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Sample records for guehomyces pullulans 17-1

  1. Laccases from Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  2. Estimation of pullulan by hydrolysis with pullulanase.

    PubMed

    Wu, Sheng-Jun; Kim, Jin-Moon; Zhou, Chao; Jin, Zheng-Yu; Tong, Qun-Yi

    2010-08-01

    A novel method for the estimation of pullulan was developed in which pullulan was hydrolysed by pullulanase. The hydrolysed product was mainly maltotriose and was determined colorimetrically using 3,5-dimethylsalicylic acid. This gave good linearity with respect to the concentration of pullulan in the fermentation broth. The content of pullulan determined in this way was less than that determined by a coupled enzyme assay and was comparable to that determined by an HPLC method. The new method was specific for estimation of pullulan, demonstrated high accuracy, and could assay pullulan from up to 3.2 mg/ml. PMID:20386956

  3. 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.

  4. Effect of uracil on pullulan production by Aureobasidium pullulans CGMCC1234.

    PubMed

    Sheng, Long; Zhu, Guilan; Tong, Qunyi

    2014-01-30

    Effect of uracil on the pullulan production, biomass and uridine phosphorylase (UPase) activity was studied in this research. Uracil was found to enhance pullulan accumulation and the addition time of uracil was crucial to pullulan production. Pullulan yield of 49.07 g/L was achieved by adding 5mM uracil at 48 h, by comparison to 37.72 g/L obtained with the control. UPase activity could not be detected at early growth stage of Aureobasidium pullulans, but stimulated by added uracil at logarithmic phase and stationary phase. The time course study on the fermentation of pullulan demonstrates that pullulan production was not closely associated with biomass accumulation. Results indicate that the increased pullulan yield brought by uracil was correlated with UPase activity. PMID:24299794

  5. Color Variability, Pullulan Production, and Pullulan Degrading Enzyme Activities in Tropical Isolates of Aureobasidium Pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Forty-six isolates from fifteen provinces in Thailand of the polymorphic fungus Aureobasidium pullulans were cultured. They were isolated from leaves, painted wall and wood surfaces. Each isolate was cultured in production medium (PM) containing sucrose and peptone as carbon and nitrogen sources, ...

  6. Phylogenetic classification of Aureobasidium pullulans strains for production of pullulan and xylanase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study tests the hypothesis that phylogenetic classification can predict whether A. pullulans strains will produce useful levels of the commercial polysaccharide, pullulan, or the valuable enzyme, xylanase. To test this hypothesis, 19 strains of A. pullulans with previously described phenotypes...

  7. a-Amylase activity during pullulan production and a-Amylase gene analyses of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungus Aureobasidium pullulans is the source of commercially produced pullulan, a high molecular weight polysaccharide that is used in the manufacture of edible films. It has been proposed that alpha-amylase negatively affects the molecular weight of pullulan in late cultures. Based on a recen...

  8. 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. PMID:26478402

  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. PMID:27542744

  10. 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.

  11. Evaluation of medium composition and fermentation parameters on pullulan production by Aureobasidium pullulans.

    PubMed

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

    2011-04-01

    The goal of this study was to enhance pullulan production by evaluating the effects of different fermentation parameters. Various carbon sources and their concentrations, yeast extract (YE) concentrations, fermentation temperatures and various pH profiles were examined. The optimal growth condition for pullulan production by Aureobasidium pullulans has been found as 75 g/L of sucrose as carbon source, 3 g/L of YE and cultivation temperature at 30 (°)C. Under these conditions with an initial pH at 5, 20.7 g/L of final pullulan concentration and 0.22 g/L/h maximum production rate were obtained. Later on, various pH profiles, agitation speeds, aerations and fed-batch fermentation were evaluated. The results demonstrated that pullulan production was enhanced to 25.8 g/L after 7-day cultivation with a 0.68 -g/L/h maximum production rate. There was no significant improvement of pullulan production from fed-batch fermentation. The optimal kinetics parameters were as follows: initial pH at 2.0, switched to pH 5.0 after 72 h and kept constant; agitation speed at 200 rpm; aeration at 1.5 vvm. The quality analysis demonstrated that the pullulan content produced from optimal conditions was 94.5% and its viscosity was 2.3 centipoise (cP). Fourier transform infrared spectroscopy also suggested that pullulan dominated the produced exopolysaccharide. PMID:21421674

  12. Production of pullulan by Aureobasidium pullulans from Asian palm kernel: a novel substrate.

    PubMed

    Sugumaran, K R; Gowthami, E; Swathi, B; Elakkiya, S; Srivastava, S N; Ravikumar, R; Gowdhaman, D; Ponnusami, V

    2013-01-30

    Production of a commercially important biodegradable polymer, pullulan, by Aureobasidium pullulans from four agricultural wastes namely wheat bran, rice bran, coconut kernel and palm kernel was evaluated in solid state fermentation. Under the experimental conditions, palm kernel resulted in highest concentration of pullulan (16 g/L) among the four solid substrates. Optimum initial pH and moisture content for pullulan production were found out to be 6.5 and 50% respectively. 18.43 g/L of pullulan was produced from Asian palm kernel with initial pH 6.5 after 7 days of fermentation and yeast like morphology was predominant under this condition. Among different nitrogen sources tried in this study, yeast extract was found to the best. The pullulan produced from palm kernel was characterized by FTIR and (1)H NMR. The results were matching with that of commercial pullulan. Thus, Asian palm kernel appears to be an attractive low cost carbon source for the production of pullulan. PMID:23218356

  13. Multilocus Phylogenetic Analyses, Pullulan Production and Xylanase Activity of Tropical Isolates of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aureobasidium pullulans is the source of the commercial polysaccharide, pullulan, and the enzyme, xylanase (EC 3.2.1.8). Isolates are typically off-white to black on solid media, while some tropical isolates have been described as "color variants" with bright pigments of red, yellow, or purple. In...

  14. Pullulan Production by Aureobasidium pullulans ATCC 201253 Cells Adsorbed onto Cellulose Anion and Cation Exchangers

    PubMed Central

    West, Thomas P.

    2012-01-01

    The anion exchanger phosphocellulose and the cation exchanger triethylaminoethyl cellulose were used to immobilize cells of the fungus Aureobasidium pullulans ATCC 201253 and the adsorbed cells were subsequently investigated for their ability to produce the polysaccharide pullulan using batch fermentation. The cells adsorbed on the triethylaminoethyl cellulose at pH 7.5 produced higher pullulan levels than those cells immobilized on phosphocellulose at pH 4.0 for 2 cycles of 168 h at 30 °C. Relative to the initial cycle of 168 h, pullulan production by the cells immobilized on the triethylaminoethyl cellulose decreased slightly after 168 h of the second production cycle while pullulan production by the phosphocellulose-immobilized cells remained about the same after 168 h of the second production cycle. PMID:23762749

  15. Relationship Between a-amylase Activity and Pullulan Profiles, and a-amylase Gene Analyses of the Fungus Aureobasidium Pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tropical isolates of Aureobasidium pullulans isolated from various habitats in Thailand were classified based on multilocus phylogenetic analyses using concordance analysis of DNA sequences. This fungus is the major source of commercially produced pullulan, a high molecular weight polysaccharide th...

  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)). PMID:12052094

  17. Copper sulfate improves pullulan production by bioconversion using whole cells of Aureobasidium pullulans as the catalyst.

    PubMed

    Wang, Dahui; Ju, Xiaomin; Zhang, Gaochuan; Wang, Donghua; Wei, Gongyuan

    2016-10-01

    The effects of mineral salts on pullulan production by bioconversion using whole cells of Aureobasidium pullulans CCTCC M 2012259 as the catalyst were investigated. Copper sulfate (CuSO4) improved pullulan production by 36.2% and 42.3% when added at the optimum concentration of 0.2mg/L to the bioconversion broth or seed medium, respectively, as compared with controls without CuSO4 addition. Pullulan production was further enhanced when CuSO4 was added to both seed medium and bioconversion broth simultaneously. In order to probe the mechanism of CuSO4 improvement, cell viability, membrane integrity, intracellular adenosine triphosphate (ATP) levels and the activities of key enzymes involved in pullulan biosynthesis were determined. As a result, CuSO4 increased the activities of key biosynthetic enzymes, maintained intracellular ATP at a higher level, and accelerated the rate of pullulan secretion, all of which contributed to improved pullulan production by bioconversion. PMID:27312631

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  19. 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. PMID:26428132

  20. 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. PMID:26434522

  1. 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.

  2. Biotechnology of Aureobasidium pullulans: A phylogenetic perspective

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  3. Peat Hydrolysate Medium Optimization for Pullulan Production

    PubMed Central

    Boa, Jacques M.; LeDuy, Anh

    1984-01-01

    Peat hydrolysate, a diluted acid-autoclaved extract of peat, was used as a substrate for the production of the extracellular polysaccharide pullulan by three strains of Aureobasidium pullulans, 140B, 142, and 2552. It was found that the addition of (NH4)2SO4 and K2HPO4 as sources of nitrogen and phosphate, respectively, is not necessary for the polysaccharide production. The economically optimized culture medium for large-scale production of pullulan contains peat hydrolysate, 0.05% NaCl, 0.02% MgSO4, and 0.01% antifoam FG-10. The initial pH of peat hydrolysate medium is adjusted to its optimum value of 6.0 with Ca(OH)2. The total ingredient cost for the production of each kilogram of pullulan with optimized medium is only 1/10 of that with the nonoptimized medium. In this study, a zero cost for peat hydrolysate was assumed, since it is an effluent of the peat and peat processing industries. PMID:16346596

  4. Heavy Oils Produced by Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  5. Dichromated pullulan as a novel photosensitive holographic material.

    PubMed

    Panteli, D; Savi, S; Jakovljevic, D

    1998-05-15

    We found that pullulan [an a -glucan with (1?4) and (1?6) linkages] doped with chromium ions posseses promising holographic properties. It is environmentally stable and is capable of registering high spatial frequencies (as many as 1500 lines/mm) and producing surface relief. At 1000 lines/mm pullulan's diffraction efficiency was 4%. Chemical treatment of pullulan with a mixture of water and isopropyl alcohol is simple. However, as expected from analogy with dichromated gelatin, pullulan is sensitive in only the blue-green region of the spectrum. PMID:18087349

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

    NASA Astrophysics Data System (ADS)

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

    2016-05-01

    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.

  7. Effect of nitrogen source on pullulan production by Aureobasidium pullulans grown in a batch bioreactor.

    PubMed

    West, T P; Strohfus, B

    1999-01-01

    Pullulan production by Aureobasidium pullulans ATCC 201253 using selected nitrogen sources was studied in a medium using corn syrup as a carbon source. Independent of the corn syrup concentration present, the use of corn steep liquor or hydrolysed soy protein as a nitrogen source instead of ammonium sulphate did not elevate polysaccharide production by ATCC 201253 cells grown in an aerated, batch bioreactor containing 4 litres of medium. Pullulan production on corn steep liquor or hydrolysed soy protein as a nitrogen source became more comparable as the concentration of corn syrup was increased. Cell weights after 7 days of growth on any of the nitrogen sources were similar. The viscosity of the polysaccharide on day 7 was highest for cells grown on ammonium sulphate and 12.5% corn syrup. The pullulan content of the polysaccharide elaborated by ammonium sulphate-grown cells on day 7 decreased as the corn syrup level rose in the medium while the pullulan content of polysaccharide produced by cells grown on corn steep liquor or soytone generally increased. PMID:10581727

  8. Pullulan-hyperproducing color variant strain of Aureobasidium pullulans FB-1 newly isolated from phylloplane of Ficus sp.

    PubMed

    Singh, R S; Saini, G K

    2008-06-01

    The studies were carried out for the isolation of efficient pullulan producing strains of Aureobasidium pullulans. Five strains were isolated from phylloplane of different plants. Amongst these, three were producing black pigment melanin, while the remaining two produced pink pigment. These two color variant isolates of A. pullulans were designated as FB-1 and FG-1, and obtained from phylloplane of Ficus benjamina and Ficus glometa, respectively. The parameters employed for the identification of the isolates included morphology, nutritional assimilation patterns and exopolysaccharide (EPS) production. Isolates were compared with standard cultures for EPS production. A. pullulans FB-1 was the best producer of pullulan giving up to 1.9, 1.4 and 1.7 times more pullulan than the control of A. pullulans NCIM 976, NCIM 1048 and NCIM 1049, respectively. The IR spectra of the isolates and standard strains revealed that the polysaccharide was pullulan, but not aubasidan. The study also supported the fact that A. pullulans is a ubiquitous organism and phylloplane being the important niche of the organism. PMID:17869506

  9. Multilocus phylogenetic analyses, pullulan production and xylanase activity of tropical isolates of Aureobasidium pullulans.

    PubMed

    Manitchotpisit, Pennapa; Leathers, Timothy D; Peterson, Stephen W; Kurtzman, Cletus P; Li, Xin-Liang; Eveleigh, Douglas E; Lotrakul, Pongtharin; Prasongsuk, Sehanat; Dunlap, Christopher A; Vermillion, Karl E; Punnapayak, Hunsa

    2009-10-01

    Aureobasidium pullulans is the source of the commercially valuable polysaccharide pullulan and the enzyme xylanase. Isolates are typically off-white to pale pink or black on solid media, while some tropical isolates have been described as 'color variants' with bright pigments of red, yellow or purple. We sequenced 5 loci (internal transcribed spacer, intergenic spacer 1, translation elongation factor-1 alpha, beta tubulin, and RNA polymerase II) from 45 new isolates from Thailand. Based on the phylogenetic analyses, isolates were classified into 12 clades. Each clade showed different colors on different culture media including two clades with 'color variants' and some clades exhibited high levels of pullulan production or xylanase activity. Colony characteristics do not correlate perfectly with DNA sequence phylogeny or the physiological characters, but DNA sequence differences rapidly identify isolates with genetic novelty. PMID:19619651

  10. Pullulan degrading enzymes of bacterial origin.

    PubMed

    Domań-Pytka, Monika; Bardowski, Jacek

    2004-01-01

    Pullulan degrading enzymes belong to a group of glycosylhydrolases that are widely distributed in nature and are produced by an extremely wide variety of species. Among them the thermophilic and mesophilic bacteria are a rich source of these enzymes. There are many biotechnological applications for these enzymes and a rapidly growing amount of information about their diversity, genetic as well as biochemical and biophysical characteristics. The properties of these enzymes vary and are somewhat linked to the natural environment inhabited by the producing organisms. Genes for these enzymes have been cloned from several strains and their amino acid sequences show highly conserved regions common to the enzymes of the amylase family. Molecular studies have greatly extended our knowledge on pullulan degrading enzymes and their biosynthesis. However, enzyme production levels have usually not been as high as had been assumed possible, and the properties of some enzymes are less than optimal for their industrial applications. Some of these problems can be overcome with the use of good producer organisms, optimized expression/secretion vectors, and site-directed mutagenesis. The molecular biology of pullulan degrading enzymes has been and continues to be a valuable system for studying basic questions of cell biology, such as mechanisms of gene regulation and secretion, and the structure-function relationships of proteins. PMID:15239382

  11. Redefinition of Aureobasidium pullulans and its varieties.

    PubMed

    Zalar, P; Gostincar, C; de Hoog, G S; Ursic, 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 beta-tubulin (TUB), translation elongation factor (EF1alpha) 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

  12. 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

  13. Pullulan production by Aureobasidium pullulans grown on ethanol stillage as a nitrogen source.

    PubMed

    West, T P; Strohfus, B

    1996-01-01

    Pullulan production by Aureobasidium pullulans strain RP-1 using thin stillage from fuel ethanol production as a nitrogen source was studied in a medium using corn syrup as a carbon source. The use of 1% thin stillage as a nitrogen source instead of ammonium sulphate elevated polysaccharide production by strain RP-1 cells when grown on a concentration of up to 7.5% corn syrup, independent of yeast extract supplementation. Dry weights of cells grown in medium containing ammonium sulphate as the nitrogen source were higher than the stillage-grown cells after 7 days of growth. The viscosity of the polysaccharide on day 7 was higher for cells grown on thin stillage rather than ammonium sulphate as a nitrogen source. The pullulan content of the polysaccharide elaborated by ammonium sulphate-grown cells on day 7 was higher than the pullulan content of polysaccharide produced by stillage-grown cells regardless of whether yeast extract was added to the culture medium. PMID:9121381

  14. Characterization of UGPase from Aureobasidium pullulans NRRL Y-12974 and Application in Enhanced Pullulan Production.

    PubMed

    Li, Haifeng; Zhang, Yunyi; Gao, Yunyun; Lan, Yuanyang; Yin, Xiaopu; Huang, Lifeng

    2016-03-01

    UDPG pyrophosphatase (UGPase) plays an important role in carbohydrate metabolism, catalyzing a reversible production of uridine diphosphate glucose (UDPG) and pyrophosphate (PPi) from Glc-1-P and UTP. UGPase gene from Aureobasidium pullulans NRRL Y-12974 was cloned, overexpressed in Escherichia coli. The recombinant UGPase possess molecular mass of 55 KDa and specific activity of 7.33 U/mg protein. The K m values of rUGPase were 5.045 μM against UTP and 3.333 μM against Glc-1-P. The V max values of rUGPase were 3.467 μM min(-1)against UTP and 2.817 μM min(-1) against Glc-1-P. And, it does not catalyze Glc-1-P and ATP, nor galactose-1-P and UTP. Homolgous expression of UGPase in native organism can improve the intracellular UDPG concentration by 4.7-fold time. The yield of pullulan in engineering strain A4 was improved to 18.2 g g(-1) cell dry weight which is 1.3-fold time of parent strain. No obvious change of growth was found between engineering strain and parent strain. To the best of our knowledge, this is the first report of improving pullulan yield in A. pullulans using metabolic engineering technique. PMID:26613730

  15. An extremely thermostable amylopullulanase from Staphylothermus marinus displays both pullulan- and cyclodextrin-degrading activities.

    PubMed

    Li, Xiaolei; Li, Dan; Park, Kwan-Hwa

    2013-06-01

    A gene encoding an amylopullulanase of the glycosyl hydrolase (GH) family 57 from Staphylothermus marinus (SMApu) was heterologously expressed in Escherichia coli. SMApu consisted of 639 amino acids with a molecular mass of 75.3 kDa. It only showed maximal amino acid identity of 17.1 % with that of Pyrococcus furiosus amylopullulanase in all identified amylases. Not like previously reported amylopullulanases, SMApu has no signal peptide but contains a continuous GH57N_Apu domain. It had the highest catalytic efficiency toward pullulan (k cat/K m , 342.34 s(-1) mL mg(-1)) and was extremely thermostable with maximal pullulan-degrading activity (42.1 U/mg) at 105 °C and pH 5.0 and a half-life of 50 min at 100 °C. Its activity increased to 116 % in the presence of 5 mM CaCl2. SMApu could also degrade cyclodextrins, which are resistant to the other amylopullulanases. The initial hydrolytic products from pullulan, γ-CD, and 6-O-maltooligosyl-β-CD were [6)-α-D-Glcp-(1 → 4)-α-D-Glcp-(1 → 4)-α-D-Glcp-(1→]n, maltooctaose, and single maltooligosaccharide plus β-CD, respectively. The final hydrolytic products from above-mentioned substrates were maltose and glucose. These results confirm that SMApu is a novel amylopullulanase of the family GH57 possessing the cyclodextrin-degrading activity of cyclomaltodextrinase. PMID:23001056

  16. [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. PMID:25345061

  17. Injectable pullulan hydrogel for the prevention of postoperative tissue adhesion.

    PubMed

    Bang, Sumi; Lee, Eungjae; Ko, Young-Gwang; Kim, Won Il; Kwon, Oh Hyeong

    2016-06-01

    Methods for reducing and preventing postoperative abdominal adhesions have been researched for decades; however, despite these efforts, the formation of postoperative peritoneal adhesions is continuously reported. Adhesions cause serious complications such as postoperative pain, intestinal obstruction, and infertility. Tissue adhesion barriers have been developed as films, membranes, knits, sprays, and hydrogels. Hydrogels have several advantages when used as adhesion barriers, including flexibility, low tissue adhesiveness, biodegradability, and non-toxic degraded products. Furthermore, compared with preformed hydrogels, injectable hydrogels can fill and cover spaces of any shape and do not require a surgical procedure for implantation. In this study, pullulan was modified through reaction with 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) to introduce carboxyl and phenyl groups as crosslinking sites. The grafting of tyramine on pullulan allows crosslinking branches on pullulan backbone. We successfully fabricated pullulan hydrogel with an enzymatic reaction using horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). The chemical structure of modified pullulan was analyzed with ATR-FTIR and (1)H NMR spectroscopies. Rheological properties were tested by measuring storage modulus with varying H2O2, HRP, polymer solution concentrations and tyramine substitution rates. Cell viability and animal tests were performed. The modified pullulan hydrogel is an invaluable advance in anti-adhesion agents. PMID:26879910

  18. Structural characterization of novel extracellular liamocins (mannitol oils) produced by Aureobasidium pullulans strain NRRL 50380

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aureobasidium pullulans is a common, ubiquitous fungus, which is used industrially to produce the polysaccharide pullulan. We have previously shown that A. pullulans produces various heavier-than-water oils, first named here as liamocins, that accumulate in fermentations. Here we report the struc...

  19. 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...

  20. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 1 2013-04-01 2013-04-01 false Scope. 17.1 Section 17.1 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL CIVIL MONEY PENALTIES HEARINGS § 17.1 Scope. This part sets forth practices and procedures for hearings concerning the administrative imposition of civil money penalties by...

  1. 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...

  2. 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,...

  3. 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...

  4. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 1 2011-04-01 2011-04-01 false Scope. 17.1 Section 17.1 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL CIVIL MONEY PENALTIES HEARINGS § 17.1 Scope. This part sets forth practices and procedures for hearings concerning the administrative imposition of civil money penalties by...

  5. 41 CFR 302-17.1 - Authority.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 41 Public Contracts and Property Management 4 2011-07-01 2011-07-01 false Authority. 302-17.1 Section 302-17.1 Public Contracts and Property Management Federal Travel Regulation System RELOCATION ALLOWANCES MISCELLANEOUS ALLOWANCES 17-RELOCATION INCOME TAX (RIT) ALLOWANCE § 302-17.1 Authority. Payment...

  6. 41 CFR 302-17.1 - Authority.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 41 Public Contracts and Property Management 4 2010-07-01 2010-07-01 false Authority. 302-17.1 Section 302-17.1 Public Contracts and Property Management Federal Travel Regulation System RELOCATION ALLOWANCES MISCELLANEOUS ALLOWANCES 17-RELOCATION INCOME TAX (RIT) ALLOWANCE § 302-17.1 Authority. Payment...

  7. 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,...

  8. 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...

  9. 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,...

  10. 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...

  11. 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,...

  12. 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,...

  13. 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...

  14. Recent insights on applications of pullulan in tissue engineering.

    PubMed

    Singh, Ram Sarup; Kaur, Navpreet; Rana, Vikas; Kennedy, John F

    2016-11-20

    Tissue engineering is a recently emerging line of act which assists the regeneration of damaged tissues, unable to self-repair themselves and in turn, enhances the natural healing potential of patients. The repair of injured tissue can be induced with the help of some artificially created polymer scaffolds for successful tissue regeneration. The pullulan composite scaffolds can be used to enhance the proliferation and differentiation of cells for tissue regeneration. The unique pattern of pullulan with α-(1→4) and α-(1→6) linkages along with the presence of nine hydroxyl groups on its surface, endows the polymer with distinctive physical features required for tissue engineering. Pullulan can be used for vascular engineering, bone repair and skin tissue engineering. Pullulan composite scaffolds can also be used for treatment of injured femoral condyle bone, skull bone and full thickness skin wound of murine models, transversal mandibular and tibial osteotomy in goat, etc. This review article highlights the latest developments on applications of pullulan and its derivatives in tissue engineering. PMID:27561517

  15. Biosynthesis of Novel Exopolymers by Aureobasidium pullulans

    PubMed Central

    Lee, Jin W.; Yeomans, Walter G.; Allen, Alfred L.; Deng, Fang; Gross, Richard A.; Kaplan, David L.

    1999-01-01

    Aureobasidium pullulans ATCC 42023 was cultured under aerobic conditions with glucose, mannose, and glucose analogs as energy sources. The exopolymer extracts produced under these conditions were composed of glucose and mannose. The molar ratio of glucose to mannose in the exopolymer extract and the molecular weight of the exopolymer varied depending on the energy source and culture time. The glucose content of exopolymer extracts formed with glucose and mannose as the carbon sources was between 91 and 87%. The molecular weight decreased from 3.5 × 106 to 2.12 × 106 to 0.85 × 106 to 0.77 × 106 with culture time. As the culture time increased, the glucose content of the exopolymer extract formed with glucosamine decreased from 55 ± 3 to 29 ± 2 mol%, and the molecular weight increased from 2.73 × 106 to 4.86 × 106. There was no evidence that glucosamine was directly incorporated into exopolymers. The molar ratios of glucose to mannose in exopolymer extracts ranged from 87 ± 3:13 ± 3 to 28 ± 2:72 ± 2 and were affected by the energy source added. On the basis of the results of an enzyme hydrolysis analysis of the exopolymer extracts and the compositional changes observed, mannose (a repeating unit) was substituted for glucose, which gave rise to a new family of exopolymer analogs. PMID:10583975

  16. Pullulan microcarriers for bone tissue regeneration.

    PubMed

    Aydogdu, Hazal; Keskin, Dilek; Baran, Erkan Turker; Tezcaner, Aysen

    2016-06-01

    Microcarrier systems offer a convenient way to repair bone defects as injectable cell carriers that can be applied with small incisions owing to their small size and spherical shape. In this study, pullulan (PULL) microspheres were fabricated and characterized as cell carriers for bone tissue engineering applications. PULL was cross-linked by trisodium trimetaphosphate (STMP) to enhance the stability of the microspheres. Improved cytocompatibility was achieved by silk fibroin (SF) coating and biomimetic mineralization on the surface by incubating in simulated body fluid (SBF). X-ray diffraction (XRD), scanning electron microscopy (SEM) and fluorescent microscopy analysis confirmed biomimetic mineralization and SF coating on microspheres. The degradation analysis revealed that PULL microspheres had a slow degradation rate with 8% degradation in two weeks period indicating that the microspheres would support the formation of new bone tissue. Furthermore, the mechanical tests showed that the microspheres had a high mechanical stability that was significantly enhanced with the biomimetic mineralization. In vitro cell culture studies with SaOs-2 cells showed that cell viability was higher on SF and SBF coated microspheres on 7th day compared to PULL ones under dynamic conditions. Alkaline phosphatase activity was higher for SF coated microspheres in comparison to uncoated microspheres when dynamic culture condition was applied. The results suggest that both organic and inorganic surface modifications can be applied on PULL microspheres to prepare a biocompatible microcarrier system with suitable properties for bone tissue engineering. PMID:27040238

  17. 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. PMID:27427741

  18. Bioproducts from diverse phylogenetic clades of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  19. Lipase production by diverse phylogenetic clades of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thirty-nine strains representing 12 diverse phylogenetic clades of Aureobasidium pullulans were surveyed for lipase production using a quantitative assay. Strains in clades 4 and 10 produced 0.2-0.3 U lipase/ml, while color variant strain NRRL Y-2311-1 in clade 8 produced 0.54 U lipase/ml. Strains i...

  20. Lipase production by diverse phylogenetic clades of Aureobasidium pullulans.

    PubMed

    Leathers, Timothy D; Rich, Joseph O; Anderson, Amber M; Manitchotpisit, Pennapa

    2013-10-01

    Thirty-nine strains representing 12 diverse phylogenetic clades of Aureobasidium pullulans were surveyed for lipase production using a quantitative assay. Strains in clades 4 and 10 produced 0.2-0.3 U lipase/ml, while color variant strain NRRL Y-2311-1 in clade 8 produced 0.54 U lipase/ml. Strains in clade 9, which exhibit a dark olivaceous pigment, produced the highest levels of lipase, with strain NRRL 62034 yielding 0.57 U lipase/ml. By comparison, Candida cylindracea strain NRRL Y-17506 produced 0.05 U lipase/ml under identical conditions. A. pullulans strain NRRL 62034 reached maximal lipase levels in 5 days on lipase induction medium, while A. pullulans strain NRRL Y-2311-1 and strains in clades 4 and 10 were highest after 6 days. A. pullulans strain NRRL Y-2311-1 and strains in clade 9 produced two extracellular proteins in common, at >50 and <37 kDa. PMID:23801121

  1. Laccase production by diverse phylogenetic clades of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  2. Fabrication of pullulan and pectin submicron fibers by electrospinning

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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. 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

  5. 14 CFR 17.1 - Applicability.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Applicability. 17.1 Section 17.1 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PROCEDURAL RULES... Office of Dispute Resolution for Acquisition (ODRA) on or after October 7, 2011, with the exception...

  6. 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...

  7. 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...

  8. 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...

  9. 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...

  10. 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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  12. Pullulan encapsulation of labile biomolecules to give stable bioassay tablets.

    PubMed

    Jahanshahi-Anbuhi, Sana; Pennings, Kevin; Leung, Vincent; Liu, Meng; Carrasquilla, Carmen; Kannan, Balamurali; Li, Yingfu; Pelton, Robert; Brennan, John D; Filipe, Carlos D M

    2014-06-10

    A simple and inexpensive method is reported for the long-term stabilization of enzymes and other unstable reagents in premeasured quantities in water-soluble tablets (cast, not compressed) made with pullulan, a nonionic polysaccharide that forms an oxygen impermeable solid upon drying. The pullulan tablets dissolve in aqueous solutions in seconds, thereby facilitating the easy execution of bioassays at remote sites with no need for special reagent handling and liquid pipetting. This approach is modular in nature, thus allowing the creation of individual tablets for enzymes and their substrates. Proof-of-principle demonstrations include a Taq polymerase tablet for DNA amplification through PCR and a pesticide assay kit consisting of separate tablets for acetylcholinesterase and its chromogenic substrate, indoxyl acetate, both of which are highly unstable. The encapsulated reagents remain stable at room temperature for months, thus enabling the room-temperature shipping and storage of bioassay components. PMID:24764260

  13. Betaine conjugated cationic pullulan as effective gene carrier.

    PubMed

    Ambattu, Lizebona August; Rekha, M R

    2015-01-01

    Polyethyleneimne (PEI) is a very efficient transfecting agent but is toxic due to high charge density. To generate a vector which is efficient and less cytotoxic, PEI was conjugated with pullulan (PPEI). Further conjugation was done on PPEI with zwitter ionic betaine which possess antifouling property. PEI of molecular weight 1.2, 2, and 10 kDa were used in the study. Buffering capacity of pullulan-PEI-betaine (PPB) conjugates was found to be sufficient enough for the polymers to make endosomal escape. The polymers proved to be less cytotoxic and highly hemocompatible than PEI. Nuclear localization of YOYO tagged DNA was observed with the nanoplexes developed using PPEI and PPBs of PEI 10 kDa. Transfection efficiency was evaluated using p53 expressing gene and the live dead assay demonstrated very high transfection efficiency with PPB conjugates of PEI 10 kDa. PMID:25304750

  14. Real-time and postprocessing holographic effects in dichromated pullulan.

    PubMed

    Savić, Svetlana; Pantelić, Dejan; Jakovijević, Dragica

    2002-08-01

    Experimental results concerning both real-time and postprocessing (after-development) behavior of a novel photosensitive material, dichromate-sensitized pullulan (DCP), are investigated. The exposure mechanism and possibilities for controlling holographic grating properties are discussed. We have shown that it is possible to maximize the diffraction efficiency of interference gratings after development by controlling diffraction efficiency in real time. Stronger real-time effects of DCP compared with those of dichromated gelatin are achieved. PMID:12153075

  15. Pullulan-based hydrogel for smooth muscle cell culture.

    PubMed

    Autissier, Aude; Letourneur, Didier; Le Visage, Catherine

    2007-08-01

    A hydrogel was prepared from pullulan and evaluated as a novel biomaterial for vascular engineering. Using a crosslinking process with sodium trimetaphosphate in aqueous solution, homogeneous, transparent, and easy-to-handle pullulan gels were obtained with water-content higher than 90%. A circular punch was used to cut 6-mm diameter and 2-mm thickness discs for cell culture. Environmental scanning electron microscopy analysis of hydrated gels revealed a smooth surface, on which rabbit vascular smooth muscle cells were successfully seeded. The absence of cytotoxicity was evidenced by a live/dead assay. Fluorescence-labeled cells were observed adhering and progressively spreading out on the surface of the material. Cellular proliferation was followed for up to 1 week using an MTT assay. In addition, a complete in vitro degradation of the gels was achieved in 3 h upon incubation in a pullulanase solution (44 U/mL). In conclusion, we have shown the feasibility of preparing a biocompatible pullulan-based hydrogel that could support vascular cell culture. Based on these promising results, future studies will focus on the seeding of vascular cells on tubular-shaped hydrogels and the in vivo implantation of these new biomaterials. PMID:17295223

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  18. Pullulan-complexed α-amylase and glucosidase in alginate beads: enhanced entrapment and stability.

    PubMed

    Jadhav, Swati B; Singhal, Rekha S

    2014-05-25

    Enhanced entrapment of the enzymes, α-amylase and glucoamylase, was found in alginate beads on addition of pullulan in the enzyme mixture. Under optimized process conditions of entrapment, enzymes-pullulan complex showed an entrapment of 85% in the alginate beads as opposed to 25% for the free enzymes. Beads of enzymes-pullulan complex showed lower inactivation rate constant and higher half life than corresponding beads of free enzymes. Activation energy of beads of enzymes-pullulan was increased by 6.81kJ/mole compared to beads of free enzymes. This implies better stability the enzymes in enzymes-pullulan beads along with increased immobilization yield. Moreover, enzymes-pullulan beads also showed pH stability at extreme acidic and alkaline pH. Addition of pullulan in the enzymes mixture lowered the Km and increased the Vmax as compared to beads of free enzymes. Hydrolysis of starch and reusability study showed better applicability of beads of enzymes-pullulan as compared to free enzymes. PMID:24708951

  19. Genetic Diversity of Tropical Isolates and Color Variants of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The polymorphic fungus Aureobasidium pullulans was isolated from many provinces in tropical Thailand and cultured in pullulan production medium (PM) containing sucrose and peptone as carbon and nitrogen sources, respectively. Liquid PM cultures varied in color from cream to pale pink, light burgund...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  1. 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. PMID:26775974

  2. 7 CFR 17.1 - General.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... practicable, under the authority of title I of the Agricultural Trade Development and Assistance Act of 1954... Secretary of Agriculture SALES OF AGRICULTURAL COMMODITIES MADE AVAILABLE UNDER TITLE I OF THE AGRICULTURAL TRADE DEVELOPMENT AND ASSISTANCE ACT OF 1954, AS AMENDED § 17.1 General. (a) What this part covers....

  3. 7 CFR 17.1 - General.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... practicable, under the authority of title I of the Agricultural Trade Development and Assistance Act of 1954... Secretary of Agriculture SALES OF AGRICULTURAL COMMODITIES MADE AVAILABLE UNDER TITLE I OF THE AGRICULTURAL TRADE DEVELOPMENT AND ASSISTANCE ACT OF 1954, AS AMENDED § 17.1 General. (a) What this part covers....

  4. 24 CFR 17.1 - Scope; definitions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Tort Claims Act issued by the Attorney General (31 FR 16616; 28 CFR part 14). (c) The terms Department and Organizational unit are defined in 24 CFR part 5. Procedures ... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Scope; definitions. 17.1 Section...

  5. 24 CFR 17.1 - Scope; definitions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Tort Claims Act issued by the Attorney General (31 FR 16616; 28 CFR part 14). (c) The terms Department and Organizational unit are defined in 24 CFR part 5. Procedures ... 24 Housing and Urban Development 1 2012-04-01 2012-04-01 false Scope; definitions. 17.1 Section...

  6. 24 CFR 17.1 - Scope; definitions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Tort Claims Act issued by the Attorney General (31 FR 16616; 28 CFR part 14). (c) The terms Department and Organizational unit are defined in 24 CFR part 5. Procedures ... 24 Housing and Urban Development 1 2013-04-01 2013-04-01 false Scope; definitions. 17.1 Section...

  7. 24 CFR 17.1 - Scope; definitions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... Tort Claims Act issued by the Attorney General (31 FR 16616; 28 CFR part 14). (c) The terms Department and Organizational unit are defined in 24 CFR part 5. Procedures ... 24 Housing and Urban Development 1 2011-04-01 2011-04-01 false Scope; definitions. 17.1 Section...

  8. 24 CFR 17.1 - Scope; definitions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Tort Claims Act issued by the Attorney General (31 FR 16616; 28 CFR part 14). (c) The terms Department and Organizational unit are defined in 24 CFR part 5. Procedures ... 24 Housing and Urban Development 1 2014-04-01 2014-04-01 false Scope; definitions. 17.1 Section...

  9. 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. PMID:25290967

  10. Hydrolysis of xylan by an immobilized xylanase from Aureobasidium pullulans

    SciTech Connect

    Allenza, P.; Scherl, D.S.; Detroy, R.W.; Leathers, T.D.; Scott, C.D. .

    1986-01-01

    The beta-(1,4)-linked xylose residues that comprise the backbone of the abundant plant polymer xylan can be released by enzymic hydrolysis. Xylanase, which is produced in exceptionally high levels by the color-variant strain Y-2311-1 of A. pullulans, was immobilized onto a macroporous ceramic carrier. Despite a low coupling efficiency, it was possible to run the reactor under a wide range of conditions with flow rates of 3-10 bed volumes/minute of 1% soluble xylan with no detectable leaching of enzyme. The size distribution of products and rate of xylan hydrolysis were very similar for the immobilized and soluble enzymes. (Refs. 13).

  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. PMID:26939662

  12. 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. PMID:23066220

  13. 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. PMID:24750929

  14. 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. PMID:23376560

  15. 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). PMID:23987421

  16. Effect of pullulan on the short-term and long-term retrogradation of rice starch.

    PubMed

    Chen, Long; Ren, Fei; Zhang, Zipei; Tong, Qunyi; Rashed, Marwan M A

    2015-01-22

    The effect of pullulan (PUL) on the retrogradation of rice starch (RS) was investigated by means of rapid visco-analyzer (RVA), rotational rheometer, differential scanning calorimetry (DSC), and X-ray diffraction (XRD). RVA results showed that addition of pullulan significantly decreased the breakdown and setback values, which meant that the short-term retrogradation of RS was inhibited. The dynamic time sweep of samples also proved the retarding effect of pullulan on the retrogradation of RS. DSC curves showed clearly that pullulan significantly reduced the retrogradation enthalpy of amylopectin, and the kinetics of retrogradation was analyzed using the Avrami model. XRD results showed that recrystallinity of RS was reduced from 11.565% to 8.841% with the addition of pullulan and this was in line with the DSC results. It could be concluded that the addition of pullulan apparently influenced not only the short-term retrogradation of amylose, but also the long-term retrogradation of amylopectin. PMID:25439913

  17. The immunomodulatory activities of pullulan and its derivatives in human pDC-like CAL-1 cell line.

    PubMed

    Wang, Fang; Qiao, Linan; Chen, Liwei; Zhang, Cong; Wang, Yan; Wang, Yinsong; Liu, Yuanyuan; Zhang, Ning

    2016-05-01

    In this study, acidic and alkaline pullulan derivates were synthesized and their immunomodulatory activities compared to pullulan were investigated in human pDC-like CAL-1 cell line. Pullulan was reacted respectively with succinic anhydride and N-(-2-aminoethyl)-1,3-propanediamine/N,N-carbonyl diimidazole to form acidic pullulan monosuccinate (SUPL) and alkaline pullulan-g-N-(-2-aminoethyl)-1,3-propanediamine (AMPL). In CAL-1 cells, pullulan, SUPL and AMPL up-regulated the mRNA expressions of type I interferons (IFN), including IFN-α and IFN-β1, and some other proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-23 (IL-23), and also significantly enhanced the protein expressions of IFN-α and TNF-α. The activation of nuclear factor kappa B (NF-κB) and the nuclear translocations of interferon regulation factors (IRFs), including IRF-3 and IRF-5, exhibited pivotal roles in the immune responses induced by pullulan, SUPL and AMPL. By comparison, pullulan and SUPL displayed weak effects on the activation of CAL-1 cells, but AMPL showed remarkably enhanced immunomodulatory activities, which were comparable to that induced by R848, an agonist for Toll-like receptor (TLR) 7/8. Our results suggested that AMPL, as an alkaline pullulan derivative, could be used as a potent immunomodulatory agent in the food and pharmacological fields. PMID:26854885

  18. 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. PMID:26165967

  19. 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

  20. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 1 2011-07-01 2011-07-01 false Purpose of these rules. 17.1 Section 17.1 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...

  1. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Purpose of these rules. 17.1 Section 17.1 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...

  2. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false When required. 96.17-1 Section 96.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS VESSEL CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean...

  3. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false When required. 96.17-1 Section 96.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS VESSEL CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean...

  4. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 1 2013-10-01 2013-10-01 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...

  5. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 1 2012-10-01 2012-10-01 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...

  6. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 1 2014-10-01 2014-10-01 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...

  7. 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...

  8. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 1 2011-10-01 2011-10-01 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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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,...

  11. 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. PMID:25935266

  12. 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. PMID:26480962

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

    PubMed

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

    2015-07-10

    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. PMID:26080998

  14. 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.

  15. β-Xylanase production by Aureobasidium pullulans grown on sugars agricultural residues.

    PubMed

    Karni, M; Deopurkar, R L; Rale, V B

    1993-07-01

    Aureobasidium pullulans grew well in media containing glucose, fructose, xylan or xylose but β-xylanase was only produced with xylan or xylose. Lactose and maltose were poor substrates for growth. β-Xylanase production was repressed in media containing glucose or fructose along with xylose. Agricultural residues, such as wheat bran, paddy husk and rice straw, could be used as carbon sources for growth and β-xylanase production of Aureobasidium pullulans. Tween 80 at 0.5% (v/v) increased the yield of β-xylanase by up to 20%. PMID:24420115

  16. 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. PMID:27507528

  17. 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 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  18. 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 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  19. 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 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  20. 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 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  1. 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 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  8. QUANTITATIVE IMAGING AND STATISTICAL ANALYSIS OF FLUORESCENCE IN SITU HYBRIDIZATION (FISH) OF AUREOBASIDIUM PULLULANS. (R823845)

    EPA Science Inventory

    Abstract

    Image and multifactorial statistical analyses were used to evaluate the intensity of fluorescence signal from cells of three strains of A. pullulans and one strain of Rhodosporidium toruloides, as an outgroup, hybridized with either a universal o...

  9. Hydrolysis of pullulan by entrapped pullulanase in Ca/alginate beads.

    PubMed

    Gasmi, Sarah Nawel; Rihouey, Christophe; Picton, Luc; Le Cerf, Didier

    2014-09-01

    Pullulanase from Klebsiella pneumoniae was entrapped into calcium alginate beads. Its activity was estimated by the determination of number-average molar masses using two different methods: a colorimetric assay of reducing ends (REs) and a size-exclusion chromatography/multiangle light scattering/differential refractive index. The second method also provided weight-average molar masses of hydrolyzed pullulan and the quantity of maltotriose (DP3) and its multiples (DP6 and DP9) produced by the enzymatic treatment. The alginate beads showed a good retention of the loaded pullulanase (30%), and the system showed a downturn of hydrolysis kinetics in comparison with free pullulanase due to the limiting access of substrate-enzyme. On the contrary with the results obtained from free enzyme hydrolysis, for which a large distribution of pullulan fragments is observed during the treatment, the immobilized enzyme system has evidenced, during the enzymatic treatment, the coexistence of native or only slightly degraded pullulan chains together with maltotriose units. Complete hydrolysis of pullulan chains was achieved once diffused into the gel. PMID:24633755

  10. Novel maltotriose-hydrolyzing thermoacidophilic type III pullulan hydrolase from Thermococcus kodakarensis.

    PubMed

    Ahmad, Nasir; Rashid, Naeem; Haider, Muhammad Saleem; Akram, Mehwish; Akhtar, Muhammad

    2014-02-01

    A novel thermoacidophilic pullulan-hydrolyzing enzyme (PUL) from hyperthermophilic archaeon Thermococcus kodakarensis (TK-PUL) that efficiently hydrolyzes starch under industrial conditions in the absence of any additional metal ions was cloned and characterized. TK-PUL possessed both pullulanase and α-amylase activities. The highest activities were observed at 95 to 100°C. Although the enzyme was active over a broad pH range (3.0 to 8.5), the pH optima for both activities were 3.5 in acetate buffer and 4.2 in citrate buffer. TK-PUL was stable for several hours at 90°C. Its half-life at 100°C was 45 min when incubated either at pH 6.5 or 8.5. The Km value toward pullulan was 2 mg ml(-1), with a Vmax of 109 U mg(-1). Metal ions were not required for the activity and stability of recombinant TK-PUL. The enzyme was able to hydrolyze both α-1,6 and α-1,4 glycosidic linkages in pullulan. The most preferred substrate, after pullulan, was γ-cyclodextrin, which is a novel feature for this type of enzyme. Additionally, the enzyme hydrolyzed a variety of polysaccharides, including starch, glycogen, dextrin, amylose, amylopectin, and cyclodextrins (α, β, and γ), mainly into maltose. A unique feature of TK-PUL was the ability to hydrolyze maltotriose into maltose and glucose. PMID:24296501

  11. Galactosylated pullulan-curcumin conjugate micelles for site specific anticancer activity to hepatocarcinoma cells.

    PubMed

    Sarika, P R; James, Nirmala Rachel; Nishna, N; Anil Kumar, P R; Raj, Deepa K

    2015-09-01

    Galactosylated pullulan-curcumin conjugate (LANH2-Pu Ald-Cur SA) is developed for target specific delivery of curcumin to hepatocarcinoma cells by five step synthetic strategy, which includes oxidation of pullulan (Pu Ald), introduction of amino group to the targeting ligand (LANH2), grafting of the LANH2 to Pu Ald, modification of curcumin (Cur SA) and conjugation of Cur SA to pullulan. Nongalactosylated pullulan-curcumin conjugate (Pu-Cur SA) is also prepared to compare the enhancement in cytotoxicity offered by the targeting group. Both LANH2-Pu Ald-Cur SA and Pu-Cur SA conjugates could self assemble to micelle in water with hydrodynamic diameters of 355±9nm and 363±10nm, respectively. Both conjugates show spherical morphology and enhance stability of curcumin in physiological pH. Compared to Pu-Cur SA, LANH2-Pu Ald-Cur SA exhibits higher toxicity and internalization towards HepG2 cells. This indicates the enhanced uptake of LANH2-Pu Ald-Cur SA conjugate via ASGPR (asialoglycoprotein receptor) mediated endocytosis into HepG2 cells. PMID:26133239

  12. 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. PMID:27142298

  13. 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. PMID:24659335

  14. Biosynthesis and hyper production of pullulan by a newly isolated strain of Aspergillus japonicus-VIT-SB1.

    PubMed

    Mishra, Bishwambhar; Suneetha, V

    2014-07-01

    The main focus of this study was to screen and characterize novel microbial strains isolated from culinary leaf samples, capable of producing high concentrations of pullulan. Hundred isolates were screened from the phylloplane of different plants. The results revealed that eight strains had the capability to produce exopolysaccharide (EPS) and only one potential strain (designated as VIT-SB1) could produce the significant amount of EPS (3.9 ± 0.02%) on the 6th day of the fermentation without optimisation. The EPS synthesized by VIT-SB1 strain was confirmed to be pullulan on the basis of the results of FT-IR, HPLC and the enzymatic (Pullulanase) analysis. More than 91% hydrolysis of pullulan by pullulanase enzyme also indicated the presence of α (1 → 6) glycosidic linkages of α (1 → 4) linked maltotriose units. This VIT-SB1 strain was identified as Aspergillus japonicus based on the nucleotide sequence of the D1/D2 domain of Large-Subunit rRNA gene. The sequence was submitted to the GenBank Nucleotide sequence database with Accession No: KC128815. This study has confirmed that pullulan production capacity of this novel strain and Aureobasidium pullulans are comparable. Hence Aspergillus japonicus-VIT-SB1 strain can be commercially exploited as a potential pullulan producing strain. PMID:24609496

  15. 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. PMID:26800902

  16. Effect of carboxymethyl groups on degradation of modified pullulan by pullulanase from Klebsiella pneumoniae.

    PubMed

    Ali, Ghina; Rihouey, Christophe; Le Cerf, Didier; Picton, Luc

    2013-03-01

    Pullulanase is an enzyme that hydrolyses the α-1,6 linkages of pullulan (Pull) to produce maltotriose units. We studied the capacity of pullulanase to cleave its modified substrate: carboxymethylpullulan (CMPull), synthesized with two different degrees of substitution (DS=0.16 and 0.8). Size exclusion chromatography with on line multi angle light scattering and differential refractive index detection (SEC/MALS/DRI) was used to estimate both number and weight average molar masses, respectively, Mn and Mw, of pullulan and CMPulls together with the percentage of maltotriose formed during hydrolysis. Determination of reduced sugars gave also a Mn that is compared to data obtained by SEC. It revealed that CMPull is partially degraded by pullulanase and the rate of hydrolysis decreased with increased DS. At the end of the hydrolysis, Mn is decreased by a factor of 23 and 1.7 for CMPull with a DS of 0.16 and 0.8 respectively. The percentage of produced maltotriose decreased also when increasing DS (24% and 7% for CMPull DS 0.16 and 0.8 respectively). The kinetic properties of pullulanase were also investigated with Pull and CMPulls by isothermal titration calorimetry (ITC) using simple injection method. Based on Michaelis-Menten kinetics, Vmax (maximal velocity) decreased and KM (Michaelis constant) increased when DS of modified pullulan CMPull increased. PMID:23465908

  17. Effect of pullulan/poly(vinyl alcohol) blend system on the montmorillonite structure with property characterization of electrospun pullulan/poly(vinyl alcohol)/montmorillonite nanofibers.

    PubMed

    Islam, Md Shahidul; Yeum, Jeong Hyun; Das, Ajoy Kumar

    2012-02-15

    Nanofibers of the composite of pullulan (PULL), poly(vinyl alcohol) (PVA), and montmorillonite clay (MMT) were prepared using electrospinning method in aqueous solutions. Pullulan is an interesting natural polymer for many of its merits and good properties. Because of biocompatibility and non-toxicity of PVA, it could be used in numerous fields. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared (FTIR), X-ray diffraction (XRD), and thermal gravimetric analysis (TGA) were done to characterize the PULL/PVA/MMT nanofibers morphology and properties. XRD patterns and FTIR data demonstrated that there were good interactions between PULL and PVA caused by possibly hydrogen bonds. Moreover, XRD data and TEM images indicated that intercalated and exfoliated MMT nanoplatelets can be obtained within the PULL/PVA/MMT nanofibers depending on the PULL/PVA blend ratios. Furthermore, the thermal stability and mechanical property (tensile strength) of PULL/PVA/MMT nanofibers could be enhanced more by exfoliated MMT nanoplatelets than intercalated structures of that nanoplatelets. PMID:22172694

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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...

  3. 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...

  4. 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....

  5. 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....

  6. 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....

  7. 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....

  8. 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....

  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. PMID:25985744

  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

    2016-07-01

    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. PMID:26567632

  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. 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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  16. Removal of anionic dyes from aqueous solutions by an ion-exchanger based on pullulan microspheres.

    PubMed

    Constantin, Marieta; Asmarandei, Ionela; Harabagiu, Valeria; Ghimici, Luminita; Ascenzi, Paolo; Fundueanu, Gheorghe

    2013-01-01

    Pullulan-graft-poly(3-acrylamidopropyl trimethylammonium chloride) (P-g-pAPTAC) microspheres were prepared by suspension cross-linking of the pullulan previously grafted with cationic moieties. Adsorption of Azocarmine B by the P-g-pAPTAC microspheres was used as a model to demonstrate the removal of anionic dyes from aqueous solutions. Batch adsorption studies concerning the effect of the contact time, pH, initial dye concentration, temperature, grafting, and the nature of sulfonated anionic dyes on the adsorption kinetics were investigated. Adsorption was shown to be independent of pH. The experimental data best fitted to the pseudo-second order model which provided values of the rate constant k(2) of 1.4×10(-4) g mg(-1) min(-1) for 100 mg L(-1) solution and of 3.7×10(-4) g mg(-1) min(-1) for 500 mg L(-1) solution. From the Langmuir isotherm linear equation, the maximum adsorption capacity determined was 113.63 mg of Azocarmine B per gram of adsorbent; the negative value of the free energy change indicated the spontaneous nature of the adsorption process. PMID:23044107

  17. 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

    2016-01-01

    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. PMID:26817622

  18. 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. PMID:24702912

  19. Natural antifouling compounds produced by a novel fungus Aureobasidium pullulans HN isolated from marine biofilm.

    PubMed

    Gao, Min; Su, Rongguo; Wang, Ke; Li, Xuzhao; Lu, Wei

    2013-12-15

    A fungus, Aureobasidium pullulans, was isolated from marine biofilm and identified. A bioassay-guided fractionation procedure was developed to isolate and purify antifouling compounds from A. pullulans HN. The procedure was: fermentation broth-aeration and addition of sodium thiosulfate-graduated pH and liquid-liquid extraction-SPE purification-GC-MS analysis. Firstly, the fermentation broth was tested for its toxicity. Then it was treated with aeration and addition of sodium thiosulfate, and its toxicity was almost not changed. Lastly, antifouling compounds were extracted at different pH, the extract had high toxicity at pH 2 but almost no toxicity at pH 10, which suggested the toxicants should be fatty acids. The EC50 of the extract against Skeletonema costatum was 90.9 μg ml(-1), and its LC50 against Balanus amphitrete larvae was 22.2 μg ml(-1). After purified by HLB SPE column, the EC50 of the extract against S. costatum was 49.4 μg ml(-1). The myristic and palmitic acids were found as the main toxicants by GC-MS. PMID:24210009

  20. Superabsorbent polysaccharide hydrogels based on pullulan derivate as antibacterial release wound dressing.

    PubMed

    Li, Huanan; Yang, Jing; Hu, Xiaona; Liang, Jie; Fan, Yujiang; Zhang, Xingdong

    2011-07-01

    To accomplish ideal wound dressing, hydrogels based on a natural polysaccharide, pullulan were synthesized by chemical cross-linking. The tensile strengths of the hydrogel films (1 mm thick) were determined to range from 0.663 to 1.097 MPa in proportion to cross-linking degrees and water contents. The swelling study of the hydrogels in water showed remarkable water absorption property with swelling ratio up to 4000%, which provided the hydrogel with quick hemostatic ability and prevent the wound bed from accumulation of exudates. The water vapor transmission rate and water retention of the hydrogels were found to be in the range of 2213-3498 g/m²/day and 34.74-45.81% (after 6 days), indicating that the hydrogel can maintain a moist environment over wound bed, which could prevent the dehydration of the wound bed and prevent the scab formation. Biocompatibility test revealed that the hydrogels were not cytotoxic. The hydrogel could load antimicrobial agents and effectively suppress bacterial proliferation to protect the wound from bacterial invasion. These results suggest that the pullulan hydrogels prepared in this study may have high potential as new ideal wound-dressing materials. PMID:21523902

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

    PubMed Central

    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. PMID:26817622

  2. 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.

  3. Versatile Types of MRI-Visible Cationic Nanoparticles Involving Pullulan Polysaccharides for Multifunctional Gene Carriers.

    PubMed

    Huang, Yajun; Hu, Hao; Li, Rui-Quan; Yu, Bingran; Xu, Fu-Jian

    2016-02-17

    Owing to the low cytotoxicity and excellent biocompatibility, polysaccharides are good candidates for the development of promising biomaterials. In this paper, a series of magnetic resonance imaging (MRI)-visible cationic polymeric nanoparticles involving liver cell-targeting polysaccharides were flexibly designed for multifunctional gene delivery systems. The pullulan-based vector (PuPGEA) consisting of one liver cell-targeting pullulan backbone and ethanolamine-functionalized poly(glycidyl methacrylate) (denoted by BUCT-PGEA) side chains with abundant hydroxyl units and secondary amine was first prepared by atom transfer radical polymerization. The resultant cationic nanoparticles (PuPGEA-GdL or PuPGEA-GdW) with MRI functions were produced accordingly by assembling PuPGEA with aminophenylboronic acid-modified Gd-DTPA (GdL) or GdW10O36(9-) (GdW) via the corresponding etherification or electrostatic interaction. The properties of the PuPGEA-GdL and PuPGEA-GdW nanoparticles including pDNA condensation ability, cytotoxicity, gene transfection, cellular uptake, and in vitro and in vivo MRI were characterized in details. Such kinds of cationic nanoparticles exhibited good performances in gene transfection in liver cells. PuPGEA-GdW demonstrated much better MRI abilities. The present design of PuPGEA-based cationic nanoparticles with the liver cell-targeting polysaccharides and MRI contrast agents would shed light on the exploration of tumor-targetable multifunctional gene delivery systems. PMID:26841955

  4. 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. PMID:24702922

  5. Covalent immobilization of pullulanase on alginate and study of its hydrolysis of pullulan.

    PubMed

    Ali, Ghina; Dulong, Virginie; Gasmi, Sarah N; Rihouey, Christophe; Picton, Luc; Le Cerf, Didier

    2015-01-01

    The immobilization of pullulanase from Klebsiella pneumoniae by grafting was investigated. Pullulanase was linked after activation of alginate via a covalent bond between the amine groups of the enzyme and the carboxylic acid groups of alginate. The immobilization yield was 60%. The activity of free pullulanase and immobilized pullulanase was followed by the quantification of reducing ends by colorimetric assay and the determination of the molar masses of the hydrolyzed pullulan by SEC/MALS/DRI. Compared to free pullulanase, the kinetics is largely slowed. The evolution of the weight average molar mass of pullulan leading to high production of shorter oligosaccharides during hydrolysis is not the same as that obtained with free enzyme. Immobilized pullulanase retained 75% and 30% of its initial activity after 24 h and 14 days of incubation at 60°C, respectively while free pullulanase lost its activity after 5 h of hydrolysis at the same temperature. The kinetic parameters of immobilized pullulanase were also investigated by isothermal titration calorimetry (ITC). The affinity of immobilized enzyme to its substrate was reduced compared to the free pullulanase due to steric hindrance and chemical links. PMID:25919860

  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. 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.

  8. 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. PMID:17498729

  9. Modification of functional properties of pullulan-whey protein bionanocomposite films with nanoclay.

    PubMed

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

    2016-02-01

    In this study, biodegradable nanocomposite film composed of pullulan - whey protein isolate (WPI) - montmorillonite (MMT) were developed and characterized as a function of incorporating various amounts of MMT nanoparticles (0, 1, 3 and 5 % wt). Results showed that the water-vapor permeability, moisture content, moisture absorption and water solubility decreased when the nano-MMT content was increased. Tensile strength improved and elongation at break simultaneously decreased with increasing MMT content. The glass transition temperature (Tg(and melting-point temperature (Tm) increased with increasing nano-MMT content. Scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis revealed uniform distribution of MMT into the polymer matrix. Atomic force microscopy (AFM) showed enhancement of films' roughness with increasing MMT content. PMID:27162410

  10. β-Glucan Derived from Aureobasidium pullulans Is Effective for the Prevention of Influenza in Mice

    PubMed Central

    Muramatsu, Daisuke; Iwai, Atsushi; Aoki, Shiho; Uchiyama, Hirohumi; Kawata, Koji; Nakayama, Yosuke; Nikawa, Yasuhiro; Kusano, Kisato; Okabe, Mitsuyasu; Miyazaki, Tadaaki

    2012-01-01

    β-(1→3)-D-glucans with β-(1→6)-glycosidic linked branches produced by mushrooms, yeast and fungi are known to be an immune activation agent, and are used in anti-cancer drugs or health-promoting foods. In this report, we demonstrate that oral administration of Aureobasidium pullulans-cultured fluid (AP-CF) enriched with the β-(1→3),(1→6)-D-glucan exhibits efficacy to protect mice infected with a lethal titer of the A/Puerto Rico/8/34 (PR8; H1N1) strain of influenza virus. The survival rate of the mice significantly increased by AP-CF administration after sublethal infection of PR8 virus. The virus titer in the mouse lung homogenates was significantly decreased by AP-CF administration. No significant difference in the mRNA expression of inflammatory cytokines, and in the population of lymphocytes was observed in the lungs of mice administered with AP-CF. Interestingly, expression level for the mRNA of virus sensors, RIG-I (retinoic acid-inducible gene-I) and MDA5 (melanoma differentiation-associated protein 5) strongly increased at 5 hours after the stimulation of A. pullulans-produced purified β-(1→3),(1→6)-D-glucan (AP-BG) in murine macrophage-derived RAW264.7 cells. Furthermore, the replication of PR8 virus was significantly repressed by pre-treatment of AP-BG. These findings suggest the increased expression of virus sensors is effective for the prevention of influenza by the inhibition of viral replication with the administration of AP-CF. PMID:22844473

  11. pH-sensitive pullulan-based nanoparticle carrier for adriamycin to overcome drug-resistance of cancer cells.

    PubMed

    Guo, Hua; Liu, Yuanyuan; Wang, Yan; Wu, Jing; Yang, Xiaoying; Li, Rongshan; Wang, Yinsong; Zhang, Ning

    2014-10-13

    Urocanic acid was conjugated to pullulan to synthesize O-urocanyl pullulan (URPA) with degree of substitution (DS) of 8.2%. URPA nanoparticles prepared by dialysis method had spherical shapes and a mean diameter of 156.8 ± 16.8 nm. Adriamycin (ADR) was successfully loaded into URPA nanoparticles and exhibited pH-sensitive in vitro release property. MTT assay showed that ADR-loaded URPA (ADR/URPA) nanoparticles had a significant higher toxicity against drug resistant MCF-7/ADR cells than free ADR, and the reversal index reached up to 9.6. The results of flow cytometry and confocal microscopy showed that URPA nanoparticles efficiently enhanced accumulation and retention of ADR in MCF-7/ADR cells and successfully delivered ADR into cell nucleus. The reversal effect of ADR/URPA nanoparticles on the drug resistance of MCF-7/ADR cells was perhaps related with their cell entry and intracellular drug release mechanisms. PMID:25037431

  12. 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. PMID:24621108

  13. 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.

  14. 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

    2014-01-01

    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. PMID:25157463

  15. Expression of Aureobasidium pullulans xynA in, and secretion of the xylanase from, Saccharomyces cerevisiae.

    PubMed Central

    Li, X L; Ljungdahl, L G

    1996-01-01

    A previous report dealt with the cloning in Escherichia coli and sequencing of both the cDNA and genomic DNA encoding a highly active xylanase (XynA) of Aureobasidium pullulans (X.-L. Li and L. G. Ljungdahl, Appl. Environ. Microbiol. 60:3160-3166, 1994). Now we show that the gene was expressed in Saccharomyces cerevisiae under the GAL1 promoter in pYES2 and that its product was secreted into the culture medium. S. cerevisiae clone pCE4 with the whole open reading frame of xynA, including the part coding for the signal peptide, had xylanase activity levels of 6.7 U ml-1 in the cell-associated fraction and 26.2 U ml-1 in the culture medium 4 h after galactose induction. Two protein bands with sizes of 25 and 27 kDa and N-terminal amino acid sequences identical to that of APX-II accounted for 82% of the total proteins in the culture medium of pCE4. These proteins were recognized by anti-APX-II antibody. The results suggest that the XynA signal peptide supported the posttranslational processing of xynA product and the efficient secretion of the active xylanase from S. cerevisiae. Clones pCE3 and pGE3 with inserts of cDNA and genomic DNA, respectively, containing only the mature enzyme region attached by a Met codon had low levels of xylanase activity in the cell-associated fractions (1.6 U ml-1) but no activity in the culture media. No xylanase activity was detected in clone pGE4, which was the same as pCE4, except that pGE4 had a 59-bp intron in the signal peptide region. A comparison of the A. pullulans and S. cerevisiae signal peptides demonstrated that the XynA signal peptide was at least three times more efficient than those of S. cerevisiae invertase or mating alpha-factor pheromone in secreting the heterologous xylanase from S. cerevisiae cells. PMID:8572698

  16. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 5 2013-04-01 2013-04-01 false Limitation on annual compensation. 1.401(a)(17)-1 Section 1.401(a)(17)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.401(a)(17)-1 Limitation on...

  17. Disulphide cross linked pullulan based cationic polymer for improved gene delivery and efflux pump inhibition.

    PubMed

    S, Priya S; R, Rekha M

    2016-10-01

    Multidrug resistance is a hurdle to successful cancer chemotherapy. Over expression of P-glycoprotein (P-gp) is a prime contributing factor for drug resistance. In this study, a disulphide cross-linked pullulan-based cationic polymer (PPSS) was synthesized to act simultaneously as gene delivery vehicle and efflux pump inhibitor. The PPSS nanoplexes were of size <200nm with the zeta potential of +15 to +20mV. The cytotoxicity studies using C6 and L929 cells showed that PPSS polymers are non-toxic even at high polymer concentrations. The PPSS/pDNA nanoplex showed superior uptake in confocal microscopy with 97% uptake by flow cytometry. The efficacy of efflux pump inhibition by the PPSS nanoplex was established by the enhanced intracellular retention of DOX. The enhanced cell death by p53/PPSS/DOX nanoplexes was attributed to the synergistic effect of P-gp inhibition and p53 transfection efficiency. Therefore, this multifunctional polymeric system may have significant promise for therapeutic application against cancer drug resistance. PMID:27459414

  18. 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.

  19. Effect of sperminated pullulans on drug permeation through isolated rabbit cornea and determination of ocular irritation.

    PubMed

    Yu, N; Xun, Y; Jin, D; Yang, H; Hang, T; Cui, H

    2010-01-01

    The aim of this study was to investigate the effect of two sperminated pullulans (SP) with a different number of amino groups (SP-L, amino group content 0.124 mmol/g polymer; and SP-H, amino group content 0.578 mmol/g polymer) on the permeation of drugs through isolated rabbit corneas. Determination of corneal hydration levels and Draize eye tests were performed to assess the safety of SP both in vitro and in vivo. For 0.2% (w/v) SP-L and 0.2% (w/v) SP-H, the enhancement ratios (ERs) with dexamethasone of 1.34 and 1.42, respectively, were not statistically significant. For ofloxacin, tobramycin and sodium fluorescein, the ERs with 0.2% SP-L were 1.37, 2.02 and 2.12, respectively, and with 0.2% SP-H the ERs were 1.84, 4.69 and 6.87, respectively; these ERs were all statistically significant. Enhancement increased with increasing amino group content of the SP. The improved transcorneal drug absorption via the paracellular route indicated opening of the tight junctions in the corneal epithelium. Irritation tests indicated that 0.2% SP-L and 0.2% SP-H did not damage the corneal tissues. PMID:20515566

  20. Cell-laden microengineered pullulan methacrylate hydrogels promote cell proliferation and 3D cluster formation

    PubMed Central

    Bae, Hojae; Ahari, Amir F.; Shin, Hyeongho; Nichol, Jason W.; Hutson, Che B.; Masaeli, Mahdokht; Kim, Su-Hwan; Aubin, Hug; Yamanlar, Seda; Khademhosseini, Ali

    2011-01-01

    The ability to encapsulate cells in three-dimensional (3D) environments is potentially of benefit for tissue engineering and regenerative medicine. In this paper, we introduce pullulan methacrylate (PulMA) as a promising hydrogel platform for creating cell-laden microscale tissues. The hydration and mechanical properties of PulMA were demonstrated to be tunable through modulation of the degree of methacrylation and gel concentration. Cells encapsulated in PulMA exhibited excellent viability. Interestingly, while cells did not elongate in PulMA hydrogels, cells proliferated and organized into clusters, the size of which could be controlled by the hydrogel composition. By mixing with gelatin methacrylate (GelMA), the biological properties of PulMA could be enhanced as demonstrated by cells readily attaching to, proliferating, and elongating within the PulMA/GelMA composite hydrogels. These data suggest that PulMA hydrogels could be useful for creating complex, cell-responsive microtissues, especially for applications that require controlled cell clustering and proliferation. PMID:21415929

  1. 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. PMID:26640053

  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. Cell-laden microengineered pullulan methacrylate hydrogels promote cell proliferation and 3D cluster formation.

    PubMed

    Bae, Hojae; Ahari, Amir F; Shin, Hyeongho; Nichol, Jason W; Hutson, Che B; Masaeli, Mahdokht; Kim, Su-Hwan; Aubin, Hug; Yamanlar, Seda; Khademhosseini, Ali

    2011-01-01

    The ability to encapsulate cells in three-dimensional (3D) environments is potentially of benefit for tissue engineering and regenerative medicine. In this paper, we introduce pullulan methacrylate (PulMA) as a promising hydrogel platform for creating cell-laden microscale tissues. The hydration and mechanical properties of PulMA were demonstrated to be tunable through modulation of the degree of methacrylation and gel concentration. Cells encapsulated in PulMA exhibited excellent viability. Interestingly, while cells did not elongate in PulMA hydrogels, cells proliferated and organized into clusters, the size of which could be controlled by the hydrogel composition. By mixing with gelatin methacrylate (GelMA), the biological properties of PulMA could be enhanced as demonstrated by cells readily attaching to, proliferating, and elongating within the PulMA/GelMA composite hydrogels. These data suggest that PulMA hydrogels could be useful for creating complex, cell-responsive microtissues, especially for applications that require controlled cell clustering and proliferation. PMID:21415929

  4. 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. PMID:26774376

  5. 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. PMID:23957414

  6. Hemocompatible pullulan-polyethyleneimine conjugates for liver cell gene delivery: In vitro evaluation of cellular uptake, intracellular trafficking and transfection efficiency.

    PubMed

    Rekha, M R; Sharma, Chandra P

    2011-01-01

    Polyethyleneimine (PEI; 25 kDa)-conjugated pullulans (PPE1, PPE2 and PPE3) were developed and investigated for possible use in gene delivery applications. The cytotoxicity, blood component interactions such as red blood cell/white blood cell aggregation, platelet and complement activation, and protein interaction of the pullulan-conjugated PEI was drastically reduced in comparison to PEI-based nanocomplexes. Based on the blood compatibility studies, PPE1 was selected for further study. The buffering capacity of this derivative was similar to that of PEI, which plays an important role in efficient gene transfection. The particle size, zeta potential, stability in the presence of plasma and resistance to nuclease degradation were evaluated. In addition, cellular uptake and localization of plasmid, as well as transgene expression, were evaluated following in vitro transfection of HepG2 cells. Endocytosis inhibitors, confocal laser scanning microscopy and fluorescent labeling techniques were used to visualize the nanoplex uptake mechanism, cellular distribution and nuclear localization. The results from inhibitor experiments in the presence of asialofetuin indicated that the asialoglycoprotein receptor is involved in transfection of hepatocytes with pullulan-PEI complexes. The conjugation of pullulan with PEI did not hinder the plasmid nuclear localization ability of PEI. The transfection efficiency of pullulan conjugate was similar to PEI, with the added advantage of hemocompatibility and non-cytotoxicity. The transfection efficiency of PEI and PPE1 was 1.6- and 2-fold more, respectively, in the presence of serum than in the absence of serum. Therefore, the pullulan-PEI conjugate seems to be a promising gene delivery vector with good hemocompatibility and low toxicity but without compromising the transfection efficacy of PEI. PMID:20659595

  7. 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.

  8. An indigenous hyperproductive species of Aureobasidium pullulans RYLF-10: influence of fermentation conditions on exopolysaccharide (EPS) production.

    PubMed

    Yadav, Kanchan L; Rahi, Deepak K; Soni, Sanjeev K

    2014-02-01

    In recent years, a significant interest has been generated in discovering and developing exopolysaccharides (EPS) produced by microorganisms, especially fungi due to their multifaceted industrial and pharmacological applications. A number of filamentous and cellular fungi have been explored for this; however, according to the existing literature, the work on exopolysaccharide production by indigenous culture on this aspect is still very less and requires a serious attention. The present work is an attempt in this regard and aims to optimize the submerged culture conditions to produce the exopolysaccharides from an indigenous yeast Aureobasidium pullulans RYLF-10 with respect to several operating parameters in shake flask fermentation. The yeast A. pullulans RYLF-10 was identified by 18s RNA sequencing and detailed study on its nutritional requirements, and environmental conditions for submerged culture have been optimized. The optimal temperature and pH for both the vegetative growth and EPS production were found to be 28 ± 1 °C and 5.0, respectively, while the agitation speed and inoculum size were reported to be 150 rpm and 1 % (v/v), respectively. Sucrose (50 g/l) and yeast extract (1 g/l) were found to be the most suitable carbon and nitrogen sources which worked best in the ratio of 60:1 and resulted in the maximum EPS yield. Similarly, the other variables like growth regulator (riboflavin) and minerals (NaCl + K2HPO4 + MgSO4) altogether resulted in a noteworthy EPS yield of 45.24 g/l which is the maximum yield from this indigenous isolate of A. pullulans RYLF-10. PMID:24293276

  9. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 5 2012-04-01 2011-04-01 true Limitation on annual compensation. 1.401(a)(17)-1...(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...

  10. 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). PMID:22765289

  11. 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. PMID:27612808

  12. Tocopheryl pullulan-based self assembling nanomicelles for anti-cancer drug delivery.

    PubMed

    Wang, Jingyun; Cui, Shuang; Bao, Yongming; Xing, Jishuang; Hao, Wenbo

    2014-10-01

    Amphiphilic α-tocopherol pullulan polymers (PUTC1, PUTC2, and PUTC3) with different degrees of substitution were synthesized as new carriers for anticancer drugs. The polymers easily self-assembled into nanomicelles through dialysis method. The critical micelle concentrations (CMCs) were 38.0, 8.0, and 4.3mg/L for PUTC1, PUTC2, and PUTC3, respectively. 10-Hydroxycamptothecin (HCPT) used as a model drug was successfully loaded into the PUTC nanomicelles. Transmission electron microscopy images demonstrated that HCPT-loaded PUTC nanomicelles were almost spherical and had sizes ranging within 171.5-257.8 nm that increased with increased HCPT-loading content, as determined by dynamic laser scattering. The highest encapsulation efficiency of HCPT in PUTC nanomicelles reached 98.3%. The in vitro release of HCPT from PUTC micelles demonstrated sustained release for over 80 h. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays showed that blank PUTC micelles were nontoxic to normal cells and that the HCPT-loaded PUTC2 nanomicelles showed higher cytotoxicity than the free drug, which was attributed to the enhanced cellular uptake of drug-loaded nanomicelles. Biodistribution experiments showed that PUTC micelles provided an excellent approach to rapid drug transport into cell nuclei. Moreover, the cellular uptake of micelles was found to be an energy-dependent and actin polymerization-associated endocytic process by endocytosis inhibition experiments. These results suggested that PUTC nanomicelles had considerable potential as a drug carrier for drug intracellular delivery in cancer therapy. PMID:25175256

  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

    2015-12-01

    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. PMID:26586655

  14. 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

  15. 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). PMID:26189365

  16. 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

    2016-06-01

    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°). PMID:27083358

  17. Collagen synthesis promoting pullulan-PEI-ascorbic acid conjugate as an efficient anti-cancer gene delivery vector.

    PubMed

    Ambattu, Lizebona August; Rekha, M R

    2015-08-01

    Cationized pullulan (pullulan-PEI; PP) was synthesized and further modified with an anti-oxidant molecule, ascorbic acid (PPAA) at various ratios. The nanoplexes formed at an optimum ratio of 4:1 was within a size of 150nm and had a zeta potential of 9-14mV. The nanoplexes at this ratio was used for further investigations. The cell internalization and transfection efficiency of these nanoplexes were determined in presence of serum. The internalization and transfection efficiency were found to be unaffected by the presence of fetal bovine serum. Another interesting observation was that this polymer was found to have collagen synthesis promoting property. The collagen synthesis effect of these polymers was quantified and observed that PPAA3 promoted the highest. Transfection efficiency was evaluated by assessing the p53 gene expression in C6 rat glioma cells and cell death was quantified to be 96% by flow cytometry, thus establishing the high efficacy of this polymer. PMID:25933522

  18. 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

  19. 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

  20. 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.

  1. Reversal of tumor growth by gene modification of mesenchymal stem cells using spermine-pullulan/DNA nanoparticles.

    PubMed

    Hu, Yu-Lan; Miao, Pei-Hong; Huang, Bing; Zhang, Tian-Yuan; Hu, Zhong-Jie; Tabata, Yasuhiko; Gao, Jian-Qing

    2014-02-01

    Mesenchymal stem cells (MSCs) are a promising tool for delivering of therapeutic agents in cancer treatment. In the present study, our findings suggested that both i.v. and intratumoral injection of MSCs could favor tumor growth under physiologic conditions. However, the anti-tumor effects of MSC-IL-12 were achieved using our strategy. Unlike the previously reported method, the genetic engineering of MSCs was conducted by non-viral transfection using the new vector, spermine-pullulan. The transfection, cytotoxicity, and the cellular internalization of this vector were evaluated. Then, the therapeutical gene, IL-12, was delivered to the MSCs using this vector. The in vitro secretions of IL-12 by MSC-IL-12 confirmed the success of using spermine-pullulan/DNA nanoparticles for the gene transfection. We used the MSC-IL-12 for the in vivo treatment of both B16F10 metastasis tumor and the established subcutaneous B16BL6 tumor. For the B16F10 metastasis tumor, treatment with MSC-IL-12 significantly reduced lung metastases. For the established subcutaneous B16BL6 tumor, intratumoral injected MSC-IL-12 cells considerably retarded tumor growth. Prolonged survival was observed when MSC-IL-12 cells were injected through the tail vein or intratumorally, indicating that the MSCs engineered with the therapeutic gene could reverse the tumor-promoting effects of MSCs using the nonviral transduction method. However, the intravenous injected MSC-IL-12 did not prevent the tumor growth of the established subcutaneous B16BL6 tumor. Thus, we examined the the in vivo distribution of MSCs in different organs and it was found that MSCs were mainly distributed in the lungs, which may explain the inability of intravenously injected MSC-IL-12 to inhibit the growth of the established subcutaneous tumor. PMID:24738338

  2. 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

  3. 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...

  4. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 5 2014-04-01 2014-04-01 false Limitation on annual compensation. 1.401(a)(17.... § 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...

  5. Succinate-bonded pullulan: An efficient and reusable super-sorbent for cadmium-uptake from spiked high-hardness groundwater.

    PubMed

    Abbas, Azhar; Hussain, Muhammad Ajaz; Amin, Muhammad; Sher, Muhammad; Tahir, Muhammad Nawaz; Tremel, Wolfgang

    2015-11-01

    Chemically modified pullulan was evaluated for its sorption efficiency and selectivity to remove cadmium (Cd) from spiked high-hardness groundwater (GW). Pullulan esterified with succinic anhydride using dimethylaminopyridine showed a fairly high degree of substitution value as confirmed by (1)H NMR spectroscopy. Pullulan succinate (Pull-Suc) was converted into the sodium salt (Pull-Suc-Na). The effect of contact time (5-200min) and pH (2-8) on Cd-uptake by the sorbent (Pull-Suc-Na) was investigated. The sorbent showed more than 90% Cd-removal in first 15min from distilled water (DW) and GW solution, respectively. Comparison of Pull-Suc-Na with other polysaccharidal sorbents suggested its high efficiency (DW 476.2mg/g and GW 454.5mg/g) and selectivity for the removal of Cd by an ion exchange mechanism, which is further supported by the negative Gibbs free energy values calculated from Langmuir isotherms. A Langmuir isotherm kinetic model provided the best fit for the sorption of Cd using Pull-Suc-Na. The sorbent showed a negligible decrease in Cd-uptake over three regeneration cycles. The thermal stability testing of the sorbents indicated that Pull-Suc-Na (sorbent) is more stable than Pull-Suc. PMID:26574087

  6. 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. PMID:27055191

  7. The biology of the 17-1A antigen (Ep-CAM).

    PubMed

    Balzar, M; Winter, M J; de Boer, C J; Litvinov, S V

    1999-10-01

    The glycoprotein recognized by the monoclonal antibody (mAb) 17-1A is present on most carcinomas, which makes it an attractive target for immunotherapy. Indeed, adjuvant treatment with mAb 17-1A did successfully reduce the 5 years mortality among colorectal cancer patients with minimal residual disease. Currently the antibody is approved for clinical use in Germany, and is on its way to approval in a number of other countries. New immunotherapeutic strategies targeting the 17-1A antigen are in development or even in early-phase clinical trials. Therefore, a better understanding of the biology of the 17-1A antigen may result in improved strategies for the treatment and diagnosis of human carcinomas. In this review the properties of the 17-1A antigen are discussed concerning tumor biology and the function of the molecule. This 40-kDa glycoprotein functions as an Epithelial Cell Adhesion Molecule, therefore the name Ep-CAM was suggested. Ep-CAM mediates Ca2+-independent homotypic cell-cell adhesions. Formation of Ep-CAM-mediated adhesions has a negative regulatory effect on adhesions mediated by classic cadherins, which may have strong effects on the differentiation and growth of epithelial cells. Indeed, in vivo expression of Ep-CAM is related to increased epithelial proliferation and negatively correlates with cell differentiation. A regulatory function of Ep-CAM in the morphogenesis of epithelial tissue has been demonstrated for a number of tissues, in particular pancreas and mammary gland. The function of Ep-CAM should be taken into consideration when developing new therapeutic approaches targeting this molecule. PMID:10606205

  8. 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

  9. 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

    2014-11-01

    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. PMID:25129721

  10. 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

  11. 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

  12. Molecular size characterization and kinetics studies on hydrolysis of pullulan by pullulanase in an entangled alginate medium.

    PubMed

    Ali, Ghina; Rihouey, Christophe; Larreta-Garde, Véronique; Le Cerf, Didier; Picton, Luc

    2013-07-01

    The behavior of a hydrolytic enzyme (pullulanase) toward its substrate (pullulan) in the presence of a nonsubstrate (alginate), both below and above the critical entanglement concentration (C*), was studied. The hydrolysis kinetics were studied with the enzyme and alginate concentrations varied using two main methods: a colorimetric assay of the reducing extremities (RE), which allowed the number-average molar masses (Mn) of the oligosaccharides to be determined, and size exclusion chromatography with on-line, multiangle light scattering, viscometer, and differential refractive index detectors, which allowed the average molar masses, Mn and Mw, of the oligosaccharides during hydrolysis to be determined. Free pullulanase acts via an "endo" process. The presence of alginate slows the hydrolysis kinetics, particularly when the alginate concentration is greater than the C*. These results were confirmed by the evolution of the kinetic parameters (KM, Vmax) obtained via isothermal titration calorimetry (ITC). The amount of oligosaccharides produced is not dependent on the alginate concentration, and the endo enzyme behavior is not modified by the entanglement in the medium. These observations were also confirmed by ITC analysis in the presence of degraded alginate (without entanglement). Our results correlated with the substrate diffusion in entangled media. The pullulanase reaction in the presence of alginate is shown to be diffusion-dependent. PMID:23713899

  13. 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...

  14. 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...

  15. 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

    ... materials or heat are recovered, the waste disposal function includes the processing of such materials or... used in solid waste disposal. The heat can be used by A to produce steam. Company B operates an... solid waste disposal facilities; temporary rules. 17.1 Section 17.1 Internal Revenue INTERNAL...

  16. 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

    ... materials or heat are recovered, the waste disposal function includes the processing of such materials or... used in solid waste disposal. The heat can be used by A to produce steam. Company B operates an... solid waste disposal facilities; temporary rules. 17.1 Section 17.1 Internal Revenue INTERNAL...

  17. 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

    ... 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...

  18. 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

    ... 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...

  19. 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

    ... 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...

  20. 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. PMID:22225984

  1. Partial Optimization of Endo-1, 4-Β-Xylanase Production by Aureobasidium pullulans Using Agro-Industrial Residues

    PubMed Central

    Nasr, Shaghayegh; Soudi, Mohammad Reza; Hatef Salmanian, Ali; Ghadam, Parinaz

    2013-01-01

    Objective(s) : Although bacteria and molds are the pioneering microorganisms for production of many enzymes, yet yeasts provide safe and reliable sources of enzymes with applications in food and feed. Materials and Methods: Single xylanase producer yeast was isolated from plant residues based on formation of transparent halo zones on xylan agar plates. The isolate showed much greater endo-1, 4-β-xylanase activity of 2.73 IU/ml after optimization of the initial extrinsic conditions. It was shown that the strain was also able to produce β-xylosidase (0.179 IU/ml) and α-arabinofuranosidase (0.063 IU/ml). Identification of the isolate was carried out and the endo-1, 4-β-xylanaseproduction by feeding the yeast cells on agro-industrial residues was optimized using one factor at a time approach. Results: The enzyme producer strain was identified as Aureobasidiumpullulans. Based on the optimization approach, an incubation time of 48 hr at 27°C, inoculum size of 2% (v/v), initial pH value of 4 and agitation rate of 90 rpm were found to be the optimal conditions for achieving maximum yield of the enzyme. Xylan, containing agricultural residues, was evaluated as low-cost alternative carbon source for production of xylanolytic enzymes. The production of xylanase enzyme in media containing wheat bran as the sole carbon source was very similar to that of the medium containing pure beechwoodxylan. Conclusion:This finding indicates the feasibility of growing of A. pullulans strain SN090 on wheat bran as an alternate economical substrate in order for reducing the costs of enzyme production and using this fortified agro-industrial byproduct in formulation of animal feed. PMID:24570830

  2. Pullulan/dextran/nHA Macroporous Composite Beads for Bone Repair in a Femoral Condyle Defect in Rats

    PubMed Central

    Schlaubitz, Silke; Derkaoui, Sidi Mohammed; Marosa, Lydia; Miraux, Sylvain; Renard, Martine; Catros, Sylvain; Le Visage, Catherine; Letourneur, Didier; Amédée, Joëlle; Fricain, Jean-Christophe

    2014-01-01

    The repair of bone defects is of particular interest for orthopedic, oral, maxillofacial, and dental surgery. Bone loss requiring reconstruction is conventionally addressed through bone grafting. Depending on the size and the location of the defect, this method has limits and risks. Biomaterials can offer an alternative and have features supporting bone repair. Here, we propose to evaluate the cellular penetration and bone formation of new macroporous beads based on pullulan/dextran that has been supplemented with nanocrystalline hydroxyapatite in a rat model. Cross-linked beads of 300–500 µm diameters were used in a lateral femoral condyle defect and analyzed by magnetic resonance imaging, micro-computed tomography, and histology in comparison to the empty defects 15, 30, and 70 days after implantation. Inflammation was absent for both conditions. For empty defects, cellularisation and mineralization started from the periphery of the defect. For the defects containing beads, cellular structures filling out the spaces between the scaffolds with increasing interconnectivity and trabecular-like organization were observed over time. The analysis of calcified sections showed increased mineralization over time for both conditions, but was more pronounced for the samples containing beads. Bone Mineral Density and Bone Mineral Content were both significantly higher at day 70 for the beads in comparison to empty defects as well as compared with earlier time points. Analysis of newly formed tissue around the beads showed an increase of osteoid tissue, measured as percentage of the defect surface. This study suggests that the use of beads for the repair of small size defects in bone may be expanded on to meet the clinical need for a ready-to-use fill-up material that can favor bone formation and mineralization, as well as promote vessel ingrowth into the defect site. PMID:25330002

  3. 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.

  4. Enzymatic synthesis of novel branched sugar alcohols mediated by the transglycosylation reaction of pullulan-hydrolyzing amylase II (TVA II) cloned from Thermoactinomyces vulgaris R-47.

    PubMed

    Shimura, Yoichiro; Oh, Keimei; Kon, Misaki; Yamamoto, Eri; Mizuno, Yoshinori; Adachi, Takashi; Abe, Tomomi; Tamogami, Shigeru; Fukushima, Jun; Inamoto, Tamio; Tonozuka, Takashi

    2011-09-27

    Transglycosylation reactions are useful for preserving a specific sugar structure during the synthesis of branched oligosaccharides. We have previously reported a panosyl unit transglycosylation reaction by pullulan-hydrolyzing amylase II (TVA II) cloned from Thermoactinomyces vulgaris R-47 (Tonozuka et al., Carbohydr. Res., 1994, 261, 157-162). The acceptor specificity of the TVA II transglycosylation reaction was investigated using pullulan as the donor and sugar alcohols as the acceptor. TVA II transferred the α-panosyl unit to the C-1 hydroxyl group of meso-erythritol, C-1 and C-2 of xylitol, and C-1 and C-6 of d-sorbitol. TVA II differentiated between the sugar alcohols' hydroxyl groups to produce five novel non-reducing branched oligosaccharides, 1-O-α-panosylerythritol, 1-O-α-panosylxylitol, 2-O-α-panosylxylitol, 1-O-α-panosylsorbitol, and 6-O-α-panosylsorbitol. The Trp(356)→Ala mutant showed similar transglycosylation reactions; however, panose production by the mutant was 4.0-4.5-fold higher than that of the wild type. This suggests that Trp(356) is important for recognizing both water and the acceptor molecules in the transglycosylation and the hydrolysis reaction. PMID:21722879

  5. 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. PMID:26001063

  6. Core-shell nanoparticles based on pullulan and poly(β-amino) ester for hepatoma-targeted codelivery of gene and chemotherapy agent.

    PubMed

    Liu, Yuanyuan; Wang, Yan; Zhang, Cong; Zhou, Ping; Liu, Yang; An, Tong; Sun, Duxin; Zhang, Ning; Wang, Yinsong

    2014-01-01

    This study designs a novel nanoparticle system with core-shell structure based on pullulan and poly(β-amino) ester (PBAE) for the hepatoma-targeted codelivery of gene and chemotherapy agent. Plasmid DNA expressing green fluorescent protein (pEGFP), as a model gene, was fully condensed with cationic PBAE to form the inner core of PBAE/pEGFP polycomplex. Methotrexate (MTX), as a model chemotherapy agent, was conjugated to pullulan by ester bond to synthesize polymeric prodrug of MTX-PL. MTX-PL was then adsorbed on the surface of PBAE/pEGFP polycomplex to form MTX-PL/PBAE/pEGFP nanoparticles with a classic core-shell structure. MTX-PL was also used as a hepatoma targeting moiety, because of its specific binding affinity for asialoglycoprotein receptor (ASGPR) overexpressed by human hepatoma HepG2 cells. MTX-PL/PBAE/pEGFP nanoparticles realized the efficient transfection of pEGFP in HepG2 cells and exhibited significant inhibitory effect on the cell proliferation. In HepG2 tumor-bearing nude mice, MTX-PL/PBAE/pEGFP nanoparticles were mainly distributed in the tumor after 24 h postintravenous injection. Altogether, this novel codelivery system with a strong hepatoma-targeting property achieved simultaneous delivery of gene and chemotherapy agent into tumor at both cellular and animal levels. PMID:25289563

  7. 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

  8. 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. PMID:18460124

  9. Stimulation with the Aureobasidium pullulans-produced β-glucan effectively induces interferon stimulated genes in macrophage-like cell lines

    PubMed Central

    Muramatsu, Daisuke; Kawata, Koji; Aoki, Shiho; Uchiyama, Hirofumi; Okabe, Mitsuyasu; Miyazaki, Tadaaki; Kida, Hiroshi; Iwai, Atsushi

    2014-01-01

    A β-(1,3),(1,6)-D-glucan produced by A. pullulans (AP-PG) is known to be an immune stimulating agent. In this study, we demonstrate that the stimulation with AP-PG effectively induces the interferon (IFN) stimulated genes (ISGs) in macrophage-like cell lines. The ISGs, Mx1, ISG15, and viperin mRNAs were significantly increased in RAW264.7 cells after stimulation with AP-PG. The stimulation with AP-PG transiently induced IFN-β mRNA. However, the expression of viperin mRNA was also increased after stimulation with AP-PG even when new protein synthesis was completely blocked by treatment with cycloheximide. Further, in IFN-α receptor knockdown RAW264.7 cells, AP-PG stimulation more effectively induced viperin mRNA compared with that of IFN-α stimulation. The phosphorylation of Ser 727 in STAT1 involved in the enhancement of STAT1 activation was immediately increased after stimulation with AP-PG. In addition, viperin mRNA expression induced after stimulation with IFN-α was significantly increased by combined stimulation with AP-PG. These results suggest that stimulation with AP-PG effectively induces the ISGs through the induction of IFN and the enhancement of STAT1-mediated transcriptional activation. PMID:24759061

  10. Biodistribution of charged 17.1A photoimmunoconjugates in a murine model of hepatic metastasis of colorectal cancer

    PubMed Central

    Hamblin, M R; Governatore, M Del; Rizvi, I; Hasan, T

    2000-01-01

    Optimizing photodynamic therapy involves attempting to increase both the absolute tumour content of photosensitizer and the selectivity between tumour and surrounding normal tissue. One reason why photodynamic therapy has not been considered suitable for treatment of metastatic tumours in the liver, is the poor selectivity of conventional photosensitizers for tumour compared to normal liver. This report details an alternative approach to increasing this selectivity by the use of antibody-targeted photosensitizers (or photoimmunoconjugates) to target intrahepatic tumours caused by human colorectal cancer cells in the nude mouse, and explores the role of molecular charge on the tumour-targeting efficiency of macromolecules. The murine monoclonal antibody 17.1A (which recognizes an antigen expressed on HT 29 cells) was used to prepare site-specific photoimmunoconjugates with the photosensitizer chlorine6. The conjugates had either a predominant cationic or anionic charge and were injected i.v. into tumour-bearing mice. Biodistribution 3 or 24 h later was measured by extraction of tissue samples and quantitation of chlorine6 content by fluorescence spectroscopy. The photoimmunoconjugates were compared to the polylysine conjugates in an attempt to define the effect of molecular charge as well as antibody targeting. The anionic 17.1A conjugate delivered more than twice as much photosensitizer to the tumour at 3 h than other species (5 times more than the cationic 17.1A conjugate) and had a tumour:normal liver ratio of 2.5. Tumour-to-liver ratios were greater than one for most compounds at 3 h but declined at 24 h. Tumour-to-skin ratios were high (> 38) for all conjugates but not for free chlorine6. Cationic species had a high uptake in the lungs compared to anionic species. The photoimmunoconjugates show an advantage over literature reports of other photosensitizers, which can result in tumour:normal liver ratios of less than 1. © 2000 Cancer Research Campaign http

  11. 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

  12. Modulating the Growth and Imatinib Sensitivity of Chronic Myeloid Leukemia Stem/Progenitor Cells with Pullulan/MicroRNA Nanoparticles In Vitro.

    PubMed

    Ma, Wenjuan; Liu, Jian; Xie, Jundan; Zhang, Xiuyan; Zhou, Haixia; Yao, Hong; Zhang, Weiqi; Guo, Dawei; Zhu, Lingying; Xiao, Lun; Wu, Depei; Xu, Haiyan; Chen, Suning; Zhao, Yun

    2015-11-01

    Chronic myeloid leukemia (CML) originates from normal hematopoietic stem cells acquiring Philadelphia chromosome (Ph) to generate BCR-ABL fusion gene whose protein product has deregulated tyrosine kinase activity. Specific inhibitors against BCR-ABL, such as Imatinib mesylate (IM), have greatly improved CML management; however, no single agent is a cure yet. Delivery of microRNA (miRNA) using non-viral vectors has been utilized to inhibit various cancer cells; however, the efficacy of this approach to target CML stem/progenitor cells has not been elucidated. In this study, we firstly validated that spermine-introduced pullulan (Ps) was a robust non-viral vector for delivery of miRNA to CML cells, including the CD34+ cells from clinical isolates. We then found that the miR-181a/RALA (V-ral simian leukemia viral oncogene homolog A) axis was aberrantly expressed in the CML CD34+ cells. The delivery of miR-181a specifically inhibited the growth of CML CD34+ cells, possibly via the inhibition of RALA. In contrast, miR-181a did not evidently affect the normal hematopoietic CD34+ cells. In addition, miR-181a increased IM sensitivity of the CD34+ CML cells. Taken together, we have therefore demonstrated that the delivery of miR-181a using Ps to CML stem/progenitor cells leads to their growth inhibition and enhancement of IM sensitivity, which will possibly be beneficial to CML treatment. PMID:26554155

  13. Efficacy test of Polycan, a beta-glucan originated from Aureobasidium pullulans SM-2001, on anterior cruciate ligament transection and partial medial meniscectomy-induced-osteoarthritis rats.

    PubMed

    Kim, Joo-Wan; Cho, Hyung-Rae; Ku, Sae-Kwang

    2012-02-01

    The object of this study was to assess the efficacy of Polycan from Aureobasidium pullulans SM-2001, which is composed mostly of beta-1,3-1,6-glucan, on osteoarthritis (OA)-induced by anterior cruciate ligament transection and partial medial meniscectomy (ACLT&PMM). Three different dosages of Polycan (85, 42.5, and 21.25 mg/kg) were orally administered once a day for 84 days to male rats a week after ACLT&PMM surgery. Changes in the circumference and maximum extension angle of each knee, and in cartilage histopathology were assessed using Mankin scores 12 weeks after Polycan administration. In addition, cartilage proliferation was evaluated using bromodeoxyuridine (BrdU). As the result of ACLT&PMM, classic OA was induced with increases in maximum extension angles, edematous knees changes, and capsule thickness, as well as decreases in chondrocyte proliferation, cartilages degenerative changes, and loss of articular cartilage. However, these changes (except for capsule thickness) were markedly inhibited in all Polycan- and diclofenac sodium-treated groups compared with OA control. Although diclofenac sodium did not influence BrdU uptake, BrdU-immunoreactive cells were increased with all dosages of Polycan, which means that Polycan treatment induced proliferation of chondrocytes in the surface articular cartilage of the tibia and femur. The results obtained in this study suggest that 84 days of continuous oral treatment of three different dosages of Polycan led to lesser degrees of articular stiffness and histological cartilage damage compared with OA controls 91 days after OA inducement, suggesting that the optimal Polycan dosage to treat OA is 42.5 mg/kg based on the present study. PMID:22370362

  14. 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. PMID:26467005

  15. 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.

  16. 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

  17. 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. PMID:23144412

  18. Genome Sequence of “Candidatus Microthrix parvicella” Bio17-1, a Long-Chain-Fatty-Acid-Accumulating Filamentous Actinobacterium from a Biological Wastewater Treatment Plant

    PubMed Central

    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.

    2012-01-01

    “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. PMID:23144412

  19. The systematic regulation of oyster CgIL17-1 and CgIL17-5 in response to air exposure.

    PubMed

    Xin, Lusheng; Zhang, Huan; Du, Xinyu; Li, Yiqun; Li, Meijia; Wang, Lingling; Wang, Hao; Qiu, Limei; Song, Linsheng

    2016-10-01

    As a proinflammatory cytokine, vertebrate interleukin 17 (IL17) plays a vital role in the balance of inflammation and homeostasis, and is involved in a systemic regulation of glucose homeostasis. In the present study, a remarkable increase of glucose concentration was observed in oyster serum after 2 d air exposure, which was followed by a rapid up-regulation of CgIL17-1 and CgIL17-5. After oysters was received an injection of extra glucose, the mRNA expressions of CgIL17-1 and CgIL17-5 were also significantly up-regulated. The histopathological changes of hepatopancreas were observed after the oysters were treated by the recombinant proteins of CgIL17-1 and CgIL17-5 in vivo or subjected to air exposure. A significant decrease of GSK3β (Glycogen synthase kinase-3β) protein was also observed after the injection of CgIL17-1 and CgIL17-5 recombinant proteins in vivo. When the oysters with CgIL17-1 and CgIL17-5 genes knocked down were subjected to air exposure, the decline of GSK3β concentration was slowed down and it could still be obviously detected after 7 d compared with that in the control. Meanwhile, the expression of CgDefensin and CgDFFA was inhibited, while CgIAP was up-regulated when CgIL17-1 and CgIL17-5 genes were knocked down, and the oysters exhibited higher mortality (p < 0.05) at 3 d, whereas lower at the late stage of air exposure compared with that in the controls. The results collectively suggested that once oysters were exposed to air, the synthesis of proinflammatory cytokines CgIL17-1 and CgIL17-5 was induced by the up-regulated glucose concentration in oyster serum, which would be not only a negative feedback to the high glucose concentration through mediating the regulation of GSK3β, but also an inducer on tissue damage and immunocompetence as well as the adaptability to stresses. PMID:27268575

  20. 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. PMID:21569942

  1. 77 FR 8731 - Aureobasidium pullulans

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-15

    ...-5805. II. Background and Statutory Findings In the Federal Register of March 10, 2010 (75 FR 11171..., entitled Regulatory Planning and Review (58 FR 51735, October 4, 1993). Because this final rule has been... FR 28355, May 22, 2001) or Executive Order 13045, entitled Protection of Children from...

  2. 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.

  3. Yeasts from sub-Antarctic region: biodiversity, enzymatic activities and their potential as oleaginous microorganisms.

    PubMed

    Martinez, A; Cavello, I; Garmendia, G; Rufo, C; Cavalitto, S; Vero, S

    2016-09-01

    Various microbial groups are well known to produce a range of extracellular enzymes and other secondary metabolites. However, the occurrence and importance of investment in such activities have received relatively limited attention in studies of Antarctic soil microbiota. Sixty-one yeasts strains were isolated from King George Island, Antarctica which were characterized physiologically and identified at the molecular level using the D1/D2 region of rDNA. Fifty-eight yeasts (belonging to the genera Cryptococcus, Leucosporidiella, Rhodotorula, Guehomyces, Candida, Metschnikowia and Debaryomyces) were screened for extracellular amylolytic, proteolytic, esterasic, pectinolytic, inulolytic xylanolytic and cellulolytic activities at low and moderate temperatures. Esterase activity was the most common enzymatic activity expressed by the yeast isolates regardless the assay temperature and inulinase was the second most common enzymatic activity. No cellulolytic activity was detected. One yeast identified as Guehomyces pullulans (8E) showed significant activity across six of seven enzymes types tested. Twenty-eight yeast isolates were classified as oleaginous, being the isolate 8E the strain that accumulated the highest levels of saponifiable lipids (42 %). PMID:27469174

  4. 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.

  5. Histological improvement in salivary gland along with effector memory Th17-1 cell reduction in a primary Sjogren's syndrome patient with dermatomyositis and diffuse large B-cell lymphoma by R-CHOP therapy.

    PubMed

    Koga, Tomohiro; Mizokami, Akinari; Nakashima, Masahiro; Shimizu, Toshimasa; Nakashima, Yoshikazu; Nakamura, Hideki; Chiwata, Masahiko; Daisuke, Niino; Kawakami, Atsushi

    2016-04-01

    We treated a 45-year-old Japanese woman with primary Sjogren's syndrome (SS) complicated with dermatomyositis (DM) followed by diffuse large B-cell lymphoma. She was admitted to our hospital for further evaluation of fever, weight loss and peritoneal lymphadenopathy. The histological examination of her lymph node revealed diffuse large B-cell lymphoma. The patient was then treated with 8 cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) therapy, which resulted in histological and functional improvement of her salivary glands. Of note, the percentage of effector memory Th17-1 (CD3+CD4+CD45RA-CCR7-CXCR3+CCR6+) cells in the peripheral blood was decreased after the R-CHOP treatment. This case suggests that an altered Th17-1 cell subset by B-cell depletion therapy is critical for the improvement of tissue damage in patients with SS, and the case suggests that clinicians should consider measuring the effector memory Th-subsets to predict the disease activity in SS patients. PMID:26960952

  6. 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... authorizing civil money penalties for any person who violates a requirement of the Family Smoking...

  7. 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... authorizing civil money penalties for any person who violates a requirement of the Family Smoking...

  8. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Regulation (45 CFR part 5). ... 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...

  9. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Regulation (45 CFR part 5). ... 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...

  10. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Regulation (45 CFR part 5). ... 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...