scholarship over the past thirty years. My study is also informed methodologically by the growing interest in rhetorical criticism among biblical scholars, although the emphasis of this dissertation bears more of a resemblance to the approach of the New Rhetoric than the categories of classical, Greco-Roman rhetoric. The textual component of this work falls into two stages. The first contains a full examination of Paul's "conversion passages" in Galatians 1:15--17 and 1 Corinthians 15:8, attempting to situate these seemingly unusual self-descriptions in their cultural contexts. The second involves an examination of F. C. Baur's presentation of Paul, and the reception of Baur's views among biblical scholars throughout the years following his scholarly activity. This dissertation makes two claims, each of which can stand on its own as an important contribution to scholarship. My first claim is that components of Baur's work support my proposal concerning Paul's Gentile mission and his experience of apostolic rejection, and that this proposal has much to commend it as an explanation of a perennial scholarly puzzle. My second claim is methodological, as I demonstrate that scholarly writings about Paul and his modern interpreters are themselves exercises in argumentation, and thus are not to be accepted uncritically, or without close attention to the rhetorical practices they utilize.

  17. Surface Plasmon Resonance Studies of Polysaccharide Self-Assembly on Cellulose

    NASA Astrophysics Data System (ADS)

    Kaya, Abdulaziz; Esker, Alan R.; Glasser, Wolfgang G.

    2006-03-01

    Wood is a multiphase material consisting of cellulose crystals embedded within a non-crystalline hetereopolysaccharide (hemicellulose) and lignin rich phase. The hierarchial arrangement of these three chief components in wood produces excellent properties like resistance to fracture and toughness. Through the study of polysaccharide self-assembly onto a model cellulose surface, further insight into the interactions between hemicelluloses and cellulose can be gained. In our study, we synthesized pullulan cinnamates with different degrees of substitution of cinnamoyl groups as a model for a hemicellulose with lignin-like moieties. Surface plasmon resonance measurements probe the self-assembly behavior of pullulan and pullulan cinnamate onto a cellulose coated gold surface. Our results suggest that pullulan does not adsorb onto the model cellulose surface, whereas pullulan cinnamate does. These preliminary results signify the important role that lignin-like substituents play on hemicellulose self-assembly onto cellulose surfaces.

  18. Preparation of cationized polysaccharides as gene transfection carrier for bone marrow-derived mesenchymal stem cells.

    PubMed

    Jo, Jun-ichiro; Okazaki, Arimichi; Nagane, Kentaro; Yamamoto, Masaya; Tabata, Yasuhiko

    2010-01-01

    The objective of this study is to prepare a non-viral carrier of gene transfection from various polysaccharides and evaluate the feasibility in gene expression for mesenchymal stem cells (MSCs). Various amounts of spermine were chemically introduced into pullulan, dextran and mannan with a molecular weight of around 40 000 or pullulan with different molecular weights to prepare cationized polysaccharides with different extents of spermine introduced (spermine-polysaccharide). Each cationized polysaccharide was complexed with a plasmid DNA at various ratios and in vitro gene transfection was investigated for rat bone marrow-derived MSCs. The level of gene expression depended on the type of cationized polysaccharide. The highest level was observed for the complex of spermine-pullulan and plasmid DNA. Additionally, the level also depended on the molecular weight of pullulan and the extent of spermine introduced to pullulan. Suppression of gene expression with chlorpromazine and methyl-beta-cyclodextrin of endocytosis inhibitors demonstrated that the cellular uptake of spermine-pullulan-plasmid DNA complexes was mediated by clathrin- and raft/caveolae-dependent endocytic pathways. The cationized pullulan is a promising non-viral carrier of plasmid DNA for MSCs.

  19. Influence of abiotic variables on culturable yeast diversity in two distinct Alpine glaciers.

    PubMed

    Turchetti, Benedetta; Goretti, Marta; Branda, Eva; Diolaiuti, Guglielmina; D'Agata, Carlo; Smiraglia, Claudio; Onofri, Andrea; Buzzini, Pietro

    2013-11-01

    The influence of some abiotic variables (pH, dry weight, organic carbon, nitrogen and phosphorous) on culturable yeast diversity in two distinct, but adjacent Alpine glaciers (Glacier du Géant, France, and Miage Glacier, Italy) was investigated. In all, 682 yeast strains were isolated and identified by D1/D2 and ITS sequencing as belonging to species of the genera Aureobasidium, Candida, Bulleromyces, Cryptococcus, Cystofilobasidium, Dioszegia, Guehomyces, Holtermanniella, Leucosporidiella, Mrakia, Mrakiella, Rhodotorula, Sporidiobolus, Sporobolomyces and Udenyomyces. Overall, the most represented genera were Cryptococcus (55% of isolates), Rhodotorula (17%) and Mrakia (10%). About 10% of strains, presumably belonging to new species (yet to be described), were preliminarily identified at the genus level. Principal component analysis (PCA) revealed that organic carbon, nitrogen and phosphorous are apparently mostly related to culturable yeast abundance and diversity. In this context, the hypothesis that the frequency of isolation of certain species may be correlated with some organic nutrients (with special emphasis for phosphorous) is discussed.

  20. Earthworms modify microbial community structure and accelerate maize stover decomposition during vermicomposting.

    PubMed

    Chen, Yuxiang; Zhang, Yufen; Zhang, Quanguo; Xu, Lixin; Li, Ran; Luo, Xiaopei; Zhang, Xin; Tong, Jin

    2015-11-01

    In the present study, maize stover was vermicomposted with the epigeic earthworm Eisenia fetida. The results showed that, during vermicomposting process, the earthworms promoted decomposition of maize stover. Analysis of microbial communities of the vermicompost by high-throughput pyrosequencing showed more complex bacterial community structure in the substrate treated by the earthworms than that in the control group. The dominant microbial genera in the treatment with the earthworms were Pseudoxanthomonas, Pseudomonas, Arthrobacter, Streptomyces, Cryptococcus, Guehomyces, and Mucor. Compared to the control group, the relative abundance of lignocellulose degradation microorganisms increased. The results indicated that the earthworms modified the structure of microbial communities during vermicomposting process, activated the growth of lignocellulose degradation microorganisms, and triggered the lignocellulose decomposition.

  1. A reverse transfection technology to genetically engineer adult stem cells.

    PubMed

    Okazaki, Arimichi; Jo, Jun-Ichiro; Tabata, Yasuhiko

    2007-02-01

    A new non-viral method of gene transfection was designed to enhance the level of gene expression for rat mesenchymal stem cells (MSCs). Pullulan was cationized using chemical introduction of spermine to prepare cationized pullulan of non-viral carrier (spermine-pullulan). The spermine-pullulan was complexed with a plasmid deoxyribonucleic acid (DNA) of luciferase and coated on the surface of culture substrate together with Pronectin of artificial cell adhesion protein. MSCs were cultured and transfected on the complex-coated substrate (reverse transfection), and the level and duration of gene expression were compared with those of MSCs transfected by culturing in the medium containing the plasmid DNA-spermine-pullulan complex (conventional method). The reverse transfection method enhanced and prolonged gene expression significantly more than did the conventional method. The reverse method permitted the transfection culture of MSCs in the presence of serum, in contrast to the conventional method, which gave cells a good culture condition to lower cytotoxicity. The reverse transfection was carried out for a non-woven fabric of polyethylene terephthalate (PET) coated with the complex and Pronectin using agitation and stirring culture methods. The two methods enhanced the level and duration of gene expression for MSCs significantly more than did the static method. It is possible that medium circulation improves the culture conditions of cells in terms of oxygen and nutrition supply and waste excretion, resulting in enhanced gene expression.

  2. An alternative anionic bio-sustainable anti-fungal agent: Investigation of its mode of action on the fungal cell membrane.

    PubMed

    Stenbæk, Jonas; Löf, David; Falkman, Peter; Jensen, Bo; Cárdenas, Marité

    2017-07-01

    The potential of a lactylate (the sodium caproyl lactylate or C10 lactylate), a typical food grade emulsifier, as an anionic environmental friendly anti-fungal additive was tested in growth medium and formulated in a protective coating for exterior wood. Different laboratory growth tests on the blue stain fungus Aureobasidium pullulans were performed and its interactions on a model fungal cell membrane were studied. Promising short term anti-fungal effects in growth tests were observed, although significant but less dramatic effects took place in coating test on wood panels. Scanning electron microscope analysis shows clear differences in the amount of fungal slime on the mycelium of Aureobasidium pullulans when the fungus was exposed of C10 lactylate. This could indicate an effect on the pullulan and melanin production by the fungus. Moreover, the interaction studies on model fungal cell membranes show that C10 lactylate affects the phospholipid bilayer in a similar manner to other negative charged detergents.

  3. Structure, specificity and function of cyclomaltodextrinase, a multispecific enzyme of the alpha-amylase family.

    PubMed

    Park, K H; Kim, T J; Cheong, T K; Kim, J W; Oh, B H; Svensson, B

    2000-05-23

    Cyclomaltodextrinase (CDase, EC 3.2.1.54), maltogenic amylase (EC 3. 2.1.133), and neopullulanase (EC 3.2.1.135) are reported to be capable of hydrolyzing all or two of the following three types of substrates: cyclomaltodextrins (CDs); pullulan; and starch. These enzymes hydrolyze CDs and starch to maltose and pullulan to panose by cleavage of alpha-1,4 glycosidic bonds whereas alpha-amylases essentially lack activity on CDs and pullulan. They also catalyze transglycosylation of oligosaccharides to the C3-, C4- or C6-hydroxyl groups of various acceptor sugar molecules. The present review surveys the biochemical, enzymatic, and structural properties of three types of such enzymes as defined based on the substrate specificity toward the CDs: type I, cyclomaltodextrinase and maltogenic amylase that hydrolyze CDs much faster than pullulan and starch; type II, Thermoactinomyces vulgaris amylase II (TVA II) that hydrolyzes CDs much less efficiently than pullulan; and type III, neopullulanase that hydrolyzes pullulan efficiently, but remains to be reported to hydrolyze CDs. These three types of enzymes exhibit 40-60% amino acid sequence identity. They occur in the cytoplasm of bacteria and have molecular masses from 62 to 90 kDa which are slightly larger than those of most alpha-amylases. Multiple amino acid sequence alignment and crystal structures of maltogenic amylase and TVA II reveal the presence of an N-terminal extension of approximately 130 residues not found in alpha-amylases. This unique N-terminal domain as seen in the crystal structures apparently contributes to the active site structure leading to the distinct substrate specificity through a dimer formation. In aqueous solution, most of these enzymes show a monomer-dimer equilibrium. The present review discusses the multiple specificity in the light of the oligomerization and the molecular structures arriving at a clarified enzyme classification. Finally, a physiological role of the enzymes is proposed.

  4. Fungitoxicity of chemical analogs with heartwood toxins.

    PubMed

    Grohs, B M; Kunz, B

    1998-07-01

    Trans-stilbene and tropolone as chemical analogs with naturally occurring fungitoxic heartwood compounds were studied with respect to their fungitoxic potency. While stilbene showed no fungitoxic activity towards the fungi Aureobasidium pullulans var. melanogenum, Penicillium glabrum, and Trichoderma harzianum in the concentrations tested, the minimal inhibiting concentration of tropolone was 10(-3) M for Penicillium glabrum and Trichoderma harzianum, and 10(-5) M for Aureobasidium pullulans var. melanogenum. In all cases, the effect of tropolone was a fungistatic one. Using chemical analogs for assessing the chemical basis of the fungitoxicity of tropolone, this substance proved to be the only compound tested which possesses fungitoxic properties.

  5. Pulsed Laser Processing of Functionalized Polysaccharides for Controlled Release Drug Delivery Systems

    NASA Astrophysics Data System (ADS)

    Cristescu, R.; Popescu, C.; Popescu, A. C.; Socol, G.; Mihailescu, I.; Caraene, G.; Albulescu, R.; Buruiana, T.; Chrisey, D.

    We report on the deposition of triacetate-pullulan polysaccharide thin films on drug pellets (diclofenac sodium) by matrix assisted pulsed laser evaporation method. The radiation generated by a pulsed excimer KrF* laser source (λ = 248 nm, τ = 20 ns) operated at 2 Hz repetition rate was used for ice targets evaporation. The timed - controlled drug delivery was proved by spectroscopic in vitro studies and in vivo anti-inflammatory investigations on rabbits. We showed that the coating of drug pellets with triacetate-pullulan thin films resulted in the delayed delivery of the drug for up to 30 min.

  6. Functional and structural microbial diversity in organic and conventional viticulture: organic farming benefits natural biocontrol agents.

    PubMed

    Schmid, Florian; Moser, Gerit; Müller, Henry; Berg, Gabriele

    2011-03-01

    Statistically significant differences in the structure and function of above-ground grapevine-associated microorganisms from organically and conventionally managed vineyards were found. Aureobasidium pullulans, a copper-detoxifying fungus and biocontrol agent, plays a key role in explaining these differences. The black fungus was strongly enriched in the communities of organically managed plants and yielded a higher indigenous antiphytopathogenic potential.

  7. Effect of carbohydrate digestibility on appetite and its relationship to postprandial blood glucose and insulin levels.

    PubMed

    Peters, H P F; Ravestein, P; van der Hijden, H T W M; Boers, H M; Mela, D J

    2011-01-01

    'Slowly digestible' carbohydrates have been claimed to reduce appetite through their effects on postprandial glucose and insulin levels, but literature is inconsistent. The inconsistencies between studies might be explained by factors other than glycemic effects per se, for example, nutritional or physical properties. We tested this possibility by examining postprandial glucose, insulin and appetite responses to drinks differing only in rate and extent of digestibility of carbohydrates. This was accomplished by comparing different glucose polymers: maltodextrin (rapidly digestible) versus medium-chain pullulan (slowly but completely digestible) versus long-chain pullulan (indigestible). In a randomized double-blind balanced crossover design, 35 subjects received drinks with 15 g test carbohydrate polymers. Key outcome measures were appetite scores, digestibility (in vitro test and breath hydrogen), and (in a subset) glucose and insulin levels. Digestibility, glucose and insulin data confirmed the rapid, slow and nondigestible nature of the test carbohydrates. Despite its low digestibility, only long-chain pullulan reduced appetite compared with the maltodextrin control, whereas the medium-chain pullulan did not. We conclude that glycemic responses per se have minimal effects on appetite, when tested in products differing in only carbohydrate digestibility rate and extent.

  8. The yeast flora of the coast redwood, Sequoia sempervirens.

    PubMed

    Middelhoven, W J

    2003-01-01

    Only four yeast species could be isolated from young and perannual shoots of the coast redwood tree, Sequoia sempervirens, and from soil beneath the trees, viz. both varieties of Debaryomyces hansenii, Trichosporon pullulans, T. porosum and an unidentified red basidiomycetous yeast.

  9. Sweet substrate: a polysaccharide nanocomposite for conformal electronic decals.

    PubMed

    Daniele, Michael A; Knight, Adrian J; Roberts, Steven A; Radom, Kathryn; Erickson, Jeffrey S

    2015-03-04

    A conformal electronic decal based on a polysaccharide circuit board (PCB) is fabricated and characterized. The PCBs are laminates composed of bioderived sugars - nanocellulose and pullulan. The PCB and decal transfer are a bioactive material system for supporting electronic devices capable of conforming to bio-logical surfaces. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Functional and Structural Microbial Diversity in Organic and Conventional Viticulture: Organic Farming Benefits Natural Biocontrol Agents ▿ †

    PubMed Central

    Schmid, Florian; Moser, Gerit; Müller, Henry; Berg, Gabriele

    2011-01-01

    Statistically significant differences in the structure and function of above-ground grapevine-associated microorganisms from organically and conventionally managed vineyards were found. Aureobasidium pullulans, a copper-detoxifying fungus and biocontrol agent, plays a key role in explaining these differences. The black fungus was strongly enriched in the communities of organically managed plants and yielded a higher indigenous antiphytopathogenic potential. PMID:21278278

  11. A survey of insect assemblages responding to volatiles from a ubiquitous fungus in an agricultural landscape

    USDA-ARS?s Scientific Manuscript database

    We report here a first survey of insect orientation to fungal cultures and and fungal volatiles from a community ecology perspective. We tested whether volatiles from a ubiquitous yeast-like fungus (Aureobasidium pullulans) are broadly attractive to insects in an agricultural landscape. We evaluated...

  12. Recovery of melanized yeasts from Eastern Mediterranean beach sand associated with the prevailing geochemical and marine flora patterns.

    PubMed

    Efstratiou, Maria A; Velegraki, Aristea

    2010-03-01

    The melanized opportunistic pathogens Exophiala dermatitidis (Chaetothyriales) and Aureobasidium pullulans (Dothideales) were sporadically isolated from beach sand. This is the first time they are reported from bathing beach sand, thus providing updates on the conditions influencing the in situ black yeast community structure and raising public health concerns.

  13. Extracellular enzyme activity in anaerobic bacterial cultures: evidence of pullulanase activity among mesophilic marine bacteria.

    PubMed Central

    Arnosti, C; Repeta, D J

    1994-01-01

    The extracellular enzymatic activity of a mixed culture of anaerobic marine bacteria enriched on pullulan [alpha(1,6)-linked maltotriose units] was directly assessed with a combination of gel permeation chromatography (GPC) and nuclear magnetic resonance spectroscopy (NMR). Hydrolysis products of pullulan were separated by GPC into three fractions with molecular weights of > or = 10,000, approximately 5,000, and < or = 1,200. NMR spectra of these fractions demonstrated that pullulan was rapidly and specifically hydrolyzed at alpha(1,6) linkages by pullulanase enzymes, most likely type II pullulanase. Although isolated pullulanase enzymes have been shown to hydrolyze pullulan completely to maltotriose (S. H. Brown, H. R. Costantino, and R. M. Kelly, Appl. Environ. Microbiol. 56:1985-1991, 1990; M. Klingeberg, H. Hippe, and G. Antranikian, FEMS Microbiol. Lett. 69:145-152, 1990; R. Koch, P. Zablowski, A. Spreinat, and G. Antranikian, FEMS Microbiol. Lett. 71:21-26, 1990), the smallest carbohydrate detected in the bacterial cultures consisted of two maltotriose units linked through one alpha(1,6) linkage. Either the final hydrolysis step was closely linked to substrate uptake, or specialized porins similar to maltoporin might permit direct transport of large oligosaccharides into the bacterial cell. This is the first report of pullulanase activity among mesophilic marine bacteria. The combination of GPC and NMR could easily be used to assess other types of extracellular enzyme activity in bacterial cultures. PMID:8161177

  14. TGF-β1 gene-engineered mesenchymal stem cells induce rat cartilage regeneration using nonviral gene vector.

    PubMed

    He, Cai-Xia; Zhang, Tian-Yuan; Miao, Pei-Hong; Hu, Zhong-Jie; Han, Min; Tabata, Yasuhiko; Hu, Yu-Lan; Gao, Jian-Qing

    2012-01-01

    This study evaluated the potential of utilizing transfected pTGFβ-1 gene-engineered rat mesenchymal stem cells (MSCs) using nonviral vector to promote cartilage regeneration. Pullulan-spermine was used as the nonviral gene vector and gelatin sponge was used as the scaffold. MSCs were engineered with TGF-β1 gene with either the three-dimensional (3D) reverse transfection system or the two-dimensional (2D) conventional transfection system. For the 3D reverse transfection system, pullulan-spermine/pTGF-β1 gene complexes were immobilized to the gelatin sponge, followed by the seeding of MSCs. Pullulan-spermine/pTGF-β1 gene complexes were delivered to MSCs cultured in the plate to perform the 2D conventional transfection system, and then MSCs were seeded to the gelatin sponge. Then, TGF-β1 gene-transfected MSC seeded gelatin sponge was implanted to the full-thickness cartilage defect. Compared with the control group, both groups of TGF-β1 gene-engineered MSCs improved cartilage regeneration through optical observation and histology staining. So, with pullulan-spermine as the nonviral vector, TGF-β1-gene engineered MSCs can induce cartilage regeneration in vivo.

  15. Fungal Colonization and Biodeterioration of Plasticized Polyvinyl Chloride

    PubMed Central

    Webb, Jeremy S.; Nixon, Marianne; Eastwood, Ian M.; Greenhalgh, Malcolm; Robson, Geoffrey D.; Handley, Pauline S.

    2000-01-01

    Significant substratum damage can occur when plasticized PVC (pPVC) is colonized by microorganisms. We investigated microbial colonization of pPVC in an in situ, longitudinal study. Pieces of pPVC containing the plasticizers dioctyl phthalate and dioctyl adipate (DOA) were exposed to the atmosphere for up to 2 years. Fungal and bacterial populations were quantified, and colonizing fungi were identified by rRNA gene sequencing and morphological characteristics. Aureobasidium pullulans was the principal colonizing fungus, establishing itself on the pPVC between 25 and 40 weeks of exposure. A group of yeasts and yeast-like fungi, including Rhodotorula aurantiaca and Kluyveromyces spp., established themselves on the pPVC much later (after 80 weeks of exposure). Numerically, these organisms dominated A. pullulans after 95 weeks, with a mean viable count ± standard error of 1,000 ± 200 yeast CFU cm−2, compared to 390 ± 50 A. pullulans CFU cm−2. No bacterial colonization was observed. We also used in vitro tests to characterize the deteriogenic properties of fungi isolated from the pPVC. All strains of A. pullulans tested could grow with the intact pPVC formulation as the sole source of carbon, degrade the plasticizer DOA, produce extracellular esterase, and cause weight loss of the substratum during growth in vitro. In contrast, several yeast isolates could not grow on pPVC or degrade DOA. These results suggest that microbial succession may occur during the colonization of pPVC and that A. pullulans is critical to the establishment of a microbial community on pPVC. PMID:10919769

  16. Molecular mobility and microscopic structure changes in κ-carrageenan solutions studied by gradient NMR.

    PubMed

    Zhao, Qiuhua; Brenner, Tom; Matsukawa, Shingo

    2013-06-05

    Changes in the molecular mobility of κ-carrageenan were observed by the pulsed field gradient stimulated echo (PGSTE) and Carr-Purcell-Meiboom-Gill (CPMG) methods for elucidating the molecular aspect of the sol-to-gel transition. The echo signal intensity of κ-carrageenan without a gradient, Ikap(0), decreased steeply near the sol-to-gel temperature (Tsg), suggesting that κ-carrageenan chains formed aggregates and a network structure. Below Tsg, the spin-spin relaxation time T2 and the diffusion coefficient of κ-carrageenan (Dkap) increased with decreasing temperature, indicating that the solute κ-carrageenan chains have a lower molecular weight Mw than chains involved in the aggregation. The diffusion coefficient of pullulan (Dpul) added as a probe molecule in κ-carrageenan solutions was measured, and the characteristic hydrodynamic screening length, ξ, was then estimated from the degree of diffusion restriction. Below a certain temperature, Dkap reached a higher value than that of Dpul, suggesting that the Mw of solute κ-carrageenan became lower than that of pullulan. GPC measurements confirmed the presence of κ-carrageenan chains with a lower Mw than that of pullulan. A simple physical model of the structural change in κ-carrageenan solution was proposed with a bimodal distribution of κ-carrageenan with higher and lower Mw than the pullulan probe. The higher Mw chains form the gel network restricting the probe's diffusion, and the lower Mw chains increase the effective viscosity. The concentration of the high Mw solute κ-carrageenan chains in 1%, 2% and 4% κ-carrageenan solutions was estimated from Ikap(0) and the total κ-carrageenan concentration, and the relation with pullulan diffusion was studied. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Identification of Medically Relevant Species of Arthroconidial Yeasts by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

    PubMed Central

    Kolecka, Anna; Khayhan, Kantarawee; Groenewald, Marizeth; Theelen, Bart; Arabatzis, Michael; Velegraki, Aristea; Kostrzewa, Markus; Mares, Mihai; Taj-Aldeen, Saad J.

    2013-01-01

    Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) was used for an extensive identification study of arthroconidial yeasts, using 85 reference strains from the CBS-KNAW yeast collection and 134 clinical isolates collected from medical centers in Qatar, Greece, and Romania. The test set included 72 strains of ascomycetous yeasts (Galactomyces, Geotrichum, Saprochaete, and Magnusiomyces spp.) and 147 strains of basidiomycetous yeasts (Trichosporon and Guehomyces spp.). With minimal preparation time, MALDI-TOF MS proved to be an excellent diagnostic tool that provided reliable identification of most (98%) of the tested strains to the species level, with good discriminatory power. The majority of strains were correctly identified at the species level with good scores (>2.0) and seven of the tested strains with log score values between 1.7 and 2.0. The MALDI-TOF MS results obtained were consistent with validated internal transcribed spacer (ITS) and/or large subunit (LSU) ribosomal DNA sequencing results. Expanding the mass spectrum database by increasing the number of reference strains for closely related species, including those of nonclinical origin, should enhance the usefulness of MALDI-TOF MS-based diagnostic analysis of these arthroconidial fungi in medical and other laboratories. PMID:23678074

  18. Identification of medically relevant species of arthroconidial yeasts by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    PubMed

    Kolecka, Anna; Khayhan, Kantarawee; Groenewald, Marizeth; Theelen, Bart; Arabatzis, Michael; Velegraki, Aristea; Kostrzewa, Markus; Mares, Mihai; Taj-Aldeen, Saad J; Boekhout, Teun

    2013-08-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was used for an extensive identification study of arthroconidial yeasts, using 85 reference strains from the CBS-KNAW yeast collection and 134 clinical isolates collected from medical centers in Qatar, Greece, and Romania. The test set included 72 strains of ascomycetous yeasts (Galactomyces, Geotrichum, Saprochaete, and Magnusiomyces spp.) and 147 strains of basidiomycetous yeasts (Trichosporon and Guehomyces spp.). With minimal preparation time, MALDI-TOF MS proved to be an excellent diagnostic tool that provided reliable identification of most (98%) of the tested strains to the species level, with good discriminatory power. The majority of strains were correctly identified at the species level with good scores (>2.0) and seven of the tested strains with log score values between 1.7 and 2.0. The MALDI-TOF MS results obtained were consistent with validated internal transcribed spacer (ITS) and/or large subunit (LSU) ribosomal DNA sequencing results. Expanding the mass spectrum database by increasing the number of reference strains for closely related species, including those of nonclinical origin, should enhance the usefulness of MALDI-TOF MS-based diagnostic analysis of these arthroconidial fungi in medical and other laboratories.

  19. Characterization of starch-debranching enzymes in pea embryos

    PubMed

    Zhu; Hylton; Rossner; Smith

    1998-10-01

    Two distinct types of debranching enzymes have been identified in developing pea (Pisum sativum L.) embryos using native gel analysis and tests of substrate preference on purified or partially purified activities. An isoamylase-like activity capable of hydrolyzing amylopectin and glycogen but not pullulan is present throughout development and is largely or entirely confined to the plastid. Activities capable of hydrolyzing pullulan are present both inside and outside of the plastid, and extraplastidial activity increases relative to the plastidial activity during development. Both types of debranching enzyme are also present in germinating embryos. We argue that debranching enzymes are likely to have a role in starch metabolism in the plastid of the developing embryo and in starch degradation during germination.

  20. Childhood hypersensitivity pneumonitis associated with fungal contamination of indoor hydroponics.

    PubMed

    Engelhart, Steffen; Rietschel, Ernst; Exner, Martin; Lange, Lars

    2009-01-01

    Childhood hypersensitivity pneumonitis (HP) is often associated with exposure to antigens in the home environment. We describe a case of HP associated with indoor hydroponics in a 14-year-old girl. Water samples from hydroponics revealed Aureobasidium pullulans as the dominant fungal micro-organism (10(4)CFU/ml). The diagnosis is supported by the existence of serum precipitating antibodies against A. pullulans, lymphocytic alveolitis on bronchoalveolar lavage (BAL) fluid, a corresponding reaction on a lung biopsy, and the sustained absence of clinical symptoms following the removal of hydroponics from the home. We conclude that hydroponics should be considered as potential sources of fungal contaminants when checking for indoor health complaints.

  1. Molecular entanglement and electrospinnability of biopolymers.

    PubMed

    Kong, Lingyan; Ziegler, Gregory R

    2014-09-03

    Electrospinning is a fascinating technique to fabricate micro- to nano-scale fibers from a wide variety of materials. For biopolymers, molecular entanglement of the constituent polymers in the spinning dope was found to be an essential prerequisite for successful electrospinning. Rheology is a powerful tool to probe the molecular conformation and interaction of biopolymers. In this report, we demonstrate the protocol for utilizing rheology to evaluate the electrospinnability of two biopolymers, starch and pullulan, from their dimethyl sulfoxide (DMSO)/water dispersions. Well-formed starch and pullulan fibers with average diameters in the submicron to micron range were obtained. Electrospinnability was evaluated by visual and microscopic observation of the fibers formed. By correlating the rheological properties of the dispersions to their electrospinnability, we demonstrate that molecular conformation, molecular entanglement, and shear viscosity all affect electrospinning. Rheology is not only useful in solvent system selection and process optimization, but also in understanding the mechanism of fiber formation on a molecular level.

  2. Highly versatile nanohydrogel platform based on riboflavin-polysaccharide derivatives useful in the development of intrinsically fluorescent and cytocompatible drug carriers.

    PubMed

    Di Meo, Chiara; Montanari, Elita; Manzi, Lucio; Villani, Claudio; Coviello, Tommasina; Matricardi, Pietro

    2015-01-22

    In this work we describe a new nanohydrogel platform, based on polysaccharides modified with the hydrophobic and fluorescent molecule riboflavin tetrabutyrate, which leads to innovative structures useful for drug delivery applications. Hyaluronic acid and pullulan were chosen as representative of anionic and neutral polysaccharides, respectively, and the bromohexyl derivative of riboflavin tetrabutyrate was chemically linked to these polymer chains. Because of such derivatization, polymer chains were able to self-assemble in aqueous environment thus forming nanohydrogels, with mean diameters of about 312 and 210 nm, for hyaluronan and pullulan, respectively. These new nanohydrogels showed low polydispersity index, and negative ζ-potential. Moreover, the nanohydrogels, which can be easily loaded with model drugs, showed long-term stability in water and physiological conditions and excellent cytocompatibility. All these properties allow to consider these intrinsically fluorescent nanohydrogels suitable for the formulation of innovative drug dosage forms. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Yeasts from native Brazilian Cerrado plants: Occurrence, diversity and use in the biocontrol of citrus green mould.

    PubMed

    Sperandio, Eugenio Miranda; do Vale, Helson Mario Martins; Moreira, Geisianny Augusta Monteiro

    2015-11-01

    Yeasts are some of the most important postharvest biocontrol agents. Postharvest oranges frequently deteriorate due to green mould (Penicillium digitatum), which causes significant losses. The aims of this study were to determine the composition and diversity of yeasts on plants of the Brazilian Cerrado and to explore their potential for inhibiting citrus green mould. Leaves and fruit of Byrsonima crassifolia and Eugenia dysenterica were collected from Cerrado conservation areas, and thirty-five yeasts were isolated and identified by sequencing the D1-D2 domain of the rDNA large subunit (26S). The isolates represented the Aureobasidium, Meyerozyma, Candida, and Pichia genera. Three isolates identified as Aureobasidium pullulans exhibited potential for the control of P. digitatum in both in vitro and in vivo tests; these isolates reduced the incidence of disease and increased the storage time of fruit. Aureobasidium. pullulans has immense potential for the biological control of filamentous fungi.

  4. The influence of different polymers on viability of Bifidobacterium lactis 300b during encapsulation, freeze-drying and storage.

    PubMed

    Pop, Oana Lelia; Brandau, Thorsten; Schwinn, Jens; Vodnar, Dan Cristian; Socaciu, Carmen

    2015-07-01

    Seven different types of natural polymers namely hydroxypropyl methylcellulose (HPMC), sodium-carboxymethyl cellulose (Na-CMC), microcrystalline cellulose (MCC), starch BR-07, starch BR-08, dextrin and pullulan were used in order to develop the optimal formula for the entrapment of Bifidobacterium lactis 300B in Ca-alginate based granules. Laminar flow drip casting with Brace-Encapsulator was used in order to prepare the granules. The results showed that alginate/pullulan and alginate/HPMC formulation provide high protection for the bacterial strain used for encapsulation. These two formulations were further used to obtain freeze dried granules, for which the viability in time and at different temperatures was tested. The final results showed a higher viability than the level of the therapeutic minimum (>10(7) CFU/g) after 15 days of storage. Other parameters like entrapment efficiency, production rate, sphericity, flowability were also discussed.

  5. Characterization of Starch-Debranching Enzymes in Pea Embryos1

    PubMed Central

    Zhu, Zhi-Ping; Hylton, Christopher M.; Rössner, Ute; Smith, Alison M.

    1998-01-01

    Two distinct types of debranching enzymes have been identified in developing pea (Pisum sativum L.) embryos using native gel analysis and tests of substrate preference on purified or partially purified activities. An isoamylase-like activity capable of hydrolyzing amylopectin and glycogen but not pullulan is present throughout development and is largely or entirely confined to the plastid. Activities capable of hydrolyzing pullulan are present both inside and outside of the plastid, and extraplastidial activity increases relative to the plastidial activity during development. Both types of debranching enzyme are also present in germinating embryos. We argue that debranching enzymes are likely to have a role in starch metabolism in the plastid of the developing embryo and in starch degradation during germination. PMID:9765544

  6. Further application of size-exclusion chromatography combined with small-angle X-ray scattering optics for characterization of biological macromolecules.

    PubMed

    Watanabe, Yasushi; Inoko, Yoji

    2011-02-01

    Size-exclusion chromatography (gel filtration chromatography or gel permeation chromatography) in conjunction with online synchrotron radiation solution small-angle X-ray scattering optics, absorbance, and/or refractive index detectors was further assessed by application of biological macromolecules, such as the hollow sphere protein complex, apoferritin, and a linear polysaccharide, pullulan. The net X-ray scattering patterns of the eluted 24-mer molecule of apoferritin showed the specific character for the hollow spherical shape. The chromatographic (time-resolved) X-ray scattering data of the linear polysaccharide pullulan revealed the flexible chain structure during the chromatographic separation in an aqueous solution. These further applications demonstrated that the present measurement technique will be useful for not only the determination of the radius of gyration value of less than about 10 nm and molecular weight below several hundred thousand but also for the structural characterization of the various macromolecules during the chromatography.

  7. Materials Biotechnology Symposium Proceedings Held in Natick, Massachusetts on June 23 and 24, 1987

    DTIC Science & Technology

    1987-11-05

    Foundation (DMB84- 01811), the National Institutes of Health (GM-14312), and the National Foundation for Cancer Research. REFERENCES 1. G. Nemethy...which might contribute substantially to our understanding of such important biological functions as growth, differentiation, immunology, cancer and the...Arnaud during the yeast-like phase of the growth cycle, but it is not used during routine metabolism. ’ Pullulan is a linear a-D- glucan

  8. Extracellular Proteinases of Yeasts and Yeastlike Fungi1

    PubMed Central

    Ahearn, D. G.; Meyers, S. P.; Nichols, R. A.

    1968-01-01

    Approximately 800 yeasts and other fungi, representing over 70 species, were tested for extracellular caseinolysis. Isolates of a variety of genera, including Aureobasidium, Cephalosporium, Endomycopsis, Kluyveromyces, and numerous sporobolomycetes, demonstrated significant proteolytic activity. Caseinolysis was not necessarily correlated with gelatin liquefaction or with albuminolysis. Numerous fungi showed significant proteolysis at 5 C. The most active organisms were isolates of Candida lipolytica, Aureobasidium pullulans, Candida punicea, and species of Cephalosporium. Taxonomic and ecological implications of proteolytic activity are discussed. Images Fig. 1 PMID:5692110

  9. Relief hologram replication using a dental composite as an embossing tool

    NASA Astrophysics Data System (ADS)

    Savić Šević, Svetlana; Pantelić, Dejan

    2005-04-01

    A simplified method for holographic embossing tool production is presented. Surface relief diffraction gratings are holographically recorded in pullulan sensitized with ammonium dichromate (DCP). The surface structure is copied into dental photopolymer composite by direct contact and subsequent photo-polymerization. It was found that arbitrary surface micropattern can be replicated. Due to its excellent mechanical and thermal properties, micro-patterned dental composite can be further used as an embossing tool for mass production of holograms.

  10. Relief hologram replication using a dental composite as an embossing tool.

    PubMed

    Savić Sević, Svetlana; Pantelić, Dejan

    2005-04-04

    A simplified method for holographic embossing tool production is presented. Surface relief diffraction gratings are holographically recorded in pullulan sensitized with ammonium dichromate (DCP). The surface structure is copied into dental photopolymer composite by direct contact and subsequent photo-polymerization. It was found that arbitrary surface micropattern can be replicated. Due to its excellent mechanical and thermal properties, micro-patterned dental composite can be further used as an embossing tool for mass production of holograms.

  11. Volatile emissions from an epiphytic fungus are semiochemicals for eusocial wasps.

    PubMed

    Davis, Thomas Seth; Boundy-Mills, Kyria; Landolt, Peter J

    2012-11-01

    Microbes are ubiquitous on plant surfaces. However, interactions between epiphytic microbes and arthropods are rarely considered as a factor that affects arthropod behaviors. Here, volatile emissions from an epiphytic fungus were investigated as semiochemical attractants for two eusocial wasps. The fungus Aureobasidium pullulans was isolated from apples, and the volatile compounds emitted by fungal colonies were quantified. The attractiveness of fungal colonies and fungal volatiles to social wasps (Vespula spp.) were experimentally tested in the field. Three important findings emerged: (1) traps baited with A. pullulans caught 2750 % more wasps on average than unbaited control traps; (2) the major headspace volatiles emitted by A. pullulans were 2-methyl-1-butanol, 3-methyl-1-butanol, and 2-phenylethyl alcohol; and (3) a synthetic blend of fungal volatiles attracted 4,933 % more wasps on average than unbaited controls. Wasps were most attracted to 2-methyl-1-butanol. The primary wasp species attracted to fungal volatiles were the western yellowjacket (Vespula pensylvanica) and the German yellowjacket (V. germanica), and both species externally vectored A. pullulans. This is the first study to link microbial volatile emissions with eusocial wasp behaviors, and these experiments indicate that volatile compounds emitted by an epiphytic fungus can be responsible for wasp attraction. This work implicates epiphytic microbes as important components in the community ecology of some eusocial hymenopterans, and fungal emissions may signal suitable nutrient sources to foraging wasps. Our experiments are suggestive of a potential symbiosis, but additional studies are needed to determine if eusocial wasp-fungal associations are widespread, and whether these associations are incidental, facultative, or obligate.

  12. A survey of insect assemblages responding to volatiles from a ubiquitous fungus in an agricultural landscape.

    PubMed

    Davis, Thomas Seth; Landolt, Peter J

    2013-07-01

    We report here a first survey of insect orientation to fungal cultures and fungal volatiles from a community ecology perspective. We tested whether volatiles from a ubiquitous yeast-like fungus (Aureobasidium pullulans) are broadly attractive to insects in an agricultural landscape. We evaluated insect attraction to fungal cultures and synthetic compounds identified in fungal headspace (2-methyl-1-butanol, 3-methyl-1-butanol, 2-phenylethanol) in a spearmint (Mentha spicata L.) plantation. Three findings emerged: (1) 1,315 insects representing seven orders and 39 species oriented to traps, but 65 % of trapped insects were Dipterans, of which 80 % were hoverflies (Diptera: Syrphidae); (2) traps baited with A. pullulans caught 481 % more insects than unbaited control traps on average, and contained more diverse (Shannon's H index) and species rich assemblages than control traps, traps baited with Penicillium expansum, or uninoculated media; and (3) insects oriented in greatest abundance to a 1:1:1 blend of A. pullulans volatiles, but mean diversity scores were highest for traps baited with only 2-phenylethanol or 2-methyl-1-butanol. Our results show that individual components of fungal headspace are not equivalent in terms of the abundance and diversity of insects that orient to them. The low abundance of insects captured with P. expansum suggests that insect assemblages do not haphazardly orient to fungal volatiles. We conclude that volatiles from a common fungal species (A. pullulans) are attractive to a variety of insect taxa in an agricultural system, and that insect orientation to fungal volatiles may be a common ecological phenomenon.

  13. Pullulanase from rice endosperm.

    PubMed

    Yamasaki, Yoshiki; Nakashima, Susumu; Konno, Haruyoshi

    2008-01-01

    Pullulanase (EC 3.2.1.41) in non-germinating seeds was compared with that in germinating seeds. Moreover, pullulanase from the endosperm of rice (Oryza sativa L., cv. Hinohikari) seeds was isolated and its properties investigated. The pI value of pullulanase from seeds after 8 days of germination was almost equal to that from non-germinating seeds, which shows that these two enzymes are the same protein. Therefore, the same pullulanase may play roles in both starch synthesis during ripening and starch degradation during germination in rice seeds. The enzyme was isolated by a procedure that included ammonium sulfate fractionation, DEAE-cellulofine column chromatography, preparative isoelectric focusing, and preparative disc gel electrophoresis. The enzyme was homogeneous by SDS/PAGE. The molecular weight of the enzyme was estimated to be 100 000 based on its mobility on SDS/PAGE and 105 000 based on gel filtration with TSKgel super SW 3000, which showed that it was composed of a single unit. The isoelectric point of the enzyme was 4.7. The enzyme was strongly inhibited by beta-cyclodextrin. The enzyme was not activated by thiol reagents such as dithiothreitol, 2-mercaptoethanol or glutathione. The enzyme most preferably hydrolyzed pullulan and liberated only maltotriose. The pullulan hydrolysis was strongly inhibited by the substrate at a concentration higher than 0.1%. The degree of inhibition increased with an increase in the concentration of pullulan. However, the enzyme hydrolyzed amylopectin, soluble starch and beta-limit dextrin more rapidly as their concentrations increased. The enzyme exhibited alpha-glucosyltransfer activity and produced an alpha-1,6-linked compound of two maltotriose molecules from pullulan.

  14. Development of a biodegradable iron oxide nanoparticle gel for tumor bed therapy

    NASA Astrophysics Data System (ADS)

    Cunkelman, B. P.; Chen, E. Y.; Petryk, A. A.; Tate, J. A.; Thappa, S. G.; Collier, R. J.; Hoopes, P. J.

    2013-02-01

    Treatments of the post-operative surgical bed have proven appealing as the majority of cancer recurrence following tumor resection occurs at the tumor margin. A novel, biodegradable pullulan-based gel infused with magnetic iron oxide nanoparticles (IONP) is presented here for surgical bed administration followed by hyperthermia therapy via alternating magnetic field (AMF) activation. Pullulan is a water soluble, film-forming starch polymer that degrades at the postoperative wound site to deliver the IONP payload, targeting the remaining cancer cells. Different gel formulations containing various % wt of pullulan were tested for IONP elution. Elution levels and amount of gel degradation were measured by immersing the gel in de-ionized water for one hour then measuring particle concentrations in the supernatant and the mass of the remaining gel formulation. The most promising gel formulations will be tested in a murine model of surgical bed resection to assess in vivo gel dissolution, IONP cell uptake kinetics via histology and TEM analysis, and heating capability of the gel with AMF exposure.

  15. An innovative polysaccharide nanobased nail formulation for improvement of onychomycosis treatment.

    PubMed

    Flores, Fernanda C; Rosso, Roberta S; Cruz, Letícia; Beck, Ruy C R; Silva, Cristiane B

    2017-03-30

    Tioconazole-loaded nanocapsule suspensions and its coating with a cationic polymer were developed for nail drug delivery. The colloidal systems presented a nanometric size around 155nm for uncoated nanoparticles and 162nm for those with the cationic coating, with negative and positive zeta potential values, respectively. Both nanosuspensions showed drug content close to theoretical values (1mgmL(-1)), association efficiency close to 100% (HPLC) and were able to control tioconazol release. The developed formulations showed in vitro antifungal activity (agar diffusion method) against C. albicans. The cationic nanocapsules were considered bioadhesive, showed higher viscosity and were chosen to be incorporated into an ungueal formulation. Pullulan nanobased nail formulation showed adequate viscosity for nail application and drug content close to the theoretical values. It was equivalent to the commercial formulation Trosid(®) in preventing nail infection by T. rubrum in an in vitro onychomycosis model. The nanocapsule suspensions and Pullulan nanobased nail formulation showed lower irritant potential than the commercial formulation and than free drug in an in vitro evaluation. Pullulan nanobased nail formulation is promising for the treatment of onychomycosis.

  16. Performance evaluation of volatile organic compounds by antagonistic yeasts immobilized on hydrogel spheres against gray, green and blue postharvest decays.

    PubMed

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2017-05-01

    Wickerhamomyces anomalus, Metschnikowia pulcherrima, Aureobasidium pullulans and Saccharomyces cerevisiae yeasts were tested for their ability to survive and synthesize antifungal volatile organic compounds (VOCs) both in vitro and in vivo conditions when immobilized on commercial hydrogel spheres. The results showed a good survival of all yeasts on hydrogel spheres up to 10 days of incubation. Moreover, VOCs produced in vitro by tested yeasts inhibited Botrytis cinerea, Penicillium digitatum and P. italicum radial growth and conidial germination, with the highest antagonistic activity reported for W. anomalus and A. pullulans strains. Experimental in vivo trials performed on strawberry and mandarin fruits proved the ability of VOCs to reduce significantly postharvest decays on artificially wounded tissues. Comprehensively, the best efficacy was detected for W. anomalus, which totally inhibited gray mold decay on strawberry fruits and significantly reduced green mold infections on mandarin fruits. On the other hand, blue mold decay on mandarin fruits was more effectively managed by A. pullulans VOCs. Accordingly, hydrogel spheres used as a support for VOC-generating yeasts could open a new way for the employment of this polymeric material as a bio-emitter in postharvest packaging. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Preservation of bacteria in natural polymers.

    PubMed

    Krumnow, April A; Sorokulova, Iryna B; Olsen, Eric; Globa, Ludmila; Barbaree, James M; Vodyanoy, Vitaly J

    2009-08-01

    A new inexpensive and simple method for preserving microorganisms has been developed. Natural polymers of acacia gum and pullulan were used to preserve model bacteria Escherichia coli and Bacillus subtilis via immobilization and storage under various conditions. Formulation of E. coli and B. subtilis in acacia gum significantly increased the viability of both cultures during desiccation at 40 degrees C as well as during the storage at various temperatures and relative humidity. In the ranges of temperatures and humidity used in experiments, the high humidity affected the viability of E. coli more than high temperature. Thermodynamic parameters for E. coli thermal degradation were used for quantification of results and characterization of the preservation process. Viability of B. subtilis in acacia gum polymer was not significantly changed during the storage in the temperature and humidity experiments. The number of viable B. subtilis recovered after storage in pullulan, and in PBS under various humidity conditions was 1-2 logs less in comparison with the number of cells before storage. It was found that acacia gum provides better protection than pullulan for both bacteria during the preservation process.

  18. Isolation and characterization of a novel thermostable neopullulanase-like enzyme from a hot spring in Thailand.

    PubMed

    Tang, Kittapong; Kobayashi, Rutchadaporn Sriprang; Champreda, Verawat; Eurwilaichitr, Lily; Tanapongpipat, Sutipa

    2008-06-01

    A gene encoding a thermostable pullulan-hydrolyzing enzyme was isolated from environmental genomic DNA extracted from soil sediments of Bor Khleung hot spring in Thailand. Sequence comparison with related enzymes suggested that the isolated enzyme, designated Env Npu193A, was most likely a neopullulanase-like enzyme. Env Npu193A was expressed in Pichia pastoris as a monomeric recombinant protein. The purified Env Npu193A exhibited pH stability ranging from 3 to 9. More than 60% of enzyme activity was retained after incubation at 60 degrees C for 1 h. Env Npu193A was found to hydrolyze various substrates, including pullulan, starch, and gamma-cyclodextrin. The optimal working condition for Env Npu193A was at pH 7 at 75 degrees C with K(m) and V(max) toward pullulan of 1.22+/-0.3% and 23.24+/-1.7 U/mg respectively. Env Npu193A exhibited distinct biochemical characteristics as compared with the previously isolated enzyme from the same source. Thus, a culture-independent approach with sequence-basing was found to be an effective way to discover novel enzymes displaying unique substrate specificity and high thermostability from natural bioresources.

  19. Psychrophilic and Mesophilic Fungi in Fruit-Filled Pastries

    PubMed Central

    Kuehn, Harold H.; Gunderson, Millard F.

    1962-01-01

    Surveys of the mold flora of frozen blueberry and cherry pastries were undertaken. Molds were enumerated by preparing pour plates of the blended product and incubating the plates at 0, 5, 10, and 20 C. In this manner, the total fungal content of the product could be ascertained from the 10 and 20 C plates, and the psychrophilic fungal population was represented by those fungi which grew at 0 and 5 C. The pastry portion, or crust, of the blueberry material was sampled separately from the filling portion. Certain differences in fungal flora were apparent. Aureobasidium pullulans was the dominant fungus in crust at all temperatures of isolation. However, Penicillium thomii proved to be the most common mesophilic fungus in the filling portion, and A. pullulans was the most common psychrophile in the filling. Aspergilli were quite common in the crust, but, in general, were absent from the fruit filling. Cherry pastries had a much smaller total fungal flora than did the blueberry product. However, A. pullulans again was the most prevalent fungus in cherry pastries at all temperatures of isolation. Certain differences in fungal flora were apparent in the two fruit products. Phoma spp. were almost completely absent in blueberries, but represented the second most common fungus in cherry pastries. Blueberry filling had 440 psychrophilic fungi per gram of sample (at 0 C), blueberry crust had 65 per gram, and cherry pastries had 77 per gram. Images FIG. 1 PMID:14460237

  20. An acetylated polysaccharide-PTFE membrane-covered stent for the delivery of gemcitabine for treatment of gastrointestinal cancer and related stenosis.

    PubMed

    Moon, Sumi; Yang, Su-Geun; Na, Kun

    2011-05-01

    Gemcitabine (Gem) eluting metal stents were prepared for potential application as drug delivery systems for localized treatment of malignant tumors. Pullulan, a natural polysaccharide, was chemically acetylated (pullulan acetate; PA) by different degrees (1.18, 1.71, and 2.10 acetyl groups per glucose unit of pullulan), layered on polytetrafluoroethylene (PTFE), and applied as part of a Gem-loaded controlled-release membrane for drug-eluting non-vascular stents. PA with a higher degree of acetylation had greater drug-loading capacity with more extended release of Gem over 30 days. The released Gem accumulated in CT-26 colon cancer without systemic exposure inducing total regression of tumors. The long-term biological activity of the released Gem and apoptosis of tumor tissues following localized delivery were confirmed by annexin V binding assays and histology. The controlled release of Gem from PA-PTFE covered drug-eluting stents (DES) may increase the patency of these stents for the treatment of malignant gastrointestinal cancer as well as cancer-related stenosis.

  1. Strategies for the production of high-content fructo-oligosaccharides through the removal of small saccharides by co-culture or successive fermentation with yeast.

    PubMed

    Nobre, C; Castro, C C; Hantson, A-L; Teixeira, J A; De Weireld, G; Rodrigues, L R

    2016-01-20

    Fructo-oligosaccharides (FOS) obtained by fermentation of sucrose may be purified at large-scale by continuous chromatography (Simulated Moving Bed: SMB). In order to improve the efficiency of the subsequent SMB purification, the optimization of the fermentative broth composition in salts and sugars was investigated. Fermentations conducted at reduced amount of salts, using Aureobasidium pullulans whole cells, yielded 0.63 ± 0.03 g of FOS per gram of initial sucrose. Additionally, a microbial treatment was proposed to reduce the amount of small saccharides in the mixture. Two approaches were evaluated, namely a co-culture of A. pullulans with Saccharomyces cerevisiae; and a two-step fermentation in which FOS were first synthesized by A. pullulans and then the small saccharides were metabolized by S. cerevisiae. Assays were performed in 100mL shaken flasks and further scaled-up to a 3 L working volume bioreactor. Fermentations in two-step were found to be more efficient than the co-culture ones. FOS were obtained with a purity of 81.6 ± 0.8% (w/w), on a dry weight basis, after the second-step fermentation with S. cerevisiae. The sucrose amount was reduced from 13.5 to 5.4% in total sugars, which suggests that FOS from this culture broth will be more efficiently separated by SMB. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Multiple rare opportunistic and pathogenic fungi in persistent foot skin infection.

    PubMed

    Chan, Giek Far; Sinniah, Sivaranjini; Idris, Tengku Idzzan Nadzirah Tengku; Puad, Mohamad Safwan Ahmad; Abd Rahman, Ahmad Zuhairi

    2013-03-01

    Persistent superficial skin infection caused by multiple fungi is rarely reported. Recently, a number of fungi, both opportunistic and persistent in nature were isolated from the foot skin of a 24-year old male in Malaysia. The fungi were identified as Candida parapsilosis, Rhodotorula mucilaginosa, Phoma spp., Debaryomyces hansenii, Acremonium spp., Aureobasidium pullulans and Aspergillus spp., This is the first report on these opportunistic strains were co-isolated from a healthy individual who suffered from persistent foot skin infection which was diagnosed as athlete's foot for more than 12 years. Among the isolated fungi, C. parapsilosis has been an increasingly common cause of skin infections. R. mucilaginosa and D. hansenii were rarely reported in cases of skin infection. A. pullulans, an emerging fungal pathogen was also being isolated in this case. Interestingly, it was noted that C. parapsilosis, R. mucilaginosa, D. hansenii and A. pullulans are among the common halophiles and this suggests the association of halotolerant fungi in causing persistent superficial skin infection. This discovery will shed light on future research to explore on effective treatment for inhibition of pathogenic halophiles as well as to understand the interaction of multiple fungi in the progress of skin infection.

  3. Psychrophilic and Mesophilic Fungi in Frozen Food Products

    PubMed Central

    Kuehn, Harold H.; Gunderson, Millard F.

    1963-01-01

    The mold flora of certain frozen pastries and chicken pies was investigated. Molds were determined qualitatively or quantitatively, or both, by preparing pour plates of the blended product and incubating the plates at various temperatures. The mesophilic fungal flora developed on plates incubated at 10 and 20 C, whereas psychrophilic fungi were obtained on plates incubated at 0 and 5 C. About 2,000 cultures of fungi, representing about 100 different species, were isolated from various products. Four different brands of blueberry, two brands of cherry pastries, two brands of apple, and one brand of raspberry pastries were examined. In addition, two brands of chicken pies were studied. Blueberry pastries had a much higher total fungal population than the other products, although different brands of blueberry pastries varied considerably. Blueberry pastries had from 347 to 1,586 psychrophilic fungi per g. Cherry pastries had about 70 to 110 psychrophiles per g, and apple pastries had 19 to 92 psychrophiles per g. Chicken pies contained very few psychrophilic fungi, about 15 per g. Aureobasidium pullulans was recovered most frequently. About 90% of the psychrophilic fungi found in blueberry products was A. pullulans. Depending upon the brand of cherry pastry, either Phoma spp. or A. pullulans was the most common fungus present. Apple pastries also displayed brand variation, but were unique in having many mesophilic aspergilli. This genus was generally absent from other products. The Penicillium content of apple pastries was also rather high; 50% of the psychrophilic flora was represented by this genus. The psychrophilic fungal flora of chicken pies was composed primarily of penicillia (50%) and Chrysosporium pannorum (46%). PMID:13927344

  4. Polysaccharide based Copolymers as Supramolecular Systems in Biomedical Applications.

    PubMed

    Célia Monteiro de Paula, Regina; Andrade Feitosa, Judith Pessoa; Beserra Paula, Haroldo César

    2015-01-01

    Polysaccharides are natural polymers, obtained from a large variety of sources ranging from fungi to more complex organisms such as birds and whales. Their use for pharmaceutical and biomedical applications has been the subject of numerous researches by the world´s academia. Polysaccharide chemical/physical modifications leading to graft copolymers are discussed in this review, focusing on those nanosystems that are potential candidates for drug delivery applications. Therefore, this review focuses on the biomedical application of polysaccharide based copolymers, particularly as nanocarriers. Copolymer of polysaccharides such as alginate, cellulose, chitosan, dextran, guar, hyaluronic acid, pullulan and starch as drug delivery nanocarriers will be discussed.

  5. Fungi Associated with Softening of Bisulfite-Brined Cherries.

    PubMed

    Lewis, J C; Pierson, C F; Powers, M J

    1963-03-01

    Softening of sound, calcium bisulfite-brined cherries was induced fairly quickly by brining them with cherries rotted by Aspergillus niger, Cytospora leucostoma, and Penicillium expansum, but not with cherries rotted by a variety of other microorganisms, including Alternaria sp., Aspergillus oryzae, Aureobasidium pullulans, Botrytis cinerea, Cladosporium sp., Mucor racemosus, Rhizopus stolonifer, and Sclerotinia fructicola. Rapid softening was correlated with the presence of a bisulfite-stable polygalacturonase, as demonstrated by a cup-plate test. A survey of naturally rotted cherries suggests the involvement of a bark-canker fungus, C. leucostoma, in softening of commercially brined cherries in the Pacific Northwest.

  6. Nanogel-quantum dot hybrid nanoparticles for live cell imaging

    SciTech Connect

    Hasegawa, Urara; Nomura, Shin-ichiro M.; Kaul, Sunil C.; Hirano, Takashi; Akiyoshi, Kazunari; E-mail: akiyoshi.org@tmd.ac.jp

    2005-06-17

    We report here a novel carrier of quantum dots (QDs) for intracellular labeling. Monodisperse hybrid nanoparticles (38 nm in diameter) of QDs were prepared by simple mixing with nanogels of cholesterol-bearing pullulan (CHP) modified with amino groups (CHPNH{sub 2}). The CHPNH{sub 2}-QD nanoparticles were effectively internalized into the various human cells examined. The efficiency of cellular uptake was much higher than that of a conventional carrier, cationic liposome. These hybrid nanoparticles could be a promising fluorescent probe for bioimaging.

  7. Isolation and characterization of bacterial endophytes of Chelidonium majus L.

    PubMed

    Goryluk, Agata; Rekosz-Burlaga, Hanna; Błaszczyk, Mieczysław

    2009-01-01

    The aim of this study was to isolate and identify endophytic bacteria from stems of Chelidonium majus L. (greater celandine) and to evaluate their antifungal properties. In total, 34 bacterial endophyte strains were isolated. The fungistatic effects of these bacteria on the growth of five moulds (Alternaria alternata, Chaetonium sp., Paecilomyces variotti, Byssochlamys fulva, Aureobasidium pullulans) and one species of black yeast (Exophiala mesophila) were tested. The majority of the bacterial isolates were found to inhibit the growth of fungi and those with the strongest antifungal properties were further characterized. Of the twelve isolates examined, 11 were species of Bacillus thuringiensis and one was Bacillus amyloliquefaciens.

  8. Changes in IR spectra of polysaccharides induced by CW CO2-laser radiation

    NASA Astrophysics Data System (ADS)

    Dlugunovich, Viacheslav A.; Zhbankov, R. G.; Zhdanovskii, Vladimir A.; Nasennik, L. N.; Puhnarevich, S. A.; Firsov, S. P.

    2003-04-01

    By IR spectroscopy methods the structural changes of high molecular polymers irradiated by CW CO2-laser radiation was investigated. Some changes in the structural sensitive regions at 1250 - 950 and 950 - 850 cm-1 of the IR spectra of the investigated polysaccharides [pullulan (molecular mass of 14500) and microcrystalline cellulose (structural modifications I and II)] were exhibit. These changes indicated that the degree of conformational order of polysaccharide molecules increases under the laser irradiation, while its structural order always decreases as a result of heating by traditional thermal sources.

  9. Determination of molecular weight of heparin by size exclusion chromatography with universal calibration.

    PubMed

    Guo, X; Condra, M; Kimura, K; Berth, G; Dautzenberg, H; Dubin, P L

    2003-01-01

    The molecular weight (MW) of heparin can be accurately determined by size exclusion chromatography using "universal calibration." A universal calibration curve was constructed for Superose 12 with standard pullulan samples and verified using characterized ficoll fractions. This calibration yielded the correct MW of heparin as determined by light scattering, when the ionic strength of the mobile phase was maintained over 1.0M. Sodium poly(styrenesulfonate) samples were not suitable standards because of adsorption at high salt concentration and repulsion from the packing material at low ionic strength. The extraordinarily high charge density of heparin leads to the need for high salt concentration to screen such repulsions.

  10. Screening of micro-organisms for decolorization of melanins produced by bluestain fungi.

    PubMed

    Rättö, M; Chatani, M; Ritschkoff, A C; Viikari, L

    2001-03-01

    A total of 17 fungi and four bacteria were screened for their ability to decolorize melanin, using isolated extracellular melanin of the bluestain fungus Aureobasidium pullulans as substrate. On agar media, decolorization was observed by four fungal strains: Bjerkandera adusta VTT-D-99746, Galactomyces geotrichum VTT-D-84228, Trametes hirsuta VTT-D-95443 and Trametes versicolor VTT-D-99747. The four fungi were more efficient on nitrogen-limited medium than on complete medium. The melanin-decolorizing activity of G. geotrichum appeared to be located on the mycelium and could be liberated into the medium enzymatically.

  11. Target-Induced and Equipment-Free DNA Amplification with a Simple Paper Device.

    PubMed

    Liu, Meng; Hui, Christy Y; Zhang, Qiang; Gu, Jimmy; Kannan, Balamurali; Jahanshahi-Anbuhi, Sana; Filipe, Carlos D M; Brennan, John D; Li, Yingfu

    2016-02-18

    We report on a paper device capable of carrying out target-induced rolling circle amplification (RCA) to produce massive DNA amplicons that can be easily visualized. Interestingly, we observed that RCA was more proficient on paper than in solution, which we attribute to a significantly higher localized concentration of immobilized DNA. Furthermore, we have successfully engineered a fully functional paper device for sensitive DNA or microRNA detection via printing of all RCA-enabling molecules within a polymeric sugar film formed from pullulan, which was integrated with the paper device. This encapsulation not only stabilizes the entrapped reagents at room temperature but also enables colorimetric bioassays with minimal steps.

  12. Isolation and selection of yeasts from wine grape ecosystem secreting cold-active pectinolytic activity.

    PubMed

    Merín, María Gabriela; Mendoza, Lucía M; Farías, Marta E; Morata de Ambrosini, Vilma Inés

    2011-05-27

    The present study was undertaken with the purpose of selecting yeasts from wine grapes that are able to produce extracellular cold-active pectinases. After two consecutive selections yeast isolates were identified by pheno- and genotyping, and pectinolytic activity was preliminarily characterised at proximate winemaking conditions. Out of 1023 indigenous microorganisms isolated from grape skins of D.O. San Rafael (Mendoza, Argentina) viticulture region, 565 (55%) showed pectinolytic activity on plates and, among them, 96 (17%) were chosen in a primary selection. Ten isolates were finally selected for exhibiting the greatest activity at low temperature (12 °C) and identified as Aureobasidium pullulans. GM-R-22 strain demonstrated the highest pectinolytic activity (0.751 U/mL) at pH 3.5 and 12 °C. Yeast pectinases were constitutively produced. This study is the first report about strains of A. pullulans producing pectinases which are able to show good activity at low temperature. These pectinolytic strains could be of interest in wine production. Copyright © 2011 Elsevier B.V. All rights reserved.

  13. Photomediated Reactive Oxygen Species-Generable Nanoparticles for Triggered Release and Endo/Lysosomal Escape of Drug upon Attenuated Single Light Irradiation.

    PubMed

    Seo, Eun Ha; Lee, Chung-Sung; Na, Kun

    2015-12-30

    Nanoparticles with "smart" stimuli-responsive materials and multiple therapeutic strategies in a single delivery platform have emerged for highly efficient cancer therapy. Here, photomediated reactive oxygen species (ROS)-generable nanoparticles are designed that can trigger drug release and endo/lysosomal escape upon attenuated single light irradiation, simultaneously, for synergistic chemo-photodynamic ablation. In this study, the self-ROS-generable nanoparticles (SRNs) are prepared from the polymer based on polysaccharide, chlorin e6 as ROS generator and lipoic acid as ROS scavenger covalently conjugated pullulan with anticancer drug (doxorubicin, DOX) through self-assembly, and can disassemble via the ROS-mediated reduction of lipoyl group in response to low level exogenous single light switch. After cellular internalization in hepatic cancer through asialoglycoprotein receptor (ASGPR, as pullulan receptor)-mediated endocytosis, once irradiated, SRNs are able to produce ROS that can simultaneously induce drug release triggering and endo/lysosomal escape of DOX into cytoplasm as well as directly photodynamic therapy for highly efficient chemo-photodynamic cancer therapy. This promising delivery system, which has huge potential in biomedical applications, may be optimal for smart delivery platform.

  14. Pressate from Peat Dewatering as a Substrate for Bacterial Growth

    PubMed Central

    Mulligan, Catherine N.; Cooper, David G.

    1985-01-01

    This study considered the possibility of using water expressed during the drying of fuel-grade peat as a substrate for microbial growth. Highly humified peat pressed for 2.5 min at 1.96 MPa produced water with a chemical oxygen demand of 690 mg/liter. Several biological compounds could be produced by using the organic matter in expressed peat water as a substrate. These included polymers such as chitosan, contained in the cell wall of Rhizopus arrhizus, and two extracellular polysaccharides, xanthan gum and pullulan, produced by Xanthomonas campestris and Aureobasidium pullulans, respectively. A very effective surfactant was produced by Bacillus subtilis grown in the expressed water. Small additions of nutrients to the peat pressate were necessary to obtain substantial yields of products. The addition of peptone, yeast extract, and glucose improved production of the various compounds. Biological treatment improved the quality of the expressed water to the extent that in an industrial process it could be returned to the environment. PMID:16346834

  15. Molecular Entanglement and Electrospinnability of Biopolymers

    PubMed Central

    Kong, Lingyan; Ziegler, Gregory R.

    2014-01-01

    Electrospinning is a fascinating technique to fabricate micro- to nano-scale fibers from a wide variety of materials. For biopolymers, molecular entanglement of the constituent polymers in the spinning dope was found to be an essential prerequisite for successful electrospinning. Rheology is a powerful tool to probe the molecular conformation and interaction of biopolymers. In this report, we demonstrate the protocol for utilizing rheology to evaluate the electrospinnability of two biopolymers, starch and pullulan, from their dimethyl sulfoxide (DMSO)/water dispersions. Well-formed starch and pullulan fibers with average diameters in the submicron to micron range were obtained. Electrospinnability was evaluated by visual and microscopic observation of the fibers formed. By correlating the rheological properties of the dispersions to their electrospinnability, we demonstrate that molecular conformation, molecular entanglement, and shear viscosity all affect electrospinning. Rheology is not only useful in solvent system selection and process optimization, but also in understanding the mechanism of fiber formation on a molecular level. PMID:25226274

  16. Yeasts associated with nectarines and their potential for biological control of brown rot.

    PubMed

    Janisiewicz, W J; Kurtzman, C P; Buyer, J S

    2010-07-01

    Resident fruit microflora has been the source of biocontrol agents for the control of postharvest decay of fruits and the active ingredient in commercialized biocontrol products. With the exception of grapes and apples, information on the resident microflora of other fruits is only fragmentary, but greater knowledge in this area can be very helpful in developing biocontrol strategies. We characterized the yeast microflora of nectarines ('Croce del Sud') from the early stages of fruit development until harvest. The fruit samples were collected from trees in an unmanaged orchard. The resident fruit microflora was separated from the occasionally deposited microorganisms by discarding initial fruit washings before the final wash, followed by sonication and plating on NYDA medium. The isolated yeasts were identified by BIOLOG and by sequencing the D1/D2 domain of a large subunit of the rRNA gene and, where available, the ITS sequence. BIOLOG identified 19 and the genetic analysis 23 species of yeasts. Although the identification by these two systems was not always the same, the predominant yeasts were Rhodotorula spp., Sporodiobolus spp., Cryptococcus spp., Pichia spp., Candida spp. and yeast-like Aureobasidium pullulans. Several of the taxa appear to represent new species. The preliminary biocontrol tests against brown rot of nectarine fruit caused by Monilinia fructicola indicates significant decay control potential of some of the identified yeast species, namely Cryptococcus magnus, Cryptococcus sp. nov., Sporidiobolus pararoseus, A. pullulans and Rhodotorula sp. nov.

  17. Recombinant bacterial amylopullulanases

    PubMed Central

    Nisha, M; Satyanarayana, T

    2013-01-01

    Pullulanases are endo-acting enzymes capable of hydrolyzing α-1, 6-glycosidic linkages in starch, pullulan, amylopectin, and related oligosaccharides, while amylopullulanases are bifunctional enzymes with an active site capable of cleaving both α-1, 4 and α-1, 6 linkages in starch, amylose and other oligosaccharides, and α-1, 6 linkages in pullulan. The amylopullulanases are classified in GH13 and GH57 family enzymes based on the architecture of catalytic domain and number of conserved sequences. The enzymes with two active sites, one for the hydrolysis of α-1, 4- glycosidic bond and the other for α-1, 6-glycosidic bond, are called α-amylase-pullulanases, while amylopullulanases have only one active site for cleaving both α-1, 4- and α-1, 6-glycosidic bonds. The amylopullulanases produced by bacteria find applications in the starch and baking industries as a catalyst for one step starch liquefaction-saccharification for making various sugar syrups, as antistaling agent in bread and as a detergent additive. PMID:23645215

  18. Pullulanase: Role in Starch Hydrolysis and Potential Industrial Applications

    PubMed Central

    Hii, Siew Ling; Tan, Joo Shun; Ling, Tau Chuan; Ariff, Arbakariya Bin

    2012-01-01

    The use of pullulanase (EC 3.2.1.41) has recently been the subject of increased applications in starch-based industries especially those aimed for glucose production. Pullulanase, an important debranching enzyme, has been widely utilised to hydrolyse the α-1,6 glucosidic linkages in starch, amylopectin, pullulan, and related oligosaccharides, which enables a complete and efficient conversion of the branched polysaccharides into small fermentable sugars during saccharification process. The industrial manufacturing of glucose involves two successive enzymatic steps: liquefaction, carried out after gelatinisation by the action of α-amylase; saccharification, which results in further transformation of maltodextrins into glucose. During saccharification process, pullulanase has been used to increase the final glucose concentration with reduced amount of glucoamylase. Therefore, the reversion reaction that involves resynthesis of saccharides from glucose molecules is prevented. To date, five groups of pullulanase enzymes have been reported, that is, (i) pullulanase type I, (ii) amylopullulanase, (iii) neopullulanase, (iv) isopullulanase, and (v) pullulan hydrolase type III. The current paper extensively reviews each category of pullulanase, properties of pullulanase, merits of applying pullulanase during starch bioprocessing, current genetic engineering works related to pullulanase genes, and possible industrial applications of pullulanase. PMID:22991654

  19. Automating multi-step paper-based assays using integrated layering of reagents.

    PubMed

    Jahanshahi-Anbuhi, Sana; Kannan, Balamurali; Pennings, Kevin; Monsur Ali, M; Leung, Vincent; Giang, Karen; Wang, Jingyun; White, Dawn; Li, Yingfu; Pelton, Robert H; Brennan, John D; Filipe, Carlos D M

    2017-02-28

    We describe a versatile and simple method to perform sequential reactions on paper analytical devices by stacking dry pullulan films on paper, where each film contains one or more reagents or acts as a delay layer. Exposing the films to an aqueous solution of the analyte leads to sequential dissolution of the films in a temporally controlled manner followed by diffusive mixing of the reagents, so that sequential reactions can be performed. The films can be easily arranged for lateral flow assays or for spot tests (reactions take place sequentially in the z-direction). We have tested the general feasibility of the approach using three different model systems to demonstrate different capabilities: 1) pH ramping from low to high and high to low to demonstrate timing control; 2) rapid ready-to-use two-step Simon's assays on paper for detection of drugs of abuse utilizing a 2-layer stack containing two different reagents to demonstrate the ability to perform assays in the z-direction; and 3) sequential cell lysing and colorimetric detection of an intracellular bacterial enzyme, to demonstrate the ability of the method to perform sample preparation and analysis in the form of a spot assay. Overall, these studies demonstrate the potential of stacked pullulan films as useful components to enable multi-step assays on simple paper-based devices.

  20. Xylo-oligosaccharides and lactitol promote the growth of Bifidobacterium lactis and Lactobacillus species in pure cultures.

    PubMed

    Mäkeläinen, H; Saarinen, M; Stowell, J; Rautonen, N; Ouwehand, A C

    2010-06-01

    The current screening study aimed at identifying promising prebiotic and synbiotic candidates. The fermentation of xylo-oligosaccharides, xylan, galacto-oligosaccharide, fructo-oligosaccharide, polydextrose, lactitol, gentiobiose and pullulan was investigated in vitro. The ability of these established and potential prebiotic candidates to function as a sole carbon source for probiotic (Bifidobacterium and Lactobacillus), intestinal and potential pathogenic microbes (Eubacterium, Bacteroides, Clostridium, Escherichia coli, Salmonella, and Staphylococcus) was assessed in pure cultures. Xylo-oligosaccharides were fermented with high specificity by the tested Bifidobacterium lactis strains and lactitol by lactobacilli, whereas galacto-oligosaccharides, fructo-oligosaccharides and gentiobiose were utilised by a larger group of microbes. Xylan, polydextrose and pullulan were utilised to a limited extent by only a few of the tested microbes. The results of this screening study indicate that xylo-oligosaccharides and lactitol support the growth of a limited number of beneficial microbes in pure cultures. Such a high degree of specificity has not been previously reported for established prebiotics. Based on these results, the most promising prebiotics and synbiotic combinations can be selected for further testing.

  1. The effect of application of micromycetes on plant growth, as well as soybean and barley yields.

    PubMed

    Ignatova, Lyudmila; Brazhnikova, Yelena; Berzhanova, Ramza; Mukasheva, Togzhan

    2015-01-01

    The possibility of application of micromycetes (strains Penicillium bilaiae Pb14, Aureobasidium pullulans YA05 and Rhodotorula mucilaginosa YR07) to increase yields of soybean (Glycine max cv Almaty) and barley (Hordeum vulgare cv Arna) was estimated. It was shown that the most positive effect on germination energy and seed germination after seed treatment with liquid culture, supernatant and filtrate, is achieved at 1:5 dilution. In studying the influence of cell-associated and extracellular biologically active compounds of micromycetes (liquid culture and supernatant) on biometric parameters of seedlings, the maximum stimulating effect was observed in the variants with liquid culture. These strains of micromycetes were used as a bases for various compositions of preparations - application of each strain separately and application of micromycetes mixes. In microfield experiments, the increase of soybean yield ranged from 4.5 to 9.4 quintal/ha, barley - from 2.9 to 5.9 quintal/ha. A significant increase in various parameters of structure of the yield was shown in all experimental variants when compared to the control. It was found that an increase in soybean and barley yields and yield components was higher in the variant with a mix of micromycetes when compared to the separate application of each strain. The most efficient mixture was based on the mix of fungal strains (culture filtrate of P. bilaiae Pb14 diluted 1:5 + liquid cultures of A. pullulans YA05 and Rh. mucilaginosa YR07 in a 1:5 dilution).

  2. The effect of locust bean gum (LBG)-based edible coatings carrying biocontrol yeasts against Penicillium digitatum and Penicillium italicum causal agents of postharvest decay of mandarin fruit.

    PubMed

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2016-09-01

    Strains belonging to Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans species were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic molds Penicillium digitatum and Penicillium italicum. Moreover, studies aimed at screening the antifungal activity of selected yeast strains in vivo conditions against P. digitatum and P. italicum, and investigated the efficacy of a polysaccharidic matrix, locust bean gum (LBG), enriched with the tested BCAs, in controlling postharvest decays in artificially inoculated mandarins. The population dynamics of BCAs on wounds and the magnitude of peroxidase (POD) and superoxide dismutase (SOD) in fruit tissues were also investigated after treatments of mandarins with antagonistic yeasts. W. anomalus BS91, M. pulcherrima MPR3 and A. pullulans PI1 provided excellent control of postharvest decays caused by P. digitatum and P. italicum on mandarins, both when the yeasts were used alone and in combination with LBG, which enhanced the yeast cell viability over time. Finally, the increased activity of POD and lower decrease in SOD activity in response to BCAs application in mandarin fruits confirmed their involvement in the biocontrol mechanism. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Pullulanase: role in starch hydrolysis and potential industrial applications.

    PubMed

    Hii, Siew Ling; Tan, Joo Shun; Ling, Tau Chuan; Ariff, Arbakariya Bin

    2012-01-01

    The use of pullulanase (EC 3.2.1.41) has recently been the subject of increased applications in starch-based industries especially those aimed for glucose production. Pullulanase, an important debranching enzyme, has been widely utilised to hydrolyse the α-1,6 glucosidic linkages in starch, amylopectin, pullulan, and related oligosaccharides, which enables a complete and efficient conversion of the branched polysaccharides into small fermentable sugars during saccharification process. The industrial manufacturing of glucose involves two successive enzymatic steps: liquefaction, carried out after gelatinisation by the action of α-amylase; saccharification, which results in further transformation of maltodextrins into glucose. During saccharification process, pullulanase has been used to increase the final glucose concentration with reduced amount of glucoamylase. Therefore, the reversion reaction that involves resynthesis of saccharides from glucose molecules is prevented. To date, five groups of pullulanase enzymes have been reported, that is, (i) pullulanase type I, (ii) amylopullulanase, (iii) neopullulanase, (iv) isopullulanase, and (v) pullulan hydrolase type III. The current paper extensively reviews each category of pullulanase, properties of pullulanase, merits of applying pullulanase during starch bioprocessing, current genetic engineering works related to pullulanase genes, and possible industrial applications of pullulanase.

  4. Stabilization of membranes upon interaction of amphipathic polymers with membrane proteins

    PubMed Central

    Picard, Martin; Duval-Terrié, Caroline; Dé, Emmanuelle; Champeil, Philippe

    2004-01-01

    Amphipathic polymers derived from polysaccharides, namely hydrophobically modified pullulans, were previously suggested to be useful as polymeric substitutes of ordinary surfactants for efficient and structure-conserving solubilization of membrane proteins, and one such polymer, 18C10, was optimized for solubilization of proteins derived from bacterial outer membranes (Duval-Terrié et al. 2003). We asked whether a similar ability to solubilize proteins could also be demonstrated in eukaryotic membranes, namely sarcoplasmic reticulum (SR) fragments, the major protein of which is SERCA1a, an integral membrane protein with Ca2+-dependent ATPase and Ca2+-pumping activity. We found that 18C10-mediated solubilization of these SR membranes did not occur. Simultaneously, however, we found that low amounts of this hydrophobically modified pullulan were very efficient at preventing long-term aggregation of these SR membranes. This presumably occurred because the negatively charged polymer coated the membranous vesicles with a hydrophilic corona (a property shared by many other amphipathic polymers), and thus minimized their flocculation. Reminiscent of the old Arabic gum, which stabilizes Indian ink by coating charcoal particles, the newly designed amphipathic polymers might therefore unintentionally prove useful also for stabilization of membrane suspensions. PMID:15459343

  5. Gene cloning, functional expression and characterisation of a novel type I pullulanase from Paenibacillus barengoltzii and its application in resistant starch production.

    PubMed

    Liu, Jingjing; Liu, Yu; Yan, Feng; Jiang, Zhengqiang; Yang, Shaoqing; Yan, Qiaojuan

    2016-05-01

    A novel pullulanase gene (PbPulA) from Paenibacillus barengoltzii was cloned. PbPulA has an open reading frame of 2028 bp encoding 675 amino acids. It was heterologously expressed in Escherichia coli as an intracellular soluble protein. The recombinant pullulanase (PbPulA) was purified to homogeneity with a molecular mass of about 75 kDa on SDS-PAGE. PbPulA was optimally active at pH 5.5 and 50 °C. It was stable within pH 5.5-10.5. The enzyme exhibited strict substrate specificity towards pullulan, but showed relatively low activity towards amylopectin and no activity towards other tested polysaccharides. The Km and Vmax values of the enzyme on pullulan were 2.94 mg/mL and 280.5 μmol/min/mg, respectively. The addition of PbPulA in gelatinized rice and maize starches significantly increased the resistant starch type 3 (RS3) yields. Final yields from rice and maize starches were 10.82 g/100 g and 11.41 g/100 g, respectively. These properties make PbPulA useful in starch industries.

  6. Occurrence and infection of Cladosporium, Fusarium, Epicoccum and Aureobasidium in withered rotten grapes during post-harvest dehydration.

    PubMed

    Lorenzini, Marilinda; Zapparoli, Giacomo

    2015-11-01

    Fungi like Cladosporium, Fusarium, Epicoccum and Aureobasidium can occur on withered grapes causing spoilage of passito wine. There is little or no information on the pathogenic role of these fungi. This study describes the isolation, incidence and identification of several isolates from different withered rotten grapes. Representative isolates grouped in several phenotypes were identified by phylogenetic analysis of internal transcribed spacer, actin or elongation factor gene sequences. Isolates of Cladosporium and Fusarium were ascribed to different species, of these C. ramotenellum, C. halotolerans and F. graminearum were isolated from Vitis vinifera for the first time. All Epicoccum and Aureobasidium isolates belonged to E. nigrum and A. pullulans, respectively. Random amplified DNA polymorphism analysis showed high level of heterogenicity among Epicoccum and Fusarium isolates. Infection assays were carried out to evaluate infectivity in some strains under different withering conditions. Fusarium spp. strains had similar infectivity, while significant variability was observed among Cladosporium spp. and E. nigrum strains. A. pullulans resulted particularly infective. This study provided insights into the occurrence and infection of these fungi in fruit-drying rooms with important implications towards control management during the withering.

  7. Rye grains and the soil derived from under the organic and conventional rye crops as a potential source of biological agents causing respiratory diseases in farmers

    PubMed Central

    Cholewa, Grażyna; Krasowska, Ewelina; Chmielewska-Badora, Jolanta; Zwoliński, Jacek; Sobczak, Paweł

    2013-01-01

    Introduction Introduction: Due to the specific work environment, farmers are exposed to various biological occupational hazard. Among these factors significant are fungi present in the grain and also in the soil. The fungi may be the cause of human diseases including skin infections, asthma, allergic rhinitis and many others. Aim The aim of this study was to quantify and identify species of fungi colonizing rye grain samples and the soil under cultivation. Material and methods The material consisted of grain and soil samples from two agricultural systems: organic and conventional. To determine the concentration and composition of fungi in collected samples, two media: Malt Agar (MA, Becton, Dickinson and Company) and Potato Dextrose Agar (PDA, Becton, Dickinson and Company) were used. The composition of species in fungal flora was determined using macroscopic and microscopic methods. The isolates of fungi were ranked in the appropriate classes of biosafety BSL. Results The most frequently isolated fungi from organic rye grain, regardless of the media used, were species: Aureobasidium pullulans and Alternaria alternata. In conventional farms, most species isolated from rye grain were: Aureobasidium pullulans, Cladosporium oxysporum, Alternaria alternata and yeast-like fungi. Most often species isolated from the soil was Penicillium citreo-viride. Conclusions All the results of the research demonstrate the potential hazard to the health of people working in agriculture. Significant exposure of this professional group is associated with the presence of harmful biological agents present in the grain and soil from its cultivation. PMID:24494000

  8. Yeasts colonizing the leaf surfaces.

    PubMed

    Sláviková, Elena; Vadkertiová, Renata; Vránová, Dana

    2007-08-01

    The yeasts were isolated from the leaf surfaces of ten species of trees. The study site was a forest park (Zelezná Studnicka) of the Small Carpathians mountain range. One hundred and thirty seven yeast strains belonging to 13 genera were isolated from 320 samples of leaves and needles. Seventeen yeast species were isolated, but only seven occurred regularly: Aureobasidium pullulans, Cryptococcus laurentii, Pichia anomala, Metschnikowia pulcherrima, Saccharomyces sp., Lachancea thermotolerans, and Rhodotorula glutinis. The remaining species were isolated from the leaves and needles of three or less tree species. A. pullulans, Cr. laurentii, and P. anomala were the most frequently found species and they occurred on leaves and needles of all ten tree species. Saccharomyces sp. occurred in leaf samples collected from eight kinds of trees. M. pulcherrima and L. thermotolerans were found in samples collected from six species of trees. Both these species occurred almost always on the leaves of deciduous trees. Rh. glutinis was the most frequently isolated carotenoids producing species. We have found out that the ascomycetous and basidiomycetous species were present in the leaf samples in approximately equal frequency, contrary to the soil samples taken from this forest park, where the ascomycetous species were found rarely.

  9. Stabilization of membranes upon interaction of amphipathic polymers with membrane proteins.

    PubMed

    Picard, Martin; Duval-Terrié, Caroline; Dé, Emmanuelle; Champeil, Philippe

    2004-11-01

    Amphipathic polymers derived from polysaccharides, namely hydrophobically modified pullulans, were previously suggested to be useful as polymeric substitutes of ordinary surfactants for efficient and structure-conserving solubilization of membrane proteins, and one such polymer, 18C(10), was optimized for solubilization of proteins derived from bacterial outer membranes (Duval-Terrie et al. 2003). We asked whether a similar ability to solubilize proteins could also be demonstrated in eukaryotic membranes, namely sarcoplasmic reticulum (SR) fragments, the major protein of which is SERCA1a, an integral membrane protein with Ca(2+)-dependent ATPase and Ca(2+)-pumping activity. We found that 18C(10)-mediated solubilization of these SR membranes did not occur. Simultaneously, however, we found that low amounts of this hydrophobically modified pullulan were very efficient at preventing long-term aggregation of these SR membranes. This presumably occurred because the negatively charged polymer coated the membranous vesicles with a hydrophilic corona (a property shared by many other amphipathic polymers), and thus minimized their flocculation. Reminiscent of the old Arabic gum, which stabilizes Indian ink by coating charcoal particles, the newly designed amphipathic polymers might therefore unintentionally prove useful also for stabilization of membrane suspensions.

  10. Inactivation of bacteria and yeast using high-frequency ultrasound treatment.

    PubMed

    Gao, Shengpu; Hemar, Yacine; Ashokkumar, Muthupandian; Paturel, Sara; Lewis, Gillian D

    2014-09-01

    High-frequency (850 kHz) ultrasound was used to inactivate bacteria and yeast at different growth phases under controlled temperature conditions. Three species of bacteria, Enterobacter aerogenes, Bacillus subtilis and Staphylococcus epidermidis as well as a yeast, Aureobasidium pullulans were considered. The study shows that high-frequency ultrasound is highly efficient in inactivating the bacteria in both their exponential and stationary growth phases, and inactivation rates of more than 99% were achieved. TEM observation suggests that the mechanism of bacteria inactivation is mainly due to acoustic cavitation generated free radicals and H2O2. The rod-shaped bacterium B. subtilis was also found to be sensitive to the mechanical effects of acoustic cavitation. The study showed that the inactivation process continued even after ultrasonic processing cessed due to the presence of H2O2, generated during acoustic cavitation. Compared to bacteria, the yeast A. pullulans was found to be more resistant to high-frequency ultrasound treatment.

  11. Breathing silicon anodes for durable high-power operations.

    PubMed

    Hwang, Chihyun; Joo, Sehun; Kang, Na-Ri; Lee, Ungju; Kim, Tae-Hee; Jeon, Yuju; Kim, Jieun; Kim, Young-Jin; Kim, Ju-Young; Kwak, Sang-Kyu; Song, Hyun-Kon

    2015-09-23

    Silicon anode materials have been developed to achieve high capacity lithium ion batteries for operating smart phones and driving electric vehicles for longer time. Serious volume expansion induced by lithiation, which is the main drawback of silicon, has been challenged by multi-faceted approaches. Mechanically rigid and stiff polymers (e.g. alginate and carboxymethyl cellulose) were considered as the good choices of binders for silicon because they grab silicon particles in a tight and rigid way so that pulverization and then break-away of the active mass from electric pathways are suppressed. Contrary to the public wisdom, in this work, we demonstrate that electrochemical performances are secured better by letting silicon electrodes breathe in and out lithium ions with volume change rather than by fixing their dimensions. The breathing electrodes were achieved by using a polysaccharide (pullulan), the conformation of which is modulated from chair to boat during elongation. The conformational transition of pullulan was originated from its α glycosidic linkages while the conventional rigid polysaccharide binders have β linkages.

  12. Breathing silicon anodes for durable high-power operations

    PubMed Central

    Hwang, Chihyun; Joo, Sehun; Kang, Na-Ri; Lee, Ungju; Kim, Tae-Hee; Jeon, Yuju; Kim, Jieun; Kim, Young-Jin; Kim, Ju-Young; Kwak, Sang-Kyu; Song, Hyun-Kon

    2015-01-01

    Silicon anode materials have been developed to achieve high capacity lithium ion batteries for operating smart phones and driving electric vehicles for longer time. Serious volume expansion induced by lithiation, which is the main drawback of silicon, has been challenged by multi-faceted approaches. Mechanically rigid and stiff polymers (e.g. alginate and carboxymethyl cellulose) were considered as the good choices of binders for silicon because they grab silicon particles in a tight and rigid way so that pulverization and then break-away of the active mass from electric pathways are suppressed. Contrary to the public wisdom, in this work, we demonstrate that electrochemical performances are secured better by letting silicon electrodes breathe in and out lithium ions with volume change rather than by fixing their dimensions. The breathing electrodes were achieved by using a polysaccharide (pullulan), the conformation of which is modulated from chair to boat during elongation. The conformational transition of pullulan was originated from its α glycosidic linkages while the conventional rigid polysaccharide binders have β linkages. PMID:26395407

  13. Remote detection of laser-induced autofluorescence on pure cultures of fungal and bacterial strains and their analysis with multivariate techniques

    NASA Astrophysics Data System (ADS)

    Raimondi, Valentina; Palombi, Lorenzo; Cecchi, Giovanna; Lognoli, David; Trambusti, Massimo; Gomoiu, Ioana

    2007-05-01

    Remotely sensed laser-induced autofluorescence spectra of pure cultures of fungal strains ( Aureobasidium pullulans, Verticillium sp.) and of bacterial strains ( Bacillus sp., Pseudomonas sp.) are presented. The strains were isolated from samples collected in a Roman archaeological site ( Tropaeum Traiani) near Constanta, Romania. The fluorescence spectra were detected in vivo from a distance of 25 m in the outdoor, using a high spectral resolution fluorescence LIDAR featuring a UV laser (XeCl@308 nm) as an excitation source. All the examined strains, except for the A. pullulans, showed fluorescence features such to allow their characterisation by processing data with multivariate techniques. Both Principal Component Analysis and Cluster Analysis were applied to the data set and compared to discriminate between the examined strains. Results demonstrate the feasibility of fluorescence-based detection and characterisation of fungi and bacteria in the outdoor with a high spectral resolution fluorescence LIDAR. In addition, they show that the proposed processing methods offer a means to discriminate between the fluorescence features due to the investigated samples and that of a fluorescence background of a known spectral shape, as that of the culture medium. This can be exploited for the remote fluorescence mapping of heterotrophic organisms on stone surfaces when the latter show a typical broad fluorescence band.

  14. Breathing silicon anodes for durable high-power operations

    NASA Astrophysics Data System (ADS)

    Hwang, Chihyun; Joo, Sehun; Kang, Na-Ri; Lee, Ungju; Kim, Tae-Hee; Jeon, Yuju; Kim, Jieun; Kim, Young-Jin; Kim, Ju-Young; Kwak, Sang-Kyu; Song, Hyun-Kon

    2015-09-01

    Silicon anode materials have been developed to achieve high capacity lithium ion batteries for operating smart phones and driving electric vehicles for longer time. Serious volume expansion induced by lithiation, which is the main drawback of silicon, has been challenged by multi-faceted approaches. Mechanically rigid and stiff polymers (e.g. alginate and carboxymethyl cellulose) were considered as the good choices of binders for silicon because they grab silicon particles in a tight and rigid way so that pulverization and then break-away of the active mass from electric pathways are suppressed. Contrary to the public wisdom, in this work, we demonstrate that electrochemical performances are secured better by letting silicon electrodes breathe in and out lithium ions with volume change rather than by fixing their dimensions. The breathing electrodes were achieved by using a polysaccharide (pullulan), the conformation of which is modulated from chair to boat during elongation. The conformational transition of pullulan was originated from its α glycosidic linkages while the conventional rigid polysaccharide binders have β linkages.

  15. Yeasts and yeast-like organisms associated with fruits and blossoms of different fruit trees.

    PubMed

    Vadkertiová, Renáta; Molnárová, Jana; Vránová, Dana; Sláviková, Elena

    2012-12-01

    Yeasts are common inhabitants of the phyllosphere, but our knowledge of their diversity in various plant organs is still limited. This study focused on the diversity of yeasts and yeast-like organisms associated with matured fruits and fully open blossoms of apple, plum, and pear trees, during 2 consecutive years at 3 localities in southwest Slovakia. The occurrence of yeasts and yeast-like organisms in fruit samples was 2½ times higher and the yeast community more diverse than that in blossom samples. Only 2 species (Aureobasidium pullulans and Metschnikowia pulcherrima) occurred regularly in the blossom samples, whereas Galactomyces candidus, Hanseniaspora guilliermondii, Hanseniaspora uvarum, M. pulcherrima, Pichia kluyveri, Pichia kudriavzevii, and Saccharomyces cerevisiae were the most frequently isolated species from the fruit samples. The ratio of the number of samples where only individual species were present to the number of samples where 2 or more species were found (consortium) was counted. The occurrence of individual species in comparison with consortia was much higher in blossom samples than in fruit samples. In the latter, consortia predominated. Aureobasidium pullulans, M. pulcherrima, and S. cerevisiae, isolated from both the fruits and blossoms, can be considered as resident yeast species of various fruit tree species cultivated in southwest Slovakia localities.

  16. Temperature-responsive size-exclusion chromatography using poly(N-isopropylacrylamide) grafted silica.

    PubMed

    Lakhiari, H; Okano, T; Nurdin, N; Luthi, C; Descouts, P; Muller, D; Jozefonvicz, J

    1998-03-02

    Silica-based packing materials induce non-specific interactions with proteins in aqueous media because of the nature of their surface, mainly silanol groups. Therefore, the silica surface has to be modified in order to be used as stationary phase for the High Performance Size-Exclusion Chromatography (HPSEC) of proteins. For this purpose, porous silica beads were coated with hydrophilic polymer gels (dextrans of different molecular weights) carrying a calculated amount of diethyl-aminoethyl groups (DEAE). Actually, as shown by HPSEC, these dextran modified supports minimize non-specific adsorption for proteins and pullulans in aqueous solution. Then, in order to change the pore size in response to temperature, temperature responsive polymer of poly(N-isopropylacrylamide) (PIPAAm) was introduced into the surface of dextran-DEAE on porous silica beads. The structure of these supports before and after modification was alternately studied by Scanning Electronic Microscopy (SEM) and Scanning Force Microscopy (SFM). An adsorption of radiolabelled albumin was performed to complete our study. Silica modifications by dextran-DEAE and PIPAAm improve the neutrality of the support and minimize the non-specific interactions between the solid support and proteins in solution. At low temperature, the support having PIPAAm exhibits a high resolution domain in HPSEC and finally permits a better resolution of proteins and pullulans. At higher temperature, hydrophobic properties of PIPAAm produce interactions with some proteins and trigger off a slight delay of their elution time.

  17. Graphene Oxide Bionanocomposite Coatings with High Oxygen Barrier Properties

    PubMed Central

    Uysal Unalan, Ilke; Boyacı, Derya; Ghaani, Masoud; Trabattoni, Silvia; Farris, Stefano

    2016-01-01

    In this work, we present the development of bionanocomposite coatings on poly(ethylene terephthalate) (PET) with outstanding oxygen barrier properties. Pullulan and graphene oxide (GO) were used as main polymer phase and nanobuilding block (NBB), respectively. The oxygen barrier performance was investigated at different filler volume fractions (ϕ) and as a function of different relative humidity (RH) values. Noticeably, the impermeable nature of GO was reflected under dry conditions, in which an oxygen transmission rate (OTR, mL·m−2·24 h−1) value below the detection limit of the instrument (0.01 mL·m−2·24 h−1) was recorded, even for ϕ as low as 0.0004. A dramatic increase of the OTR values occurred in humid conditions, such that the barrier performance was totally lost at 90% RH (the OTR of coated PET films was equal to the OTR of bare PET films). Modelling of the experimental OTR data by Cussler’s model suggested that the spatial ordering of GO sheets within the main pullulan phase was perturbed because of RH fluctuations. In spite of the presence of the filler, all the formulations allowed the obtainment of final materials with haze values below 3%, the only exception being the formulation with the highest loading of GO (ϕ ≈ 0.03). The mechanisms underlying the experimental observations are discussed. PMID:28335372

  18. Pressate from peat dewatering as a substrate for bacterial growth.

    PubMed

    Mulligan, C N; Cooper, D G

    1985-07-01

    This study considered the possibility of using water expressed during the drying of fuel-grade peat as a substrate for microbial growth. Highly humified peat pressed for 2.5 min at 1.96 MPa produced water with a chemical oxygen demand of 690 mg/liter. Several biological compounds could be produced by using the organic matter in expressed peat water as a substrate. These included polymers such as chitosan, contained in the cell wall of Rhizopus arrhizus, and two extracellular polysaccharides, xanthan gum and pullulan, produced by Xanthomonas campestris and Aureobasidium pullulans, respectively. A very effective surfactant was produced by Bacillus subtilis grown in the expressed water. Small additions of nutrients to the peat pressate were necessary to obtain substantial yields of products. The addition of peptone, yeast extract, and glucose improved production of the various compounds. Biological treatment improved the quality of the expressed water to the extent that in an industrial process it could be returned to the environment.

  19. Biocontrol ability and action mechanism of food-isolated yeast strains against Botrytis cinerea causing post-harvest bunch rot of table grape.

    PubMed

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2015-05-01

    Strains belonging to the species Saccharomyces cerevisiae, Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans, isolated from different food sources, were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic mold Botrytis cinerea. All yeast strains demonstrated antifungal activity at different levels depending on species and medium. Killer strains of W. anomalus and S. cerevisiae showed the highest biocontrol in vitro activity, as demonstrated by largest inhibition halos. The competition for iron and the ability to form biofilm and to colonize fruit wounds were hypothesized as the main action mechanisms for M. pulcherrima. The production of hydrolytic enzymes and the ability to colonize the wounds were the most important mechanisms for biocontrol activity in A. pullulans and W. anomalus, which also showed high ability to form biofilm. The production of volatile organic compounds (VOCs) with in vitro and in vivo inhibitory effect on pathogen growth was observed for the species W. anomalus, S. cerevisiae and M. pulcherrima. Our study clearly indicates that multiple modes of action may explain as M. pulcherrima provide excellent control of postharvest botrytis bunch rot of grape.

  20. Carbohydrate Availability Regulates Virulence Gene Expression in Streptococcus suis

    PubMed Central

    Ferrando, M. Laura; van Baarlen, Peter; Orrù, Germano; Piga, Rosaria; Bongers, Roger S.; Wels, Michiel; De Greeff, Astrid; Smith, Hilde E.; Wells, Jerry M.

    2014-01-01

    Streptococcus suis is a major bacterial pathogen of young pigs causing worldwide economic problems for the pig industry. S. suis is also an emerging pathogen of humans. Colonization of porcine oropharynx by S. suis is considered to be a high risk factor for invasive disease. In the oropharyngeal cavity, where glucose is rapidly absorbed but dietary α-glucans persist, there is a profound effect of carbohydrate availability on the expression of virulence genes. Nineteen predicted or confirmed S. suis virulence genes that promote adhesion to and invasion of epithelial cells were expressed at higher levels when S. suis was supplied with the α-glucan starch/pullulan compared to glucose as the single carbon source. Additionally the production of suilysin, a toxin that damages epithelial cells, was increased more than ten-fold when glucose levels were low and S. suis was growing on pullulan. Based on biochemical, bioinformatics and in vitro and in vivo gene expression studies, we developed a biological model that postulates the effect of carbon catabolite repression on expression of virulence genes in the mucosa, organs and blood. This research increases our understanding of S. suis virulence mechanisms and has important implications for the design of future control strategies including the development of anti-infective strategies by modulating animal feed composition. PMID:24642967

  1. Thermostable amylolytic enzymes from a new Clostridium isolate

    SciTech Connect

    Madi, E.; Antranikian, G.; Ohmiya, K.; Gottschalk, G.

    1987-07-01

    A new Clostridium strain was isolated on starch at 60 degrees C. Starch, pullulan, maltotriose, and maltose induced the synthesis of alpha-amylase and pullulanase, while glucose, ribose, fructose, and lactose did not. The formation of the amylolytic enzymes was dependent on growth and occurred predominantly in the exponential phase. The enzymes were largely cell bound during growth of the organism with 0.5% starch, but an increase of the starch concentration in the growth medium was accompanied by the excretion of alpha-amylase and pullulanase into the culture broth; but also by a decrease of total activity. Alpha-amylase, pullulanase, and alpha-glucosidase were active in a broad temperature range (40 to 85 degrees C) and displayed temperature optima for activity at 60 to 70 degrees C. During incubation with starch under aerobic conditions at 75 degrees C for 2 hours, the activity of both enzymes decreased to only 90 or 80%. The apparent Km values of alpha-amylase, pullulanase, and alpha-glucosidase for their corresponding substrates, starch, pullulan, and maltose were 0.35 mg/ml, 0.63 mg/ml, and 25 mM, respectively. (Refs. 31).

  2. Isolation and characterization of Schwanniomyces alluvius amylolytic enzymes

    SciTech Connect

    Wilson, J.J.; Ingledew, W.M.

    1982-08-01

    The extracellular amylolytic enzymes of Schwanniomyces alluvius were studied to determine future optimization of this yeast for the production of industrial ethanol from starch. Both alpha-amylase and glucoamylase were isolated and purified. Alpha-amylase has an optimum pH of 6.3 and was stable from pH 4.5 to 7.5. The optimum temperature for the enzyme was 40 degrees C, but it was quickly inactivated at temperatures above 40 degrees C. The Km for soluble starch was 0.364 mg/ml. The molecular weight was calculated to be 61,900 + or - 700. Alpha-amylase was capable of releasing glucose from starch, but not from pullulan. Glucoamylase had an optimum pH of 5.0 and was stable from pH 4.0 to more than 8.0. The optimum temperature for the enzyme was 50 degrees C, and although less heat sensitive than alpha-amylase, it was quickly inactivated at 60 degrees C. Km values were 12.67 mg/ml for soluble starch and 0.72 mM for maltose. The molecular weight was calculated to be 155,000 + or - 3,000. Glucoamylase released only glucose from both soluble starch and pullulan. Schwanniomyces alluvius is one of the very few yeasts to possess both alpha-amylase and glucoamylase as well as some fermentative capacity to produce ethanol. (Refs. 9).

  3. Fungal exopolysaccharide: production, composition and applications.

    PubMed

    Mahapatra, Subhadip; Banerjee, Debdulal

    2013-01-01

    Fungal exopolysaccharides (EPSs) have been recognized as high value biomacromolecules for the last two decades. These products, including pullulan, scleroglucan, and botryosphaeran, have several applications in industries, pharmaceuticals, medicine, foods etc. Although fungal EPSs are highly relevant, to date information concerning fungal biosynthesis is scarce and an extensive search for new fugal species that can produce novel EPSs is still needed. In most cases, the molecular weight variations and sugar compositions of fungal EPSs are dependent to culture medium composition and different physical conditions provided during fermentation. An inclusive and illustrative review on fungal EPS is presented here. The general outline of the present work includes fungal EPS production, their compositions and applications. An emphasis is also given to listing out different fungal strains that can produce EPSs.

  4. Glycoproteins functionalized natural and synthetic polymers for prospective biomedical applications: A review.

    PubMed

    Tabasum, Shazia; Noreen, Aqdas; Kanwal, Arooj; Zuber, Mohammad; Anjum, Muhammad Naveed; Zia, Khalid Mahmood

    2017-05-01

    Glycoproteins have multidimensional properties such as biodegradability, biocompatibility, non-toxicity, antimicrobial and adsorption properties; therefore, they have wide range of applications. They are blended with different polymers such as chitosan, carboxymethyl cellulose (CMC), polyvinyl pyrrolidone (PVP), polycaprolactone (PCL), heparin, polystyrene fluorescent nanoparticles (PS-NPs) and carboxyl pullulan (PC) to improve their properties like thermal stability, mechanical properties, resistance to pH, chemical stability and toughness. Considering the versatile charateristics of glycoprotein based polymers, this review sheds light on synthesis and characterization of blends and composites of glycoproteins, with natural and synthetic polymers and their potential applications in biomedical field such as drug delivery system, insulin delivery, antimicrobial wound dressing uses, targeting of cancer cells, development of anticancer vaccines, development of new biopolymers, glycoproteome research, food product and detection of dengue glycoproteins. All the technical scientific issues have been addressed; highlighting the recent advancement.

  5. Production of Cellulolytic and Hemicellulolytic Enzymes From Aureobasidium pulluans on Solid State Fermentation

    NASA Astrophysics Data System (ADS)

    Leite, Rodrigo Simões Ribeiro; Bocchini, Daniela Alonso; da Silva Martins, Eduardo; Silva, Dênis; Gomes, Eleni; da Silva, Roberto

    This article investigates a strain of the yeast Aureobasidium pullulans for cellulase and hemicellulase production in solid state fermentation. Among the substrates analyzed, the wheat bran culture presented the highest enzymatic production (1.05 U/mL endoglucanase, 1.3 U/mL β-glucosidase, and 5.0 U/mL xylanase). Avicelase activity was not detected. The optimum pH and temperature for xylanase, endoglucanase and β-glucosidase were 5.0 and 50, 4.5 and 60, 4.0 and 75°C, respectively. These enzymes remained stable between a wide range of pH. The β-glucosidase was the most thermostable enzyme remaining 100% active when incubated at 75°C for 1 h.

  6. Therapeutic Trials Based on Combination of Drug Delivery System and Ultrasound

    NASA Astrophysics Data System (ADS)

    Tabata, Yasuhiko

    2005-03-01

    The objective of this study is to show how therapeutically effective a combination of a drug delivery system (DDS) and ultrasound (US) is by introducing data from two related experiments. Fullerene (C60) was chemically modified by polyethylene glycol (PEG) for water-solubilization and tumour targeting. When the PEG-modified C60 was injected intravenously into tumour bearing mice, followed by US irradiation to the tumour site, a synergistic anti-tumour effect was observed. Following the intravenous injection of interferon (IFN) conjugated with pullulan which has an inherent affinity for the liver, and subsequent US irradiation to the liver, the activity level of an IFN-specific enzyme was significantly enhanced compared with that of the conjugate injection alone.

  7. Purification of extrachloroplastic. beta. -amylase from leaves of starchless and wild type Arabidopsis

    SciTech Connect

    Somerville, C.; Monroe, J.; Preiss, J. )

    1989-04-01

    Amylase activity in crude leaf extracts from starchless mutants of Arabidopsis thaliana is 5 to 10 fold higher than in the wild type (WT) when plants are grown under a 12 h photoperiod. Visualized on native PAGE, the increased activity is attributed primarily to a previously characterized extrachloroplastic {beta}-(exo)amylase. The {beta}-amylases from phosoglucomutase deficient (starchless) and WT leaves were purified to homogeneity in two steps utilizing polyethylene glycol fractionation, and cyclohexaamylose affinity chromatography. The enzyme from both mutant and WT leaves had negligible activity toward either {beta}-limit dextrin or pullulan. The specific activities of both purified enzymes were similar indicating that the protein is over-expressed in the mutant. Preliminary antibody neutralization experiments suggest that the two {beta}-amylases are not different.

  8. Design and analysis of a low actuation voltage electrowetting-on-dielectric microvalve for drug delivery applications.

    PubMed

    Samad, Mst Fateha; Kouzani, Abbas Z

    2014-01-01

    This paper presents a low actuation voltage microvalve with optimized insulating layers that manipulates a conducting ferro-fluid droplet by the principle of electrowetting-on-dielectric (EWOD). The proposed EWOD microvalve contains an array of chromium (Cr) electrodes on the soda-lime glass substrate, covered by both dielectric and hydrophobic layers. Various dielectric layers including Su-8 2002, Polyvinylidenefluoride (PVDF) and Cyanoethyl pullulan (CEP), and thin (50 nm) hydrophobic Teflon and Cytonix are used to analyze the EWOD microvalves at different voltages. The Finite Element Method (FEM) based software, Coventorware is used to carry out the simulation analysis. It is observed that the EWOD microvalve having a CEP dielectric layer with dielectric constant of about 20 and thickness of 1 μm, and a Cytonix hydrophobic layer with thickness of 50 nm operated the conducting ferro-fluid droplet at the actuation voltage as low as 7.8 V.

  9. Effect of soluble polysaccharides addition on rheological properties and microstructure of chitin nanocrystal aqueous dispersions.

    PubMed

    Tzoumaki, Maria V; Moschakis, Thomas; Biliaderis, Costas G

    2013-06-05

    Mixtures of chitin nanocrystal aqueous dispersions (at pH 3.0) with soluble polysaccharides of varying molecular features were examined rheologically and microscopically, under different conditions of biopolymer concentration, ionic strength, pH and temperature. The addition of non-adsorbing polysaccharides (guar gum, locust bean gum and xanthan) as well as oppositely charged (κ-carrageenan) to a chitin nanocrystal dispersion, resulted in a network formation and the gel strength increased with the chitin nanocrystal concentration. In contrast, the chitin nanocrystal - chitosan or - pullulan mixed dispersions did not show any network formation (tanδ>1) at the concentration range examined. An increase in ionic strength and pH also resulted in an enhanced elasticity of the chitin nanocrystal-guar gum dispersions. Furthermore, an increase in the elastic modulus, which was irreversible upon cooling, was observed upon heating the chitin nanocrystal-polysaccharide mixed dispersions.

  10. Expression of multiple complex polysaccharide-degrading enzyme systems by marine bacterium strain 2-40.

    PubMed

    Ensor; Stosz; Weiner

    1999-08-01

    Saprophytic marine bacterium strain 2-40 (2-40) can degrade numerous complex polysaccharides (CP) including agar, alginic acid, carrageenan, carboxymethylcellulose, chitin, beta-glucan, laminarin, pectin, pullulan, starch, and xylan. The growth of 2-40 was assessed in minimal media containing one of 16 CP or simple carbohydrates, with the result that all supported growth. Each of the carbohydrase systems was elicited at highest levels by the homologous substrate. Each, excluding amylase, was repressed when 2-40 was cultured in glucose minimal synthetic media. Cyclic adenosine monophosphate alleviated the repression. Agarose as sole carbon source supported the synthesis of the most heterologous complex carbohydrase systems, although, generally, at a lower level of activity than the homologous CP.

  11. Fungal endophytes in germinated seeds of the common bean, Phaseolus vulgaris

    PubMed Central

    Parsa, Soroush; García-Lemos, Adriana M.; Castillo, Katherine; Ortiz, Viviana; López-Lavalle, Luis Augusto Becerra; Braun, Jerome; Vega, Fernando E.

    2016-01-01

    We conducted a survey of fungal endophytes in 582 germinated seeds belonging to 11 Colombian cultivars of the common bean (Phaseolus vulgaris). The survey yielded 394 endophytic isolates belonging to 42 taxa, as identified by sequence analysis of the ribosomal DNA internal transcribed spacer (ITS) region. Aureobasidium pullulans was the dominant endophyte, isolated from 46.7 % of the samples. Also common were Fusarium oxysporum, Xylaria sp., and Cladosporium cladosporioides, but found in only 13.4 %, 11.7 %, and 7.6 % of seedlings, respectively. Endophytic colonization differed significantly among common bean cultivars and seedling parts, with the highest colonization occurring in the first true leaves of the seedlings. PMID:27109374

  12. Purification and characterization of a thermostable raw starch digesting amylase from a Streptomyces sp. isolated in a milling factory.

    PubMed

    Kaneko, Takahiro; Ohno, Toshihisa; Ohisa, Naganori

    2005-06-01

    A raw starch utilizing microbe was isolated from mud in a milling factory. The 16S ribosomal DNA (rDNA) sequencing and morphological properties of the strain indicated that it belongs to the genus Streptomyces. A strongly raw starch digesting amylase was purified from the culture supernatant of the strain by chromatographic procedures. The specific activity of the enzyme was 11.7 U/mg, molecular mass 47 kDa, optimum pH 6.0, and optimum temperature 50 to 60 degrees C. The enzyme showed sufficient activity even at 70 degrees C. It was activated by calcium, cobaltous, and magnesium ions, and inhibited by copper, nickel, zinc, and ferrous ions. It formed maltose mainly from raw and gelatinized starch, and glycogen. No products were formed from glucose, maltose, maltotriose, pullulan, or cyclodextrins (CDs). The enzyme digested raw wheat, rice, and waxy rice starch rapidly, and raw corn, waxy corn, sweet potato, tapioca, and potato starch normally.

  13. Extracellular Polysaccharides Produced by Yeasts and Yeast-Like Fungi

    NASA Astrophysics Data System (ADS)

    van Bogaert, Inge N. A.; de Maeseneire, Sofie L.; Vandamme, Erick J.

    Several yeasts and yeast-like fungi are known to produce extracellular polysaccharides. Most of these contain D-mannose, either alone or in combination with other sugars or phosphate. A large chemical and structural variability is found between yeast species and even among different strains. The types of polymers that are synthesized can be chemically characterized as mannans, glucans, phosphoman-nans, galactomannans, glucomannans and glucuronoxylomannans. Despite these differences, almost all of the yeast exopolysaccharides display some sort of biological activity. Some of them have already applications in chemistry, pharmacy, cosmetics or as probiotic. Furthermore, some yeast exopolysaccharides, such as pullulan, exhibit specific physico-chemical and rheological properties, making them useful in a wide range of technical applications. A survey is given here of the production, the characteristics and the application potential of currently well studied yeast extracellular polysaccharides.

  14. Formation of polyelectrolyte complexes with diethylaminoethyl dextran: charge ratio and molar mass effect.

    PubMed

    Le Cerf, Didier; Pepin, Anne Sophie; Niang, Pape Momar; Cristea, Mariana; Karakasyan-Dia, Carole; Picton, Luc

    2014-11-26

    The formation of polyelectrolyte complexes (PECs) between carboxymethyl pullulan and DEAE Dextran, was investigated, in dilute solution, with emphasis on the effect of charge density (molar ratio or pH) and molar masses. Electrophoretic mobility measurements have evidenced that insoluble PECs (neutral electrophoretic mobility) occurs for charge ratio between 0.6 (excess of polycation) and 1 (stoichiometry usual value) according to the pH. This atypical result is explained by the inaccessibility of some permanent cationic charge when screened by pH dependant cationic ones (due to the Hoffman alkylation). Isothermal titration calorimetry (ITC) indicates an endothermic formation of PEC with a binding constant around 10(5) L mol(-1). Finally asymmetrical flow field flow fractionation coupled on line with static multi angle light scattering (AF4/MALS) evidences soluble PECs with very large average molar masses and size around 100 nm, in agreement with scrambled eggs multi-association between various polyelectrolyte chains.

  15. Sugarbeet as a renewable resource

    SciTech Connect

    Edye, L.A.; Clarke, M.A.

    1995-12-01

    Sugarbeet (Beta vulgaris) is produced annually on the order of 400 million tonnes, in temperate climates. The primary product is sugar (sucrose); other products include feeds (molasses and beet pulp), and raffinose, pectin and arabinan. Recently, production of paper from sugarbeet pulp has begun. A wide range of non-food products is available through microbial and chemical reactions on sugarbeet juices, molasses and sugars. Products of microbial processes (chemical transformations are discussed in the companion presentation on sugarcane) include polymers to use as biodegradable plastics (pullulans, polyhydroxyalkanoates, polylactide) and others for food and non food use (levan, dextran). Basic chemicals, including citric acid and lactic acid, and amino acids, notably lysine, are produced from sugarbeet sources. The production of ethanol, as fuel or as beverage, is well known. Products and processes are outlined, and recent developments are emphasized.

  16. Development of taste masked fast disintegrating films of levocetirizine dihydrochloride for oral use.

    PubMed

    Mahesh, A; Shastri, Nalini; Sadanandam, M

    2010-01-01

    Fast disintegrating films of levocetirizine dihydrochloride useful for the treatment of acute allergic rhinitis and chronic urticaria have been developed by using the taste masking ability of cyclodextrins. The fast disintegrating films were prepared by solvent casting method. The films contained water-soluble polymers such as Kollicoat IR or pullulan, aspartame and sucralose as sweeteners and pre-gelatinized starch as disintegrant. Levocetirizine dihydrochloride was incorporated into these films by in-situ complex formation with hydroxy propyl beta-cyclodextrin. The optimized films were evaluated for weight variation, film thickness, folding endurance, tackiness, tensile strength, assay, content uniformity, in vitro disintegration and dissolution, in vivo disintegration and taste masking ability by human gustatory sensation test. Results revealed that the organoleptic properties of levocetirizine dihydrochloride were improved by complexation with hydroxy propyl beta-cyclodextrin and the complex could be successfully formulated into a fast disintegrating film.

  17. Bacillus stearothermophilus Neopullulanase Selective Hydrolysis of Amylose to Maltose in the Presence of Amylopectin

    PubMed Central

    Kamasaka, Hiroshi; Sugimoto, Kazuhisa; Takata, Hiroki; Nishimura, Takahisa; Kuriki, Takashi

    2002-01-01

    The specificity of Bacillus stearothermophilus TRS40 neopullulanase toward amylose and amylopectin was analyzed. Although this neopullulanase completely hydrolyzed amylose to produce maltose as the main product, it scarcely hydrolyzed amylopectin. The molecular mass of amylopectin was decreased by only one order of magnitude, from approximately 108 to 107 Da. Furthermore, this neopullulanase selectively hydrolyzed amylose when starch was used as a substrate. This phenomenon, efficient hydrolysis of amylose but not amylopectin, was also observed with cyclomaltodextrinase from alkaliphilic Bacillus sp. strain A2-5a and maltogenic amylase from Bacillus licheniformis ATCC 27811. These three enzymes hydrolyzed cyclomaltodextrins and amylose much faster than pullulan. Other amylolytic enzymes, such as bacterial saccharifying α-amylase, bacterial liquefying α-amylase, β-amylase, and neopullulanase from Bacillus megaterium, did not exhibit this distinct substrate specificity at all, i.e., the preference of amylose to amylopectin. PMID:11916682

  18. Endophytic fungi associated with cacti in Arizona.

    PubMed

    Suryanarayanan, Trichur S; Wittlinger, Sally K; Faeth, Stanley H

    2005-05-01

    21 cactus species occurring in various localities within Arizona were screened for the presence of fungal endophytes. 900 endophyte isolates belonging to 22 fungal species were isolated. Cylindropuntia fulgida had the maximum endophyte species diversity, while C. ramosissima harboured the maximum number of endophyte isolates. Alternaria sp., Aureobasidium pullulans, and Phoma spp. were isolated from several cactus species. The diversity of the endophyte assemblages was low and no host specificity among endophytes was observed. However, the frequencies of colonization of the few endophyte species recovered were high and comparable to those reported for tropical plant hosts. Species of Colletotrichum, Phomopsis, and Phyllosticta, which are commonly isolated as endophytes from plants of more mesic habitats, were absent from these cacti.

  19. Pressate from peat dewatering as a substrate for bacterial growth. [Rhizopus arrhizus; Xanthomonas campestris; Aureobasidium

    SciTech Connect

    Mulligan, C.N.; Cooper, D.G.

    1985-07-01

    This study considered the possibility of using water expressed during the drying of fuel-grade peat as a substrate for microbial growth. Highly humified peat pressed for 2.5 min at 1.96 MPa produced water with a chemical oxygen demand of 690 mg/liter. Several biological compounds could be produced by using the organic matter inexpressed peat water as a substrate. These included polymers such as chitosan, contained in the cell wall of Rhizopus arrhizus, and two extracellular polysaccharides, xanthan gum and pullulan, produced by Bacillus subtilis grown in the expressed water. Small additions of nutrients to the peat pressate were necessary to obtain substantial yields of products. The addition of peptone, yeast extract, and glucose improved production of the various compounds. Biological treatment improved the quality of the expressed water to the extent that in an industrial process it could be returned to the environment.

  20. Action of Azotobacter vinelandii poly-beta-D-mannuronic acid C-5-epimerase on synthetic D-glucuronans.

    PubMed

    Chang, P S; Mukerjea, R; Fulton, D B; Robyt, J F

    2000-12-01

    Eleven different glucans (wheat starch, potato amylopectin, potato amylose, pullulan, alternan, regular comb dextran, alpha-cellulose, microcrystalline cellulose, CM-cellulose, chitin, and chitosan) that had their C-6 primary alcohol groups oxidized to carboxyl groups by reaction with 2,2,6,6-tetramethyl-1-piperidine oxoammonium ion (TEMPO), were reacted with Azotobacter vinelandii poly-beta-(1-->4)-D-mannuronic acid C-5-epimerase. All of the oxidized polysaccharides reacted with the C-5-epimerase, as evidenced by comparing: (1) differences in the relative viscosities; (2) differences in the carbazole reaction; (3) differences in their susceptibility to acid hydrolysis, and (4) differences in their ability to form calcium gels, before and after reaction. We further show the formation of L-iduronic acid from D-glucuronic acid for oxidized and epimerized amylose by 2D NOESY and COSY + 1H NMR.

  1. Fungal Exopolysaccharide: Production, Composition and Applications

    PubMed Central

    Mahapatra, Subhadip; Banerjee, Debdulal

    2013-01-01

    Fungal exopolysaccharides (EPSs) have been recognized as high value biomacromolecules for the last two decades. These products, including pullulan, scleroglucan, and botryosphaeran, have several applications in industries, pharmaceuticals, medicine, foods etc. Although fungal EPSs are highly relevant, to date information concerning fungal biosynthesis is scarce and an extensive search for new fugal species that can produce novel EPSs is still needed. In most cases, the molecular weight variations and sugar compositions of fungal EPSs are dependent to culture medium composition and different physical conditions provided during fermentation. An inclusive and illustrative review on fungal EPS is presented here. The general outline of the present work includes fungal EPS production, their compositions and applications. An emphasis is also given to listing out different fungal strains that can produce EPSs. PMID:24826070

  2. A glycoside hydrolase family 31 dextranase with high transglucosylation activity from Flavobacterium johnsoniae.

    PubMed

    Gozu, Yoshifumi; Ishizaki, Yuichi; Hosoyama, Yuhei; Miyazaki, Takatsugu; Nishikawa, Atsushi; Tonozuka, Takashi

    2016-08-01

    Glycoside hydrolase family (GH) 31 enzymes exhibit various substrate specificities, although the majority of members are α-glucosidases. Here, we constructed a heterologous expression system of a GH31 enzyme, Fjoh_4430, from Flavobacterium johnsoniae NBRC 14942, using Escherichia coli, and characterized its enzymatic properties. The enzyme hydrolyzed dextran and pullulan to produce isomaltooligosaccharides and isopanose, respectively. When isomaltose was used as a substrate, the enzyme catalyzed disproportionation to form isomaltooligosaccharides. The enzyme also acted, albeit inefficiently, on p-nitrophenyl α-D-glucopyranoside, and p-nitrophenyl α-isomaltoside was the main product of the reaction. In contrast, Fjoh_4430 did not act on trehalose, kojibiose, nigerose, maltose, maltotriose, or soluble starch. The optimal pH and temperature were pH 6.0 and 60 °C, respectively. Our results indicate that Fjoh_4430 is a novel GH31 dextranase with high transglucosylation activity.

  3. Selection and use of pectinolytic yeasts for improving clarification and phenolic extraction in winemaking.

    PubMed

    Belda, Ignacio; Conchillo, Lorena B; Ruiz, Javier; Navascués, Eva; Marquina, Domingo; Santos, Antonio

    2016-04-16

    Pectinase enzymes have shown a considerable influence in both, sensitive and technological properties of wines. They can help to improve clarification process, releasing more color and flavor compounds entrapped in grape skin, facilitating the liberation of phenolic compounds. This work aims to find yeasts that, because of their native pectinases, can be applied on combined fermentations with Saccharomyces cerevisiae obtaining significant benefits over single-inoculated traditional fermentations. 462 yeast strains isolated from wineries were identified and tested for several enzymatic activities of recognized interest for enology industry. Considering the 7 identified species, only Aureobasidium pullulans, Metschnikowia pulcherrima and Metschnikowia fructicola showed polygalacturonase activity. Because of its interest in winemaking, due to its reported incidence in wine flavor, the impact of M. pulcherrima as a source of pectinolytic enzymes was analyzed by measuring its influence in filterability, turbidity and the increase on color, anthocyanin and polyphenol content of wines fermented in combination with S. cerevisiae. Among the strains screened, M. pulcherrima NS-EM-34 was selected, due to its polygalacturonase activity, for further characterization in both, laboratory and semi-industrial scale assays. The kinetics concerning several metabolites of enological concern were followed during the entire fermentation process at microvinification scale. Improved results were obtained in the expected parameters when M. pulcherrima NS-EM-34 was used, in comparison to wines fermented with S. cerevisiae alone and combined with other pectinolytic and non-pectinolytic yeasts (A. pullulans and Lachancea thermotolerans, respectively), even working better than commercial enzymes preparations in most parameters. Additionally, M. pulcherrima NS-EM-34 was used at a semi-industrial scale combined with three different S. cerevisiae strains, confirming its potential application for

  4. Novel Symbiotic Protoplasts Formed by Endophytic Fungi Explain Their Hidden Existence, Lifestyle Switching, and Diversity within the Plant Kingdom

    PubMed Central

    Atsatt, Peter R.; Whiteside, Matthew D.

    2014-01-01

    Diverse fungi live all or part of their life cycle inside plants as asymptomatic endophytes. While endophytic fungi are increasingly recognized as significant components of plant fitness, it is unclear how they interact with plant cells; why they occur throughout the fungal kingdom; and why they are associated with most fungal lifestyles. Here we evaluate the diversity of endophytic fungi that are able to form novel protoplasts called mycosomes. We found that mycosomes cultured from plants and phylogenetically diverse endophytic fungi have common morphological characteristics, express similar developmental patterns, and can revert back to the free-living walled state. Observed with electron microscopy, mycosome ontogeny within Aureobasidium pullulans may involve two organelles: double membrane-bounded promycosome organelles (PMOs) that form mycosomes, and multivesicular bodies that may form plastid-infecting vesicles. Cultured mycosomes also contain a double membrane-bounded organelle, which may be homologous to the A. pullulans PMO. The mycosome PMO is often expressed as a vacuole-like organelle, which alternatively may contain a lipoid body or a starch grain. Mycosome reversion to walled cells occurs within the PMO, and by budding from lipid or starch-containing mycosomes. Mycosomes discovered in chicken egg yolk provided a plant-independent source for analysis: they formed typical protoplast stages, contained fungal ITS sequences and reverted to walled cells, suggesting mycosome symbiosis with animals as well as plants. Our results suggest that diverse endophytic fungi express a novel protoplast phase that can explain their hidden existence, lifestyle switching, and diversity within the plant kingdom. Importantly, our findings outline “what, where, when and how”, opening the way for cell and organelle-specific tests using in situ DNA hybridization and fluorescent labels. We discuss developmental, ecological and evolutionary contexts that provide a robust

  5. The effect of surface properties on the strength of attachment of fungal spores using AFM perpendicular force measurements.

    PubMed

    Whitehead, Kathryn A; Deisenroth, Ted; Preuss, Andrea; Liauw, Christopher M; Verran, Joanna

    2011-02-01

    Polymeric substrata may be biodegraded by fungal species resulting in damaged, weakened and unsightly materials. This process typically begins with fungal spore attachment to the surface. In order to better understand the processes that precedes a biofouling event, fungal spore attachment to a range of surfaces, was determined using perpendicular force measurements. This was carried out using atomic force microscope cantilevers modified with fungal spores from Aspergillus niger 1957 (5μm diameter, non-wettable, spherical), Aspergillus niger 1988 (5μm diameter non-wettable, spikey) or Aureobasidium pullulans (5μm-10μm sized, wettable, ellipsoidal). The strength of attachment of the spores was determined in combination with seven surfaces (nitric acid cleaned glass, cast poly(methylmethacrylate) sheet [c-PMMA], polytetrafluoroethylene [PTFE], silicon wafers spin coated with poly(3-methacryloxypropyltrimethoxy silane (γ-MPS)-co-methylmethacrylate (MMA)) [p(γ-MPS-co-MMA)], poly (γ-MPS-co-lauryl methacrylate) [p(γ-MPS-co-LMA)] [both in a ratio of 10-90], PMMA dissolved in a solvent [PMMAsc] and silicon wafers). Perpendicular force measurements could not be related to the R(a) values of the surfaces, but surface wettability was shown to have an effect. All three spore types interacted comparably with the surfaces. All spores attached strongly to c-PMMA and glass (wettable surfaces), and weakly to PTFE, (p(γ- MPS-co-LMA)) (non-wettable) and (p(γ-MPS-co-MMA)). Spore shape also affected the strength of attachment. Aureobasidium pullulans spores attached with the widest range of forces whilst A. niger 1957 attached with the smallest. Findings will inform the selection of surfaces for use in environments where biofouling is an important consideration.

  6. Colonization of a Submersed Aquatic Plant, Eurasian Water Milfoil (Myriophyllum spicatum), by Fungi under Controlled Conditions

    PubMed Central

    Smith, Craig S.; Chand, Tara; Harris, Robin F.; Andrews, John H.

    1989-01-01

    A laboratory assay to assess colonization of a submersed aquatic plant, Eurasian water milfoil (Myriophyllum spicatum), by fungi was developed and used to evaluate the colonization potential of Colletotrichum gloeosporioides, Acremonium curvulum, Cladosporium herbarum, Aureobasidium pullulans, a Paecilomyces sp., and an unidentified sterile, septate fungus. Stem segments of plants were first immersed in suspensions of fungal propagules for 24 h and then washed to remove all but the tightly attached component of the population. Inoculation was followed by two growth cycles of 3 days each. At the start of each cycle, washed plants were transferred to a mineral salts medium to provide an opportunity for the attached fungal populations to grow. After each growth period, plants were again washed, and fungal populations in the medium (nonattached), loosely attached and tightly attached to the plant, and within the plant (endophytic) were assayed by dilution plating. The fungi differed in the extent to which they attached to water milfoil and in their ability to grow in association with it. There were relatively few significant differences among the tightly attached fungal populations after 24 h, but growth of the better colonizers led to a greater number of significant differences after 4 and 7 days. In addition, the better colonizers showed sustained regrowth of loosely and nonattached fungal propagules in the face of intermittent removal by washing. A milfoil pathogen, C. gloeosporioides, was the only endophytic colonizer; it was also among the best epiphytic colonizers but was not demonstrably better than A. curvulum, a fungus commonly found as an epiphyte on watermilfoil. The yeastlike hyphomycete Aureobasidium pullulans was the only fungus that consistently failed to establish an increasing population on the plant. Images PMID:16348013

  7. Size and shape of soil humic acids estimated by viscosity and molecular weight.

    PubMed

    Kawahigashi, Masayuki; Sumida, Hiroaki; Yamamoto, Kazuhiko

    2005-04-15

    Ultrafiltration fractions of three soil humic acids were characterized by viscometry and high performance size-exclusion chromatography (HPSEC) in order to estimate shapes and hydrodynamic sizes. Intrinsic viscosities under given solute/solvent/temperature conditions were obtained by extrapolating the concentration dependence of reduced viscosities to zero concentration. Molecular mass (weight average molecular weight (M (w)) and number average molecular weight (M (n))) and hydrodynamic radius (R(H)) were determined by HPSEC using pullulan as calibrant. Values of M (w) and M (n) ranged from 15 to 118 x 10(3) and from 9 to 50 x 10(3) (g mol(-1)), respectively. Polydispersity, as indicated by M (w)/M (n), increased with increasing filter size from 1.5 to 2.4. The hydrodynamic radii (R(H)) ranged between 2.2 and 6.4 nm. For each humic acid, M (w) and [eta] were related. Mark-Houwink coefficients calculated on the basis of the M (w)-[eta] relationships suggested restricted flexible chains for two of the humic acids and a branched structure for the third humic acid. Those structures probably behave as hydrated sphere colloids in a good solvent. Hydrodynamic radii of fractions calculated from [eta] using Einstein's equation, which is applicable to hydrated sphere colloids, ranged from 2.2 to 7.1 nm. These dimensions are fit to the size of nanospaces on and between clay minerals and micropores in soil particle aggregates. On the other hand, the good agreement of R(H) values obtained by applying Einstein's equation with those directly determined by HPSEC suggests that pullulan is a suitable calibrant for estimation of molecular mass and size of humic acids by HPSEC.

  8. Capillary Force Seeding of Hydrogels for Adipose-Derived Stem Cell Delivery in Wounds

    PubMed Central

    Garg, Ravi K.; Rennert, Robert C.; Duscher, Dominik; Sorkin, Michael; Kosaraju, Revanth; Auerbach, Lauren J.; Lennon, James; Chung, Michael T.; Paik, Kevin; Nimpf, Johannes; Rajadas, Jayakumar; Longaker, Michael T.

    2014-01-01

    Effective skin regeneration therapies require a successful interface between progenitor cells and biocompatible delivery systems. We previously demonstrated the efficiency of a biomimetic pullulan-collagen hydrogel scaffold for improving bone marrow-derived mesenchymal stem cell survival within ischemic skin wounds by creating a “stem cell niche” that enhances regenerative cytokine secretion. Adipose-derived mesenchymal stem cells (ASCs) represent an even more appealing source of stem cells because of their abundance and accessibility, and in this study we explored the utility of ASCs for hydrogel-based therapies. To optimize hydrogel cell seeding, a rapid, capillary force-based approach was developed and compared with previously established cell seeding methods. ASC viability and functionality following capillary hydrogel seeding were then analyzed in vitro and in vivo. In these experiments, ASCs were seeded more efficiently by capillary force than by traditional methods and remained viable and functional in this niche for up to 14 days. Additionally, hydrogel seeding of ASCs resulted in the enhanced expression of multiple stemness and angiogenesis-related genes, including Oct4, Vegf, Mcp-1, and Sdf-1. Moving in vivo, hydrogel delivery improved ASC survival, and application of both murine and human ASC-seeded hydrogels to splinted murine wounds resulted in accelerated wound closure and increased vascularity when compared with control wounds treated with unseeded hydrogels. In conclusion, capillary seeding of ASCs within a pullulan-collagen hydrogel bioscaffold provides a convenient and simple way to deliver therapeutic cells to wound environments. Moreover, ASC-seeded constructs display a significant potential to accelerate wound healing that can be easily translated to a clinical setting. PMID:25038246

  9. Yeast biocontrol of fungal spoilage of pears stored at low temperature.

    PubMed

    Robiglio, Andrea; Sosa, M Cristina; Lutz, M Cecilia; Lopes, Christian A; Sangorrín, Marcela P

    2011-06-30

    To reduce the use of fungicides, biological control with yeasts has been proposed in postharvest pears. Most studies of antagonists selection have been carried out at room temperature. However, in regions like North Patagonia where fruits are stored at -1/0 °C during 5-7 months the selection of potential antagonist agents must be carried out at low temperature. In this study, 75 yeast cultures were isolated from healthy pears from two Patagonian cold-storage packinghouses. Aureobasidium pullulans, Cryptococcus albidus, Cryptococcus difluens, Pichia membranifaciens, Pichia philogaea, Rhodotorula mucilaginosa and Saccharomyces cerevisiae yeast species were identified. Additionally, 13 indigenous isolates of Penicillium expansum and 10 isolates of Botrytis cinerea were obtained from diseased pears, characterized by aggressiveness and tested for sensitivity to postharvest fungicides. The yeasts were pre-selected for their ability to grow at low temperature. In a first biocontrol assay using the most aggressive and the most sensitive isolate of each pathogen, two epiphytic isolates of A. pullulans and R. mucilaginosa were the most promising isolates to be used as biocontrol agents. They reduced the decay incidence by P. expansum to 33% and the lesion diameter in 88% after 60 days of incubation in cold. Foreign commercial yeast used as a reference in assays, only reduced 30% of lesion diameter in the same conditions. Yeasts were not able to reduce the incidence of B. cinerea decay. The control activity of the best two yeasts was compared with the control caused by the fungicides in a second bioassay, obtaining higher levels of protection against P. expansum by the yeasts. These two regional yeasts isolates could be promising tools for the future development of commercial products for biological control.

  10. Genome Sequence of Microbulbifer mangrovi DD-13(T) Reveals Its Versatility to Degrade Multiple Polysaccharides.

    PubMed

    Imran, Md; Pant, Poonam; Shanbhag, Yogini P; Sawant, Samir V; Ghadi, Sanjeev C

    2017-02-01

    Microbulbifer mangrovi strain DD-13(T) is a novel-type species isolated from the mangroves of Goa, India. The draft genome sequence of strain DD-13 comprised 4,528,106 bp with G+C content of 57.15%. Out of 3479 open reading frames, functions for 3488 protein coding sequences were predicted on the basis of similarity with the cluster of orthologous groups. In addition to protein coding sequences, 34 tRNA genes and 3 rRNA genes were detected. Analysis of nucleotide sequence of predicted gene using a Carbohydrate-Active Enzymes (CAZymes) Analysis Toolkit indicates that strain DD-13 encodes a large set of CAZymes including 255 glycoside hydrolases, 76 carbohydrate esterases, 17 polysaccharide lyases, and 113 carbohydrate-binding modules (CBMs). Many genes from strain DD-13 were annotated as carbohydrases specific for degradation of agar, alginate, carrageenan, chitin, xylan, pullulan, cellulose, starch, β-glucan, pectin, etc. Some of polysaccharide-degrading genes were highly modular and were appended at least with one CBM indicating the versatility of strain DD-13 to degrade complex polysaccharides. The cell growth of strain DD-13 was validated using pure polysaccharides such as agarose or alginate as carbon source as well as by using red and brown seaweed powder as substrate. The homologous carbohydrase produced by strain DD-13 during growth degraded the polysaccharide, ensuring the production of metabolizable reducing sugars. Additionally, several other polysaccharides such as carrageenan, xylan, pullulan, pectin, starch, and carboxymethyl cellulose were also corroborated as growth substrate for strain DD-13 and were associated with concomitant production of homologous carbohydrase.

  11. Novel symbiotic protoplasts formed by endophytic fungi explain their hidden existence, lifestyle switching, and diversity within the plant kingdom.

    PubMed

    Atsatt, Peter R; Whiteside, Matthew D

    2014-01-01

    Diverse fungi live all or part of their life cycle inside plants as asymptomatic endophytes. While endophytic fungi are increasingly recognized as significant components of plant fitness, it is unclear how they interact with plant cells; why they occur throughout the fungal kingdom; and why they are associated with most fungal lifestyles. Here we evaluate the diversity of endophytic fungi that are able to form novel protoplasts called mycosomes. We found that mycosomes cultured from plants and phylogenetically diverse endophytic fungi have common morphological characteristics, express similar developmental patterns, and can revert back to the free-living walled state. Observed with electron microscopy, mycosome ontogeny within Aureobasidium pullulans may involve two organelles: double membrane-bounded promycosome organelles (PMOs) that form mycosomes, and multivesicular bodies that may form plastid-infecting vesicles. Cultured mycosomes also contain a double membrane-bounded organelle, which may be homologous to the A. pullulans PMO. The mycosome PMO is often expressed as a vacuole-like organelle, which alternatively may contain a lipoid body or a starch grain. Mycosome reversion to walled cells occurs within the PMO, and by budding from lipid or starch-containing mycosomes. Mycosomes discovered in chicken egg yolk provided a plant-independent source for analysis: they formed typical protoplast stages, contained fungal ITS sequences and reverted to walled cells, suggesting mycosome symbiosis with animals as well as plants. Our results suggest that diverse endophytic fungi express a novel protoplast phase that can explain their hidden existence, lifestyle switching, and diversity within the plant kingdom. Importantly, our findings outline "what, where, when and how", opening the way for cell and organelle-specific tests using in situ DNA hybridization and fluorescent labels. We discuss developmental, ecological and evolutionary contexts that provide a robust

  12. Nanogel-crosslinked nanoparticles increase the inhibitory effects of W9 synthetic peptide on bone loss in a murine bone resorption model

    PubMed Central

    Sato, Toshimi; Alles, Neil; Khan, Masud; Nagano, Kenichi; Takahashi, Mariko; Tamura, Yukihiko; Shimoda, Asako; Ohya, Keiichi; Akiyoshi, Kazunari; Aoki, Kazuhiro

    2015-01-01

    We investigated the biological activity of W9, a bone resorption inhibitor peptide, using NanoClik nanoparticles as an injectable carrier, where acryloyl group-modified cholesterol-bearing pullulan (CHPOA) nanogels were crosslinked by pentaerythritol tetra (mercaptoethyl) polyoxyethylene. Thirty 5-week-old male C57BL/6J mice were fed a low calcium diet and received once-daily subcutaneous injections of the carrier alone, W9 24 mg/kg/day alone, W9 24 mg/kg/day incorporated in cholesterol bearing pullulan (CHP) nanogels, or W9 (8 and 24 mg/kg/day) incorporated in NanoClik nanoparticles for 4 days (n=5). Mice that received a normal calcium diet with NanoClik nanoparticle injections without W9 were used as a control group. Radiological analyses showed that administration of W9 24 mg/kg/day significantly prevented low calcium-induced reduction of bone mineral density in the long bones and lumbar vertebrae, but only when the NanoClik nanoparticles were used as a carrier. Histomorphometric analyses of the proximal tibiae revealed that W9 24 mg/kg/day incorporated in NanoClik nanoparticles prevented the increase in bone resorption indices induced by a low calcium diet, which was confirmed by measurement of serum bone resorption markers. These data suggest that NanoClik nanoparticles could be a useful carrier for peptide therapeutics, and also demonstrate that daily subcutaneous injections of the W9 peptide with the nanoparticles were able to inhibit bone loss in vivo. An osteoclastogenesis inhibition assay performed in vitro confirmed a slower release profile of W9 from NanoClik nanoparticles compared with conventional CHP nanogels. PMID:25999711

  13. Enzyme activities in the Delaware Estuary affected by elevated suspended sediment load

    NASA Astrophysics Data System (ADS)

    Ziervogel, K.; Arnosti, C.

    2009-09-01

    Extracellular enzyme activities were compared among surface water, bottom water, and sediments of the Delaware Estuary using six fluorescently labeled, structurally distinct polysaccharides to determine the effects of suspended sediment transport on water column hydrolytic activities. Potential hydrolysis rates in surface waters were also measured for the nearby shelf. Samples were taken in December 2006, 6 months after a major flood event in the Delaware Basin that was followed by high freshwater run-off throughout the fall of 2006. All substrates were hydrolyzed in sediments and in the water column, including two (pullulan and fucoidan) that previously were not hydrolyzed in surface waters of the Delaware estuary. At the time of sampling, total particulate matter (TPM) in surface waters at the lower bay, bay mouth, and shelf ranged between 31 mg l -1 and 48 mg l -1 and were 2 to 20 times higher than previously reported. The presence of easily resuspended sediments at the lower bay and bay mouth indicated enhanced suspended sediment transport in the estuary prior to our sampling. Bottom water hydrolysis rates at the two sites affected by sediment resuspension were generally higher than those in surface waters from the same site. Most notably, fucoidan and pullulan hydrolysis rates in bay mouth bottom waters were 22.6 and 6.2 nM monomer h -1, respectively, and thus three and five times higher than surface water rates. Our data suggest that enhanced mixing processes between the sediment and the overlying water broadened the spectrum of water column hydrolases activity, improving the efficiency of enzymatic degradation of high molecular weight organic matter in the water with consequences for organic matter cycling in the Delaware estuary.

  14. House Fly (Musca domestica L.) Attraction to Insect Honeydew

    PubMed Central

    Hung, Kim Y.; Michailides, Themis J.; Millar, Jocelyn G.; Wayadande, Astri; Gerry, Alec C.

    2015-01-01

    House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house

  15. Specific Antibodies for the Detection of Alternaria Allergens and the Identification of Cross-Reactive Antigens in Other Fungi

    PubMed Central

    Twaroch, Teresa E.; Curin, Mirela; Sterflinger, Katja; Focke-Tejkl, Margit; Swoboda, Ines; Valenta, Rudolf

    2017-01-01

    Background The mould Alternaria alternata is an important source of respiratory allergens. A. alternata extracts show great variations regarding allergenic potency. The aim of this study was to generate antibody probes specific for important Alternaria allergens and to use them to study allergen expression, depending on different culture conditions, as well as to search for cross-reactive allergens in other mould species. Methods Synthetic peptides from antigenic regions of A. alternata allergens (Alt a 1, Alt a 2, Alt a 3, Alt a 6 and Alt a 8) were used to raise highly specific rabbit antibodies. These antibodies and IgE from allergic patients were used to detect allergens by immunoblotting in extracts of 4 A. alternata strains grown under varying culturing conditions, in commercial skin-prick extracts and in closely (Cladosporium herbarum and Aureobasidium pullulans) or distantly related (Aspergillus niger and Penicillium chrysogenum) mould species. Results There was a wide variation of expression of the individual A. Alternata allergens, depending on the strain and culture conditions, but the antibody probes allowed us to distinguish strains and culture conditions with low and high allergen expression. In the commercial skin-prick solutions, varying levels of Alt a 1 were found, but no other allergens were detectable. Alt a 1 was identified as species-specific A. Alternata allergen, whereas Alt a 3, 6- and Alt a 8-cross-reactive antigens were found in C. herbarum and/or A. pullulans. Conclusions and Clinical Relevance Peptide-specific antibodies are useful to analyze diagnostic and therapeutic mould extracts, to study the presence of A. Alternata allergens in biological samples and to search for cross-reactive allergens in other mould species. PMID:27780168

  16. Improving Department of Defense Global Distribution Performance Through Network Analysis

    DTIC Science & Technology

    2016-06-01

    Segment Days .................................................................. 16 2. Standards...17 C. METHODOLOGY ......................................................................... 17 1. Segment ...Independence ................................................... 17 2. Statistical Analysis of IPGs by Segment ........................ 18 3. Network

  17. Application of exopolysaccharides to improve the performance of ceramic bodies in the unidirectional dry pressing process

    NASA Astrophysics Data System (ADS)

    Caneira, Inês; Machado-Moreira, Bernardino; Dionísio, Amélia; Godinho, Vasco; Neves, Orquídia; Dias, Diamantino; Saiz-Jimenez, Cesareo; Miller, Ana Z.

    2015-04-01

    Ceramic industry represents an important sector of economic activity in the European countries and involves complex and numerous manufacturing processes. The unidirectional dry pressing process includes milling and stirring of raw materials (mainly clay and talc minerals) in aqueous suspensions, followed by spray drying to remove excess water obtaining spray-dried powders further subjected to dry pressing process (conformation). However, spray-dried ceramic powders exhibit an important variability in their performance when subjected to the dry pressing process, particularly in the adhesion to the mold and mechanical strength, affecting the quality of the final conformed ceramic products. Therefore, several synthetic additives (deflocculants, antifoams, binders, lubricants and plasticizers) are introduced in the ceramic slips to achieve uniform and homogeneous pastes, conditioning their rheological properties. However, an important variability associated with the performance of the conformed products is still reported. Exopolysaccharides or Extracellular Polymeric Substances (EPS) are polymers excreted by living organisms, such as bacteria, fungi and algae, which may confer unique and potentially interesting properties with potential industrial uses, such as viscosity control, gelation, and flocculation. Polysaccharides, such as pullulan, gellan, carrageenan and xanthan have found a wide range of applications in food, pharmaceutical, petroleum, and in other industries. The aim of this study was the assessment of exopolysaccharides as natural additives to optimize the performance of spray-dried ceramic powders during the unidirectional dry pressing process, replacing the synthetic additives used in the ceramic production process. Six exopolysaccharides, namely pullulan, gellan, xanthan gum, κappa- and iota-carrageenan, and guar gum were tested in steatite-based spray-dried ceramic powders at different concentrations. Subsequently, these ceramic powders were

  18. Cloning, Sequencing, and Characterization of a Heat- and Alkali-Stable Type I Pullulanase from Anaerobranca gottschalkii

    PubMed Central

    Bertoldo, Costanzo; Armbrecht, Martin; Becker, Fiona; Schäfer, Thomas; Antranikian, Garabed; Liebl, Wolfgang

    2004-01-01

    The gene encoding a type I pullulanase was identified from the genome sequence of the anaerobic thermoalkaliphilic bacterium Anaerobranca gottschalkii. In addition, the homologous gene was isolated from a gene library of Anaerobranca horikoshii and sequenced. The proteins encoded by these two genes showed 39% amino acid sequence identity to the pullulanases from the thermophilic anaerobic bacteria Fervidobacterium pennivorans and Thermotoga maritima. The pullulanase gene from A. gottschalkii (encoding 865 amino acids with a predicted molecular mass of 98 kDa) was cloned and expressed in Escherichia coli strain BL21(DE3) so that the protein did not have the signal peptide. Accordingly, the molecular mass of the purified recombinant pullulanase (rPulAg) was 96 kDa. Pullulan hydrolysis activity was optimal at pH 8.0 and 70°C, and under these physicochemical conditions the half-life of rPulAg was 22 h. By using an alternative expression strategy in E. coli Tuner(DE3)(pLysS), the pullulanase gene from A. gottschalkii, including its signal peptide-encoding sequence, was cloned. In this case, the purified recombinant enzyme was a truncated 70-kDa form (rPulAg′). The N-terminal sequence of purified rPulAg′ was found 252 amino acids downstream from the start site, presumably indicating that there was alternative translation initiation or N-terminal protease cleavage by E. coli. Interestingly, most of the physicochemical properties of rPulAg′ were identical to those of rPulAg. Both enzymes degraded pullulan via an endo-type mechanism, yielding maltotriose as the final product, and hydrolytic activity was also detected with amylopectin, starch, β-limited dextrins, and glycogen but not with amylose. This substrate specificity is typical of type I pullulanases. rPulAg was inhibited by cyclodextrins, whereas addition of mono- or bivalent cations did not have a stimulating effect. In addition, rPulAg′ was stable in the presence of 0.5% sodium dodecyl sulfate, 20% Tween

  19. A Test for Measuring Gustatory Function

    PubMed Central

    Smutzer, Gregory; Lam, Si; Hastings, Lloyd; Desai, Hetvi; Abarintos, Ray A.; Sobel, Marc; Sayed, Nabil

    2010-01-01

    Objectives The purpose of this study is to determine the usefulness of edible taste strips for measuring human gustatory function. Research Design The physical properties of edible taste strips were examined in order to determine their potential for delivering threshold and suprathreshold amounts of taste stimuli to the oral cavity. Taste strips were then assayed by fluorescence to analyze the uniformity and distribution of bitter tastant in the strips. Finally, taste recognition thresholds for sweet taste were examined in order to determine whether or not taste strips would produce recognition thresholds that were equal to or better than those obtained from aqueous tests. Methodology Edible strips were prepared from pullulan-hydroxypropyl methylcellulose solutions that were dried to a thin film. The maximal amount of a tastant that could be incorporated in a 2.54 × 2.54 cm taste strip was identified by including representative taste stimuli for each class of tastant (sweet, sour, salty, bitter, and umami) during strip formation. Distribution of the bitter tastant quinine hydrochloride in taste strips was assayed by fluorescence emission spectroscopy. The efficacy of taste strips for evaluating human gustatory function was examined by using a single series ascending method of limits protocol. Sucrose taste recognition threshold data from edible strips was then compared to results that were obtained from a standard “sip and spit” recognition threshold test. Results Edible films that formed from a pullulan-hydroxypropyl methylcellulose polymer mixture can be used to prepare clear, thin strips that have essentially no background taste and leave no physical presence after release of tastant. Edible taste strips could uniformly incorporate up to five percent of their composition as tastant. Taste recognition thresholds for sweet taste were over one order of magnitude lower with edible taste strips when compared to an aqueous taste test. Conclusion Edible taste

  20. Optimised method for the analysis of phenolic compounds from caper (Capparis spinosa L.) berries and monitoring of their changes during fermentation.

    PubMed

    Francesca, Nicola; Barbera, Marcella; Martorana, Alessandra; Saiano, Filippo; Gaglio, Raimondo; Aponte, Maria; Moschetti, Giancarlo; Settanni, Luca

    2016-04-01

    In this work, an ad hoc method to identify and quantify polyphenols from caper berries was developed on high-performance liquid chromatography/electrospray ionisation source/mass spectrometry (HPLC-ESI-MS). The method was applied during fermentation carried out with Lactobacillus pentosus OM13 (Trial S) and without starter (Trial C). A total of five polyphenols were identified. All samples contained high concentrations of rutin. Epicatechin was found in untreated fruits, on the contrary quercetin was detected during fermentation. Trial S was characterised by a more rapid acidification and lower levels of spoilage microorganisms than Trial C. L. pentosus dominated among the microbial community of both trials and the highest biodiversity, in terms of strains, was displayed by Trial C. Aureobasidium pullulans was the only yeast species found. The analytical method proposed allowed a high polyphenolic compound recovery from untreated and processed caper berries in short time. The starter culture reduced the bitter taste of the final product. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Characterization of pectinase activity for enology from yeasts occurring in Argentine Bonarda grape

    PubMed Central

    Merín, María Gabriela; Martín, María Carolina; Rantsiou, Kalliopi; Cocolin, Luca; de Ambrosini, Vilma Inés Morata

    2015-01-01

    Pectinolytic enzymes are greatly important in winemaking due to their ability to degrade pectic polymers from grape, contributing to enhance process efficiency and wine quality. This study aimed to analyze the occurrence of pectinolytic yeasts during spontaneous fermentation of Argentine Bonarda grape, to select yeasts that produce extracellular pectinases and to characterize their pectinolytic activity under wine-like conditions. Isolated yeasts were grouped using PCR-DGGE and identified by partial sequencing of 26S rRNA gene. Isolates comprised 7 genera, with Aureobasidium pullulans as the most predominant pectinolytic species, followed by Rhodotorula dairenensis and Cryptococcus saitoi. No pectinolytic activity was detected among ascomycetous yeasts isolated on grapes and during fermentation, suggesting a low occurrence of pectinolytic yeast species in wine fermentation ecosystem. This is the first study reporting R. dairenensis and Cr. saitoi species with pectinolytic activity. R. dairenensis GM-15 produced pectinases that proved to be highly active at grape pH, at 12 °C, and under ethanol and SO2 concentrations usually found in vinifications (pectinase activity around 1.1 U/mL). This strain also produced cellulase activity at 12 °C and pH 3.5, but did not produce β-glucosidase activity under these conditions. The strain showed encouraging enological properties for its potential use in low-temperature winemaking. PMID:26413065

  2. 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.

  3. Asialoglycoprotein receptor targeted delivery of doxorubicin nanoparticles for hepatocellular carcinoma.

    PubMed

    Pranatharthiharan, Sandhya; Patel, Mitesh D; Malshe, Vinod C; Pujari, Vaishali; Gorakshakar, Ajit; Madkaikar, Manisha; Ghosh, Kanjaksha; Devarajan, Padma V

    2017-11-01

    We report asialoglycoprotein receptor (ASGPR)-targeted doxorubicin hydrochloride (Dox) nanoparticles (NPs) for hepatocellular carcinoma (HCC). Polyethylene sebacate (PES)-Gantrez® AN 119 Dox NPs of average size 220 nm with PDI < 0.62 and ∼20% Dox loading were prepared by modified nanoprecipitation. ASGPR ligands, pullulan (Pul), arabinogalactan (AGn), and the combination (Pul-AGn), were anchored by adsorption. Ligand anchoring enabled high liver uptake with a remarkable hepatocyte:nonparenchymal cell ratio of 85:15. Furthermore, Pul-AGn NPs exhibited an additive effect implying incredibly high hepatocyte accumulation. Galactose-mediated competitive inhibition confirmed ASGPR-mediated uptake of ligand-anchored NPs in HepG2 cell lines. Subacute toxicity in rats confirmed the safety of the NP groups. However, histopathological evaluation suggested mild renal toxicity of AGn. Pul NPs revealed sustained reduction in tumor volume in PLC/PRF/5 liver tumor-bearing Nod/Scid mice up to 46 days. Extensive tumor necrosis, reduced collagen content, reduction in the HCC biomarker serum α-fetoprotein (p < 0.05), a mitotic index of 1.135 (day 46), and tumor treated/tumor control (T/C) values of <0.42 signified superior efficacy of Pul NPs. Furthermore, weight gain in the NP groups, and no histopathological alterations indicated that they were well tolerated by the mice. The high efficacy coupled with greater safety portrayed Pul Dox NPs as a promising nanocarrier for improved therapy of HCC.

  4. Evaluation of certain food additives and contaminants.

    PubMed

    2011-01-01

    This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, with a view to recommending acceptable daily intakes (ADIs) and to preparing specifications for identity and purity. The Committee also evaluated the risk posed by two food contaminants, with the aim of deriving tolerable intakes where appropriate and advising on risk management options for the purpose of public health protection. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of and assessment of dietary exposure to food additives and contaminants. A summary follows of the Committee's evaluations of technical, toxicological and dietary exposure data for certain food additives (aluminium-containing food additives, Benzoe Tonkinensis, glycerol ester of gum rosin, glycerol ester of tall oil rosin, glycerol ester of wood rosin, octenyl succinic acid modified gum arabic, polydimethyl siloxane, Ponceau 4R, pullulan, pullulanase from Bacillus deromificans expressed in Bacillus licheniformis, Quinoline Yellow and Sunset Yellow FCF) and two food contaminants (cyanogenic glycosides and fumonisins). Specifications for the following food additives were revised: aluminium lakes of colouring matters; beta-apo-8'-carotenal; beta-apo-8'-carotenoic acid ethyl ester; beta-carotene, synthetic; hydroxypropyl methyl cellulose; magnesium silicate, synthetic; modified starches; nitrous oxide; sodium carboxymethyl cellulose; and sucrose monoesters of lauric, palmitic or stearic acid. Annexed to the report are tables summarizing the Committee's recommendations for dietary exposures to and toxicological evaluations of the food additives and contaminants considered.

  5. Different utilization of alginate and other algal polysaccharides by marine Alteromonas macleodii ecotypes.

    PubMed

    Neumann, Anna M; Balmonte, John P; Berger, Martine; Giebel, Helge-Ansgar; Arnosti, Carol; Voget, Sonja; Simon, Meinhard; Brinkhoff, Thorsten; Wietz, Matthias

    2015-10-01

    The marine bacterium Alteromonas macleodii is a copiotrophic r-strategist, but little is known about its potential to degrade polysaccharides. Here, we studied the degradation of alginate and other algal polysaccharides by A. macleodii strain 83-1 in comparison to other A. macleodii strains. Cell densities of strain 83-1 with alginate as sole carbon source were comparable to those with glucose, but the exponential phase was delayed. The genome of 83-1 was found to harbour an alginolytic system comprising five alginate lyases, whose expression was induced by alginate. The alginolytic system contains additional CAZymes, including two TonB-dependent receptors, and is part of a 24 kb genomic island unique to the A. macleodii 'surface clade' ecotype. In contrast, strains of the 'deep clade' ecotype contain only a single alginate lyase in a separate 7 kb island. This difference was reflected in an eightfold greater efficiency of surface clade strains to grow on alginate. Strain 83-1 furthermore hydrolysed laminarin, pullulan and xylan, and corresponding polysaccharide utilization loci were detected in the genome. Alteromonas macleodii alginate lyases were predominantly detected in Atlantic Ocean metagenomes. The demonstrated hydrolytic capacities are likely of ecological relevance and represent another level of adaptation among A. macleodii ecotypes.

  6. Filling the gap: Calibration of the low molar-mass range of cellulose in size exclusion chromatography with cello-oligomers.

    PubMed

    Oberlerchner, J T; Vejdovszky, P; Zweckmair, T; Kindler, A; Koch, S; Rosenau, T; Potthast, A

    2016-11-04

    Degraded celluloses are becoming increasingly important as part of product streams coming from various biorefinery scenarios. Analysis of the molar mass distribution of such fractions is a challenge, since neither established methods for mono- or disaccharides nor common methods for polysaccharide characterization cover the intermediate oligomer range appropriately. Size exclusion chromatography (SEC) with multi-angle laser light scattering (MALLS), the standard approach for celluloses, suffers from decreased scattering intensities in the lower-molar mass range. The limitation in the low-molecular range can, in principle, be overcome by calibration, but calibration standards for such "short" celluloses are either not readily available or structurally remote and thus questionable. In this paper, we present the calibration of a SEC system- for the first time - with monodisperse cellooligomer standards up to about 3400gmol(-1). These cellooligomers are "short-chain celluloses" and can be seen as the "true" standard compounds, by contrast to commonly used standards that are chemically different from cellulose, such as pullulan, dextran, polystyrene, or poly(methyl methacrylate). The calibration is compared against those commercial standards and correction factors are calculated. Calibrations with non-cellulose standards can now be adjusted to yield better fitting results, and data already available can be corrected retrospectively.

  7. Characteristics of Yeasts Isolated from Pacific Crab Meat

    PubMed Central

    Eklund, M. W.; Spinelli, J.; Miyauchi, D.; Groninger, H.

    1965-01-01

    A total of 202 cultures of yeasts were isolated and characterized from king crab and Dungeness crab meat. A yeastlike organism, resembling Aureobasidium pullulans, and 15 different species distributed among the genera Rhodotorula, Cryptococcus, Torulopsis, Candida, and Trichosporon were represented. Nine of the species grew at 5 C or lower. Although two of the species grew at 37 C, none of the isolates had the characteristics of pathogenic species. Members of the Cryptococcus and Candida failed to grow at 37 C. Furthermore, species of the former genus were not pathogenic to mice. The pigmentation of the Rhodotorula cultures decreased in intensity as the incubation temperature was decreased. Biochemical activities of the different species were studied by use of triglycerides, lecithin, and proteins (casein, gelatin, and crab-meat protein) as substrates. Eight of the species could attack triglycerides; eight, lecithin; five, gelatin; one, casein; and one, crab protein. An organism, tentatively identified as Trichosporon sp., was very active in attacking each of the substrates tested and grew well at 0.5 C. PMID:5866045

  8. Endophytic fungi from Vitis labrusca L. ('Niagara Rosada') and its potential for the biological control of Fusarium oxysporum.

    PubMed

    Brum, M C P; Araújo, W L; Maki, C S; Azevedo, J L

    2012-12-06

    We investigated the diversity of endophytic fungi found on grape (Vitis labrusca cv. Niagara Rosada) leaves collected from Salesópolis, SP, Brazil. The fungi were isolated and characterized by amplified ribosomal DNA restriction analysis, followed by sequencing of the ITS1-5.8S-ITS2 rDNA. In addition, the ability of these endophytic fungi to inhibit the grapevine pathogen Fusarium oxysporum f. sp herbemontis was determined in vitro. We also observed that the climatic factors, such as temperature and rainfall, have no effect on the frequency of infection by endophytic fungi. The endophytic fungal community that was identified included Aporospora terricola, Aureobasidium pullulans, Bjerkandera adusta, Colletotrichum boninense, C. gloeosporioides, Diaporthe helianthi, D. phaseolorum, Epicoccum nigrum, Flavodon flavus, Fusarium subglutinans, F. sacchari, Guignardia mangiferae, Lenzites elegans, Paraphaeosphaeria pilleata, Phanerochaete sordida, Phyllosticta sp, Pleurotus nebrodensis, Preussia africana, Tinctoporellus epiniltinus, and Xylaria berteri. Among these isolates, two, C. gloeosporioides and F. flavus, showed potential antagonistic activity against F. oxysporum f. sp herbemontis. We suggest the involvement of the fungal endophyte community of V. labrusca in protecting the host plant against pathogenic Fusarium species. Possibly, some endophytic isolates could be selected for the development of biological control agents for grape fungal disease; alternatively, management strategies could be tailored to increase these beneficial fungi.

  9. Phenolic Responses to Esca-Associated Fungi in Differently Decayed Grapevine Woods from Different Trunk Parts of 'Cabernet Sauvignon'.

    PubMed

    Rusjan, Denis; Persic, Martina; Likar, Matevž; Biniari, Katerina; Mikulic-Petkovsek, Maja

    2017-08-09

    Alterations in phenolic contents were studied in Esca symptomatic (Sym) and asymptomatic (Asym) vines of 'Cabernet Sauvignon' based on wood conditions (healthy, HLT; necrotic, Nec; and rotten, Rot) and vine parts (head, trunk, and rootstock). In Asym vines, only Alternaria alternate was identified in Nec wood, while the HLT wood of Sym vines was colonized by Botryospaeriaceae sp. and Aureobasidium pullulans, Nec wood by Fomitiporia mediterranea, and Rot wood by Fomitiporia mediterranea and Phaeomoniella chlamydospora. Esca infection caused a significant accumulation of gallic acid, total flavanols, stilbenes (STB), and total analyzed phenolics (TAP) in all studied woods, especially in Nec wood. In Asym vines, TAP in the head increased with necrosis, but in Sym it decreased, while TAP in the trunk and rootstock of Sym showed an opposite response. The significantly highest contents of procyanidins (Pcys), catechin, epicatechin, epicatechin gallates, and Pcys dimers and tetramers were measured in HLT wood in the head and in Nec wood in the trunk of Sym vines. The significant increase of STB content was not caused only by Esca infection in HLT wood but also by necrosis in Asym vines, especially of ε-viniferin glucoside, resveratrol glycosides, and astringin. The obtained results suggest that the alteration in phenolics differed not only due to Esca infection but also due to the wood conditions and vine part, which might reflect the impact of the duration of the presence of the pathogen in different parts of the vine.

  10. Analysis of Fungal Flora in Indoor Dust by Ribosomal DNA Sequence Analysis, Quantitative PCR, and Culture▿ †

    PubMed Central

    Pitkäranta, M.; Meklin, T.; Hyvärinen, A.; Paulin, L.; Auvinen, P.; Nevalainen, A.; Rintala, H.

    2008-01-01

    In recent years increasing attention has been given to the potential health effects of fungal exposure in indoor environments. We used large-scale sequencing of the fungal internal transcribed spacer region (ITS) of nuclear ribosomal DNA to describe the mycoflora of two office buildings over the four seasons. DNA sequencing was complemented by cultivation, ergosterol determination, and quantitative PCR analyses. Sequences of 1,339 clones were clustered into 394 nonredundant fungal operational taxonomical units containing sequences from 18 fungal subclasses. The observed flora differed markedly from that recovered by cultivation, the major differences being the near absence of several typical indoor mold genera such as Penicillium and Aspergillus spp. and a high prevalence of basidiomycetes in clone libraries. A total of 55% of the total diversity constituted of unidentifiable ITS sequences, some of which may represent novel fungal species. Dominant species were Cladosporium cladosporioides and C. herbarum, Cryptococcus victoriae, Leptosphaerulina americana and L. chartarum, Aureobasidium pullulans, Thekopsora areolata, Phaeococcomyces nigricans, Macrophoma sp., and several Malassezia species. Seasonal differences were observed for community composition, with ascomycetous molds and basidiomycetous yeasts predominating in the winter and spring and Agaricomycetidae basidiomycetes predominating in the fall. The comparison of methods suggested that the cloning, cultivation, and quantitative PCR methods complemented each other, generating a more comprehensive picture of fungal flora than any of the methods would give alone. The current restrictions of the methods are discussed. PMID:17981947

  11. SEDFIT-MSTAR: Molecular weight and molecular weight distribution analysis of polymers by sedimentation equilibrium in the ultracentrifuge

    PubMed Central

    Schuck, Peter; Gillis, Richard B.; Besong, Tabot M.D.; Almutairi, Fahad; Adams, Gary G.; Rowe, Arthur J.; Harding, Stephen E.

    2014-01-01

    Sedimentation equilibrium (analytical ultracentrifugation) is one of the most inherently suitable methods for the determination of average molecular weights and molecular weight distributions of polymers, because of its absolute basis (no conformation assumptions) and inherent fractionation ability (without the need for columns or membranes and associated assumptions over inertness). With modern instrumentation it is also possible to run up to 21 samples simultaneously in a single run. Its application has been severely hampered because of difficulties in terms of baseline determination (incorporating estimation of the concentration at the air/solution meniscus) and complexity of the analysis procedures. We describe a new method for baseline determination based on a smart-smoothing principle and built into the highly popular platform SEDFIT for the analysis of the sedimentation behavior of natural and synthetic polymer materials. The SEDFIT-MSTAR procedure – which takes only a few minutes to perform - is tested with four synthetic data sets (including a significantly non-ideal system) a naturally occurring protein (human IgG1) and two naturally occurring carbohydrate polymers (pullulan and λ–carrageenan) in terms of (i) weight average molecular weight for the whole distribution of species in the sample (ii) the variation in “point” average molecular weight with local concentration in the ultracentrifuge cell and (iii) molecular weight distribution. PMID:24244936

  12. The Structural Basis of Alpha-Glucan Recognition by a Family 41 Carbohydrate-Binding Module from Therotoga Maritima

    SciTech Connect

    van Bueren,A.; Boraston, A.

    2006-01-01

    Starch recognition by carbohydrate-binding modules (CBMs) is important for the activity of starch-degrading enzymes. The N-terminal family 41 CBM, TmCBM41 (from pullulanase PulA secreted by Thermotoga maritima) was shown to have {alpha}-glucan binding activity with specificity for {alpha}-1, 4-glucans but was able to tolerate the {alpha}-1, 6-linkages found roughly every three or four glucose units in pullulan. Using X-ray crystallography, the structures were solved for TmCBM41 in an uncomplexed form and in complex with maltotetraose and 63-{alpha}-d-glucosyl-maltotriose (GM3). Ligand binding was facilitated by stacking interactions between the {alpha}-faces of the glucose residues and two tryptophan side-chains in the two main subsites of the carbohydrate-binding site. Overall, this mode of starch binding is quite well conserved by other starch-binding modules. The structure in complex with GM3 revealed a third binding subsite with the flexibility to accommodate an {alpha}-1, 4- or an {alpha}-1, 6-linked glucose.

  13. Biotechnological Applications of Dimorphic Yeasts

    NASA Astrophysics Data System (ADS)

    Doiphode, N.; Joshi, C.; Ghormade, V.; Deshpande, M. V.

    The dimorphic yeasts have the equilibrium between spherical growth (budding) and polarized (hyphal or pseudohyphal tip elongation) which can be triggered by change in the environmental conditions. The reversible growth phenomenon has made dimorphic yeasts as an useful model to understand fungal evolution and fungal differentiation, in general. In nature dimorphism is clearly evident in plant and animal fungal pathogens, which survive and most importantly proliferate in the respective hosts. However, number of organisms with no known pathogenic behaviour also show such a transition, which can be exploited for the technological applications due to their different biochemical make up under different morphologies. For instance, chitin and chitosan production using dimorphic Saccharomyces, Mucor, Rhizopus and Benjaminiella, oil degradation and biotransformation with yeast-form of Yarrowia species, bioremediation of organic pollutants, exopolysac-charide production by yeast-phase of Aureobasidium pullulans, to name a few. Myrothecium verrucaria can be used for seed dressing in its yeast form and it produces a mycolytic enzyme complex in its hyphal-form for the biocontrol of fungal pathogens, while Beauveria bassiana and other entomopathogens kill the insect pest by producing yeast- like cells in the insect body. The form-specific expression of protease, chitinase, lipase, ornithine decarboxylase, glutamate dehydrogenases, etc. make Benjaminiella poitrasii, Basidiobolus sp., and Mucor rouxii strains important in bioremediation, nanobiotechnology, fungal evolution and other areas.

  14. Bio-relevant dissolution testing of hard capsules prepared from different shell materials using the dynamic open flow through test apparatus.

    PubMed

    Garbacz, Grzegorz; Cadé, Dominique; Benameur, Hassan; Weitschies, Werner

    2014-06-16

    Current compendial dissolution and disintegrating testing is unable to mimic physiological conditions affecting gastric drug release from immediate release dosage forms. In order to obtain more realistic data, a novel test setup was developed that we term a 'dynamic open flow through test apparatus'. It is based on the previously described dissolution stress test device and attempts to simulate the intra-gastric dissolution conditions pertinent to immediate release dosage forms administered under fasting conditions with respect to flow rates, intra-gastric temperature profiles and gastric motility. The concept of the dynamic open flow through test apparatus has been tested using five different types of hard capsules: conventional hard gelatin capsules (HGC), three hypromellose based capsules (Vcaps, Vcaps Plus and DRcaps) and pullulan based capsules (Plantcaps). These were of different sizes but all contained 100mg caffeine in each formulation, adjusted to avoid buoyancy by addition of excipient. When the capsules were stressed in the apparatus under the dynamic flow conditions applying mild pressure simulating gastric motility, release from release from Vcaps Plus, Vcaps and Plantcaps capsules was very well comparable to HGC. Capsules are usually swallowed with cold water and the temperature dependency of release from gelatin was noted as a significant factor, since heat exchange in the stomach is slow. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Status, Antimicrobial Mechanism, and Regulation of Natural Preservatives in Livestock Food Systems

    PubMed Central

    Lee, Na-Kyoung; Paik, Hyun-Dong

    2016-01-01

    This review discusses the status, antimicrobial mechanisms, application, and regulation of natural preservatives in livestock food systems. Conventional preservatives are synthetic chemical substances including nitrates/nitrites, sulfites, sodium benzoate, propyl gallate, and potassium sorbate. The use of artificial preservatives is being reconsidered because of concerns relating to headache, allergies, and cancer. As the demand for biopreservation in food systems has increased, new natural antimicrobial compounds of various origins are being developed, including plant-derived products (polyphenolics, essential oils, plant antimicrobial peptides (pAMPs)), animal-derived products (lysozymes, lactoperoxidase, lactoferrin, ovotransferrin, antimicrobial peptide (AMP), chitosan and others), and microbial metabolites (nisin, natamycin, pullulan, ε-polylysine, organic acid, and others). These natural preservatives act by inhibiting microbial cell walls/membranes, DNA/RNA replication and transcription, protein synthesis, and metabolism. Natural preservatives have been recognized for their safety; however, these substances can influence color, smell, and toxicity in large amounts while being effective as a food preservative. Therefore, to evaluate the safety and toxicity of natural preservatives, various trials including combinations of other substances or different food preservation systems, and capsulation have been performed. Natamycin and nisin are currently the only natural preservatives being regulated, and other natural preservatives will have to be legally regulated before their widespread use. PMID:27621697

  16. Purification and biochemical characterization of a thermostable extracellular glucoamylase produced by the thermotolerant fungus Paecilomyces variotii.

    PubMed

    Michelin, Michele; Ruller, Roberto; Ward, Richard J; Moraes, Luiz Alberto B; Jorge, João A; Terenzi, Héctor F; Polizeli, Maria de Lourdes T M

    2008-01-01

    An extracellular glucoamylase produced by Paecilomyces variotii was purified using DEAE-cellulose ion exchange chromatography and Sephadex G-100 gel filtration. The purified protein migrated as a single band in 7% PAGE and 8% SDS-PAGE. The estimated molecular mass was 86.5 kDa (SDS-PAGE). Optima of temperature and pH were 55 degrees C and 5.0, respectively. In the absence of substrate the purified glucoamylase was stable for 1 h at 50 and 55 degrees C, with a t (50) of 45 min at 60 degrees C. The substrate contributed to protect the enzyme against thermal denaturation. The enzyme was mainly activated by manganese metal ions. The glucoamylase produced by P. variotii preferentially hydrolyzed amylopectin, glycogen and starch, and to a lesser extent malto-oligossacarides and amylose. Sucrose, p-nitrophenyl alpha-D-maltoside, methyl-alpha-D-glucopyranoside, pullulan, alpha- and beta-cyclodextrin, and trehalose were not hydrolyzed. After 24 h, the products of starch hydrolysis, analyzed by thin layer chromatography, showed only glucose. The circular dichroism spectrum showed a protein rich in alpha-helix. The sequence of amino acids of the purified enzyme VVTDSFR appears similar to glucoamylases purified from Talaromyces emersonii and with the precursor of the glucoamylase from Aspergillus oryzae. These results suggested the character of the enzyme studied as a glucoamylase (1,4-alpha-D-glucan glucohydrolase).

  17. SEDFIT-MSTAR: molecular weight and molecular weight distribution analysis of polymers by sedimentation equilibrium in the ultracentrifuge.

    PubMed

    Schuck, Peter; Gillis, Richard B; Besong, Tabot M D; Almutairi, Fahad; Adams, Gary G; Rowe, Arthur J; Harding, Stephen E

    2014-01-07

    Sedimentation equilibrium (analytical ultracentrifugation) is one of the most inherently suitable methods for the determination of average molecular weights and molecular weight distributions of polymers, because of its absolute basis (no conformation assumptions) and inherent fractionation ability (without the need for columns or membranes and associated assumptions over inertness). With modern instrumentation it is also possible to run up to 21 samples simultaneously in a single run. Its application has been severely hampered because of difficulties in terms of baseline determination (incorporating estimation of the concentration at the air/solution meniscus) and complexity of the analysis procedures. We describe a new method for baseline determination based on a smart-smoothing principle and built into the highly popular platform SEDFIT for the analysis of the sedimentation behavior of natural and synthetic polymer materials. The SEDFIT-MSTAR procedure - which takes only a few minutes to perform - is tested with four synthetic data sets (including a significantly non-ideal system), a naturally occurring protein (human IgG1) and two naturally occurring carbohydrate polymers (pullulan and λ-carrageenan) in terms of (i) weight average molecular weight for the whole distribution of species in the sample (ii) the variation in "point" average molecular weight with local concentration in the ultracentrifuge cell and (iii) molecular weight distribution.

  18. Characterization of a Novel Amylolytic Enzyme Encoded by a Gene from a Soil-Derived Metagenomic Library

    PubMed Central

    Yun, Jiae; Kang, Seowon; Park, Sulhee; Yoon, Hyunjin; Kim, Myo-Jeong; Heu, Sunggi; Ryu, Sangyeol

    2004-01-01

    It has been estimated that less than 1% of the microorganisms in nature can be cultivated by conventional techniques. Thus, the classical approach of isolating enzymes from pure cultures allows the analysis of only a subset of the total naturally occurring microbiota in environmental samples enriched in microorganisms. To isolate useful microbial enzymes from uncultured soil microorganisms, a metagenome was isolated from soil samples, and a metagenomic library was constructed by using the pUC19 vector. The library was screened for amylase activity, and one clone from among approximately 30,000 recombinant Escherichia coli clones showed amylase activity. Sequencing of the clone revealed a novel amylolytic enzyme expressed from a novel gene. The putative amylase gene (amyM) was overexpressed and purified for characterization. Optimal conditions for the enzyme activity of the AmyM protein were 42°C and pH 9.0; Ca2+ stabilized the activity. The amylase hydrolyzed soluble starch and cyclodextrins to produce high levels of maltose and hydrolyzed pullulan to panose. The enzyme showed a high transglycosylation activity, making α-(1, 4) linkages exclusively. The hydrolysis and transglycosylation properties of AmyM suggest that it has novel characteristics and can be regarded as an intermediate type of maltogenic amylase, α-amylase, and 4-α-glucanotransferase. PMID:15574921

  19. Encapsulation of volatiles in nanofibrous polysaccharide membranes for humidity-triggered release.

    PubMed

    Mascheroni, Erika; Fuenmayor, Carlos Alberto; Cosio, Maria Stella; Di Silvestro, Giuseppe; Piergiovanni, Luciano; Mannino, Saverio; Schiraldi, Alberto

    2013-10-15

    A single-step electrospinning process will be applied to a blend of edible carbohydrate polymers (pullulan and β-cyclodextrin) to encapsulate bioactive aroma compounds and allow a humidity-triggered release. The encapsulation is rapid and efficient and the final product is an active nanofibrous membrane that can be directly used for food or active packaging applications. The membrane hosts small and homogeneously dispersed crystals of cyclodextrin-aroma complexes which are formed during the electrospinning. With this type of structure, the release of aroma compound is negligible at ambient conditions (23 °C and 55% UR) even at high temperature (up to 230 °C), and it occurs beyond a given relative humidity threshold (90%), useful for food packaging applications. The mass fraction of free aroma released is directly related to the water activity of the system, namely, φ=aW(n)/(aW(n)+Kapp) explaining the observed key role played by the relative humidity on the release of the aroma compounds.

  20. Asialoglycoprotein receptor mediated hepatocyte targeting - strategies and applications.

    PubMed

    D'Souza, Anisha A; Devarajan, Padma V

    2015-04-10

    Hepatocyte resident afflictions continue to affect the human population unabated. The asialoglycoprotein receptor (ASGPR) is primarily expressed on hepatocytes and minimally on extra-hepatic cells. This makes it specifically attractive for receptor-mediated drug delivery with minimum concerns of toxicity. ASGPR facilitates internalization by clathrin-mediated endocytosis and exhibits high affinity for carbohydrates specifically galactose, N-acetylgalactosamine and glucose. Isomeric forms of sugar, galactose density and branching, spatial geometry and galactose linkages are key factors influencing ligand-receptor binding. Popular ligands for ASGPR mediated targeting are carbohydrate polymers, arabinogalactan and pullulan. Other ligands include galactose-bearing glycoproteins, glycopeptides and galactose modified polymers and lipids. Drug-ligand conjugates provide a viable strategy; nevertheless ligand-anchored nanocarriers provide an attractive option for ASGPR targeted delivery and are widely explored. The present review details various ligands and nanocarriers exploited for ASGPR mediated delivery of drugs to hepatocytes. Nanocarrier properties affecting ASGPR mediated uptake are discussed at length. The review also highlights the clinical relevance of ASGPR mediated targeting and applications in diagnostics. ASGPR mediated hepatocyte targeting provides great promise for improved therapy of hepatic afflictions.