WorldWideScience

Sample records for aureobasidium pullulans y-2311-1

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

    Science.gov (United States)

    Yegin, Sirma

    2017-04-15

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

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

    Science.gov (United States)

    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)).

  3. Utility of Aureobasidium pullulans and pullulan in food biotechnology

    International Nuclear Information System (INIS)

    Gniewosz, M.; Sobczak, E.

    1999-01-01

    Fungus Aureobasidium pullulans is the producer of numerous valuable substances (enzymes, melanins, antibiotics). Pullulan - the product of metabolism of A. pullulans strains has a great potential utility in food industry. Since chemical synthesis of pullulan is expensive, a great effort has been made to improve the productivity of A. pullulans strains. The application of mutagenesis (UV light, several chemical mutagenes) allowed to receive mutants with high productivity of that compound. The most promising mutants are those which produce pullulan without melanin. (author)

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

    Science.gov (United States)

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

    1975-01-01

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

  5. Biotechnology of Aureobasidium pullulans: A phylogenetic perspective

    Science.gov (United States)

    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...

  6. Pullulan production from coconut by-products by Aureobasidium ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-01-19

    Jan 19, 2009 ... Key words: Pullulan, Aureobasidium pullulans, coconut water, coconut milk, characterization, FT-IR. ... sugars such as sucrose, glucose, fructose, maltose, starch, or ... sate, cornmeal hydrolysates, corn syrup, fuel ethanol.

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

    Science.gov (United States)

    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.

  8. Isolation and Characterization of Pollulan Produced by a Local Isolate of Aureobasidium Pullulans

    OpenAIRE

    Kassim, Mohammad Bashir Ismail [محمد بشيراسماعيل قاسم

    2000-01-01

    A local isolate of Aureobasidium pullulans was obtained from fig leaves surface. The isolate was identified as A. pullulans on the basis of appearance of polymorphism in the colonies, melanin pigment and pullulan production in shake flask cultures. Pullulan production by the local isolate of A pullulans was two thirds of that produced by ATCC 42023. The studies of physico — chemical properties, chemical analysis, sugar composition periodate oxidation and smith degradation showed that the pull...

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

    Science.gov (United States)

    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.

  10. The mechanism of improved pullulan production by nitrogen limitation in batch culture of Aureobasidium pullulans.

    Science.gov (United States)

    Wang, Dahui; Chen, Feifei; Wei, Gongyuan; Jiang, Min; Dong, Mingsheng

    2015-08-20

    Batch culture of Aureobasidium pullulans CCTCC M 2012259 for pullulan production at different concentrations of ammonium sulfate and yeast extract was investigated. Increased pullulan production was obtained under nitrogen-limiting conditions, as compared to that without nitrogen limitation. The mechanism of nitrogen limitation favoring to pullulan overproduction was revealed by determining the activity as well as gene expression of key enzymes, and energy supply for pullulan biosynthesis. Results indicated that nitrogen limitation increased the activities of α-phosphoglucose mutase and glucosyltransferase, up-regulated the transcriptional levels of pgm1 and fks genes, and supplied more ATP intracellularly, which were propitious to further pullulan biosynthesis. The economic analysis of batch pullulan production indicated that nitrogen limitation could reduce more than one third of the cost of raw materials when glucose was supplemented to a total concentration of 70 g/L. This study also helps to understand the mechanism of other polysaccharide overproduction by nitrogen limitation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Aureobasidium pullulans infection in a patient with chronic lymphocytic leukemia

    Directory of Open Access Journals (Sweden)

    Leonardo Rodrigues de Oliveira

    2013-09-01

    Full Text Available Saprophytic fungi are being increasingly recognized as etiologic agents of mycoses in immunosuppressed patients. We report a case of subcutaneous infiltration by Aureobasidium pullulans, likely due to traumatic inoculation, in a neutropenic patient during chemotherapy for chronic lymphocytic leukemia. The patient was treated with amphotericin B deoxycholate but was subsequently switched to itraconazole, which improved the lesion. This case highlights the importance of considering unusual fungal infections in critically ill patients such as those who are immunosuppressed due to chemotherapy. Diagnostic techniques and effective antifungal therapy have improved the prognosis of these cases.

  12. High-level pullulan production by Aureobasidium pullulans var. melanogenium P16 isolated from mangrove system.

    Science.gov (United States)

    Ma, Zai-Chao; Fu, Wen-Juan; Liu, Guang-Lei; Wang, Zhi-Peng; Chi, Zhen-Ming

    2014-06-01

    After over 100 strains of Aureobasidium spp. isolated from mangrove system were screened for their ability to produce exopolysaccharide (EPS), it was found that Aureobasidium pullulans var. melanogenium P16 strain among them could produce high level of EPS. Under the optimal conditions, 65.3 g/L EPS was produced by the P16 strain within 120 h at flask level. During 10-L batch fermentation, when the medium contained 120.0 g/L sucrose, 67.4 g/L of EPS and 23.1 g/L of cell dry weight in the culture were obtained within 120 h, leaving 0.78 g/L of reducing sugar and 11.4 g/L of total sugar in the fermented medium. It should be stressed that during the fermentation, no melanin was observed. After purification, the purified EPS was confirmed to be pullulan. This is the first time to report that A. pullulans var. melanogenium P16 strain isolated from the mangrove system can produce high level of pullulan.

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

    OpenAIRE

    Gostinčar, Cene; Ohm, Robin A; Kogej, Tina; Sonjak, Silva; Turk, Martina; Zajc, Janja; Zalar, Polona; Grube, Martin; Sun, Hui; Han, James; Sharma, Aditi; Chiniquy, Jennifer; Ngan, Chew Yee; Lipzen, Anna; Barry, Kerrie

    2014-01-01

    Background Aureobasidium pullulans is a black-yeast-like fungus used for production of the polysaccharide pullulan and the antimycotic aureobasidin A, and as a biocontrol agent in agriculture. It can cause opportunistic human infections, and it inhabits various extreme environments. To promote the understanding of these traits, we performed de-novo genome sequencing of the four varieties of A. pullulans. Results The 25.43-29.62 Mb genomes of these four varieties of A. pullulans encode between...

  14. Production of pullulan by a thermotolerant Aureobasidium pullulans strain in non-stirred fed batch fermentation process

    OpenAIRE

    Singh, Ranjan; Gaur, Rajeeva; Tiwari, Soni; Gaur, Manogya Kumar

    2012-01-01

    Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l) at 42ºC, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with th...

  15. Statistical Optimization of Medium for Pullulan Production by Aureobasidium pullulans NCPS2016 Using Fructose and Soybean Meal Hydrolysates

    Directory of Open Access Journals (Sweden)

    Jinyu Yang

    2018-06-01

    Full Text Available Pullulan, with its excellent characteristics of film-forming, water solubility, and biodegradability, is attracting more and more attention in agricultural products preservation. However, high pullulan production cost largely restricts its widely application due to its low production. In order to improve pullulan production by Aureobasidium pullulans NCPS2016, the medium was optimized using single factor experiment and response surface methodology. Based on the single factor experiments, the contents of soybean meal hydrolysates (SMHs, (NH42SO4, and K2HPO4·3H2O were considered to be main factors influencing the extracellular polysaccharide (EPS production, and were further optimized by Box–Behnken design. The optimal content of SMHs of 7.71 g/L, (NH42SO4 of 0.35 g/L, and K2HPO4·3H2O of 8.83 g/L were defined. Finally, EPS production of 59.8 g/L was obtained, 39% higher in comparison with the production in the basal medium. The purified EPS produced by NCPS2016 was confirmed to be pullulan. This is the first time fructose is reported to be the optimal carbon source for pullulan production by Aureobasidium pullulans, which is of great significance for the further study of the mechanism of the synthesis of pullulan by NCPS2016. Also, the results here have laid a foundation for reducing the industrial production cost of pullulan.

  16. [Amylase production by Aureobasidium pullulans in liquid and solid media].

    Science.gov (United States)

    Lodato, P B; Forchiassin, F; Segovia de Huergo, M B

    1997-01-01

    Amylase production by a strain of Aureobasidium pullulans isolated in the laboratory was evaluated in liquid media (complex and synthetic) and in solid medium (wheat bran). There was an inhibitory effect in amylase production or amylase secretion by glucose. Asparagine was the best nitrogen source for amylase production (4-6 g/l). Only chlamidospores and melanin but not, amylase activity, were obtained with ammonium sulfate. Amylase production in solid culture was higher than the production obtained in the liquid media assayed. Optimum initial moisture content in solid culture ranged between 57 and 74%. No difference was observed in amylase production between solid media inoculated with cells grown in liquid or solid media.

  17. Aureobasidium pullulans Fermented Feruloyl Oligosaccharide: Optimization of Production, Preliminary Characterization, and Antioxidant Activity

    OpenAIRE

    Xiaohong Yu; Zhenxin Gu

    2013-01-01

    Wheat bran (WB) was subjected to processing with Aureobasidium pullulans (A. pullulans) under selected conditions to partially break down the xylan into soluble products (mainly feruloyl oligosaccharides, FOs). The objective of this study was to investigate the technology for one-step fermentation of WB by A. pullulans without melanin secretion to produce FOs as well as to determine their structural features and antioxidant activity. Initial pH, inoculation quantity, and fermentation temperat...

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

    Science.gov (United States)

    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.

  19. [Complete genome sequencing of polymalic acid-producing strain Aureobasidium pullulans CCTCC M2012223].

    Science.gov (United States)

    Wang, Yongkang; Song, Xiaodan; Li, Xiaorong; Yang, Sang-tian; Zou, Xiang

    2017-01-04

    To explore the genome sequence of Aureobasidium pullulans CCTCC M2012223, analyze the key genes related to the biosynthesis of important metabolites, and provide genetic background for metabolic engineering. Complete genome of A. pullulans CCTCC M2012223 was sequenced by Illumina HiSeq high throughput sequencing platform. Then, fragment assembly, gene prediction, functional annotation, and GO/COG cluster were analyzed in comparison with those of other five A. pullulans varieties. The complete genome sequence of A. pullulans CCTCC M2012223 was 30756831 bp with an average GC content of 47.49%, and 9452 genes were successfully predicted. Genome-wide analysis showed that A. pullulans CCTCC M2012223 had the biggest genome assembly size. Protein sequences involved in the pullulan and polymalic acid pathway were highly conservative in all of six A. pullulans varieties. Although both A. pullulans CCTCC M2012223 and A. pullulans var. melanogenum have a close affinity, some point mutation and inserts were occurred in protein sequences involved in melanin biosynthesis. Genome information of A. pullulans CCTCC M2012223 was annotated and genes involved in melanin, pullulan and polymalic acid pathway were compared, which would provide a theoretical basis for genetic modification of metabolic pathway in A. pullulans.

  20. Morphological changes during the life cycle of Aureobasidium pullulans (de Bary) Arnaud.

    Science.gov (United States)

    Kocková-Kratochvílová, A; Cernáková, M; Sláviková, E

    1980-01-01

    Aureobasidium pullulans (de Bary) Arnaud was isolated from different natural materials plant blossoms in particular. Elements of vegetative multiplication, structure of colonies and cultures in liquid media were analyzed in detail, leading to construction of the life cycle of this organism. Morphological polymorphism was found to be combined with the production of melanin and the polysaccharide pullulan. Morphological analysis served for a directed selection for studies of physiological properties of this organism and its practical application.

  1. Production of pullulan by a thermotolerant Aureobasidium pullulans strain in non-stirred fed batch fermentation process

    Directory of Open Access Journals (Sweden)

    Ranjan Singh

    2012-09-01

    Full Text Available Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l at 42ºC, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with the 2 liters in two successive batches at 5 h interval into the fermentor. The sterile air was supplied only for 10h at the rate of 0.5 vvm.

  2. Production of pullulan by a thermotolerant aureobasidium pullulans strain in non-stirred fed batch fermentation process.

    Science.gov (United States)

    Singh, Ranjan; Gaur, Rajeeva; Tiwari, Soni; Gaur, Manogya Kumar

    2012-07-01

    Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l) at 42(o)C, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with the 2 liters in two successive batches at 5 h interval into the fermentor. The sterile air was supplied only for 10h at the rate of 0.5 vvm.

  3. Discovering the role of the apolipoprotein gene and the genes in the putative pullulan biosynthesis pathway on the synthesis of pullulan, heavy oil and melanin in Aureobasidium pullulans.

    Science.gov (United States)

    Guo, Jian; Huang, Siyao; Chen, Yefu; Guo, Xuewu; Xiao, Dongguang

    2017-12-18

    Pullulan produced by Aureobasidium pullulans presents various applications in food manufacturing and pharmaceutical industry. However, the pullulan biosynthesis mechanism remains unclear. This work proposed a pathway suggesting that heavy oil and melanin may correlate with pullulan production. The effects of overexpression or deletion of genes encoding apolipoprotein, UDPG-pyrophosphorylase, glucosyltransferase, and α-phosphoglucose mutase on the production of pullulan, heavy oil, and melanin were examined. Pullulan production increased by 16.93 and 8.52% with the overexpression of UDPG-pyrophosphorylase and apolipoprotein genes, respectively. Nevertheless, the overexpression or deletion of other genes exerted little effect on pullulan biosynthesis. Heavy oil production increased by 146.30, 64.81, and 33.33% with the overexpression of UDPG-pyrophosphorylase, α-phosphoglucose mutase, and apolipoprotein genes, respectively. Furthermore, the syntheses of pullulan, heavy oil, and melanin can compete with one another. This work may provide new guidance to improve the production of pullulan, heavy oil, and melanin through genetic approach.

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

    Science.gov (United States)

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

    Science.gov (United States)

    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 ...

  6. Biological activities of organic extracts of four Aureobasidium pullulans varieties isolated from extreme marine and terrestrial habitats.

    Science.gov (United States)

    Botić, Tanja; Kralj-Kunčič, Marjetka; Sepčić, Kristina; Batista, Urška; Zalar, Polona; Knez, Željko; Gunde-Cimerman, Nina

    2014-01-01

    We report on the screening for biological activities of organic extracts from seven strains that represent four varieties of the fungus Aureobasidium pullulans, that is A. pullulans var. melanogenum, A. pullulans var. pullulans, A. pullulans var. subglaciale and A. pullulans var. namibiae. We monitored haemolysis, cytotoxicity, antioxidant capacity and growth inhibition against three bacterial species. The haemolytic activity of A. pullulans var. pullulans EXF-150 strain was due to five different haemolytically active fractions. Extracts from all of the other varieties contained at least one haemolytically active fraction. Short-term exposure of cell lines to these haemolytically active organic extracts resulted in more than 95% cytotoxicity. Strong antioxidant capacity, corresponding to 163.88 μg ascorbic acid equivalent per gram of total solid, was measured in the organic extract of the strain EXF-3382, obtained from A. pullulans var. melanogenum, isolated from the deep sea. Organic extracts from selected varieties of A. pullulans exhibited weak antibacterial activities.

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

    Science.gov (United States)

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

    2014-07-01

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

  8. Enhanced production of pullulan by two strains of A. pullulans with different concentrations of soybean oil in sucrose solution in batch fermentations

    Directory of Open Access Journals (Sweden)

    R. F. Sena

    2006-12-01

    Full Text Available Aureobasidium pullulans is a microorganism that produces pullulan (homopolysaccharide extracellularly through a fermentation process with sugars (maltose, d-xylose, sucrose and starch as its carbon source. Pullulan is a linear polysaccharide of D-glycopyranose containing (1 ->4-alpha and (1 -> 6-alpha linkages at a 2:1 ratio, is highly soluble in water and has various applications in the food, packaging, film and pharmaceutical industries. Lipids, primarily oils, having antifoaming properties as well as nutritional particularities, are considered an essential additional carbon source for the growth of microorganisms, especially fungi. These nutrient sources are very important for the maintenance of microorganism cells. In fact, these positive effects are only achieved when the right source is added at both the right time and the right dosage into the broth of the fermentation process. In this research on pullulan production with the strains NRRL Y-6220 and NRRL Y-2311-1, it was found that the latter strain achieved better results for undesirable pigment formation, pullulan titer, time of maximum production (96 hours and biomass yields than strain NRRL Y-6220, which also showed suitable results for biomass yields and cell morphology. However, the dark pigmentation of the strain NRRL Y-6220, formed through the process, makes its application unacceptable for foods and pharmaceuticals. Strain NRRL Y-2311-1 was shown to be a promising potential industrial microorganism, whose applications should be studied more in depth.

  9. Effects of melanin on the accumulation of exopolysaccharides by Aureobasidium pullulans grown on nitrate.

    Science.gov (United States)

    Zheng, Weifa; Campbell, Bradley S; McDougall, Barbara M; Seviour, Robert J

    2008-11-01

    Aureobasidium pullulans produced pullulan and melanin when grown in medium containing low nitrate levels. With high nitrate concentrations, however, this fungus produced a mixture of exopolysaccharides (EPS) without melanin synthesis. At 0.78 g l(-1) N as nitrate, where no melanin synthesis occurred, maximum EPS yields reached 6.92 g l(-1) and then decreased to the final yield of 2.36 g l(-1). Following melanin addition (0.1 g l(-1)), yields reached 7.02 g l(-1) at 48 h and fell to a final yield of 5.21 g l(-1). The EPS produced in high nitrate medium contained both pullulan and (1-->3)-beta-glucan, but only pullulan was produced with melanin-supplementation. With melanin addition a doubling of (1-->3)-beta-glucanase activity was observed in high nitrate medium compared to that without supplementation. On the other hand amylolytic activities disappeared in medium with melanin production or addition. Culture filtrates sustained a higher reducing capacity (RC) when melanin was present. Low RC appeared to reduce (1-->3)-beta-glucanase activity and increase amylolytic activities. Thus, higher RC appears to inhibit production/activity of amylose-degrading enzymes capable of degrading pullulan, and stimulates (1-->3)-beta-glucanase synthesis/activity, leading to a preferential accumulation of pullulan.

  10. Strain improvement and statistical optimization as a combined strategy for improving fructosyltransferase production by Aureobasidium pullulans NAC8

    Directory of Open Access Journals (Sweden)

    Adedeji Nelson Ademakinwa

    2017-12-01

    A relatively low FTase-producing strain of Aureobasidium pullulans NAC8 was enhanced for optimum production using a two-pronged approach involving mutagenesis and statistical optimization. The improved mutant strain also had remarkable biotechnological properties that make it a suitable alternative than the wild-type.

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

    Science.gov (United States)

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

    2017-04-01

    Pullulan is a polymer produced by Aureobasidium pullulans and the main bottleneck for its industrial production is the presence of melanin pigment. In this study, light-emitting diodes (LEDs) of different wavelengths were used to assist the fermentation process aiming to produce low-melanin containing pullulan by wild strain of A. pullulans LB83 with different carbon sources. Under white light using glucose-based medium, 11.75g.L -1 of pullulan with high melanin content (45.70UA 540nm .g -1 ) was obtained, this production improved in process assisted by blue LED light, that resulted in 15.77g.L -1 of pullulan with reduced content of melanin (4.46UA 540nm .g -1 ). By using sugarcane bagasse (SCB) hydrolysate as carbon source, similar concentration of pullulan (about 20g.L -1 ) was achieved using white and blue LED lights, with lower melanin contents in last. Use of LED light was found as a promising approach to assist biotechnological process for low-melanin containing pullulan production. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Aureobasidium pullulans Fermented Feruloyl Oligosaccharide: Optimization of Production, Preliminary Characterization, and Antioxidant Activity

    Directory of Open Access Journals (Sweden)

    Xiaohong Yu

    2013-11-01

    Full Text Available Wheat bran (WB was subjected to processing with Aureobasidium pullulans (A. pullulans under selected conditions to partially break down the xylan into soluble products (mainly feruloyl oligosaccharides, FOs. The objective of this study was to investigate the technology for one-step fermentation of WB by A. pullulans without melanin secretion to produce FOs as well as to determine their structural features and antioxidant activity. Initial pH, inoculation quantity, and fermentation temperature were found to be efficient for releasing FOs according to the Plackett-Burman design (PBD. Based on the D-Optimal design, a yield of 904 nmol of FOs / L of fermentation broth was obtained under optimal conditions of initial pH 6.0, inoculation quantity 4.50%, and fermentation temperature 29 oC. Purification of FOs was performed with alcohol precipitation and Amberlite XAD-2. GC, IR, and ESI-MS demonstrated that FOs consist of feruloyl arabinosyl xylopentose (FAX5, Mw986, feruloyl arabinosyl xylotetraose (FAX4, Mw854, feruloyl arabinosyl xylotriose (FAX3, Mw722, and feruloyl arabinosyl xylobiose (FAX2, Mw590. Increasing the FO dose led to increased activity of SOD and GSH-Px in serum of S180 tumor-bearing mice, while the level of MDA was reduced, thus improving its in vivo antioxidant activity.

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

    Science.gov (United States)

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

    2016-12-01

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

  14. Plasticizers increase adhesion of the deteriogenic fungus Aureobasidium pullulans to polyvinyl chloride.

    Science.gov (United States)

    Webb, J S; Van der Mei, H C; Nixon, M; Eastwood, I M; Greenhalgh, M; Read, S J; Robson, G D; Handley, P S

    1999-08-01

    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 10(8) 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 degrees 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.

  15. Production of pigment-free pullulan by swollen cell in Aureobasidium pullulans NG which cell differentiation was affected by pH and nutrition.

    Science.gov (United States)

    Li, Bing-xue; Zhang, Ning; Peng, Qing; Yin, Tie; Guan, Fei-fei; Wang, Gui-li; Li, Ying

    2009-08-01

    A black yeast strain "NG" was isolated from strawberry fruit and identified as Aureobasidium pullulans. Strain NG displayed yeast-like cell (YL), swollen cell (SC), septate swollen cell (SSC), meristematic structure (MS), and chlamydospore (CH) morphologies. pH was the key factor regulating cell morphogenesis of strain NG. Differentiation of YL controlled by extracellular pH had no relationship with nutrition level. YL was maintained at pH >6.0, but was transformed into SC at pH approximately 4.5. SC, a stable cell type of A. pullulans, could bud, septate, or transform into MS or CH, in response to nutrition level and low pH. SC produced swollen cell blastospores (SCB) at pH 2.1 with abundant nutrition, and could transform into MS at lower pH (1.5). SC was induced to form CH by low level nutrition and pH melanin) were produced by SC of strain NG. Pullulan content of the polysaccharides was very high (98.37%). Fourier-transform infrared spectroscopy confirmed that chemical structures of the polysaccharides and standard pullulan were identical. Swollen cells produced 2.08 mg/ml non-pigmented polysaccharides at 96 h in YPD medium. Controlling pH of fermentation is an effective and convenient method to harvest SC for melanin-free pullulan production.

  16. Efficient transformation of sucrose into high pullulan concentrations by Aureobasidium melanogenum TN1-2 isolated from a natural honey.

    Science.gov (United States)

    Jiang, Hong; Xue, Si-Jia; Li, Yan-Feng; Liu, Guang-Lei; Chi, Zhen-Ming; Hu, Zhong; Chi, Zhe

    2018-08-15

    A very high pullulan producing yeast-like fungus, Aureobasidium melanogenum TN1-2 isolated from a natural honey was found to be able to produce 97.0 g/L of pullulan from 140.0 g/L sucrose at a flask level while it could yield 114.0 g/L of pullulan within 134 h during a 10-liter fermentation, the yield was 0.81 g/g and the productivity was 0.86 g/L/h. The high ability to biosynthesize pullulan by this yeast-like fungal strain TN1-2 was related to high glucosyltransferase activity, high phosphofructo-2-kinase activity, high content of its cellular glycerol and low glucose repressor. The Mw of the produced pullulan was 1.42 × 10 5  g/mol. The low Mw may be due to the high α-amylase, glucoamylase and isopullulanase activities. The intracellular level of trehalose had no influence on high pullulan production by the yeast-like fungal strain TN1-2. Copyright © 2018. Published by Elsevier Ltd.

  17. Heterologous expression of Spathaspora passalidarum xylose reductase and xylitol dehydrogenase genes improved xylose fermentation ability of Aureobasidium pullulans.

    Science.gov (United States)

    Guo, Jian; Huang, Siyao; Chen, Yefu; Guo, Xuewu; Xiao, Dongguang

    2018-04-30

    Aureobasidium pullulans is a yeast-like fungus that can ferment xylose to generate high-value-added products, such as pullulan, heavy oil, and melanin. The combinatorial expression of two xylose reductase (XR) genes and two xylitol dehydrogenase (XDH) genes from Spathaspora passalidarum and the heterologous expression of the Piromyces sp. xylose isomerase (XI) gene were induced in A. pullulans to increase the consumption capability of A. pullulans on xylose. The overexpression of XYL1.2 (encoding XR) and XYL2.2 (encoding XDH) was the most beneficial for xylose utilization, resulting in a 17.76% increase in consumed xylose compared with the parent strain, whereas the introduction of the Piromyces sp. XI pathway failed to enhance xylose utilization efficiency. Mutants with superior xylose fermentation performance exhibited increased intracellular reducing equivalents. The fermentation performance of all recombinant strains was not affected when glucose or sucrose was utilized as the carbon source. The strain with overexpression of XYL1.2 and XYL2.2 exhibited excellent fermentation performance with mimicked hydrolysate, and pullulan production increased by 97.72% compared with that of the parent strain. The present work indicates that the P4 mutant (using the XR/XDH pathway) with overexpressed XYL1.2 and XYL2.2 exhibited the best xylose fermentation performance. The P4 strain showed the highest intracellular reducing equivalents and XR and XDH activity, with consequently improved pullulan productivity and reduced melanin production. This valuable development in aerobic fermentation by the P4 strain may provide guidance for the biotransformation of xylose to high-value products by A. pullulans through genetic approach.

  18. Direct production of feruloyl oligosaccharides and hemicellulase inducement and distribution in a newly isolated Aureobasidium pullulans strain.

    Science.gov (United States)

    Yu, Xiao-hong; Gu, Zhen-xin

    2014-02-01

    Studies were carried out to screen and identify strains that are able to directly produce ferulic oligosaccharides (FOs) from wheat bran (WB). The inducement and distribution of hemicellulases from strain 2012, which was identified as a non-melanin secreting strain of Aureobasidium pullulans (A. pullulans), were also determined. In a 60 g/L WB solution, A. pullulans could produce 545 nmol/L FOs, 64.12 IU/mL xylanase and 0.14 IU/mL ferulic acid esterase (FAE). A. pullulans was cultivated in media with WB, glucose, xylose, sucrose, lactose or xylan as the carbon source, and hemicellulases were mainly induced by xylan and WB and inhibited by glucose and sucrose. Xylanase and FAE were mainly present in the culture filtrate, xylosidase in the hyphal filaments and arabinofuranosidase was a membrane-bound enzyme. The yield of FOs was positively correlated to the hemicellulases activity, and significantly positively (P < 0.05) correlated to the xylanase activity (r = 0.992).

  19. Anti-skin-aging benefits of exopolymers from Aureobasidium pullulans SM2001.

    Science.gov (United States)

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

    2014-01-01

    There have been many attempts to search for affordable and effective functional cosmetic ingredients, especially from natural sources. As research into developing a functional cosmetic ingredient, we investigated whether exopolymers from Aureobasidium pullulans SM2001 (E-AP-SM2001) exert antioxidant, antiwrinkle, whitening, and skin moisturizing effects. Antioxidant effects of E-AP-SM2001 were determined by measuring free radical scavenging capacity and superoxide dismutase (SOD)-like activity. Antiwrinkle effects were assessed through the inhibition of hyaluronidase, elastase, collagenase, and matrix metalloproteinase (MMP)-1. Whitening effects were measured by tyrosinase inhibition assay, and by melanin formation test in B16/F10 melanoma cells. Skin moisturizing effects were detected by mouse skin water content test. E-AP-SM2001 showed potent DPPH radical scavenging activity and SOD-like effects. Additionally, hyaluronidase, elastase, collagenase, and MMP-1 activities were significantly inhibited by E-AP-SM2001. We also observed that E-AP-SM2001 effectively reduced melanin production by B16/F10 melanoma cells and mushroom tyrosinase activities. Furthermore, significant increases in skin water content were detected in E-AP-SM2001- treated mouse skin, as compared with vehicle-treated control skin. Notably, a mask pack containing E-AP-SM2001 showed a >twofold more extensive moisturizing effect compared with one containing Saccharomycopsis ferment filtrate. Our results suggest that E-AP-SM2001 has adequate antiaging, antiwrinkle, and whitening benefits and skin moisturizing effect. These effects involve reducing hyaluronidase, elastase, collagenase, and MMP-1 activities, as well as inhibition of melanin production and tyrosinase activities. Therefore, the antioxidant E-AP-SM2001 may serve as a predictable functional ingredient.

  20. Effects of sugar and amino acid supplementation on Aureobasidium pullulans NRRL 58536 antifungal activity against four Aspergillus species.

    Science.gov (United States)

    Prasongsuk, Sehanat; Ployngam, Saowaluck; Wacharasindhu, Sumrit; Lotrakul, Pongtharin; Punnapayak, Hunsa

    2013-09-01

    Cultured cell extracts from ten tropical strains of Aureobasidium pullulans were screened for antifungal activity against four pathogenic Aspergillus species (Aspergillus flavus, Aspergillus niger, Aspergillus fumigatus, and Aspergillus terreus) using the well diffusion and conidial germination inhibition assays. The crude cell extract from A. pullulans NRRL 58536 resulted in the greatest fungicidal activity against all four Aspergillus species and so was selected for further investigation into enhancing the production of antifungal activity through optimization of the culture medium, carbon source (sucrose and glucose) and amino acid (phenylalanine, proline, and leucine) supplementation. Sucrose did not support the production of any detectable antifungal activity, while glucose did with the greatest antifungal activity against all four Aspergillus species being produced in cells grown in medium containing 2.5 % (w/v) glucose. With respect to the amino acid supplements, variable trends between the different Aspergillus species and amino acid combinations were observed, with the greatest antifungal activities being obtained when grown with phenylalanine plus leucine supplementation for activity against A. flavus, proline plus leucine for A. terreus, and phenylalanine plus proline and leucine for A. niger and A. fumigatus. Thin layer chromatography, spectrophotometry, high-performance liquid chromatography, (1)H-nuclear magnetic resonance, and MALDI-TOF mass spectrometry analyses were all consistent with the main component of the A. pullulans NRRL 58536 extracts being aureobasidins.

  1. Purification, Characterization of Amylase from Indigenously Isolated Aureobasidium pullulans Cau 19 and Its Bioconjugates with Gold Nanoparticles.

    Science.gov (United States)

    Mulay, Y R; Deopurkar, R L

    2018-02-01

    The amylase from Aureobasidium pullulans Cau 19 was purified by ammonium sulfate precipitation and Sephadex G-100 chromatography with a 9.25-fold increase in specific activity as compared to crude enzyme. Km and turn over values of the enzyme were 6.25 mg/mL and 5.0 × 10 2 /min, respectively. Effect of different metal ions on the purified enzyme was investigated; 1 mM calcium (Ca) and cobalt (Co) enhanced enzyme activity by twofold; copper (Cu) had no effect on the activity of the enzyme. Mercury (Hg) 1 mM caused 90% inactivation whereas iron (Fe) and manganese (Mn) caused 10 to 16% inhibition. Amylase from A. pullulans Cau 19 was bioconjugated to gold nanoparticles synthesized using the biomass of A. pullulans Cau 19. Fourier transform infrared spectroscopy confirmed the conjugation of the enzyme to the gold nanoparticles. Though, only 20% of the added enzyme was adsorbed/conjugated on gold nanoparticles, 80% of the adsorbed activity could be estimated in the assay. The conjugated enzyme exhibited better tolerance to a broad pH range of 3.0-9.0 and higher temperatures compared with native enzyme.

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

    Science.gov (United States)

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

    2016-08-22

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

  3. Production of poly(β-l-malic acid) by Aureobasidium pullulans HA-4D under solid-state fermentation.

    Science.gov (United States)

    Xia, Jun; Li, Rongqing; He, Aiyong; Xu, Jiaxing; Liu, Xiaoyan; Li, Xiangqian; Xu, Jiming

    2017-11-01

    Poly(β-l-malic acid) (PMA) production by Aureobasidium pullulans HA-4D was carried out through solid-state fermentation (SSF) using agro-industrial residues. Maximum PMA production (75.4mg/g substrate) was obtained from a mixed substrate of sweet potato residue and wheat bran (1:1, w/w) supplemented with NaNO 3 (0.8%, w/w) and CaCO 3 (2%, w/w), with an initial moisture content of 70% and inoculum size of 13% (v/w) for 8days. Repeated-batch SSF was successfully conducted for 5 cycles with a high productivity. The scanning electron microscopy showed that the yeast-like cells of A. pullulans HA-4D could grow well on the solid substrate surface. Moreover, the cost analysis showed that the unit price of PMA in SSF was much lower than that of SmF. This is the first report on PMA production via SSF, and this study provided a new method to produce PMA from inexpensive agro-industrial residues. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Isolation and Characterization of melanized yeast form of Aureobasidium pullulans and physiological studies on the melanization process

    International Nuclear Information System (INIS)

    El-Gamal, M.S.; El-Bialy, H.A.; Elsayed, M.A.; Khalifa, M.A.

    2017-01-01

    Melanin pigments have immense application potentials in the fields of agriculture , cosmetics and pharmaceutical industries . Isolation and characterization of melanin producer from local resources is the main aim of the present study . Six melanized yeast isolates were selected from seventeen isolation resources including natural sources (Eucalyptus globules plant), industrial wastes (Tomato paste processing) and surface contaminants (Bathrooms). The selected yeasts were screened for melanin production in an individual experiment , results revealed the ability of yeast isolate selected from wastes of tomato paste processing (W 3 ) to produce melanin at 300 mM concentration . The selected melanized yeast was identified as Aureobasidium pullulans according to the morphological characteristics and 18S rRNA . Melanin production by the selected yeast was maximized by optimization of the cultural conditions and nutritional parameters by nearly two folds ( 550 mM ). The highest melanin yield was achieved in the fermentation medium contains 30 gl -1 sucrose , 35 gl -1 peptone and 1 mM L - DOPA concentration after nine days of incubation at 30°C and its pH value is adjusted at 7.0. Melanin characterization by FTIR and NMR resemble aliphatic and aromatic composition of A . pullulans ’ s melanin . In conclusion, the selected yeast is a promising candidate for melanin production in a semipilot scale

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

    Science.gov (United States)

    AbstractImage 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...

  6. Development of a one-step gene knock-out and knock-in method for metabolic engineering of Aureobasidium pullulans.

    Science.gov (United States)

    Guo, Jian; Wang, Yuanhua; Li, Baozhong; Huang, Siyao; Chen, Yefu; Guo, Xuewu; Xiao, Dongguang

    2017-06-10

    Aureobasidium pullulans is an increasingly attractive host for bio-production of pullulan, heavy oil, polymalic acid, and a large spectrum of extracellular enzymes. To date, genetic manipulation of A. pullulans mainly relies on time-consuming conventional restriction enzyme digestion and ligation methods. In this study, we present a one-step homologous recombination-based method for rapid genetic manipulation in A. pullulans. Overlaps measuring >40bp length and 10μg DNA segments for homologous recombination provided maximum benefits to transformation of A. pullulans. This optimized method was successfully applied to PKSIII gene (encodes polyketide synthase) knock-out and gltP gene (encodes glycolipid transfer protein) knock-in. After disruption of PKSIII gene, secretion of melanin decreased slightly. The melanin purified from disruptant showed lower reducing capacity compared with that of the parent strain, leading to a decrease in exopolysaccharide production. Knock-in of gltP gene resulted in at least 4.68-fold increase in heavy oil production depending on the carbon source used, indicating that gltP can regulate heavy oil synthesis in A. pullulans. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. A novel biosurfactant produced by Aureobasidium pullulans L3-GPY from a tiger lily wild flower, Lilium lancifolium Thunb.

    Science.gov (United States)

    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.

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

    Science.gov (United States)

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

    2017-01-01

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

  9. Cofactor and CO2 donor regulation involved in reductive routes for polymalic acid production by Aureobasidium pullulans CCTCC M2012223.

    Science.gov (United States)

    Zou, Xiang; Tu, Guangwei; Zan, Zhanquan

    2014-10-01

    Polymalic acid (PMA) is a water-soluble polyester with many attractive properties for biomedical application. Its monomer L-malic acid is widely used in the food industry and also a potential C4 platform chemical. Cofactor and CO2 donor involved in the reductive routes were investigated for PMA production by Aureobasidium pullulans. Biotin as the key cofactor of pyruvate carboxylase was favor for the PMA biosynthesis. Na2CO3 as CO2 donor can obviously improved PMA titer when compared with no CO2 supplier NaOH, and also exhibit more advantages than the other donor CaCO3 because of its water-soluble characteristic. A combinational process with addition of biotin 70 mg/L and Na2CO3 as the CO2 donor was scaled-up in 50 L fermentor, achieving the high product 34.3 g/L of PMA and productivity of 0.41 g/L h. This process provides an efficient and economical way for PMA and malic acid production, and is promising for industrial application.

  10. Production of Antimicrobial Films by Incorporation of Partially Purified Lysozyme into Biodegradable Films of Crude Exopolysaccharides Obtained from Aureobasidium pullulans Fermentation

    Directory of Open Access Journals (Sweden)

    Nilay Kandemir

    2005-01-01

    Full Text Available Antimicrobial films were produced by incorporating partially purified lysozyme into films of crude exopolysaccharides (59 % pullulan obtained from Aureobasidium pullulans fermentation. After film making, the films containing lysozyme at 100, 260, 520 and 780 μg/cm2 showed 23 to 70 % of their expected enzyme activities. The highest recovery of enzyme activity (65–70 % after the film making was obtained in films prepared by incorporating lysozyme at 260 μg/cm2 (1409 U/cm2. The incorporation of disodium EDTA×2H2O and sucrose did not affect the initial lysozyme activity of the films significantly. With or without the presence of disodium EDTA×2H2O at 52 or 520 μg/cm2, lysozyme activity showed sufficient stability in the films during 21 days of cold storage. However, the presence of sucrose at 10 mg/cm2 in the films caused the destabilization of part of enzyme activity (almost 35 % at the end of storage. The combinational incorporation of lysozyme at 780 μg/cm2 (4227 U/cm2 and disodium EDTA×2H2O at 520 μg/cm2 gave antimicrobial films effective on Escherichia coli. However, in the studied lysozyme concentration range the films did not show any antimicrobial activity against Lactobacillus plantarum. This study clearly showed that the partially purified lysozyme and crude exopolysaccharides from Aureobasidium pullulans may be used to obtain antimicrobial films to increase the safety of foods.

  11. Extracellular polysaccharides purified from Aureobasidium pullulans SM-2001 (Polycan) inhibit dexamethasone-induced muscle atrophy in mice

    Science.gov (United States)

    Cho, Hyung-Rae; Park, Dong-Chan; Jung, Go-Woon

    2018-01-01

    The present study assessed the beneficial skeletal muscle-preserving effects of extracellular polysaccharides from Aureobasidium pullulans SM-2001 (Polycan) (EAP) on dexamethasone (DEXA)-induced catabolic muscle atrophy in mice. To investigate whether EAP prevented catabolic DEXA-induced muscle atrophy, and to examine its mechanisms of action, EAP (100, 200 and 400 mg/kg) was administered orally, once a day for 24 days. EAP treatment was initiated 2 weeks prior to DEXA treatment (1 mg/kg, once a day for 10 days) in mice. Body weight alterations, serum biochemistry, calf thickness, calf muscle strength, gastrocnemius muscle thickness and weight, gastrocnemius muscle antioxidant defense parameters, gastrocnemius muscle mRNA expression, histology and histomorphometry were subsequently assessed. After 24 days, DEXA control mice exhibited muscle atrophy according to all criteria indices. However, these muscle atrophy symptoms were significantly inhibited by oral treatment with all three doses of EAP. Regarding possible mechanisms of action, EAP exhibited favorable ameliorating effects on DEXA-induced catabolic muscle atrophy via antioxidant and anti-inflammatory effects; these effects were mediated by modulation of the expression of genes involved in muscle protein synthesis (AKT serine/threonine kinase 1, phosphatidylinositol 3-kinase, adenosine A1 receptor and transient receptor potential cation channel subfamily V member 4) and degradation (atrogin-1, muscle RING-finger protein-1, myostatin and sirtuin 1). Therefore, these results indicated that EAP may be helpful in improving muscle atrophies of various etiologies. EAP at 400 mg/kg exhibited favorable muscle protective effects against DEXA-induced catabolic muscle atrophy, comparable with the effects of oxymetholone (50 mg/kg), which has been used to treat various muscle disorders. PMID:29138805

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

    Science.gov (United States)

    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...

  13. The Synergism of Chemical Herbicides and Aureobasidium pullulans for Control Cleavers (Galium aparine L.) in Wheat

    Institute of Scientific and Technical Information of China (English)

    Liang CHENG

    2015-01-01

    Aureobasidium pul ulans, a biocontrol agent for the annual weed Galium aparine L. was evaluated in vitro for its compatibility with commercial formulation of five herbicides at 1X (recommended field rate), 0.5X, 0.2X, 0.1X, 0.067X, and 0.05X concentrations. Germination of A. pul ulans with paraquat, 2, 4-D, quizalofop-p, and clethodim treatment appeared reduced compared with germination of A. pul ulans with fluroxypyr treatment at al concentrations. Stunted and shorter germ tubes in comparison with the control were observed with 2, 4-D, quizalofop-p, and clethodim at 0.2X. Al concentration of paraquat, 2, 4-D, quizalofop-p, and clethodim except 0.05X, significantly decreased radial growth of A. pul ulans compared with its growth on the untreated PDA medium. Field trials to further develop A. pul ulans as bio-control agent for control G. aparine L. was conducted to test the effectiveness of this fungus in wheat plots for 2 years at the same location in Xining. Treatments included spore suspensions of A. pul ulans alone, a mixture of both fungus and fluroxypyr in wheat. Biocontrol agent effectiveness was estimated at approximately 7 and 14 days after treatment, as disease incidence, percent weed control, and weed biomass reduction. Significant reduction in weed biomass occurred in combination treatments, and potential exists to tank mix A. pul ulans with fluroxypyr. Leaf surface moisture and air temperatures fol owing application may account for inconsistencies in field results between years. This fungal organisms show potential as bioherbicides for weeds in G. aparine L.

  14. 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).

    Science.gov (United States)

    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. Copyright © 2016. Published by Elsevier B.V.

  15. Introduction of Aureobasidium pullulans to the Phyllosphere of Organically Grown Strawberries with Focus on Its Establishment and Interactions with the Resident Microbiome

    Directory of Open Access Journals (Sweden)

    Annette Reineke

    2013-10-01

    Full Text Available Currently, there is little knowledge of the establishment of repeatedly applied biological control agents (BCAs in the phyllosphere of plants and, in particular, their interactions with the resident microbiome. Under field conditions, the BCA Aureobasidium pullulans was applied as a model organism to organically grown strawberries during two subsequent years (2011, 2012, either as single strain treatment or with the co-application of the entomopathogenic fungus Beauveria bassiana. Fungal and bacterial communities of strawberry leaves were investigated by means of plate counts and 454 pyrosequencing. The establishment of the introduced A. pullulans strains considerably differed between the two years, presumably due to distinct environmental conditions. Short-term and long-term effects of BCA applications on the composition and diversity of fungal communities could be observed as a result of successful establishment of A. pullulans, in 2011, showing, for instance, reduced diversity of fungal communities by competitive displacement shortly after BCA introduction. Due to considerable dynamics in untreated resident microbial communities in the phyllosphere in general, however, we suggest that even the effects caused by the applied BCA preparations in 2011 are negligible under practical conditions.

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

    Science.gov (United States)

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

    2016-10-29

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

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

    Directory of Open Access Journals (Sweden)

    Leila Bennamoun

    2016-10-01

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

  18. Effects of Two-stage Controlled pH and Temperature vs. One-step Process for Hemicellulase Biosynthesis and Feruloyl Oligosaccharide Fermentation using Aureobasidium pullulans

    Directory of Open Access Journals (Sweden)

    Xiaohong Yu

    2016-04-01

    Full Text Available A two-stage, pH- and temperature-controlled wheat bran fermentation method using Aureobasidium pullulans was investigated for feruloyl oligosaccharides (FOs production and the activities of xylanase, xylosidase, and ferulic acid esterase (FAE. A. pullulans secreted xylanase, xylosidase, and FAE at high levels in the initial pH of 4.0 to 5.0 and a fermentation liquid temperature of 31 °C to 33 °C. FOs production via two-stage fermentation (FOs 2 reached 1123 nmol/L after fermentation for 96 h, by controlling the initial pH at 4.0 and the initial temperature at 33 °C, and then changing the pH to 6.0 and the temperature to 29 °C at the same time at 36 h. This process was 12 h shorter and 219 nmol/L higher than a one-stage fermentation for producing FOs 1. Xylanase, xylosidase, and FAE activities were highly correlated with controlled pH and temperature and FOs biosynthesis rate. Thus, the combination of two-stage controlled pH and temperature could support mass production of FOs.

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

    Science.gov (United States)

    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. Copyright © 2011 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Shaghayegh Nasr

    2013-12-01

    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.

  1. Two-dimensional NMR data of a water-soluble β-(1→3, 1→6-glucan from Aureobasidium pullulans and schizophyllan from Schizophyllum commune

    Directory of Open Access Journals (Sweden)

    Hiroyuki Kono

    2017-12-01

    Full Text Available This article contains two-dimensional (2D NMR experimental data, obtained by the Bruker BioSpin 500 MHz NMR spectrometer (Germany which can used for the determination of primary structures of schizophyllan from Schizophyllum commune (SPG and a water-soluble β-(1→3, 1→6-glucan from Aureobasidium pullulans. Data include analyzed the 2D NMR spectra of these β-glucans, which are related to the subject of an article in Carbohydrate Polymers, entitled “NMR spectroscopic structural characterization of a water-soluble β-(1→3, 1→6-glucan from A. pullulans” (Kono et al., 2017 [1]. Data can help to assign the 1H and 13C chemical shifts of the structurally complex polysaccharides. Keywords: NMR, β-(1→3, 1→6-glucan, Aureobasidium pullulans, Schizophyllan, Spectral data

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

    Science.gov (United States)

    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.

  3. 77 FR 8731 - Aureobasidium pullulans

    Science.gov (United States)

    2012-02-15

    .... Fliessbach, L. Tamm, F.P. Weibel, G.S. Wyss. 2008. Culturable fungi of stored `Golden Delicious' apple fruits... apply to me? You may be potentially affected by this action if you are an agricultural [[Page 8732

  4. Optimization of physico-chemical and nutritional parameters for a novel pullulan-producing fungus, Eurotium chevalieri.

    Science.gov (United States)

    Gaur, R; Singh, R; Tiwari, S; Yadav, S K; Daramwal, N S

    2010-09-01

    To isolate the novel nonmelanin pullulan-producing fungi from soil and to optimize the physico-chemical and nutritional parameters for pullulan production. A selective enrichment method was followed for the isolation, along with development of a suitable medium for pullulan production, using shake flask experiments. Pullulan content was confirmed using pure pullulan and pullulanase hydrolysate. Eurotium chevalieri was able to produce maximum pullulan (38 ± 1·0 g l(-1) ) at 35°C, pH 5·5, 2·5% sucrose, 0·3% ammonium sulfate and 0·2% yeast extract in a shake flash culture medium with an agitation rate of 30 rev min(-1) for 65 h. The novel pullulan-producing fungus was identified as E. chevalieri (MTCC no. 9614), which was able to produce nonmelanin pullulan at from poorer carbon and nitrogen sources than Aureobasidium pullulans and may therefore be useful for the commercial production of pullulan. Eurotium chevalieri could produce pullulan in similar amounts to A. pullulans. Therefore, in future, this fungus could also be used for commercial pullulan production, because it is neither polymorphic nor melanin producing, hence its handling during pullulan fermentation will be easier and more economical. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.

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

    Science.gov (United States)

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

    2012-07-01

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

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

    Science.gov (United States)

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

    2010-04-01

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

  7. Aureobasidium pullulansas a biocontrol agent of blue mold in "Rocha" pear.

    Science.gov (United States)

    Ferreira-Pinto, M M; Moura-Guedes, M C; Barreiro, M G; Pais, I; Santos, M R; Silva, M J

    2006-01-01

    The blue mold of "Rocha" pear caused by Penicillium expansum is an important postharvest disease which is adequately controlled by application of synthetic fungicides. In recent years, strategies like biological control have been considered a desirable alternative to chemicals. Several studies have demonstrated the potential of the yeast-like fungus Aureobasidium pullulans for control of postharvest decay of pear. A Portuguese isolate of Aureobasidium pullulans was characterized and evaluated for its activity in reducing postharvest blue mold decay of "Rocha" pear caused by Penicillium expansum. Study of optimal conditions for antagonist growth was carried out in six different culture media. The effect of four maturity stages of fruits in the development of A. pullulans was also studied. Biocontrol studies were performed with two concentrations of the antagonist (3 x 10(8) and 4 x 10(9) CFU/ml). A. pullulans growth was significantly different (P Corn Meal Agar (CMA) and Potato Dextrose Agar (PDA) media which contains the higher concentration of glucose (20 mg/l). Medium resulted from fruits of the first harvest date presented lower colony diameter. Inoculation of A. pullulans at 3 x 10(8) and 4 x 10(9) CFU/ml reduced the incidence of the disease by 23 and 63%, and reduced the lesion diameter by 36 and 46%, respectively.

  8. Occurrence and infection of Cladosporium, Fusarium, Epicoccum and Aureobasidium in withered rotten grapes during post-harvest dehydration.

    Science.gov (United States)

    Lorenzini, Marilinda; Zapparoli, Giacomo

    2015-11-01

    Fungi like Cladosporium, Fusarium, Epicoccum and Aureobasidium can occur on withered grapes causing spoilage of passito wine. There is little or no information on the pathogenic role of these fungi. This study describes the isolation, incidence and identification of several isolates from different withered rotten grapes. Representative isolates grouped in several phenotypes were identified by phylogenetic analysis of internal transcribed spacer, actin or elongation factor gene sequences. Isolates of Cladosporium and Fusarium were ascribed to different species, of these C. ramotenellum, C. halotolerans and F. graminearum were isolated from Vitis vinifera for the first time. All Epicoccum and Aureobasidium isolates belonged to E. nigrum and A. pullulans, respectively. Random amplified DNA polymorphism analysis showed high level of heterogenicity among Epicoccum and Fusarium isolates. Infection assays were carried out to evaluate infectivity in some strains under different withering conditions. Fusarium spp. strains had similar infectivity, while significant variability was observed among Cladosporium spp. and E. nigrum strains. A. pullulans resulted particularly infective. This study provided insights into the occurrence and infection of these fungi in fruit-drying rooms with important implications towards control management during the withering.

  9. Production of Cellulolytic and Hemicellulolytic Enzymes From Aureobasidium pulluans on Solid State Fermentation

    Science.gov (United States)

    Leite, Rodrigo Simões Ribeiro; Bocchini, Daniela Alonso; da Silva Martins, Eduardo; Silva, Dênis; Gomes, Eleni; da Silva, Roberto

    This article investigates a strain of the yeast Aureobasidium pullulans for cellulase and hemicellulase production in solid state fermentation. Among the substrates analyzed, the wheat bran culture presented the highest enzymatic production (1.05 U/mL endoglucanase, 1.3 U/mL β-glucosidase, and 5.0 U/mL xylanase). Avicelase activity was not detected. The optimum pH and temperature for xylanase, endoglucanase and β-glucosidase were 5.0 and 50, 4.5 and 60, 4.0 and 75°C, respectively. These enzymes remained stable between a wide range of pH. The β-glucosidase was the most thermostable enzyme remaining 100% active when incubated at 75°C for 1 h.

  10. Effect of Meadowsweet Flower Extract-Pullulan Coatings on Rhizopus Rot Development and Postharvest Quality of Cold-Stored Red Peppers

    Directory of Open Access Journals (Sweden)

    Alicja Synowiec

    2014-08-01

    Full Text Available 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Li, Xian [College of chemistry & materials science, Northwest University, Taibai North Road 229, Xi’an, Shaanxi 710069 (China); Shaanxi Key Laboratory of Degradable Biomedical Materials, Department of Chemical Engineering, Northwest University, Taibai North Road 229, Xi’an, Shaanxi 710069 (China); Xue, Wenjiao [Shannxi provincial institute of microbiology, Xi’ an 710043 (China); Liu, Yannan; Li, Weina [Shaanxi Key Laboratory of Degradable Biomedical Materials, Department of Chemical Engineering, Northwest University, Taibai North Road 229, Xi’an, Shaanxi 710069 (China); Fan, Daidi, E-mail: fandaidi@nwu.edu.cn [Shaanxi Key Laboratory of Degradable Biomedical Materials, Department of Chemical Engineering, Northwest University, Taibai North Road 229, Xi’an, Shaanxi 710069 (China); Zhu, Chenhui [Shaanxi Key Laboratory of Degradable Biomedical Materials, Department of Chemical Engineering, Northwest University, Taibai North Road 229, Xi’an, Shaanxi 710069 (China); Wang, Yaoyu, E-mail: wyaoyu@nwu.edu.cn [College of chemistry & materials science, Northwest University, Taibai North Road 229, Xi’an, Shaanxi 710069 (China)

    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 NaIO{sub 4} to form a pullulan hydrogel and then may coupled with HLC using the reaction between the –NH{sub 2} end-group of HLC and the –CHO group present on the aldehyde pullulan to form the HLC/pullulan hydrogel, wherein the NaIO{sub 4} 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. - Graphical abstract: The HLC/pullulan hydrogels were prepared by dialysis, wet granulation and UV radiation after various MWs of pullulan and HLC were crosslinked with NaIO{sub 4}, and injected subcutaneously into Kunming mouse. The formation of HLC/pullulan hydrogels is due to the amide bond linkage with the amino group of HLC and the aldehyde groups in pullulan aqueous media after crosslinking by NaIO{sub 4}. HLC/pullulan hydrogels may have therapeutic value as efficient soft tissue fillers, with reduced inflammation, improved cell adhesion and

  12. QUANTITATIVE FLUORESCENCE IN SITU HYBRIDIZATION OF AUREOBASIDIUM PULLULANS ON MICROSCOPE SLIDES AND LEAF SURFACES. (R823845)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

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

    Science.gov (United States)

    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. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

    Shivananda, C. S.; Rao, B. Lakshmeesha; Madhukumar, R.; Asha, S. [Department of Studies in Physics, Mangalore University, Mangalagangotri – 574 199 (India); Sarojini, B. K. [Department of Industrial Chemistry, Mangalore University, Mangalagangotri, Mangalore –574 199 (India); Somashekhar, R. [Department of Studies in Physics, University of Mysore, Manasagangotri, Mysore – 570 006 (India); Sangappa, Y., E-mail: syhalabhavi@yahoo.co.in [Department of Studies in Physics, Mangalore University, Mangalagangotri – 574 199 (India); School of Material Science and Engineering, Georgia Institute of Technology, Atlanta, GA 30332 (United States)

    2016-05-23

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

  15. Oxidative stability of pullulan electrospun fibers containing fish oil

    DEFF Research Database (Denmark)

    García Moreno, Pedro Jesús; Damberg, Cecilie; Chronakis, Ioannis S.

    2017-01-01

    The effect of oil content and addition of natural antioxidants on the morphology and oxidative stability of pullulan ultra-thin fibers loaded with fish oil and obtained by electrospinning was investigated. Pullulan sub-micron fibers containing 10 and 30wt% fish oil were prepared and both presented...... into food matrices. These results show the feasibility to encapsulate fish oil in pullulan ultra-thin fibers and to improve their oxidative stability by adding natural antioxidants such as δ-tocopherol and rosemary extract. Therefore, this study might open up new opportunities for further technological...... development in the production of omega-3 nanodelivery systems, which have potential applications in different types of fortified foods. Encapsulation of fish oil in electrospun pullulan fibers stabilized by natural antioxidants....

  16. Aureobasidium melanogenum: a native of dark biofinishes on oil treated wood.

    Science.gov (United States)

    van Nieuwenhuijzen, Elke J; Houbraken, Jos A M P; Meijer, Martin; Adan, Olaf C G; Samson, Robert A

    2016-05-01

    The genus Aureobasidium, which is known as a wood staining mould, has been detected on oil treated woods in the specific stain formation called biofinish. This biofinish is used to develop a new protective, self-healing and decorative biotreatment for wood. In order to understand and control biofinish formation on oil treated wood, the occurrence of different Aureobasidium species on various wood surfaces was studied. Phenotypic variability within Aureobasidium strains presented limitations of morphological identification of Aureobasidium species. PCR amplification and Sanger sequencing of ITS and RPB2 were used to identify the culturable Aureobasidium species composition in mould stained wood surfaces with and without a biofinish. The analysed isolates showed that several Aureobasidium species were present and that Aureobasidium melanogenum was predominantly detected, regardless of the presence of a biofinish and the type of substrate. A. melanogenum was detected on wood samples exposed in the Netherlands, Cameroon, South Africa, Australia and Norway. ITS-specific PCR amplification, cloning and sequencing of DNA extracted from biofinish samples confirmed results of the culturing based method: A. melanogenum is predominant within the Aureobasidium population of biofinishes on pine sapwood treated with raw linseed oil and the outdoor placement in the Netherlands.

  17. Pullulan microcarriers for bone tissue regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Aydogdu, Hazal [Middle East Technical University, Department of Biomedical Engineering, Ankara 06800 (Turkey); Keskin, Dilek [Middle East Technical University, Department of Biomedical Engineering, Ankara 06800 (Turkey); Middle East Technical University, Department of Engineering Sciences, Ankara 06800 (Turkey); METU BIOMATEN Center of Excellence in Biomaterials and Tissue Engineering, Ankara 06800 (Turkey); Baran, Erkan Turker, E-mail: erkanturkerbaran@gmail.com [METU BIOMATEN Center of Excellence in Biomaterials and Tissue Engineering, Ankara 06800 (Turkey); Tezcaner, Aysen, E-mail: tezcaner@metu.edu.tr [Middle East Technical University, Department of Biomedical Engineering, Ankara 06800 (Turkey); Middle East Technical University, Department of Engineering Sciences, Ankara 06800 (Turkey); METU BIOMATEN Center of Excellence in Biomaterials and Tissue Engineering, Ankara 06800 (Turkey)

    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. - Highlights: • Porous PULL microspheres were prepared as cell carrier for the first time. • Mineralization on the microspheres improved their mechanical properties. • Mineralization and SF coating enhanced cell proliferation on PULL microspheres.

  18. Pullulan microcarriers for bone tissue regeneration

    International Nuclear Information System (INIS)

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

    2016-01-01

    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. - Highlights: • Porous PULL microspheres were prepared as cell carrier for the first time. • Mineralization on the microspheres improved their mechanical properties. • Mineralization and SF coating enhanced cell proliferation on PULL microspheres.

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

    Directory of Open Access Journals (Sweden)

    Choudhury Anirban

    2012-03-01

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

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

    Science.gov (United States)

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

    2017-01-01

    Pullulan based films possess several advantages, including high transparency, low toxicity, good biodegradability, good mechanical properties, and low oxygen permeability, are preferable for food packaging. The application of pullulan films on food packaging, however, has inherent disadvantage of high water solubility. In this study, glutaraldehyde and glycerol were used as the cross-linking reagent and the plasticizer respectively to improve water resistance and physical properties of the pullulan films. Effects of cross-linking degree on physical properties, including water absorptions, swelling behaviors, water vapor permeability and tensile strengths of films were evaluated. FTIR results demonstrated that the pullulan films were successfully cross-linked by glutaraldehyde. The tensile strength of pullulan films could be enhanced significantly (P permeability. © 2016 Institute of Food Technologists®.

  1. Affect the Cross Linking Degree and Polymer Composition on the Mechanical Properties of Poly (vinyl alcohol/ Pullu-lan Films

    Directory of Open Access Journals (Sweden)

    Shemaa Abdul Satar Soud

    2018-04-01

    Full Text Available In this study Cross-linked PVA/Pullulan film was prepared. Cross-linking reaction done by addi-tion of gluteraldehyde at different reaction time (10,30and 60 min. Chemical interaction, me-chanical, thermal properties, water solubility and film morphology was studied for cross-linked PVA/Pullulan, PVA and Pullulan only. Thus FTIR investigated formation of hydrogen bonding between pullulan and PVA with (GA. Tensile strength, tensile modulus and elongation (% at break for PVA/Pullulan film was improved with addition of (GA as the reaction time proceed equivalent with increasing PVA content

  2. Preactivated thiolated pullulan as a versatile excipient for mucosal drug targeting.

    Science.gov (United States)

    Leonaviciute, Gintare; Suchaoin, Wongsakorn; Matuszczak, Barbara; Lam, Hung Thanh; Mahmood, Arshad; Bernkop-Schnürch, Andreas

    2016-10-20

    The purpose of the present study was to generate a novel mucoadhesive thiolated pullulan with protected thiol moieties and to evaluate its suitability as mucosal drug delivery system. Two different synthetic pathways: bromination-nucleophilic substitution and reductive amination including periodate cleavage were utilized to synthesize such thiolated pullulans. The thiomer (pullulan-cysteamine) with the highest amount of free thiol groups was further enrolled in a reaction with 6-mercaptonicotinamide and its presence in pullulan structure was confirmed via NMR analysis. Furthermore, unmodified, thiolated and preactivated thiolated pullulan were investigated in terms of mucoadhesion via rotating cylinder studies and rheological synergism method as well as their toxicity potential over Caco-2 cells. Comparing both methods the reductive amination seems to be the method of choice resulting in comparatively higher coupling rates. Using this procedure pullulan-cysteamine conjugate displayed 1522±158μmol immobilized thiol groups and 280±70μmol free thiol groups per gram polymer. Furthermore, 82% of free thiol groups on this conjugate were linked with 6-mercaptonicotinamide (6-MNA). The adhesion time on the rotating cylinder was up to 46-fold prolonged in case of the thiolated polymer and up to 75-fold in case of the preactivated polymer. Rheological measurements of modified pullulan samples showed 98-fold and 160-fold increase in dynamic viscosity upon the addition of mucus within 60min, whereas unmodified pullulan did not show an increase in viscosity at all. Both conjugates had a minor effect on Caco-2 cell viability. Because of these features preactivated thiolated pullulan seems to represent a promising type of mucoadhesive polymers for the development of various mucosal drug delivery systems. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

    OpenAIRE

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

    2012-01-01

    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, X1), superdisintigrant (sodium starch glycolate, X2), and direct compression aid ...

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

    Science.gov (United States)

    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

  5. Incorporation of Rutin in Electrospun Pullulan/PVA Nanofibers for Novel UV-Resistant Properties.

    Science.gov (United States)

    Qian, Yongfang; Qi, Mengjie; Zheng, Laijiu; King, Martin W; Lv, Lihua; Ye, Fang

    2016-06-23

    This study aimed to investigate the incorporation of rutin into electrospun pullulan and poly(vinyl alcohol) (PVA) nanofibers to obtain ultraviolet (UV)-resistant properties. The effect of weight ratios between pullulan and PVA, and the addition of rutin on the nanofibers' morphology and diameters were studied and characterized by scanning electron microscopy (SEM). Fourier transform infrared (FTIR) analysis was utilized to investigate the interaction between pullulan and PVA, as well as with rutin. The results showed that the inclusion of PVA results in the increase in the fiber's diameter. The addition of rutin had no obvious effect on the fibers' average diameters when the content of rutin was less than 7.41%. FTIR results indicated that a hydrogen bond formed between pullulan and PVA, also between these polymers and rutin. Moreover, the addition of rutin could enhance the mechanical properties due to its stiff structure and could decrease the transmittance of UVA and UVB to be fewer than 5%; meanwhile, the value of ultraviolet protection factor (UPF) reached more than 40 and 50 when the content of rutin was 4.46% and 5.67%, respectively. Therefore, the electrospun pullulan/PVA/rutin nanofibrous mats showed excellent UV resistance and have potential applications in anti-ultraviolet packaging and dressing materials.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

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

    International Nuclear Information System (INIS)

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

    2015-01-01

    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. (paper)

  9. Synthesis and Characterization of Self-Assembled Nanogels Made of Pullulan

    Directory of Open Access Journals (Sweden)

    Sílvia A. Ferreira

    2011-03-01

    Full Text Available Self-assembled nanogels made of hydrophobized pullulan were obtained using a versatile, simple, reproducible and low-cost method. In a first reaction pullulan was modified with hydroxyethyl methacrylate or vinyl methacrylate, further modified in the second step with hydrophobic 1-hexadecanethiol, resulting as an amphiphilic material, which self-assembles in water via the hydrophobic interaction among alkyl chains. Structural features, size, shape, surface charge and stability of the nanogels were studied using hydrogen nuclear magnetic resonance, fluorescence spectroscopy, cryo-field emission scanning electron microscopy and dynamic light scattering. Above the critical aggregation concentration spherical polydisperse macromolecular micelles revealed long-term colloidal stability in aqueous medium, with a nearly neutral negative surface charge and mean hydrodynamic diameter in the range 100–400 nm, depending on the polymer degree of substitution. Good size stability was observed when nanogels were exposed to potential destabilizing pH conditions. While the size stability of the nanogel made of pullulan with vinyl methacrylate and more hydrophobic chains grafted was affected by the ionic strength and urea, nanogel made of pullulan with hydroxyethyl methacrylate and fewer hydrophobic chains grafted remained stable.

  10. Pullulan as a potent green inhibitor for corrosion mitigation of aluminum composite: Electrochemical and surface studies.

    Science.gov (United States)

    B P, Charitha; Rao, Padmalatha

    2018-06-01

    This work emphasizes the corrosion inhibition ability of pullulan, an environmentally benign fungal polysaccharide on acid corrosion of 6061Aluminum-15% (v) SiC (P) composite material (Al-CM). The electrochemical measurements such as potentiodynamic polarization (PDP) and electrochemical impedance spectroscopy (EIS) studies were carried out for the corrosion inhibition studies. Conditions were optimized to obtain maximum inhibition efficiency, by performing the experiment at varying concentrations of inhibitor, in the temperature range of 308K- 323K. Surface morphology studies were done to reaffirm the adsorption of inhibitor on the surface of composite material. Pullulan acted as mixed type of inhibitor with a maximum efficiency of 89% at 303K for the addition of 1.0 gL -1 of inhibitor. Evaluation of kinetic and thermodynamic parameters revealed that inhibitor underwent physical adsorption onto the surface of Al-CM and obeyed Freundlich adsorption isotherm. The surface characterization like SEM-EDX, AFM confirmed the adsorption of pullulan molecule. Pullulan can be considered as effective, eco friendly green inhibitor for the corrosion control of Al-CM. Copyright © 2018 Elsevier B.V. All rights reserved.

  11. Pullulan-based composite scaffolds for bone tissue engineering: Improved osteoconductivity by pore wall mineralization.

    Science.gov (United States)

    Amrita; Arora, Aditya; Sharma, Poonam; Katti, Dhirendra S

    2015-06-05

    Porous hydrogels have been explored for bone tissue engineering; however their poor mechanical properties make them less suitable as bone graft substitutes. Since incorporation of fillers is a well-accepted method for improving mechanical properties of hydrogels, in this work pullulan hydrogels were reinforced with nano-crystalline hydroxyapatite (nHAp) (5 wt% nHAp in hydrogel) and poly(3-hydroxybutyrate) (PHB) fibers (3 wt% fibers in hydrogel) containing nHAp (3 wt% nHAp in fibers). Addition of these fillers to pullulan hydrogel improved compressive modulus of the scaffold by 10 fold. However, the hydrophilicity of pullulan did not support adhesion and spreading of cells. To overcome this limitation, porous composite scaffolds were modified using a double diffusion method that enabled deposition of hydroxyapatite on pore walls. This method resulted in rapid and uniform coating of HAp throughout the three-dimensional scaffolds which not only rendered them osteoconductive in vitro but also led to an improvement in their compressive modulus. These results demonstrate the potential of mineralized pullulan-based composite scaffolds in non-load bearing bone tissue engineering. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Augmented liver targeting of exosomes by surface modification with cationized pullulan.

    Science.gov (United States)

    Tamura, Ryo; Uemoto, Shinji; Tabata, Yasuhiko

    2017-07-15

    Exosomes are membrane nanoparticles containing biological substances that are employed as therapeutics in experimental inflammatory models. Surface modification of exosomes for better tissue targetability and enhancement of their therapeutic ability was recently attempted mainly using gene transfection techniques. Here, we show for the first time that the surface modification of exosomes with cationized pullulan, which has the ability to target hepatocyte asialoglycoprotein receptors, can target injured liver and enhance the therapeutic effect of exosomes. Surface modification can be achieved by a simple mixing of original exosomes and cationized pullulan and through an electrostatic interaction of both substances. The exosomes modified with cationized pullulan were internalized into HepG2 cells in vitro to a significantly greater extent than unmodified ones and this internalization was induced through the asialoglycoprotein receptor that was specifically expressed on HepG2 cells and hepatocytes. When injected intravenously into mice with concanavalin A-induced liver injury, the modified exosomes accumulated in the liver tissue, resulting in an enhanced anti-inflammatory effect in vivo. It is concluded that the surface modification with cationized pullulan promoted accumulation of the exosomes in the liver and the subsequent biological function, resulting in a greater therapeutic effect on liver injury. Exosomes have shown potentials as therapeutics for various inflammatory disease models. This study is the first to show the specific accumulation of exosomes in the liver and enhanced anti-inflammatory effect via the surface modification of exosomes using pullulan, which is specifically recognized by the asialoglycoprotein receptor (AGPR) on HepG2 cells and hepatocytes. The pullulan was expressed on the surface of PKH-labeled exosomes, and it led increased accumulation of PKH into HepG2 cells, whereas the accumulation was canceled by AGPR inhibitor. In the mouse

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

    Science.gov (United States)

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

    2015-05-01

    There has been a growing interest in the use of natural materials as a delivery mechanism for antimicrobials and coatings in foods. The aim of the present study was to evaluate the effectiveness of pullulan coatings to improve internal quality and shelf-life of fresh eggs during 10 wk of storage at 25 and 4 °C. Three treatments of eggs were evaluated as follows; non-coated (control; C), coated with pullulan (P), and coated with pullulan containing nisin (N). The effects of the pullulan coatings on microbiological qualities, physical properties, and freshness parameters were investigated and compared with non-coated eggs. For non-coated eggs, as storage time increased, yolk index, albumen index, and Haugh unit value decreased and weight loss increased. However, pullulan coatings (P or N) minimized weight loss (eggs (with a final B grade) 3 wk longer than non-coated eggs at 25 °C. At 4 °C, both P- and N-coated eggs went from AA to A grade after 9 wk and maintained the grade for 10 wk (4 wk longer than that of non-coated eggs). This study is the first to demonstrate that pullulan coatings can preserve the internal quality, prolong the shelf-life, and minimize weight loss of fresh eggs. © 2015 Institute of Food Technologists®

  14. Role of Pullulan in preparation of ceria nanoparticles and investigation of their biological activities

    Science.gov (United States)

    Khorrami, Mohammad Bagher; Sadeghnia, Hamid Reza; Pasdar, Alireza; Ghayour-Mobarhan, Majid; Riahi-Zanjani, Bamdad; Darroudi, Majid

    2018-04-01

    Throughout this work, a facile, environmental-friendly, and "green" method is delineated for preparing ceria nanoparticles (CNPs), which utilizes nontoxic and renewable degraded polysaccharide polymer including pullulan as a natural matrix. Pullulan behaves as a suitable stabilizing (capping) agent for CNPs that are effectively formed at various high temperatures, while they are structurally analyzed through different techniques such as TGA/DTG, XRD, FESEM, and FTIR instruments. This procedure was found to be comparable to the ones that were acquired from conventional preparation methods that employ hazardous materials, which confirms this approach to be an exquisite alternative in preparing CNPs through the benefit of bioorganic materials. The in vitro cytotoxicity studies on Neuro2A cells have mentioned nontoxic particles in a range of concentrations (0.97-125 μg/ml) and thus, we reckon that the prepared particular CNPs will have persistent utilization in various fields of biology and medicine.

  15. Oxidative stability of pullulan nanofibers loaded with fish oil: effect of oil content and antioxidants addition

    DEFF Research Database (Denmark)

    García Moreno, Pedro Jesús; Damberg, C.; Stephansen, K.

    Electrospinning processing is a promising technique for the encapsulation of thermolabile bioactive compounds (e.g. fish oil) since it does not require the use of heat. Furthermore, the nano-microfibers (NMF) obtained present a reduced size, which makes them easier to disperse in food matrices...... compared to traditional encapsulates (e.g. microcapsules produced by spray-drying). Biopolymers such as proteins and polysaccharides are required for the production of food-grade NMF. In this sense, pullulan, which is a food-approved polysaccharide, is an interesting encapsulating material due to its high...... electrospinnability and low oxygen permeability. In light of the above, the aim of this work was to investigate the oxidative stability of omega-3 enriched pullulan NMF. First, the influence of fish oil content (10-20-30 %) on the properties of the electrospinning solutions (e.g. viscosity, conductivity and surface...

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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.

  18. Self-Assembled Nanoparticles of Glycyrrhetic Acid-Modified Pullulan as a Novel Carrier of Curcumin

    Directory of Open Access Journals (Sweden)

    Roufen Yuan

    2014-08-01

    Full Text Available Glycyrrhetic acid (GA-modified pullulan nanoparticles (GAP NPs were synthesized as a novel carrier of curcumin (CUR with a degree of substitution (DS of GA moieties within the range of 1.2–6.2 groups per hundred glucose units. In the present study, we investigated the physicochemical characteristics, release behavior, in vitro cytotoxicity and cellular uptake of the particles. Self-assembled GAP NPs with spherical shapes could readily improve the water solubility and stability of CUR. The CUR release was sustained and pH-dependent. The cellular uptake of CUR-GAP NPs was confirmed by green fluorescence in the cells. An MTT study showed CUR-GAP NPs with higher cytotoxicity in HepG2 cells than free CUR, but GAP NPs had no significant cytotoxicity. GAP is thus an excellent carrier for the solubilization, stabilization, and controlled delivery of CUR.

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

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jingyun, E-mail: wangjingyun67@dlut.edu.cn [School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024 (China); Cui, Shuang [School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024 (China); Bao, Yongming, E-mail: biosci@dlut.edu.cn [School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024 (China); Xing, Jishuang [School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024 (China); Hao, Wenbo [Department of Physics and Chemistry, Heihe University, Heihe 164300 (China)

    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 4.3 mg/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. - Highlights: • Tocopheryl pullulan-based (PUTC) self-assembling nanomicelles were fabricated. • These micelles showed low CMC and dispersed uniformly with regular spherical shape. • High entrapment efficiency and in vitro sustained release of HCPT in PUTC micelles • HCPT–PUTC micelles accumulated in cell nuclei and showed higher anticancer activity.

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

    International Nuclear Information System (INIS)

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

    2014-01-01

    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 4.3 mg/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. - Highlights: • Tocopheryl pullulan-based (PUTC) self-assembling nanomicelles were fabricated. • These micelles showed low CMC and dispersed uniformly with regular spherical shape. • High entrapment efficiency and in vitro sustained release of HCPT in PUTC micelles • HCPT–PUTC micelles accumulated in cell nuclei and showed higher anticancer activity.

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

    Directory of Open Access Journals (Sweden)

    Wan L

    2016-12-01

    Full Text Available Li Wan,1,* Xinglei Yao,1–3,* Francesco Faiola,3 Bojun Liu,4 Tianyuan Zhang,2 Yasuhiko Tabata,5 Hiroyuki Mizuguchi,6 Shinsaku Nakagawa,7 Jian-Qing Gao,2 Robert Chunhua Zhao1 1Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, Peking Union Medical College Hospital, Center of Excellence in Tissue Engineering Chinese Academy of Medical Sciences, Beijing, 2Institute of Pharmaceutics, Zhejiang University, Hangzhou, 3State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, 4YouAn Hospital, Capital Medical University, Beijing, People’s Republic of China; 5Department of Biomaterials, Field of Tissue Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, 6Department of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, 7Department of Biotechnology and Therapeutics, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan *These authors contributed equally to this work Abstract: Mesenchymal stem cells (MSCs are adult stem cells with multilineage potential, which makes them attractive tools for regenerative medicine applications. Efficient gene transfer into MSCs is essential not only for basic research in developmental biology but also for therapeutic applications involving gene-modification in regenerative medicine. Adenovirus vectors (Advs can efficiently and transiently introduce an exogenous gene into many cell types via their primary receptors, the coxsackievirus and adenovirus receptors, but not into MSCs, which are deficient in coxsackievirus and adenovirus receptors expression. To overcome this problem, we developed an Adv coated with a spermine-pullulan (SP cationic polymer and investigated its physicochemical properties and internalization mechanisms. We demonstrated that the SP

  2. Optimization of physico-chemical and nutritional parameters for ...

    African Journals Online (AJOL)

    Optimization of physico-chemical and nutritional parameters for pullulan production by a mutant of thermotolerant Aureobasidium pullulans, in fed batch ... minutes, having killing rate of 70% level, produced 6 g l-1 higher pullulan as compared to the wild type without loosing thermotolerant and non-melanin producing ability.

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

    Science.gov (United States)

    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. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Effect of pullulan nanoparticle surface charges on HSA complexation and drug release behavior of HSA-bound nanoparticles.

    Directory of Open Access Journals (Sweden)

    Xiaojun Tao

    Full Text Available Nanoparticle (NP compositions such as hydrophobicity and surface charge are vital to determine the presence and amount of human serum albumin (HSA binding. The HSA binding influences drug release, biocompatibility, biodistribution, and intercellular trafficking of nanoparticles (NPs. Here, we prepared 2 kinds of nanomaterials to investigate HSA binding and evaluated drug release of HSA-bound NPs. Polysaccharides (pullulan carboxyethylated to provide ionic derivatives were then conjugated to cholesterol groups to obtain cholesterol-modified carboxyethyl pullulan (CHCP. Cholesterol-modified pullulan (CHP conjugate was synthesized with a similar degree of substitution of cholesterol moiety to CHCP. CHCP formed self-aggregated NPs in aqueous solution with a spherical structure and zeta potential of -19.9 ± 0.23 mV, in contrast to -1.21 ± 0.12 mV of CHP NPs. NPs could quench albumin fluorescence intensity with maximum emission intensity gradually decreasing up to a plateau at 9 to 12 h. Binding constants were 1.12 × 10(5 M(-1 and 0.70 × 10(5 M(-1 to CHP and CHCP, respectively, as determined by Stern-Volmer analysis. The complexation between HSA and NPs was a gradual process driven by hydrophobic force and inhibited by NP surface charge and shell-core structure. HSA conformation was altered by NPs with reduction of α-helical content, depending on interaction time and particle surface charges. These NPs could represent a sustained release carrier for mitoxantrone in vitro, and the bound HSA assisted in enhancing sustained drug release.

  5. TEMPO-mediated oxidation of pullulan and influence of ionic strength and linear charge density on the dimensions of the obtained polyelectrolyte chains

    NARCIS (Netherlands)

    Nooy, A.E.J. de; Besemer, A.C.; Bekkum, H. van; Dijk, J.A.P.P. van; Smit, J.A.M.

    1996-01-01

    The applied pH during the TEMPO-mediated (2,2,6,6-tetramethylpiperidine-1-oxyl) oxidation of the polysaccharide pullulan has a substantial influence on the molar mass distribution of the products. The least degradation was found in the pH range 9.2-9.7 and especially at higher pH, significant

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

    Science.gov (United States)

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

    2016-06-05

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

  7. Oil type and cross-linking influence growth of Aureobasidium melanogenum on vegetable oils as a single carbon source.

    Science.gov (United States)

    Peeters, Loes H M; Huinink, Hendrik P; Voogt, Benjamin; Adan, Olaf C G

    2018-03-12

    Aureobasidium melanogenum is the main fungus found in a spontaneously formed biofilm on a oil-treated wood. This dark colored biofilm functions as a protective coating. To better understand biofilm formation, in this study A. melanogenum was cultured on olive oil and raw linseed oil. Metabolic activity and oil conversion were measured. The results show that A. melanogenum is able to grow on linseed oil and olive oil as a single carbon source. The fungus produces the enzyme lipase to convert the oil into fatty acids and glycerol. Metabolic activity and oil conversion were equal on linseed oil and olive oil. The fungus was not able to grow on severe cross-linked linseed oil, meaning that the degree of cross-linking of the oil is important for growth of A. melanogenum. Dark coloring of the colony was seen on linseed oil, which might be a stress response on the presence of autoxidation products in linseed oil. The colony on olive oil showed delayed melanin production indicating an inhibitory effect of olive oil on melanin production. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

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

    Energy Technology Data Exchange (ETDEWEB)

    Atila, Deniz [Department of Engineering Sciences, Middle East Technical University (Turkey); Keskin, Dilek [Department of Engineering Sciences, Middle East Technical University (Turkey); Biomaterials and Tissue Engineering Center of Excellence, Middle East Technical University (Turkey); Tezcaner, Ayşen, E-mail: tezcaner@metu.edu.tr [Department of Engineering Sciences, Middle East Technical University (Turkey); Biomaterials and Tissue Engineering Center of Excellence, Middle East Technical University (Turkey)

    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 (P{sub 80}/CA{sub 20}, P{sub 50}/CA{sub 50}, and P{sub 20}/CA{sub 80}%) 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 10 min. Fiber morphologies of P{sub 50}/CA{sub 50} were more uniform without phase separation and this group was crosslinked most efficiently among groups. It was found that mechanical properties of P{sub 20}/CA{sub 80} and P{sub 50}/CA{sub 50} were higher than that of P{sub 80}/CA{sub 20.} After crosslinking strain values of P{sub 50}/CA{sub 50} scaffolds were improved and these scaffolds became more stable. Unlike P{sub 80}/CA{sub 20,} uncrosslinked P{sub 50}/CA{sub 50} and P{sub 20}/CA{sub 80} were not lost in PBS. Among all groups, crosslinked P{sub 50}/CA{sub 50} 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 P{sub 50}/CA{sub 50} 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 P{sub 50}/CA{sub 50} scaffolds were proposed as candidate cell carriers for bone tissue engineering applications. - Highlights: • Crosslinked 3D electrospun P/CA scaffolds were prepared for the first time. • CA

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

    Science.gov (United States)

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

    2014-12-01

    The amalgamation of chemotherapy and gene therapy is promising treatment option for cancer. In this study, novel biocompatible self-assembled nanocomplexes (NCs) between carboxylmethylated pullulan t335 (CMP) with polyallylamine (CMP-PAA NCs) were developed for plasmid DNA (pDNA) and pH-sensitive doxorubicin (DOX) delivery. DOX was conjugated to CMP (DOX-CMP) via hydrazone and confirmed by FTIR and 1H-NMR. In vitro release studies of pH-sensitive DOX-CMP conjugate showed 23 and 85 % release after 48 h at pH 7.4 (physiological pH) and pH 5 (intracellular/tumoral pH), respectively. The CMP-PAA NCs or DOX-CMP-PAA NCs self-assembled into a nanosized (successful cellular uptake of DOX-CMP-PAA NCs in HEK293 cells. Thus, NCs hold great potential for targeted pDNA and pH-sensitive intratumoral drug delivery.

  10. Biodiesel production from Jatropha curcas oil catalyzed by whole ...

    African Journals Online (AJOL)

    my mord

    2013-07-03

    Jul 3, 2013 ... catalyzed by whole cells of Aureobasidium pullulans var. melanogenum ... friendly and renewable fuel that can be used directly in diesel engines ... methanol (or supercritical ethanol) transesterification is not commercially ...

  11. Iodoacetyl-functionalized pullulan: A supplemental enhancer for single-domain antibody-polyclonal antibody sandwich enzyme-linked immunosorbent assay for detection of survivin.

    Science.gov (United States)

    Matsushita, Takahiko; Arai, Hidenao; Koyama, Tetsuo; Hatano, Ken; Nemoto, Naoto; Matsuoka, Koji

    2017-11-01

    Survivin, an inhibitor of the apoptosis protein family, is a potent tumor marker for diagnosis and prognosis. The enzyme-linked immunosorbent assay (ELISA) is one of the methods that has been used for detection of survivin. However, ELISA has several disadvantages caused by the use of conventional antibodies, and we have therefore been trying to develop a novel ELISA system using camelid single-domain antibodies (VHHs) as advantageous replacements. Here we report a supplemental approach to improve the VHH-polyclonal antibody sandwich ELISA for survivin detection. Iodoacetyl-functionalized pullulan was synthesized, and its thiol reactivity was characterized by a model reaction with l-cysteine. The thiophilic pullulan was applied to an immunoassay asan additive upon coating of standard assay plates with an anti-survivin VHH fusion protein with C-terminal cysteine. The results showed that the mole ratio of the additive to VHH had a significant effect on the consequent response. Mole ratios of 0.07, 0.7, and 7 led to 90% lower, 15% higher, and 69% lower responses, respectively, than the response of a positive control in which no additive was used. The background levels observed in any additive conditions were as low as that of a negative control lacking both VHH and the additive. These results indicate the applicability of the thiol-reactive pullulan as a response enhancer to VHH-based ELISA. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    International Nuclear Information System (INIS)

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

    2014-01-01

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

  13. Optimization of physico-chemical and nutritional parameters for ...

    African Journals Online (AJOL)

    hope&shola

    2010-10-25

    Oct 25, 2010 ... industrial production in order to reduce the cost of production. ... is of great economic importance with increased appli- ... industries (Seviour et al., 1992; Leathers, 2003). .... The various process parameters influencing pullulan production ..... formation by Aureobasidium pullulans in stirred tanks. Enzyme.

  14. Cloning, recombinant expression and characterization of a new ...

    African Journals Online (AJOL)

    A new amylase gene APGA1 was cloned from Aureobasidium pullulans NRRL 12974 and expressed in Pichia pastoris. This is the first report on cloning and expression of amylolytic gene from the industrially important microorganism A. pullulans. The purified recombinant protein with MW of 66 kDa and specific activity of ...

  15. Eudragit E100 and Polysaccharide Polymer Blends as Matrices for ...

    African Journals Online (AJOL)

    Methods: LB, SCMC and E100 were blended in their dry (as purchased) state or modified by aqueous blending and subsequent lyophilization, prior to use as matrices in tablets. ... pullulan from Aureobasidium pullulans, 3-(3,4- .... the frozen polymer before sublimation and drying). Subsequently, milling generated a more.

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

    Science.gov (United States)

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

    2013-08-01

    The objective of these experiments was to measure in vitro hydrolytic digestion and glycemic and insulinemic responses of select carbohydrate blends, all containing the novel carbohydrate soluble corn fiber (SCF). Two SCF that varied in their method of production were used to formulate the carbohydrate blends. One set of blends contained a SCF that was spray dried (SCFsd) and then blended with different amounts of either pullulan, sorbitol, or fructose. The other set of blends contained a SCF produced using longer evaporation time (SCF) and then blended with different ratios of pullulan, sorbitol, and fructose. Free sugar concentrations found in the individual SCFsd and SCF substrates were low but varied. Spray-dried soluble corn fiber had a reduced free sugar concentration compared with SCF (2.8 vs. 14.2%). Glucose was the main free sugar found in both SCFsd and SCF but at different concentrations (2.7 vs. 12.7%, respectively). The majority of the SCFsd blends were completely hydrolyzed to their monosaccharide components. Glucose accounted for most of the hydrolyzed monosaccharides for SCFsd and all the SCFsd blends. Hydrolyzed monosaccharide concentrations for the SCF:pullulan:sorbitol:fructose blends followed similar trends to the SCFsd blends where greater percentages of fructose and sorbitol resulted in decreased (P sorbitol. Total released monosaccharides were high in SCFsd blends containing either 50% fructose or sorbitol, but the combination resulted in reduced concentrations of glucose released (P sorbitol:fructose blends also had intermediate to high released monosaccharides as a result of in vitro hydrolytic digestion. All SCF blends resulted in decreased glycemic and insulinemic responses compared with the maltodextrin control (P sorbitol in the blends had the greatest impact on glycemic and insulinemic responses, even at concentrations as low as 5% of the blends. Overall, SCF and their blends may prove beneficial as components of low glycemic

  17. Production, Purification, and Gene Cloning of a β-Fructofuranosidase with a High Inulin-hydrolyzing Activity Produced by a Novel Yeast Aureobasidium sp. P6 Isolated from a Mangrove Ecosystem.

    Science.gov (United States)

    Jiang, Hong; Ma, Yan; Chi, Zhe; Liu, Guang-Lei; Chi, Zhen-Ming

    2016-08-01

    After screening of over 300 yeast strains isolated from the mangrove ecosystems, it was found that Aureobasidium sp. P6 strain had the highest inulin-hydrolyzing activity. Under the optimal conditions, this yeast strain produced an inulin-hydrolyzing activity of 30.98 ± 0.8 U/ml after 108 h of a 10-l fermentation. After the purification, a molecular weight of the enzyme which had the inulin-hydrolyzing activity was estimated to be 47.6 kDa, and the purified enzyme could actively hydrolyze both sucrose and inulin and exhibit a transfructosylating activity at 30.0 % sucrose, converting sucrose into fructooligosaccharides (FOS), indicating that the purified enzyme was a β-D-fructofuranosidase. After the full length of a β-D-fructofuranosidase gene (accession number KU308553) was cloned from Aureobasidium sp. P6 strain, a protein deduced from the cloned gene contained the conserved sequences MNDPNGL, RDP, ECP, FS, and Q of a glycosidehydrolase GH32 family, respectively, but did not contain a conserved sequence SVEVF, and the amino acid sequence of the protein from Aureobasidium sp. P6 strain had a high similarity to that of the β-fructofuranosidase from any other fungal strains. After deletion of the β-D-fructofuranosidase gene, the disruptant still had low inulin hydrolyzing and invertase activities and a trace amount of the transfructosylating activity, indicating that the gene encoding an inulinase may exist in the Aureobasidium sp. P6 strain.

  18. Immobilized Burkholderia cepacia Lipase on pH-Responsive Pullulan Derivatives with Improved Enantioselectivity in Chiral Resolution

    Directory of Open Access Journals (Sweden)

    Li Xu

    2018-01-01

    Full Text Available A kind of pH-responsive particle was synthesized using modified pullulan polysaccharide. The synthesized particle possessed a series of merits, such as good dispersity, chemical stability and variability of particle size, making it a suitable carrier for enzyme immobilization. Then, Burkholderia cepacia lipase (BCL, a promising biocatalyst in transesterification reaction, was immobilized on the synthesized particle. The highest catalytic activity and immobilization efficiency were achieved at pH 6.5 because the particle size was obviously enlarged and correspondingly the adsorption surface for BCL was significantly increased. The immobilization enzyme loading was further optimized, and the derivative lipase was applied in chiral resolution. Under the optimal reaction conditions, the immobilized BCL showed a very good performance and significantly shortened the reaction equilibrium time from 30 h of the free lipase to 2 h with a conversion rate of 50.0% and ees at 99.2%. The immobilized lipase also exhibited good operational stability; after being used for 10 cycles, it still retained over 80% of its original activity. Moreover, it could keep more than 80% activity after storage for 20 days at room temperature in a dry environment. In addition, to learn the potential mechanism, the morphology of the particles and the immobilized lipase were both characterized with a scanning electron microscope and confocal laser scanning microscopy. It was found that the enlarged spherical surface of the particle in low pH values probably led to high immobilized efficiency, resulting in the improvement of enantioselectivity activity in chiral resolution.

  19. Genetics of trehalose biosynthesis in desert-derived Aureobasidium melanogenum and role of trehalose in the adaptation of the yeast to extreme environments.

    Science.gov (United States)

    Jiang, Hong; Liu, Guang-Lei; Chi, Zhe; Hu, Zhong; Chi, Zhen-Ming

    2018-04-01

    Melanin plays an important role in the stress adaptation of Aureobasidium melanogenum XJ5-1 isolated from the Taklimakan desert. A trehalose-6-phosphate synthase gene (TPS1 gene) was cloned from K5, characterized, and then deleted to determine the role of trehalose in the stress adaptation of the albino mutant K5. No stress response element and heat shock element were found in the promoter of the TPS1 gene. Deletion of the TPS1 gene in the albino mutant rendered a strain DT43 unable to synthesize any trehalose, but DT43 still could grow in glucose, suggesting that its hexokinase was insensitive to inhibition by trehalose-6-phosphate. Overexpression of the TPS1 gene enhanced trehalose biosynthesis in strain ET6. DT43 could not grow at 33 °C, whereas K5, ET6, and XJ5-1 could grow well at this temperature. Compared with K5 and ET6, DT43 was highly sensitive to heat shock treatment, high oxidation, and high desiccation, but all the three strains demonstrated the same sensitivity to UV light and high NaCl concentration. Therefore, trehalose played an important role in the adaptation of K5 to heat shock treatment, high oxidation, and high desiccation.

  20. Both a PKS and a PPTase are involved in melanin biosynthesis and regulation of Aureobasidium melanogenum XJ5-1 isolated from the Taklimakan desert.

    Science.gov (United States)

    Jiang, Hong; Liu, Guang-Lei; Chi, Zhe; Wang, Jian-Ming; Zhang, Ly-Ly; Chi, Zhen-Ming

    2017-02-20

    A PKS1 gene responsible for the melanin biosynthesis and a NPG1 gene in Aureobasidium melanogenum XJ5-1 were cloned and characterized. An ORF of the PKS1 gene encoding a protein with 2165 amino acids contained 6495bp while an ORF of the NPG1 gene encoding a protein with 340 amino acids had 1076bp. After analysis of their promoters, it was found that expression of both the PKS1 gene and the NPG1 gene was repressed by nitrogen sources and glucose, respectively. The PKS deduced from the cloned gene consisted of one ketosynthase, one acyl transferase, two acyl carrier proteins, one thioesterase and one cyclase while the PPTase belonged to the family Sfp-type. After disruption of the PKS1 gene and the NPG1 gene, expression of the PKS1 gene and the NPG1 gene and the melanin biosynthesis in the disruptants K5 and DP107 disappeared and expression of the PKS1 gene in the disruptant DP107 was also negatively influenced. However, after the NPG1 gene was complemented in the disruptant DP107, the melanin biosynthesis in the complementary strain BP17 was restored and expression of the PKS1 gene and the NPG1 gene was greatly enhanced, suggesting that the PKS was indeed activated and regulated by the PPTase and expression of the PKS1 gene and the NPG1 gene had a coordinate regulation. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Occurrence and diversity of marine yeasts in Antarctica environments

    Science.gov (United States)

    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.

  2. African Journal of Clinical and Experimental Microbiology - Vol 6, No ...

    African Journals Online (AJOL)

    Antistaphylococcal metabolite from Aureobasidium pullulans: production and characterization · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT ... Opportunistic infections and clinico-epidemiological factors in HIV/AIDS casesseen in a tertiary care hospital in Nepal · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT

  3. JPRS Report Science & Technology USSR: Life Sciences

    Science.gov (United States)

    1990-06-18

    Fractionated Irradiation [V. N. Malakhovskiy, M. I. Bokk; RADIOBIOLOGIYA, Vol 29 No 3, May-Jun 89] 41 Melanin Acts as Radioprotector [A. Patyko...Alternaria alternata, Aureobasidium pullulans, Tricho- derma viride; but Aspergillus niger, Cladosporium her- barum, Stemphylium ilicis, and Ulocladium atrum...of the above. Melanin -producing micromycetes predominated on materials subjected to storage both in open fields and sheds. Determinations of

  4. Pullulan-coated phospholipid and Pluronic F68 complex nanoparticles for carrying IR780 and paclitaxel to treat hepatocellular carcinoma by combining photothermal therapy/photodynamic therapy and chemotherapy

    Directory of Open Access Journals (Sweden)

    Wang D

    2017-12-01

    Full Text Available Dan Wang,1,* Sipei Zhang,1,* Tao Zhang,1 Guoyun Wan,1 Bowei Chen,1 Qingqing Xiong,2 Jie Zhang,3 Wenxue Zhang,4 Yinsong Wang1 1Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics, School of Pharmacy, Tianjin Medical University, 2Key Laboratory of Cancer Prevention and Therapy, National Clinical Research Center for Cancer, Tianjin Medical University Cancer Institute and Hospital, 3Surgery Department, 4Radiotherapy Department, Tianjin Medical University General Hospital, Tianjin, China *These authors contributed equally to this work Abstract: IR780, a near-infrared dye, can also be used as a photosensitizer both for photothermal therapy (PTT and photodynamic therapy (PDT. In this study, we designed a simple but effective nanoparticle system for carrying IR780 and paclitaxel, thus hoping to combine PTT/PDT and chemotherapy to treat hepatocellular carcinoma (HCC. This nanosystem, named PDF nanoparticles, consisted of phospholipid/Pluronic F68 complex nanocores and pullulan shells. IR780 and paclitaxel were loaded separately into PDF nanoparticles to form PDFI and PDFP nanoparticles, which had regular sphere shapes and relatively small sizes. Upon near-infrared laser irradiation at 808 nm, PDFI nanoparticles showed strong PTT/PDT efficacy both in vitro and in vivo. In MHCC-97H cells, the combined treatment of PDFI nanoparticles/laser irradiation and PDFP nanoparticles exhibited significant synergistic effects on inhibiting cell proliferation and inducing cell apoptosis and cell cycle arrest at G2/M phase. In MHCC-97H tumor-bearing mice, PDFI nanoparticles exhibited excellent HCC-targeting and accumulating capability after intravenous injection. Furthermore, the combined treatment of PDFI nanoparticles/laser irradiation and PDFP nanoparticles also effectively inhibited the tumor growth and the tumor angiogenesis in MHCC-97H tumor-bearing mice. In summary, we put forward a therapeutic

  5. (Cu, Zn) واﻟﺳﻣﺎد اﻟﻌﻧﺎﺻر اﻟﺻﻐرى اﻟﻧﺷﺎط اﻟﻣﯾﮐروﺑﯾوﻟوﺟﻲ ﻟﺗر

    African Journals Online (AJOL)

    Aureobasidium Pullulans. وَ. Candida ablicans ... أﯾﺿﺎً ﮐﺎﻧت أﺑﺣﺎث. [14]. اﻟﺗﻲ وﺟدا ﻓﯾﮭﺎ أن. )Auerobasidium pullulans(. ﺗﺳﺗطﯾﻊ إﻧﺗﺎج. اﻟﻣﯾﻼﻧﯾن. (Melanin). وھذا ﯾﺳﺗﺣث إﻧ. زﯾم.

  6. Microbial Influenced Corrosion (MIC) Study

    Science.gov (United States)

    2012-05-23

    fumigatus Fusarium oxysporum Fungal Consortium Penicillium oxalicum Rhodoturula sp . Trichoderma sp . Dosed with microbes known to influence Control...Hypocrea jecorina (FI-1) Penicillium oxalicum (FI-12) – Pleosporacea sp . (FI-17) Rhodoturala mucilaginosa (FI-7) – Ustilago maydis (FI-13) T t S t• es...and Dirt Accumulation • Fungal Consortium – Aspergillus sp (FI-19) Aureobasidium pullulans (FI-16) – Fusarium oxysporum (FI-6) Fusarium sp . (FI-18

  7. F-16 Microbially Influenced Corrosion (MIC) Characterization & Prevention Study

    Science.gov (United States)

    2011-05-12

    Staphylococcus epidermidis Fungal Consortium Aspergillus fumigatus Fusarium oxysporum Penicillium oxalicum Rhodoturula sp . Trichoderma sp . Control... sp . (FI-18) – Hypocrea jecorina (FI-1) Penicillium oxalicum (FI-12) – Pleosporacea sp . (FI-17) Rhodoturala mucilaginosa (FI-7) – Ustilago maydis (FI...Growth, or Soil and Dirt Accumulation • Fungal Consortium – Aspergillus sp (FI-19) Aureobasidium pullulans (FI-16) – Fusarium oxysporum (FI-6) Fusarium

  8. Antimicrobial activity of berberine--a constituent of Mahonia aquifolium.

    Science.gov (United States)

    Cernáková, M; Kostálová, D

    2002-01-01

    The antimicrobial activity of the protoberberine alkaloid, berberine, isolated from Mahonia aquifolium, was evaluated against 17 microorganisms including two Gram-negative bacteria--Pseudomonas aeruginosa and Escherichia coli (both resistant and sensitive), two Gram-positive bacteria--Bacillus subtilis and Staphylococcus aureus, Zoogloea ramigera, six filamentous fungi--Penicilium chrysogenum, Aspergillus niger, Aureobasidium pullulans (black and white strain), Trichoderma viride (original green strain and brown mutant), Fusarium nivale, Mycrosporum gypseum and two yeasts--Candida albicans and Saccharomyces cerevisiae. The IC50, minimum inhibitory concentration (MIC), minimum microbicidal concentration (MMC) and minimum microbistatic concentration (MMS) varied considerably depending on the microorganism tested, the sensitivity decreasing as follows: S. aureus > P. aeruginosa S (sensitive) > E. coli S > P. aeruginosa R (resistant) > E. coli R > B. subtilis > Z. ramigera > C. albicans > S. cerevisiae > A. pullulans B (black) > A. pullulans W (white) > T. viride Br (brown) > M. gypseum > A. niger > F. nivale > P. chrysogenum > T. viride G (green).

  9. An alternative anionic bio-sustainable anti-fungal agent: Investigation of its mode of action on the fungal cell membrane.

    Science.gov (United States)

    Stenbæk, Jonas; Löf, David; Falkman, Peter; Jensen, Bo; Cárdenas, Marité

    2017-07-01

    The potential of a lactylate (the sodium caproyl lactylate or C10 lactylate), a typical food grade emulsifier, as an anionic environmental friendly anti-fungal additive was tested in growth medium and formulated in a protective coating for exterior wood. Different laboratory growth tests on the blue stain fungus Aureobasidium pullulans were performed and its interactions on a model fungal cell membrane were studied. Promising short term anti-fungal effects in growth tests were observed, although significant but less dramatic effects took place in coating test on wood panels. Scanning electron microscope analysis shows clear differences in the amount of fungal slime on the mycelium of Aureobasidium pullulans when the fungus was exposed of C10 lactylate. This could indicate an effect on the pullulan and melanin production by the fungus. Moreover, the interaction studies on model fungal cell membranes show that C10 lactylate affects the phospholipid bilayer in a similar manner to other negative charged detergents. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Screening of micro-organisms for decolorization of melanins produced by bluestain fungi.

    Science.gov (United States)

    Rättö, M; Chatani, M; Ritschkoff, A C; Viikari, L

    2001-03-01

    A total of 17 fungi and four bacteria were screened for their ability to decolorize melanin, using isolated extracellular melanin of the bluestain fungus Aureobasidium pullulans as substrate. On agar media, decolorization was observed by four fungal strains: Bjerkandera adusta VTT-D-99746, Galactomyces geotrichum VTT-D-84228, Trametes hirsuta VTT-D-95443 and Trametes versicolor VTT-D-99747. The four fungi were more efficient on nitrogen-limited medium than on complete medium. The melanin-decolorizing activity of G. geotrichum appeared to be located on the mycelium and could be liberated into the medium enzymatically.

  11. Childhood hypersensitivity pneumonitis associated with fungal contamination of indoor hydroponics.

    Science.gov (United States)

    Engelhart, Steffen; Rietschel, Ernst; Exner, Martin; Lange, Lars

    2009-01-01

    Childhood hypersensitivity pneumonitis (HP) is often associated with exposure to antigens in the home environment. We describe a case of HP associated with indoor hydroponics in a 14-year-old girl. Water samples from hydroponics revealed Aureobasidium pullulans as the dominant fungal micro-organism (10(4)CFU/ml). The diagnosis is supported by the existence of serum precipitating antibodies against A. pullulans, lymphocytic alveolitis on bronchoalveolar lavage (BAL) fluid, a corresponding reaction on a lung biopsy, and the sustained absence of clinical symptoms following the removal of hydroponics from the home. We conclude that hydroponics should be considered as potential sources of fungal contaminants when checking for indoor health complaints.

  12. Yeasts and yeast-like organisms associated with fruits and blossoms of different fruit trees.

    Science.gov (United States)

    Vadkertiová, Renáta; Molnárová, Jana; Vránová, Dana; Sláviková, Elena

    2012-12-01

    Yeasts are common inhabitants of the phyllosphere, but our knowledge of their diversity in various plant organs is still limited. This study focused on the diversity of yeasts and yeast-like organisms associated with matured fruits and fully open blossoms of apple, plum, and pear trees, during 2 consecutive years at 3 localities in southwest Slovakia. The occurrence of yeasts and yeast-like organisms in fruit samples was 2½ times higher and the yeast community more diverse than that in blossom samples. Only 2 species (Aureobasidium pullulans and Metschnikowia pulcherrima) occurred regularly in the blossom samples, whereas Galactomyces candidus, Hanseniaspora guilliermondii, Hanseniaspora uvarum, M. pulcherrima, Pichia kluyveri, Pichia kudriavzevii, and Saccharomyces cerevisiae were the most frequently isolated species from the fruit samples. The ratio of the number of samples where only individual species were present to the number of samples where 2 or more species were found (consortium) was counted. The occurrence of individual species in comparison with consortia was much higher in blossom samples than in fruit samples. In the latter, consortia predominated. Aureobasidium pullulans, M. pulcherrima, and S. cerevisiae, isolated from both the fruits and blossoms, can be considered as resident yeast species of various fruit tree species cultivated in southwest Slovakia localities.

  13. Volatile emissions from an epiphytic fungus are semiochemicals for eusocial wasps.

    Science.gov (United States)

    Davis, Thomas Seth; Boundy-Mills, Kyria; Landolt, Peter J

    2012-11-01

    Microbes are ubiquitous on plant surfaces. However, interactions between epiphytic microbes and arthropods are rarely considered as a factor that affects arthropod behaviors. Here, volatile emissions from an epiphytic fungus were investigated as semiochemical attractants for two eusocial wasps. The fungus Aureobasidium pullulans was isolated from apples, and the volatile compounds emitted by fungal colonies were quantified. The attractiveness of fungal colonies and fungal volatiles to social wasps (Vespula spp.) were experimentally tested in the field. Three important findings emerged: (1) traps baited with A. pullulans caught 2750 % more wasps on average than unbaited control traps; (2) the major headspace volatiles emitted by A. pullulans were 2-methyl-1-butanol, 3-methyl-1-butanol, and 2-phenylethyl alcohol; and (3) a synthetic blend of fungal volatiles attracted 4,933 % more wasps on average than unbaited controls. Wasps were most attracted to 2-methyl-1-butanol. The primary wasp species attracted to fungal volatiles were the western yellowjacket (Vespula pensylvanica) and the German yellowjacket (V. germanica), and both species externally vectored A. pullulans. This is the first study to link microbial volatile emissions with eusocial wasp behaviors, and these experiments indicate that volatile compounds emitted by an epiphytic fungus can be responsible for wasp attraction. This work implicates epiphytic microbes as important components in the community ecology of some eusocial hymenopterans, and fungal emissions may signal suitable nutrient sources to foraging wasps. Our experiments are suggestive of a potential symbiosis, but additional studies are needed to determine if eusocial wasp-fungal associations are widespread, and whether these associations are incidental, facultative, or obligate.

  14. Fungi from interior organs of free-living small mammals in Czechoslovakia and Yugoslavia.

    Science.gov (United States)

    Hubálek, Z; Rosický, B; Otcenásek, M

    1980-01-01

    A total of 308 fungi was isolated from interior organs (lungs, spleen, liver) of 529 small mammals belonging to 21 species, 7 families and 3 orders (Insectivora, Chiroptera, Rodentia), some of these being potentially pathogenic to vertebrates (e.g. Aspergillus flavus, A. fumigatus, Geotrichum candidum, Mucor pusillus, Rhizopus arrhizus). In one vole (Microtus arvalis) captured in South Moravia, adiaspiromycosis (Emmonsia crescens) was demonstrated. Comparison of mycoflora of hair and that of interior organs of wild small mammals revealed that out of the total number of isolates the following fungi were represented in a higher proportion from visceral organs than from the hair: Aspergillus (A. amstelodami, A. flavus, A. repens), Aureobasidium (A. pullulans), Candida, Cladosporium (C. herbarum), Cryptococcus, Fusarium, Gliocladium (G. deliquescens), Helminthosporium, Kloeckera, Mucor (M. fragilis, M. hiemalis, M. pusillus), Paecilomyces marquandii, Penicillium (P. purpurogenum), Phoma, Rhizopus arrhizus, Scopulariopsis (S. candida, S. koningii) and Torulopsis.

  15. Дрожжи в борьбе с возбудителем заболевания серая гниль Botrytis cinerea

    OpenAIRE

    Храбрых, О.; Барайщук, Г.; Kollar, Andreas; Jelkmann, Wilhelm

    2007-01-01

    Серая гниль наиболее распространенное и вредоносное заболевание, вызываемое грибом Botrytis cinerea.The ability of yeasts to inhibit the growth and development of the fungus Botrytis cinerea causes grey mould was detected in vitro during co-culturing the micro organisms on agar plates and in liquid conditions. The radial growth of the pathogen was inhibited in the presence of many tested yeasts on agar plates. In liquid, Aureobasidium pullulans, Cryptococcus laurentii, Candida sake, Hansenias...

  16. Studium biodegradace polyhydroxyalkanoátů.

    OpenAIRE

    Wurstová, Agáta

    2014-01-01

    Diplomová práce je zaměřena na studium biodegradace polyhydroxyalkanoátů, konkrétně polymeru polyhydroxybutyrátu. První část práce se zabývá studiem biodegradace polyhydroxybutyrátu ve formě PHB prášku a PHB fólie pomocí vybraných druhů mikroorganismů z řad bakterií, kvasinek a plísní. Jako zástupce bakterií byl vybrán mikroorganismus Delftia acidovorans, z řad kvasinek Aureobasidium pullulans a ze zástupců plísní Aspergillus fumigatus. Aktivita extracelulární PHB depolymerázy byla analyzován...

  17. Biocontrol ability and action mechanism of food-isolated yeast strains against Botrytis cinerea causing post-harvest bunch rot of table grape.

    Science.gov (United States)

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2015-05-01

    Strains belonging to the species Saccharomyces cerevisiae, Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans, isolated from different food sources, were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic mold Botrytis cinerea. All yeast strains demonstrated antifungal activity at different levels depending on species and medium. Killer strains of W. anomalus and S. cerevisiae showed the highest biocontrol in vitro activity, as demonstrated by largest inhibition halos. The competition for iron and the ability to form biofilm and to colonize fruit wounds were hypothesized as the main action mechanisms for M. pulcherrima. The production of hydrolytic enzymes and the ability to colonize the wounds were the most important mechanisms for biocontrol activity in A. pullulans and W. anomalus, which also showed high ability to form biofilm. The production of volatile organic compounds (VOCs) with in vitro and in vivo inhibitory effect on pathogen growth was observed for the species W. anomalus, S. cerevisiae and M. pulcherrima. Our study clearly indicates that multiple modes of action may explain as M. pulcherrima provide excellent control of postharvest botrytis bunch rot of grape. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Oligo-DNA custom macroarray for monitoring major pathogenic and non-pathogenic fungi and bacteria in the phyllosphere of apple trees.

    Science.gov (United States)

    He, Ying-Hong; Isono, Sayaka; Shibuya, Makoto; Tsuji, Masaharu; Adkar Purushothama, Charith-Raj; Tanaka, Kazuaki; Sano, Teruo

    2012-01-01

    To monitor the richness in microbial inhabitants in the phyllosphere of apple trees cultivated under various cultural and environmental conditions, we developed an oligo-DNA macroarray for major pathogenic and non-pathogenic fungi and bacteria inhabiting the phyllosphere of apple trees. First, we isolated culturable fungi and bacteria from apple orchards by an agar-plate culture method, and detected 32 fungal and 34 bacterial species. Alternaria, Aureobasidium, Cladosporium, Rhodotorula, Cystofilobasidium, and Epicoccum genera were predominant among the fungi, and Bacillus, Pseudomonas, Sphingomonas, Methylobacterium, and Pantoea genera were predominant among the bacteria. Based on the data, we selected 29 major non-pathogenic and 12 phytopathogenic fungi and bacteria as the targets of macroarray. Forty-one species-specific 40-base pair long oligo-DNA sequences were selected from the nucleotide sequences of rDNA-internal transcribed spacer region for fungi and 16S rDNA for bacteria. The oligo-DNAs were fixed on nylon membrane and hybridized with digoxigenin-labeled cRNA probes prepared for each species. All arrays except those for Alternaria, Bacillus, and their related species, were specifically hybridized. The array was sensitive enough to detect 10(3) CFU for Aureobasidium pullulans and Bacillus cereus. Nucleotide sequencing of 100 each of independent fungal rDNA-ITS and bacterial 16S-rDNA sequences from apple tree was in agreement with the macroarray data obtained using the same sample. Finally, we analyzed the richness in the microbial inhabitants in the samples collected from apple trees in four orchards. Major apple pathogens that cause scab, Alternaria blotch, and Marssonina blotch were detected along with several non-phytopathogenic fungal and bacterial inhabitants. The macroarray technique presented here is a strong tool to monitor the major microbial species and the community structures in the phyllosphere of apple trees and identify key species

  19. Diversity and antifungal activity of the endophytic fungi associated with the native medicinal cactus Opuntia humifusa (Cactaceae) from the United States.

    Science.gov (United States)

    Silva-Hughes, Alice F; Wedge, David E; Cantrell, Charles L; Carvalho, Camila R; Pan, Zhiqiang; Moraes, Rita M; Madoxx, Victor L; Rosa, Luiz H

    2015-06-01

    The endophytic fungal community associated with the native cactus Opuntia humifusa in the United States was investigated and its potential for providing antifungal compounds. A hundred-eight endophytic fungal isolates were obtained and identified by molecular methods into 17 different taxa of the genera Alternaria, Aureobasidium, Biscogniauxia, Cladosporium, Cryptococcus, Curvularia, Diaporthe, Epicoccum, Paraconiothyrium, Pestalotiopsis and Phoma. The most frequent species associated with O. humifusa were Alternaria sp. 3, Aureobasidium pullulans and Diaporthe sp. The fungal community of O. humifusa had a high richness and diversity; additionally, the species richness obtained indicates that the sample effort was enough to recover the diversity pattern obtained. Six extracts of endophytes showed antifungal properties and (1)H NMR analyses of the extracts of Alternaria sp. 5 Ohu 8B2, Alternaria sp. 3 Ohu 30A, Cladosporium funiculosum Ohu 17C1 and Paraconiothyrium sp. Ohu 17A indicated the presence of functional groups associated with unsaturated fatty-acid olefinic protons and fatty acid methylene and methyl protons. GC-FID analysis of these extracts confirmed the presence of a mixture of different fatty acids. The (1)H NMR analyses of Biscogniauxia mediterranea Ohu 19B extracts showed the presence of aromatic compounds. From the extract of B. mediterranea we isolated the compound 5-methylmellein that displayed moderate antifungal activity against the phytopathogenic fungi Phomopsis obscurans. Our results suggest that native medicinal cacti of the United States can live symbiotically with rich and diverse endophytic communities and may be a source of bioactive molecules, including those able to inhibit or control plant disease pathogens. Copyright © 2015 Elsevier GmbH. All rights reserved.

  20. House Fly (Musca domestica L. Attraction to Insect Honeydew.

    Directory of Open Access Journals (Sweden)

    Kim Y Hung

    Full Text Available House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the

  1. House Fly (Musca domestica L.) Attraction to Insect Honeydew

    Science.gov (United States)

    Hung, Kim Y.; Michailides, Themis J.; Millar, Jocelyn G.; Wayadande, Astri; Gerry, Alec C.

    2015-01-01

    House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house

  2. House Fly (Musca domestica L.) Attraction to Insect Honeydew.

    Science.gov (United States)

    Hung, Kim Y; Michailides, Themis J; Millar, Jocelyn G; Wayadande, Astri; Gerry, Alec C

    2015-01-01

    House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house

  3. Microbial Production of Malic Acid from Biofuel-Related Coproducts and Biomass

    Directory of Open Access Journals (Sweden)

    Thomas P. West

    2017-04-01

    Full Text Available The dicarboxylic acid malic acid synthesized as part of the tricarboxylic acid cycle can be produced in excess by certain microorganisms. Although malic acid is produced industrially to a lesser extent than citric acid, malic acid has industrial applications in foods and pharmaceuticals as an acidulant among other uses. Only recently has the production of this organic acid from coproducts of industrial bioprocessing been investigated. It has been shown that malic acid can be synthesized by microbes from coproducts generated during biofuel production. More specifically, malic acid has been shown to be synthesized by species of the fungus Aspergillus on thin stillage, a coproduct from corn-based ethanol production, and on crude glycerol, a coproduct from biodiesel production. In addition, the fungus Ustilago trichophora has also been shown to produce malic acid from crude glycerol. With respect to bacteria, a strain of the thermophilic actinobacterium Thermobifida fusca has been shown to produce malic acid from cellulose and treated lignocellulosic biomass. An alternate method of producing malic acid is to use agricultural biomass converted to syngas or biooil as a substrate for fungal bioconversion. Production of poly(β-l-malic acid by strains of Aureobasidium pullulans from agricultural biomass has been reported where the polymalic acid is subsequently hydrolyzed to malic acid. This review examines applications of malic acid, metabolic pathways that synthesize malic acid and microbial malic acid production from biofuel-related coproducts, lignocellulosic biomass and poly(β-l-malic acid.

  4. Plant-microbe interactions: Plant hormone production by phylloplane fungi. Research report

    Energy Technology Data Exchange (ETDEWEB)

    Tuomi, T.; Ilvesoksa, J.; Rosenqvist, H.

    1993-06-23

    The molds Botrytis cinerea, Cladosporium cladosporioides and the yeast Aureobasidium pullulans, isolated from the leaves of three short-rotation Salix clones, were found to produce indole-3-acetic acid (a growth promoter of plants). Abscisic acid (a growth inhibitor of plants) production was detected in B. cinerea. The contents of indole-3-acetic acid and abscisic acid in the leaves of the Salix clones and the amounts of fungal propagules in these leaves were also measured, in order to evaluate whether the amounts of plant growth regulators produced by the fungi would make a significant contribution to the hormonal quantities of the leaves. The content of abscisic acid, and to a lesser degree that of indole-3-acetic acid, showed a positive correlation with the frequency of infection by the hormone producing organisms. The amounts of hormone producing fungi on leaves that bore visible colonies were, however, not sufficiently high to support the argument that neither the fungal production of abscisic nor indole-3-acetic acid would to a significant degree contribute to the hormonal contents of the leaves of the Salix clones.

  5. Fungal contaminants in man-made water systems connected to municipal water.

    Science.gov (United States)

    Kadaifciler, Duygu Göksay; Demirel, Rasime

    2018-04-01

    Water-related fungi are known to cause taste and odor problems, as well as negative health effects, and can lead to water-pipeline clogging. There is no legal regulation on the occurrence of fungi in water environments. However, much research has been performed, but further studies are needed. The main objectives of this study were to evaluate the fungal load and the presence of mycotoxigenic fungi in man-made water systems (for homes, hospitals, and shopping centers) connected to municipal water in Istanbul, Turkey. The mean fungal concentrations found in the different water samples were 98 colony-forming units (CFU)/100 mL in shopping centers, 51 CFU/100 mL in hospitals, and 23 CFU/100 mL in homes. The dominant fungal species were identified as Aureobasidium pullulans and Fusarium oxysporum. Aflatoxigenic Aspergillus flavus and ochratoxigenic Aspergillus westerdijkiae were only detected in the hospital water samples. Alternaria alternata, Aspergillus clavatus, Aspergillus fumigatus, and Cladosporium cladosporioides were also detected in the samples. The study reveals that the municipal water supplies, available for different purposes, could thus contain mycotoxigenic fungi. It was concluded that current disinfection procedures may be insufficient, and the presence of the above-mentioned fungi is important for people with suppressed immune systems.

  6. Fungi associated with black mould on baobab trees in southern Africa.

    Science.gov (United States)

    Cruywagen, Elsie M; Crous, Pedro W; Roux, Jolanda; Slippers, Bernard; Wingfield, Michael J

    2015-07-01

    There have been numerous reports in the scientific and popular literature suggesting that African baobab (Adansonia digitata) trees are dying, with symptoms including a black mould on their bark. The aim of this study was to determine the identity of the fungi causing this black mould and to consider whether they might be affecting the health of trees. The fungi were identified by sequencing directly from mycelium on the infected tissue as well as from cultures on agar. Sequence data for the ITS region of the rDNA resulted in the identification of four fungi including Aureobasidium pullulans, Toxicocladosporium irritans and a new species of Rachicladosporium described here as Rachicladosporium africanum. A single isolate of an unknown Cladosporium sp. was also found. These fungi, referred to here as black mould, are not true sooty mould fungi and they were shown to penetrate below the bark of infected tissue, causing a distinct host reaction. Although infections can lead to dieback of small twigs on severely infected branches, the mould was not found to kill trees.

  7. Cross-reactivity among antigens of different air-borne fungi detected by ELISA using five monoclonal antibodies against Penicillium notatum.

    Science.gov (United States)

    Shen, H D; Lin, W L; Chen, R J; Han, S H

    1990-10-01

    Cross-reactivity among antigens of 12 genera of air-borne fungi, 13 species of Penicillium, and 5 species of Aspergillus was studied by ELISA using five monoclonal antibodies (MoAbs) against Penicillium notatum. Epitopes recognized by all the five MoAbs were susceptible to treatment of mild periodate oxidation and may therefore be associated with carbohydrates. Furthermore, our results showed that there is cross-reactivity among antigens of Penicillium, Aspergillus, and Eurotium species. By using these MoAbs, cross reactivity was not detected between antigens of Penicillium notatum and antigens of Fusarium solani, Alternaria porri, Cladosporium cladosporoides, Curvularia species, Nigrospora species, Aureobasidium pullulans, Wallemia species, Rhizopus arrhizus, and Candida albicans. Cross-reactivity among antigens of 11 species of Penicillium and 5 species of Aspergillus could be detected by ELISA using one of the five MoAbs (MoAb P15). The fact that there may be cross-reactivity among antigens of closely related fungi species should be considered in the diagnosis and treatment of mold allergic diseases.

  8. Increased prevalence of IgG-induced sensitization and hypersensitivity pneumonitis (humidifier lung) in nonsmokers exposed to aerosols of a contaminated air conditioner.

    Science.gov (United States)

    Baur, X; Richter, G; Pethran, A; Czuppon, A B; Schwaiblmair, M

    1992-01-01

    Specific IgG antibodies against antigens of a contaminated air conditioner were estimated in serum of 134 workers of a printing company. Altogether 64% of the workers investigated revealed significantly elevated levels (> 3 U/ml) of IgG antibodies specific to these antigens as compared to a nonexposed control group. The occurrence of IgG antibodies for microbial extracts were 25% for Fusarium, 23% for Penicillium notatum, 13% for Alternaria tenuis, 12% for Aureobasidium pullulans, 9% for Sphaeropsidales species, 3% for Micropolyspora faeni, 2% for Aspergillus fumigatus and 2% for Thermoactionomyces vulgaris. Out of the 86 workers with elevated IgG antibodies for air conditioner antigens, 59 were nonsmokers. Considering a cut-off level of 10 U/ml IgG for high values, the proportion of smokers to nonsmokers becomes even more pronounced (6 to 36 respectively, binominal test p air conditioners are the best choice as antigen source for the diagnosis of humidifier lung in exposed workers. Nonsmokers are shown to have a high risk for immunological sensitization.

  9. Grain dust originating from organic and conventional farming as a potential source of biological agents causing respiratory diseases in farmers.

    Science.gov (United States)

    Zukiewicz-Sobczak, Wioletta A; Cholewa, Grażyna; Krasowska, Ewelina; Chmielewska-Badora, Jolanta; Zwoliński, Jacek; Sobczak, Paweł

    2013-12-01

    Agricultural producers are exposed to a number of different health risks associated with their work environment. The objective of the study was to assess the degree of colonization by fungi in terms of quantity and in terms of variety of species the samples taken from the settled dust from combine threshing of rye cultivation from organic and conventional farms in the Province of Lublin. This paper is a preliminary quantitative assessment of the species of fungi colonizing the samples of settled dust collected during combine threshing from organic and conventional farms in the Province of Lublin. One of the stages of the project was the classification of biosafety BSL (biosafety level) of selected isolates and API ZYM tests to evaluate the potential ability of isolates to cause adverse health effects. To determine the concentration and composition of fungi in collected samples plate dilution method was used with two media: Malt Agar and Potato Dextrose Agar. MOST COMMONLY ISOLATED FUNGI IN SETTLED DUST SAMPLES COLLECTED DURING COMBINE THRESHING FROM ORGANIC FARMS, ON PDA MEDIUM WERE: Alternaria alternata and Aureobasidium pullulans. Cultures on MA medium were dominated by Alternaria alternata, Mycelia sterilia and Fusarium poae. In samples of dust from conventional crops, the predominant species was Alternaria alternata on PDA medium and on MA medium. The obtained results show a potential risk of people involved in agricultural work.

  10. [Diversity and community structure of endophytic fungi from Taxus chinensis var. mairei].

    Science.gov (United States)

    2014-07-01

    A total of 628 endophytic fungi were isolated from 480 tissue segments of needles and branches of Taxus chinensis var. mairei. According to morphological characteristics and ITS sequences, they represented 43 taxa in 28 genera, of which 10 Hyphomycetes, 20 Coelomycetes, 12 Ascomycetes and 1 unknown fungus. Phomopsis mali was confirmed as the dominant species. In accordance with relative frequency, Alternaria alternata, Aureobasidium pullulans, Colletotrichum boninense, C. gloeosporioides, Epicoccum nigrum , Fungal sp., Fusarium lateritium, Glomerella cingulata, Magnaporthales sp. , Nigrospora oryzae, Pestalotiopsis maculiformans, P. microspora, Peyronellaea glomerata and Xylaria sp. 1 were more common in T. chinensis var. mairei. T. chinensis var. mairei were severely infected by endophytic fungi. Endophytic fungi were found in 81 percent of plant tissues with a high diversity. Distribution ranges of endophytic fungi were influenced by tissue properties. The colonization rate, richness, diversity of endophytic fungi in needles were obviously lower than in branches, and kinds of endophytic fungi between branches were more similar than those in needles, thus endophytic fungi had tissue preference. In addition, tissue age influenced the community structure of endophytic fungi. The elder branch tissues were, the higher colonization rate, richness, diversity of endophytic fungi were. Systematic studying the diversity and community structure of endophytic fungi in T. chinensis var. mairei and clarifying their distribution regularity in plant tissues would offer basic data and scientific basis for their development and utilization. Discussing the presence of fungal pathogens in healthy plant tissues would be of positive significance for source protection of T. chinensis var. mairei.

  11. Study on the effects of near-future ocean acidification on marine yeasts: a microcosm approach

    Science.gov (United States)

    Krause, Evamaria; Wichels, Antje; Erler, René; Gerdts, Gunnar

    2013-12-01

    Marine yeasts play an important role in biodegradation and nutrient cycling and are often associated with marine flora and fauna. They show maximum growth at pH levels lower than present-day seawater pH. Thus, contrary to many other marine organisms, they may actually profit from ocean acidification. Hence, we conducted a microcosm study, incubating natural seawater from the North Sea at present-day pH (8.10) and two near-future pH levels (7.81 and 7.67). Yeasts were isolated from the initial seawater sample and after 2 and 4 weeks of incubation. Isolates were classified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and representative isolates were identified by partial sequencing of the large subunit rRNA gene. From the initial seawater sample, we predominantly isolated a yeast-like filamentous fungus related to Aureobasidium pullulans, Cryptococcus sp., Candida sake, and various cold-adapted yeasts. After incubation, we found more different yeast species at near-future pH levels than at present-day pH. Yeasts reacting to low pH were related to Leucosporidium scottii, Rhodotorula mucilaginosa, Cryptococcus sp., and Debaryomyces hansenii. Our results suggest that these yeasts will benefit from seawater pH reductions and give a first indication that the importance of yeasts will increase in a more acidic ocean.

  12. Characterization of pectinase activity for enology from yeasts occurring in Argentine Bonarda grape

    Science.gov (United States)

    Merín, María Gabriela; Martín, María Carolina; Rantsiou, Kalliopi; Cocolin, Luca; de Ambrosini, Vilma Inés Morata

    2015-01-01

    Pectinolytic enzymes are greatly important in winemaking due to their ability to degrade pectic polymers from grape, contributing to enhance process efficiency and wine quality. This study aimed to analyze the occurrence of pectinolytic yeasts during spontaneous fermentation of Argentine Bonarda grape, to select yeasts that produce extracellular pectinases and to characterize their pectinolytic activity under wine-like conditions. Isolated yeasts were grouped using PCR-DGGE and identified by partial sequencing of 26S rRNA gene. Isolates comprised 7 genera, with Aureobasidium pullulans as the most predominant pectinolytic species, followed by Rhodotorula dairenensis and Cryptococcus saitoi. No pectinolytic activity was detected among ascomycetous yeasts isolated on grapes and during fermentation, suggesting a low occurrence of pectinolytic yeast species in wine fermentation ecosystem. This is the first study reporting R. dairenensis and Cr. saitoi species with pectinolytic activity. R. dairenensis GM-15 produced pectinases that proved to be highly active at grape pH, at 12 °C, and under ethanol and SO2 concentrations usually found in vinifications (pectinase activity around 1.1 U/mL). This strain also produced cellulase activity at 12 °C and pH 3.5, but did not produce β-glucosidase activity under these conditions. The strain showed encouraging enological properties for its potential use in low-temperature winemaking. PMID:26413065

  13. Application of ATR-FTIR Spectroscopy to Compare the Cell Materials of Wood Decay Fungi with Wood Mould Fungi

    Directory of Open Access Journals (Sweden)

    Barun Shankar Gupta

    2015-01-01

    Full Text Available Wood fungi create vast damage among standing trees and all types of wood materials. The objectives of this study are to (a characterize the cell materials of two major wood decay fungi (Basidiomycota, namely, Trametes versicolor and Postia placenta, and (b compare the cell materials of decay fungi with four wood mould fungi (Ascomycota, namely, Aureobasidium pullulans, Alternaria alternata, Cladosporium cladosporioides, and Ulocladium atrum. Fourier transform infrared (FTIR spectroscopy is used to characterize the microbial cellular materials. The results showed that the IR bands for the fatty acid at ∼2900 cm−1 were different for the two-decay-fungi genre. Postia placenta shows more absorbance peaks at the fatty acid region. Band ratio indices for amide I and amide II from protein amino acids were higher for the mould fungi (Ascomycota than the decay fungi (Basidiomycota. Similarly, the band ratio index calculated for the protein end methyl group was found to be higher for the mould fungi than the decay fungi. Mould fungi along with the decay fungi demonstrated a positive correlation (R2=0.75 between amide I and amide II indices. The three-component multivariate, principal component analysis showed a strong correlation of amide and protein band indices.

  14. Biotechnological Applications of Dimorphic Yeasts

    Science.gov (United States)

    Doiphode, N.; Joshi, C.; Ghormade, V.; Deshpande, M. V.

    The dimorphic yeasts have the equilibrium between spherical growth (budding) and polarized (hyphal or pseudohyphal tip elongation) which can be triggered by change in the environmental conditions. The reversible growth phenomenon has made dimorphic yeasts as an useful model to understand fungal evolution and fungal differentiation, in general. In nature dimorphism is clearly evident in plant and animal fungal pathogens, which survive and most importantly proliferate in the respective hosts. However, number of organisms with no known pathogenic behaviour also show such a transition, which can be exploited for the technological applications due to their different biochemical make up under different morphologies. For instance, chitin and chitosan production using dimorphic Saccharomyces, Mucor, Rhizopus and Benjaminiella, oil degradation and biotransformation with yeast-form of Yarrowia species, bioremediation of organic pollutants, exopolysac-charide production by yeast-phase of Aureobasidium pullulans, to name a few. Myrothecium verrucaria can be used for seed dressing in its yeast form and it produces a mycolytic enzyme complex in its hyphal-form for the biocontrol of fungal pathogens, while Beauveria bassiana and other entomopathogens kill the insect pest by producing yeast- like cells in the insect body. The form-specific expression of protease, chitinase, lipase, ornithine decarboxylase, glutamate dehydrogenases, etc. make Benjaminiella poitrasii, Basidiobolus sp., and Mucor rouxii strains important in bioremediation, nanobiotechnology, fungal evolution and other areas.

  15. Fungal melanins and their interactions with metals.

    Science.gov (United States)

    Fogarty, R V; Tobin, J M

    1996-09-01

    Fungal melanins are dark brown or black pigments located in cell walls. They also exist as extracellular polymers. Melanized fungi possess increased virulence and resistance to microbial attack as well as enhanced survival while under environmental stress. Melanins contain various functional groups which provide an array of multiple nonequivalent binding sites for metal ions. Pigmented Cladosporium cladosporoides was shown to biosorb 2.5- to four-fold more Ni, Cu, Zn, Cd, and Pb than albino Penicillium digitatum and at four- to six-fold higher rates. Metal desorption was significantly lower for extracellular melanin than from pigmented or albino biomass which indicated the strength of the melanin-metal bond. At equilibrium, tributyltin chloride (TBTC) concentrations of 2.5 mM, pigmented and albino Aureobasidium pullulans absorbed approximately 0.9 and 0.7 mumol TBTC mg -1 dry wt, respectively, whereas purified extracellular melanin exhibited uptake levels of approximately 22 mumol TBTC mg-1 dry wt at an equilibrium concentration of only 0.4 mM. Addition of melanin to the growth medium reduced the toxic effect of CuSO4 and TBTC due to melanin metal binding and sequestration.

  16. Genome sequence of the olive tree, Olea europaea.

    Science.gov (United States)

    Cruz, Fernando; Julca, Irene; Gómez-Garrido, Jèssica; Loska, Damian; Marcet-Houben, Marina; Cano, Emilio; Galán, Beatriz; Frias, Leonor; Ribeca, Paolo; Derdak, Sophia; Gut, Marta; Sánchez-Fernández, Manuel; García, Jose Luis; Gut, Ivo G; Vargas, Pablo; Alioto, Tyler S; Gabaldón, Toni

    2016-06-27

    The Mediterranean olive tree (Olea europaea subsp. europaea) was one of the first trees to be domesticated and is currently of major agricultural importance in the Mediterranean region as the source of olive oil. The molecular bases underlying the phenotypic differences among domesticated cultivars, or between domesticated olive trees and their wild relatives, remain poorly understood. Both wild and cultivated olive trees have 46 chromosomes (2n). A total of 543 Gb of raw DNA sequence from whole genome shotgun sequencing, and a fosmid library containing 155,000 clones from a 1,000+ year-old olive tree (cv. Farga) were generated by Illumina sequencing using different combinations of mate-pair and pair-end libraries. Assembly gave a final genome with a scaffold N50 of 443 kb, and a total length of 1.31 Gb, which represents 95 % of the estimated genome length (1.38 Gb). In addition, the associated fungus Aureobasidium pullulans was partially sequenced. Genome annotation, assisted by RNA sequencing from leaf, root, and fruit tissues at various stages, resulted in 56,349 unique protein coding genes, suggesting recent genomic expansion. Genome completeness, as estimated using the CEGMA pipeline, reached 98.79 %. The assembled draft genome of O. europaea will provide a valuable resource for the study of the evolution and domestication processes of this important tree, and allow determination of the genetic bases of key phenotypic traits. Moreover, it will enhance breeding programs and the formation of new varieties.

  17. Yeasts and coliform bacteria of water accumulated in bromeliads of mangrove and sand dune ecosystems of southeast Brazil.

    Science.gov (United States)

    Hagler, A N; Rosa, C A; Morais, P B; Mendonça-Hagler, L C; Franco, G M; Araujo, F V; Soares, C A

    1993-10-01

    Yeasts and coliform bacteria were isolated from water that accumulated in the central cups and adjacent leaf axilae of two bromeliads, Neoregelia cruenta of a coastal sand dune and Quesnelia quesneliana of a mangrove ecosystem near the city of Rio de Janeiro, Brazil. The mean total coliform counts were above 10,000 per 100 mL for waters of both plants, but the mean fecal coliform counts were only 74 per 100 mL for Q. quesneliana and mostly undetected in water from N. cruenta. Of 90 fecal coliform isolates, 51 were typical of Escherichia coli in colony morphology and indol, methyl red, Volges-Proskauer, and citrate (IMViC) tests. Seven representatives of the typical E. coli cultures were identified as this species, but the identifications of nine other coliform bacteria were mostly dubious. The yeast community of N. cruenta was typical of plant surfaces with basidiomycetous yeasts anamorphs, and the black yeast Aureobasidium pullulans was prevalent. Quesnelia quesneliana had a substantial proportion of ascomycetous yeasts and their anamorphs, including a probable new biotype of Saccharomyces unisporus. Our results suggested that the microbial communities in bromeliad waters are typically autochtonous and not contaminants.

  18. Addition of Pullulan to Trehalose Glasses Improves the Stability of β-Galactosidase at High Moisture Conditions

    NARCIS (Netherlands)

    Teekamp, Naomi; Tian, Yu; Visser, J. Carolina; Olinga, Peter; Frijlink, Henderik W.; Woerdenbag, Herman J.; Hinrichs, Wouter L. J.

    2017-01-01

    Incorporation of therapeutic proteins in a matrix of sugar glass is known to enhance protein stability, yet protection is often lost when exposed to high relative humidity (RH). We hypothesized that especially in these conditions the use of binary glasses of a polysaccharide and disaccharide might

  19. Bio-Sustainable Control of the Blue Stain Fungi Aureobasidium pullulans on Exterior Wood Coatings

    DEFF Research Database (Denmark)

    Stenbæk, Jonas

    on the coating has been studied. In this study, manipulation of the micro structure on the coating surface has been performed and revealed that small scratches-structure (between 4-20 μm) had an inhibitory effect on the attachment on fungal conidia from moulds while bigger structure scratches (>50 μm) did...

  20. Oligo-DNA custom macroarray for monitoring major pathogenic and non-pathogenic fungi and bacteria in the phyllosphere of apple trees.

    Directory of Open Access Journals (Sweden)

    Ying-Hong He

    Full Text Available BACKGROUND: To monitor the richness in microbial inhabitants in the phyllosphere of apple trees cultivated under various cultural and environmental conditions, we developed an oligo-DNA macroarray for major pathogenic and non-pathogenic fungi and bacteria inhabiting the phyllosphere of apple trees. METHODS AND FINDINGS: First, we isolated culturable fungi and bacteria from apple orchards by an agar-plate culture method, and detected 32 fungal and 34 bacterial species. Alternaria, Aureobasidium, Cladosporium, Rhodotorula, Cystofilobasidium, and Epicoccum genera were predominant among the fungi, and Bacillus, Pseudomonas, Sphingomonas, Methylobacterium, and Pantoea genera were predominant among the bacteria. Based on the data, we selected 29 major non-pathogenic and 12 phytopathogenic fungi and bacteria as the targets of macroarray. Forty-one species-specific 40-base pair long oligo-DNA sequences were selected from the nucleotide sequences of rDNA-internal transcribed spacer region for fungi and 16S rDNA for bacteria. The oligo-DNAs were fixed on nylon membrane and hybridized with digoxigenin-labeled cRNA probes prepared for each species. All arrays except those for Alternaria, Bacillus, and their related species, were specifically hybridized. The array was sensitive enough to detect 10(3 CFU for Aureobasidium pullulans and Bacillus cereus. Nucleotide sequencing of 100 each of independent fungal rDNA-ITS and bacterial 16S-rDNA sequences from apple tree was in agreement with the macroarray data obtained using the same sample. Finally, we analyzed the richness in the microbial inhabitants in the samples collected from apple trees in four orchards. Major apple pathogens that cause scab, Alternaria blotch, and Marssonina blotch were detected along with several non-phytopathogenic fungal and bacterial inhabitants. CONCLUSIONS: The macroarray technique presented here is a strong tool to monitor the major microbial species and the community structures in

  1. Competition assays and physiological experiments of soil and phyllosphere yeasts identify Candida subhashii as a novel antagonist of filamentous fungi.

    Science.gov (United States)

    Hilber-Bodmer, Maja; Schmid, Michael; Ahrens, Christian H; Freimoser, Florian M

    2017-01-05

    While recent advances in next generation sequencing technologies have enabled researchers to readily identify countless microbial species in soil, rhizosphere, and phyllosphere microbiomes, the biological functions of the majority of these species are unknown. Functional studies are therefore urgently needed in order to characterize the plethora of microorganisms that are being identified and to point out species that may be used for biotechnology or plant protection. Here, we used a dual culture assay and growth analyses to characterise yeasts (40 different isolates) and their antagonistic effect on 16 filamentous fungi; comprising plant pathogens, antagonists, and saprophytes. Overall, this competition screen of 640 pairwise combinations revealed a broad range of outcomes, ranging from small stimulatory effects of some yeasts up to a growth inhibition of more than 80% by individual species. On average, yeasts isolated from soil suppressed filamentous fungi more strongly than phyllosphere yeasts and the antagonistic activity was a species-/isolate-specific property and not dependent on the filamentous fungus a yeast was interacting with. The isolates with the strongest antagonistic activity were Metschnikowia pulcherrima, Hanseniaspora sp., Cyberlindnera sargentensis, Aureobasidium pullulans, Candida subhashii, and Pichia kluyveri. Among these, the soil yeasts (C. sargentensis, A. pullulans, C. subhashii) assimilated and/or oxidized more di-, tri- and tetrasaccharides and organic acids than yeasts from the phyllosphere. Only the two yeasts C. subhashii and M. pulcherrima were able to grow with N-acetyl-glucosamine as carbon source. The competition assays and physiological experiments described here identified known antagonists that have been implicated in the biological control of plant pathogenic fungi in the past, but also little characterised species such as C. subhashii. Overall, soil yeasts were more antagonistic and metabolically versatile than yeasts from

  2. Melanin production by a yeast strain XJ5-1 of Aureobasidium melanogenum isolated from the Taklimakan desert and its role in the yeast survival in stress environments.

    Science.gov (United States)

    Jiang, Hong; Liu, Nan-Nan; Liu, Guang-Lei; Chi, Zhe; Wang, Jian-Ming; Zhang, Ly-Ly; Chi, Zhen-Ming

    2016-07-01

    The yeast strain XJ5-1 isolated from the Taklimakan desert soil was identified to be a strain of Aureobasdium melanogenum and could produce a large amount of melanin when it was grown in the PDA medium, but its melanin biosynthesis and expression of the PKS gene responsible for the melanin biosynthesis was significantly repressed in the presence of (NH4)2SO4. However, A. melanogenum P5 strain isolated from a mangrove ecosystem grown in both the presence and the absence of (NH4)2SO4 did not produce any melanin. The cell size of A. melanogenum XJ5-1 strain was much higher than that of A. melanogenum P5 strain. The melanized cells of the yeast strain XJ5-1 had higher tolerance to UV radiation, oxidation (200.0 mM H2O2), heat treatment (40 °C), salt shock (200.0 g/L NaCl), desiccation and strong acid hydrolysis (6.0 M HCl) at high temperature (80 °C) than the non-melanized cells of the same yeast strain XJ5-1. At the same time, the melanized cells of the yeast strain XJ5-1 also had higher tolerance to UV radiation, oxidation (200.0 mM H2O2), desiccation and strong acid hydrolysis (6.0 M HCl) at high temperature (80 °C) than A. melanogenum P5 strain, but had similar resistance to heat treatment (40 °C) and salt shock (200.0 g/L NaCl) compared to those of A. melanogenum P5 strain. All the results revealed that many characteristics of A. melanogenum XJ5-1 isolated from the Taklimakan desert soil was different from those of A. melanogenum P5 strain isolated from the mangrove ecosystem.

  3. Análise da contaminação fúngica em amostras de Cassia acutifolia Delile (sene e Peumus boldus (Molina Lyons (boldo-do-Chile comercializadas na cidade de Campinas, Brasil Analysis of the fungal contamination in Cassia acutifolia Delile (sene and Peumus boldus (Molina Lyons (boldo-do-Chile sold in Campinas, Brazil

    Directory of Open Access Journals (Sweden)

    Liliana de O. Rocha

    2004-12-01

    Full Text Available A sociedade atual tem buscado a fitoterapia como um importante recurso terapêutico, sendo a avaliação da qualidade microbiológica destes produtos um requisito essencial, considerando a sua origem. Deste modo, o objetivo da pesquisa foi realizar a contagem e a identificação de fungos filamentosos em 20 amostras de folhas de Cassia acutifolia Delile (sene e de Peumus boldus (Molina Lyons (boldo-do-Chile comercializadas em farmácias de manipulação e mercados da cidade de Campinas, Brasil, usando as técnicas microbiológicas clássicas. Os resultados obtidos evidenciaram que 45% das amostras analisadas se situavam fora dos padrões estabelecidos pela Organização Mundial da Saúde (OMS. Não foram observadas diferenças significativas na análise quantitativa da contaminação fúngica entre amostras comercializadas em farmácias de manipulação e mercados, tanto para o boldo-do-Chile como para a sene. Os fungos identificados nestas amostras foram: Aspergillus, Penicillium, Phoma, Cladosporium, Trichoderma, Rhizopus, Mucor, Aureobasidium pullulans, Mycelia sterilia, Acremonium e Monilia sitophila. Estes resultados demonstraram o baixo nível de qualidade desses fitoterápicos, pois, além do elevado número de amostras contaminadas, foram identificados fungos de gêneros produtores de micotoxinas, como o Aspergillus e o Penicillium. Verifica-se a urgência na realização de adequado controle de qualidade microbiológico dos fitoterápicos e de implantação de fiscalização efetiva, para garantir a segurança e eficácia destes produtos.The consumption of medicinal plants has increased during the last years. The purpose of this study was to evaluate and identify fungi specimens present in samples of Cassia acutifolia Delile (20 (sene and Peumus boldus (Molina Lyons (Boldo-do-Chile (20, that were purchased in drugstores and markets of Campinas, Brazil, by usual methods. The results showed that 45% of samples did not fit the minimum

  4. Evolution of Yeast Consortia during the Fermentation of Kalamata Natural Black Olives upon Two Initial Acidification Treatments

    Directory of Open Access Journals (Sweden)

    Stamatoula Bonatsou

    2018-01-01

    Full Text Available The objective of this study was to elucidate the yeast consortia structure and dynamics during Greek-style processing of Kalamata natural black olives in different brine solutions. Olives were subjected to spontaneous fermentation in 7% (w/v NaCl brine solution (control treatment or brine acidified with (a 0.5% (v/v vinegar, and (b 0.1% (v/v lactic acid at the onset of fermentation. Changes in microbial counts, pH, acidity, organic acids, sugars, and alcohols were analyzed for a period of 187 days. Yeast consortia diversity was evaluated at days 4, 34, 90, 140, and 187 of fermentation. A total of 260 isolates were characterized at sub-species level by rep-PCR genomic fingerprinting with the oligo-nucleotide primer (GTG5. The characterization of yeast isolates at species level was performed by sequencing of the D1/D2 domain of 26S rRNA gene. Results showed that yeasts dominated the process presenting a relatively broad range of biodiversity composed of 11 genera and 21 species. No lactic acid bacteria (LAB or Enterobacteriaceae could be enumerated after 20 and 10 days of fermentation, respectively. The dominant yeast species at the beginning were Aureobasidium pullulans for control and vinegar acidification treatments, and Candida naeodendra for lactic acid treatment. Between 34 and 140 days the dominant species were Candida boidinii, Candida molendinolei and Saccharomyces cerevisiae. In the end of fermentation the dominant species in all processes were C. boidinii and C. molendinolei, followed by Pichia manshurica and S. cerevisiae in lactic acid acidification treatment, P. manshurica in vinegar acidification treatment, and Pichia membranifaciens in control fermentation.

  5. Sequence-based analysis of the Vitis vinifera L. cv Cabernet Sauvignon grape must mycobiome in three South African vineyards employing distinct agronomic systems

    Directory of Open Access Journals (Sweden)

    MATHABATHA EVODIA SETATI

    2015-11-01

    Full Text Available Recent microbiomic research of agricultural habitats has highlighted tremendous microbial biodiversity associated with such ecosystems. Data generated in vineyards have furthermore highlighted significant regional differences in vineyard biodiversity, hinting at the possibility that such differences might be responsible for regional differences in wine style and character, a hypothesis referred to as microbial terroir. The current study further contributes to this body of work by comparing the mycobiome associated with South African (SA Cabernet Sauvignon grapes in three neighboring vineyards that employ different agronomic approaches, and comparing the outcome with similar data sets from Californian vineyards. The aim of this study was to fully characterize the mycobiomes associated with the grapes from these vineyards. The data revealed approximately 10 times more fungal diversity than what is typically retrieved from culture-based studies. The Biodynamic vineyard was found to harbor a more diverse fungal community (H = 2.6 than the conventional (H = 2.1 and integrated (H = 1.8 vineyards. The data show that ascomycota are the most abundant phylum in the three vineyards, with Aureobasidium pullulans and its close relative Kabatiella microsticta being the most dominant fungi. This is the first report to reveal a high incidence of K. microsticta in the grape/wine ecosystem. Different common wine yeast species, such as Metschnikowia pulcherrima and Starmerella bacillaris dominated the mycobiome in the three vineyards. The data show that the filamentous fungi are the most abundant community in grape must although they are not regarded as relevant during wine fermentation. Comparison of metagenomic datasets from the three SA vineyards and previously published data from Californian vineyards revealed only 25% of the fungi in the SA dataset was also present in the Californian dataset, with greater variation evident amongst ubiquitous epiphytic fungi.

  6. Sequence-based Analysis of the Vitis vinifera L. cv Cabernet Sauvignon Grape Must Mycobiome in Three South African Vineyards Employing Distinct Agronomic Systems.

    Science.gov (United States)

    Setati, Mathabatha E; Jacobson, Daniel; Bauer, Florian F

    2015-01-01

    Recent microbiomic research of agricultural habitats has highlighted tremendous microbial biodiversity associated with such ecosystems. Data generated in vineyards have furthermore highlighted significant regional differences in vineyard biodiversity, hinting at the possibility that such differences might be responsible for regional differences in wine style and character, a hypothesis referred to as "microbial terroir." The current study further contributes to this body of work by comparing the mycobiome associated with South African (SA) Cabernet Sauvignon grapes in three neighboring vineyards that employ different agronomic approaches, and comparing the outcome with similar data sets from Californian vineyards. The aim of this study was to fully characterize the mycobiomes associated with the grapes from these vineyards. The data revealed approximately 10 times more fungal diversity than what is typically retrieved from culture-based studies. The Biodynamic vineyard was found to harbor a more diverse fungal community (H = 2.6) than the conventional (H = 2.1) and integrated (H = 1.8) vineyards. The data show that ascomycota are the most abundant phylum in the three vineyards, with Aureobasidium pullulans and its close relative Kabatiella microsticta being the most dominant fungi. This is the first report to reveal a high incidence of K. microsticta in the grape/wine ecosystem. Different common wine yeast species, such as Metschnikowia pulcherrima and Starmerella bacillaris dominated the mycobiome in the three vineyards. The data show that the filamentous fungi are the most abundant community in grape must although they are not regarded as relevant during wine fermentation. Comparison of metagenomic datasets from the three SA vineyards and previously published data from Californian vineyards revealed only 25% of the fungi in the SA dataset was also present in the Californian dataset, with greater variation evident amongst ubiquitous epiphytic fungi.

  7. Yeast identification in floral nectar of Mimulus aurantiacus (Invited)

    Science.gov (United States)

    Kyauk, C.; Belisle, M.; Fukami, T.

    2009-12-01

    Nectar is such a sugar-rich resource that serves as a natural habitat in which microbes thrive. As a result, yeasts arrive to nectar on the bodies of pollinators such as hummingbirds and bees. Yeasts use the sugar in nectar for their own needs when introduced. This research focuses on the identification of different types of yeast that are found in the nectar of Mimulus aurantiacus (commonly known as sticky monkey-flower). Unopened Mimulus aurantiacus flower buds were tagged at Jasper Ridge and bagged three days later. Floral nectar was then extracted and plated on potato dextrose agar. Colonies on the plates were isolated and DNA was extracted from each sample using QIAGEN DNeasy Plant Mini Kit. The DNA was amplified through PCR and ran through gel electrophoresis. The PCR product was used to clone the nectar samples into an E.coli vector. Finally, a phylogenetic tree was created by BLAST searching sequences in GenBank using the Internal Transcribed Space (ITS) locus. It was found that 18 of the 50 identified species were Candida magnifica, 14 was Candida rancensis, 6 were Crytococcus albidus and there were 3 or less of the following: Starmella bombicola, Candida floricola, Aureobasidium pullulans, Pichia kluyvera, Metschnikowa cibodaserisis, Rhodotorua colostri, and Malassezia globosa. The low diversity of the yeast could have been due to several factors: time of collection, demographics of Jasper Ridge, low variety of pollinators, and sugar concentration of the nectar. The results of this study serve as a necessary first step for a recently started research project on ecological interactions between plants, pollinators, and nectar-living yeast. More generally, this research studies the use of the nectar-living yeast community as a natural microcosm for addressing basic questions about the role of dispersal and competitive and facilitative interactions in ecological succession.

  8. Extracellular enzymatic activities and physiological profiles of yeasts colonizing fruit trees.

    Science.gov (United States)

    Molnárová, Jana; Vadkertiová, Renáta; Stratilová, Eva

    2014-07-01

    Yeasts form a significant and diverse part of the phyllosphere microbiota. Some yeasts that inhabit plants have been found to exhibit extracellular enzymatic activities. The aim of the present study was to investigate the ability of yeasts isolated from leaves, fruits, and blossoms of fruit trees cultivated in Southwest Slovakia to produce extracellular enzymes, and to discover whether the yeasts originating from these plant organs differ from each other in their physiological properties. In total, 92 strains belonging to 29 different species were tested for: extracellular protease, β-glucosidase, lipase, and polygalacturonase activities; fermentation abilities; the assimilation of xylose, saccharose and alcohols (methanol, ethanol, glycerol); and for growth in a medium with 33% glucose. The black yeast Aureobasidium pullulans showed the largest spectrum of activities of all the species tested. Almost 70% of the strains tested demonstrated some enzymatic activity, and more than 90% utilized one of the carbon compounds tested. Intraspecies variations were found for the species of the genera Cryptococcus and Pseudozyma. Interspecies differences of strains exhibiting some enzymatic activities and utilizing alcohols were also noted. The largest proportion of the yeasts exhibited β-glucosidase activity and assimilated alcohols independently of their origin. The highest number of strains positive for all activities tested was found among the yeasts associated with leaves. Yeasts isolated from blossoms assimilated saccharose and D-xylose the most frequently of all the yeasts tested. The majority of the fruit-inhabiting yeasts grew in the medium with higher osmotic pressure. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Characterization of a monoclonal antibody (P40) against the 68 kD major allergen of Penicillium notatum.

    Science.gov (United States)

    Shen, H D; Choo, K B; Chen, J H; Lin, W L; Chang, Z N; Han, S H

    1992-04-01

    A monoclonal antibody (MoAb P40) against the 68 kD major allergen of Penicillium notatum (P. notatum) was obtained by immunizing the mouse with a crude extract of P. notatum. Analysed by two-dimensional gel electrophoresis and immunoblotting, P40 reacted with two different isoforms of the 68 kD component of P. notatum with pIs of 5.4 and 5.5. In addition to P. notatum, P40 showed positive ELISA activity to Aspergillus fumigatus (A. fumigatus) but not to components of six other fungi including Alternaria porri, Cladosporium cladosporoides, Aureobasidium pullulans, Fusarium solani, Rhizopus arrhizus and Candida albicans. Analysed by ELISA, MoAb P40 also showed positive activity to two (P. frequentans and P. roseopurpureum) of the 10 other Penicillium species and two (A. terreus and A. flavus) of the four other Aspergillus species tested. SDS-PAGE and immunoblotting studies demonstrated P40 positive reactivity to components with MW of about 67 kD in all these Penicillium and Aspergillus species with positive ELISA activity to P40. Furthermore, immunoblotting activity of MoAb P40 to the 67 kD component of A. niger was also observed. The epitope of the 68 kD allergen of P. notatum recognized by MoAb P40 was resistant to treatment of periodate oxidation with concentration of NaIO4 up to 20 mM. This MoAb may thus be useful in the characterization and purification of the 68 kD allergen from crude extracts, and in the molecular cloning of allergen genes.

  10. Yeast Identification During Fermentation of Turkish Gemlik Olives.

    Science.gov (United States)

    Mujdeci, Gamze; Arévalo-Villena, María; Ozbas, Z Yesim; Briones Pérez, Ana

    2018-05-01

    Naturally fermented black table olives of the Gemlik variety are one of the most consumed fermented products in Turkey. The objective of this work was to identify yeast strains isolated during their natural fermentation by using Restriction Fragments Lengths Polymorphism-Polimerase Chain Reaction (RFLP-PCR) and DNA sequencing methods. The study also focused on determining the effect of regional differences on yeast microflora of naturally fermented Gemlik olives. A total of 47 yeast strains belonging to 12 different species which had been previously isolated from the natural brine of Akhisar and Iznik-Gemlik cv. olives were characterized by molecular methods. Forty-two of the tested strains could be identified by RFLP-PCR to species level. These yeast species were determined as Candida mycetangi, Candida hellenica, Candida membranaefaciens, Candida famata, Candida pelliculosa, Saccharomyces cerevisiae, and Zygosaccharomyces mrakii. Five strains were identified by DNA sequencing. These strains belonged to three different species: Aureobasidium pullulans, Kloeckera apiculate, and Cryptococcus saitoi. The most frequent species were C. famata and C. pelliculosa in both regions. This work studies the yeasts from Turkish table olives which could prove to be of importance to the food industry in that area. On the other hand, it compares identification by molecular and classical biochemical methods and offers an idea about the differences between the ecosystems of Gemlik olives in the Akhisar (AO) and Iznik (IO) regions. The study could be useful in characterizing a very important product and, in this way, could help to promote its marketing. © 2018 Institute of Food Technologists®.

  11. The diversity, extracellular enzymatic activities and photoprotective compounds of yeasts isolated in Antarctica

    Directory of Open Access Journals (Sweden)

    Aline B. M Vaz

    2011-09-01

    Full Text Available The diversity of yeasts collected from different sites in Antarctica (Admiralty Bay, King George Island and Port Foster Bay and Deception Island and their ability to produce extracellular enzymes and mycosporines were studied. Samples were collected during the austral summer season, between November 2006 and January 2007, from the rhizosphere of Deschampsia antarctica, ornithogenic (penguin guano soil, soil, marine and lake sediments, marine water and freshwater from lakes. A total of 89 isolates belonging to the following genera were recovered: Bensingtonia, Candida, Cryptococcus, Debaryomyces, Dioszegia, Exophiala, Filobasidium, Issatchenkia (Pichia, Kodamaea, Leucosporidium, Leucosporidiella, Metschnikowia, Nadsonia, Pichia, Rhodotorula, and Sporidiobolus, and the yeast-like fungi Aureobasidium, Leuconeurospora and Microglossum. Cryptococcus victoriae was the most frequently identified species. Several species isolated in our study have been previously reported to be Antarctic psychophilic yeasts, including Cr. antarcticus, Cr. victoriae, Dioszegia hungarica and Leucosporidium scottii. The cosmopolitan yeast species A. pullulans, C. zeylanoides, D. hansenii, I. orientalis, K. ohmeri, P. guilliermondii, Rh. mucilaginosa, and S. salmonicolor were also isolated. Five possible new species were identified. Sixty percent of the yeasts had at least one detectable extracellular enzymatic activity. Cryptococcus antarcticus, D. aurantiaca, D. crocea, D. hungarica, Dioszegia sp., E. xenobiotica, Rh. glaciales, Rh. laryngis, Microglossum sp. 1 and Microglossum sp. 2 produced mycosporines. Of the yeast isolates, 41.7% produced pigments and/or mycosporines and could be considered adapted to survive in Antarctica. Most of the yeasts had extracellular enzymatic activities at 4ºC and 20ºC, indicating that they could be metabolically active in the sampled substrates.

  12. Are Epiphytic Microbial Communities in the Carposphere of Ripening Grape Clusters (Vitis vinifera L.) Different between Conventional, Organic, and Biodynamic Grapes?

    Science.gov (United States)

    Kecskeméti, Elizabeth; Berkelmann-Löhnertz, Beate; Reineke, Annette

    2016-01-01

    Using barcoded pyrosequencing fungal and bacterial communities associated with grape berry clusters (Vitis vinifera L.) obtained from conventional, organic and biodynamic vineyard plots were investigated in two subsequent years at different stages during berry ripening. The four most abundant operational taxonomic units (OTUs) based on fungal ITS data were Botrytis cinerea, Cladosporium spp., Aureobasidium pullulans and Alternaria alternata which represented 57% and 47% of the total reads in 2010 and 2011, respectively. Members of the genera Sphingomonas, Gluconobacter, Pseudomonas, Erwinia, and Massilia constituted 67% of the total number of bacterial 16S DNA reads in 2010 samples and 78% in 2011 samples. Viticultural management system had no significant effect on abundance of fungi or bacteria in both years and at all three sampling dates. Exceptions were A. alternata and Pseudomonas spp. which were more abundant in the carposphere of conventional compared to biodynamic berries, as well as Sphingomonas spp. which was significantly less abundant on conventional compared to organic berries at an early ripening stage in 2011. In general, there were no significant differences in fungal and bacterial diversity indices or richness evident between management systems. No distinct fungal or bacterial communities were associated with the different maturation stages or management systems, respectively. An exception was the last stage of berry maturation in 2011, where the Simpson diversity index was significantly higher for fungal communities on biodynamic compared to conventional grapes. Our study highlights the existence of complex and dynamic microbial communities in the grape cluster carposphere including both phytopathogenic and potentially antagonistic microorganisms that can have a significant impact on grape production. Such knowledge is particularly relevant for development, selection and application of effective control measures against economically important

  13. Environmental Metabarcoding Reveals Contrasting Belowground and Aboveground Fungal Communities from Poplar at a Hg Phytomanagement Site.

    Science.gov (United States)

    Durand, Alexis; Maillard, François; Foulon, Julie; Gweon, Hyun S; Valot, Benoit; Chalot, Michel

    2017-11-01

    Characterization of microbial communities in stressful conditions at a field level is rather scarce, especially when considering fungal communities from aboveground habitats. We aimed at characterizing fungal communities from different poplar habitats at a Hg-contaminated phytomanagement site by using Illumina-based sequencing, network analysis approach, and direct isolation of Hg-resistant fungal strains. The highest diversity estimated by the Shannon index was found for soil communities, which was negatively affected by soil Hg concentration. Among the significant correlations between soil operational taxonomic units (OTUs) in the co-occurrence network, 80% were negatively correlated revealing dominance of a pattern of mutual exclusion. The fungal communities associated with Populus roots mostly consisted of OTUs from the symbiotic guild, such as members of the Thelephoraceae, thus explaining the lowest diversity found for root communities. Additionally, root communities showed the highest network connectivity index, while rarely detected OTUs from the Glomeromycetes may have a central role in the root network. Unexpectedly high richness and diversity were found for aboveground habitats, compared to the root habitat. The aboveground habitats were dominated by yeasts from the Lalaria, Davidiella, and Bensingtonia genera, not detected in belowground habitats. Leaf and stem habitats were characterized by few dominant OTUs such as those from the Dothideomycete class producing mutual exclusion with other OTUs. Aureobasidium pullulans, one of the dominating OTUs, was further isolated from the leaf habitat, in addition to Nakazawaea populi species, which were found to be Hg resistant. Altogether, these findings will provide an improved point of reference for microbial research on inoculation-based programs of tailings dumps.

  14. Dispersal, density dependence, and population dynamics of a fungal microbe on leaf surfaces.

    Science.gov (United States)

    Woody, Scott T; Ives, Anthony R; Nordheim, Erik V; Andrews, John H

    2007-06-01

    Despite the ubiquity and importance of microbes in nature, little is known about their natural population dynamics, especially for those that occupy terrestrial habitats. Here we investigate the dynamics of the yeast-like fungus Aureobasidium pullulans (Ap) on apple leaves in an orchard. We asked three questions. (1) Is variation in fungal population density among leaves caused by variation in leaf carrying capacities and strong density-dependent population growth that maintains densities near carrying capacity? (2) Do resident populations have competitive advantages over immigrant cells? (3) Do Ap dynamics differ at different times during the growing season? To address these questions, we performed two experiments at different times in the growing season. Both experiments used a 2 x 2 factorial design: treatment 1 removed fungal cells from leaves to reveal density-dependent population growth, and treatment 2 inoculated leaves with an Ap strain engineered to express green fluorescent protein (GFP), which made it possible to track the fate of immigrant cells. The experiments showed that natural populations of Ap vary greatly in density due to sustained differences in carrying capacities among leaves. The maintenance of populations close to carrying capacities indicates strong density-dependent processes. Furthermore, resident populations are strongly competitive against immigrants, while immigrants have little impact on residents. Finally, statistical models showed high population growth rates of resident cells in one experiment but not in the other, suggesting that Ap experiences relatively "good" and "bad" periods for population growth. This picture of Ap dynamics conforms to commonly held, but rarely demonstrated, expectations of microbe dynamics in nature. It also highlights the importance of local processes, as opposed to immigration, in determining the abundance and dynamics of microbes on surfaces in terrestrial systems.

  15. Water-soluble low-molecular-weight -(1, 3–1, 6 D-Glucan inhibit cedar pollinosis

    Directory of Open Access Journals (Sweden)

    Tomoko Jippo

    2015-02-01

    Full Text Available Background: The incidence of allergic diseases such as allergic rhinitis, atopic dermatitis, asthma, and food allergies has increased in several countries. Mast cells have critical roles in various biologic processes related to allergic diseases. Mast cells express the high-affinity receptor for immunoglobulin (Ig E on their surface. The interaction of multivalent antigens with surface-bound IgE causes the secretion of granule-stored mediators, as well as the de novosynthesis of cytokines. Those mediators and cytokines proceed the allergic diseases. We investigated the effects of water-soluble, low-molecular-weight -(1, 3–1, 6 D-glucan isolated from Aureobasidium pullulans 1A1 strain black yeast (LMW--glucan on mast cell-mediated anaphylactic reactions. We reported that LMW--glucan dose-dependently inhibited the degranulation of mast cells. Furthermore, we found that orally administered LMW--glucan inhibited the IgE-mediated passive cutaneous anaphylaxis (PCA reaction in mice. Here, we examined if LMW--glucan had effects on Japanese cedar pollinosis. Findings: In a clinical study, a randomized, single-blind, placebo-controlled, parallel group study in 65 subjects (aged 2262 was performed. This study was undertaken 3 weeks before and until the end of the cedar pollen season. During the study, all subjects consumed one bottle of placebo or LMW--glucan daily and all subjects were required to record allergic symptoms in a diary. The LMW--glucan group had a significantly lower prevalence of sneezing, nose-blowing, tears, and hindrance to the activities of daily living than the placebo group. Conclusions: These results suggested that LMW--glucan could be an effective treatment for allergic diseases

  16. Removal of Cr(VI) from aqueous solution by fungal biomass

    Energy Technology Data Exchange (ETDEWEB)

    Ahluwalia, Sarabjeet Singh [Department of Biotechnology, General Shivdev Singh Diwan Gurbachan Singh Khalsa College, Patiala, Punjab (India); Goyal, Dinesh [Department of Biotechnology and Environmental Sciences, Thapar University, Patiala, Punjab (India)

    2010-10-15

    Chromium compounds are released by industrial processes including leather production, mining, petroleum refining, in textile industry and dyeing. They are a significant threat to the environment and public health because of their toxicity. Removal of hexavalent chromium by living biomass of different fungi was effective in the order of Aspergillus terricola>Aspergillus niger>Acremonium strictum>Aureobasidium pullulans>Paecilomyces variotii>Aspergillus foetidus>Cladosporium resinae>Phanerochaete chrysosporium. Non-living dried fungal biomass showed higher potential for metal removal than living cells. Among all fungi dead biomass of P. chrysosporium, C. resinae and P. variotii had the maximum specific chromium uptake capacity, which was 11.02, 10.69 and 10.35 mg/g of dry biomass respectively at pH 4.0-5.0 in batch sorption. Removal of Cr(VI) by P. chrysosporium from multi-metallic synthetic solution as well as chrome effluent was significant by bringing down the residual concentration to 0.1 mg/L in the effluent, which falls within the permissible range and its removal was not affected by the presence of other metal ions such as Fe, Zn and Ni. Fourier transform infrared spectral analysis revealed the presence of carboxylate (C=O) and amine (-NH{sup +}{sub 3}-NH{sup +}{sub 2}) functional groups commonly present on the cell surface of all fungi, with possible involvement in chromium binding. The result indicates that non-living fungal biomass either obtained as a by-product of fermentation industry or mass produced using inexpensive culture media can be used for bioremediation of Cr(VI) from chrome effluent on large scale. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  17. Metabarcoding Analysis of Fungal Diversity in the Phyllosphere and Carposphere of Olive (Olea europaea).

    Science.gov (United States)

    Abdelfattah, Ahmed; Li Destri Nicosia, Maria Giulia; Cacciola, Santa Olga; Droby, Samir; Schena, Leonardo

    2015-01-01

    The fungal diversity associated with leaves, flowers and fruits of olive (Olea europaea) was investigated in different phenological stages (May, June, October and December) using an implemented metabarcoding approach. It consisted of the 454 pyrosequencing of the fungal ITS2 region and the subsequent phylogenetic analysis of relevant genera along with validated reference sequences. Most sequences were identified up to the species level or were associated with a restricted number of related taxa enabling supported speculations regarding their biological role. Analyses revealed a rich fungal community with 195 different OTUs. Ascomycota was the dominating phyla representing 93.6% of the total number of detected sequences followed by unidentified fungi (3.6%) and Basidiomycota (2.8%). A higher level of diversity was revealed for leaves compared to flowers and fruits. Among plant pathogens the genus Colletotrichum represented by three species (C. godetiae syn. C. clavatum, C. acutatum s.s and C. karstii) was the most abundant on ripe fruits but it was also detected in other organs. Pseudocercospora cladosporioides was detected with a high frequency in all leaf samples and to a less extent in ripe fruits. A much lower relative frequency was revealed for Spilocaea oleagina and for other putative pathogens including Fusarium spp., Neofusicoccum spp., and Alternaria spp. Among non-pathogen taxa, Aureobasidium pullulans, the species complex of Cladosporium cladosporioides and Devriesia spp. were the most represented. This study highlights the existence of a complex fungal consortium including both phytopathogenic and potentially antagonistic microorganisms that can have a significant impact on olive productions.

  18. Insertion of a specific fungal 3'-phosphoadenosine-5'-phosphatase motif into a plant homologue improves halotolerance and drought tolerance of plants.

    Science.gov (United States)

    Gašparič, Meti Buh; Lenassi, Metka; Gostinčar, Cene; Rotter, Ana; Plemenitaš, Ana; Gunde-Cimerman, Nina; Gruden, Kristina; Zel, Jana

    2013-01-01

    Soil salinity and drought are among the most serious agricultural and environmental problems of today. Therefore, investigations of plant resistance to abiotic stress have received a lot of attention in recent years. In this study, we identified the complete coding sequence of a 3'-phosphoadenosine-5'-phosphatase protein, ApHal2, from the halotolerant yeast Aureobasidium pullulans. Expression of the ApHAL2 gene in a Saccharomyces cerevisiae hal2 mutant complemented the mutant auxotrophy for methionine, and rescued the growth of the hal2 mutant in media with high NaCl concentrations. A 21-amino-acids-long region of the ApHal2 enzyme was inserted into the Arabidopsis thaliana homologue of Hal2, the SAL1 phosphatase. The inserted sequence included the META motif, which has previously been implicated in increased sodium tolerance of the Hal2 homologue from a related fungal species. Transgenic Arabidopsis plants overexpressing this modified SAL1 (mSAL1) showed improved halotolerance and drought tolerance. In a medium with an elevated salt concentration, mSAL1-expressing plants were twice as likely to have roots in a higher length category in comparison with the wild-type Arabidopsis and with plants overexpressing the native SAL1, and had 5% to 10% larger leaf surface area under moderate and severe salt stress, respectively. Similarly, after moderate drought exposure, the mSAL1-expressing plants showed 14% increased dry weight after revitalisation, with no increase in dry weight of the wild-type plants. With severe drought, plants overexpressing native SAL1 had the worst rehydration success, consistent with the recently proposed role of SAL1 in severe drought. This was not observed for plants expressing mSAL1. Therefore, the presence of this fungal META motif sequence is beneficial under conditions of increased salinity and moderate drought, and shows no drawbacks for plant survival under severe drought. This demonstrates that adaptations of extremotolerant fungi should

  19. Ultraviolet-B radiation influences the abundance and distribution of phylloplane fungi on pedunculate oak (Quercus robur)

    International Nuclear Information System (INIS)

    Newsham, K.K.; Low, M.N.R.; McLeod, A.R.; Greenslade, P.D.; Emmett, B.A.

    1997-01-01

    The effects of u.v.-B radiation (280-315 nm) on the fungi occurring on the lammas leaves of pedunculate oak (Quercus robur L.) were examined using saplings that were exposed at an outdoor facility to supplemental levels of u.v.-B radiation under treatment arrays of cellulose diacetate-filtered fluorescent lamps, which also produce u.v.-A radiation (315-400 nm). Saplings were also exposed to u.v.-A radiation alone under control arrays of polyester-filtered lamps, and to ambient levels of solar radiation under arrays of unenergized lamps. The u.v.-B treatment corresponded to a 30% elevation above the ambient level of erythemally-weighted u.v.-B radiation. The fungi were examined weekly over a 4-month-period in summer and autumn 1995 using two techniques, the spore fall and leaf impression methods, which differentiated between those fungi occurring on the upper (adaxial) and lower (abaxial) surfaces of the leaves. The abundances of Aureobasidium pullulans (De Bary) Arnaud and Sporobolomyces roseus Kluy. et van Niel, two leaf yeasts which had adaxial:abaxial ratios of < 1 under ambient levels of u.v.-B radiation, were negatively correlated with increasing ambient levels of u.v.-B radiation and were significantly reduced on adaxial leaf surfaces by supplemental levels of u.v.-B. There were few effects of supplemental u.v.-B radiation on the abundances of these yeasts on abaxial leaf surfaces. The abundances of the dematiaceous hyphomycetes, Cladosporium spp. and Epicoccum nigrum Link., species with adaxial:abaxial ratios of ⩾ 1 under ambient levels of u.v.-B radiation, were not correlated with ambient levels of u.v.-B radiation, nor were they usually affected on either leaf surface by supplemental u.v.-B radiation. Alternaria spp. and Microdochium nivale (Fr.) Samuels & Hallet showed consistent responses on adaxial leaf surfaces to u.v.-A radiation applied under control and treatment arrays. Our results suggest that current levels of shortwave radiation already

  20. The microbial ecology of wine grape berries.

    Science.gov (United States)

    Barata, A; Malfeito-Ferreira, M; Loureiro, V

    2012-02-15

    Grapes have a complex microbial ecology including filamentous fungi, yeasts and bacteria with different physiological characteristics and effects upon wine production. Some species are only found in grapes, such as parasitic fungi and environmental bacteria, while others have the ability to survive and grow in wines, constituting the wine microbial consortium. This consortium covers yeast species, lactic acid bacteria and acetic acid bacteria. The proportion of these microorganisms depends on the grape ripening stage and on the availability of nutrients. Grape berries are susceptible to fungal parasites until véraison after which the microbiota of truly intact berries is similar to that of plant leaves, which is dominated by basidiomycetous yeasts (e.g. Cryptococcus spp., Rhodotorula spp. Sporobolomyces spp.) and the yeast-like fungus Aureobasidium pullulans. The cuticle of visually intact berries may bear microfissures and softens with ripening, increasing nutrient availability and explaining the possible dominance by the oxidative or weakly fermentative ascomycetous populations (e.g. Candida spp., Hanseniaspora spp., Metschnikowia spp., Pichia spp.) approaching harvest time. When grape skin is clearly damaged, the availability of high sugar concentrations on the berry surface favours the increase of ascomycetes with higher fermentative activity like Pichia spp. and Zygoascus hellenicus, including dangerous wine spoilage yeasts (e.g. Zygosaccharomyces spp., Torulaspora spp.), and of acetic acid bacteria (e.g. Gluconobacter spp., Acetobacter spp.). The sugar fermenting species Saccharomyces cerevisiae is rarely found on unblemished berries, being favoured by grape damage. Lactic acid bacteria are minor partners of grape microbiota and while being the typical agent of malolactic fermentation, Oenococcus oeni has been seldom isolated from grapes in the vineyard. Environmental ubiquitous bacteria of the genus Enterobacter spp., Enterococcus spp., Bacillus spp

  1. Yeast Biodiversity from DOQ Priorat Uninoculated Fermentations.

    Science.gov (United States)

    Padilla, Beatriz; García-Fernández, David; González, Beatriz; Izidoro, Iara; Esteve-Zarzoso, Braulio; Beltran, Gemma; Mas, Albert

    2016-01-01

    Climate, soil, and grape varieties are the primary characteristics of terroir and lead to the definition of various appellations of origin. However, the microbiota associated with grapes are also affected by these conditions and can leave a footprint in a wine that will be part of the characteristics of terroir. Thus, a description of the yeast microbiota within a vineyard is of interest not only to provide a better understanding of the winemaking process, but also to understand the source of microorganisms that maintain a microbial footprint in wine from the examined vineyard. In this study, two typical grape varieties, Grenache and Carignan, have been sampled from four different vineyards in the DOQ Priorat winegrowing region. Afterward, eight spontaneous alcoholic fermentations containing only grapes from one sampling point and of one variety were conducted at laboratory scale. The fermentation kinetics and yeast population dynamics within each fermentation experiment were evaluated. Yeast identification was performed by RFLP-PCR of the 5.8S-ITS region and by sequencing D1/D2 of the 26S rRNA gene of the isolates. The fermentation kinetics did not indicate clear differences between the two varieties of grapes or among vineyards. Approximately 1,400 isolates were identified, exhibiting high species richness in some fermentations. Of all the isolates studied, approximately 60% belong to the genus Hanseniaspora, 16% to Saccharomyces, and 11% to Candida. Other minor genera, such as Hansenula, Issatchenkia, Kluyveromyces, Saccharomycodes, and Zygosaccharomyces, were also found. The distribution of the identified yeast throughout the fermentation process was studied, and Saccharomyces cerevisiae was found to be present mainly at the end of the fermentation process, while Aureobasidium pullulans was isolated primarily during the first days of fermentation in three of the eight spontaneous fermentations. This work highlights the complexity and diversity of the vineyard

  2. Diversidad de levaduras asociadas a chichas tradicionales de Colombia

    Directory of Open Access Journals (Sweden)

    William Andrés López-Arboleda

    2010-07-01

    , Saccharomyces martiniae, Dekkera anomala, Aureobasidium pullulans y Candida pseudointermedia. La caracterización preliminar de los aislados obtenidos, basada en pruebas de tolerancia a etanol y halo-tolerancia, permitió identificar levaduras nativas con posible utilización biotecnológica en el sector industrial. Palabras clave: Levaduras; diversidad; chicha; identificación molecular; biotecnología Abstract In Colombia, knowledge about yeast communities has been limited because most reports have focused on yeast species with clinical relevance. The spontaneous fermentation of different substrates creates important habitats for analyzing wild yeast populations; for this reason, in this study we isolated and identified yeasts associated with the “chichas” of corn, pineapple, and “arracacha,” which are traditional fermented Colombian beverages. The most representative yeasts were isolated from “chicha” during its three phases of fermentation: initial, tumultuous and final. Initially, we made a partial characterization of isolated yeasts, including macroscopic and microscopic descriptions, physiological tests, and measurement of capacity for producing spores and filaments. However, because these techniques were not sufficient for identification of isolated yeasts to the level of genus and species, the study was complemented by using molecular techniques based on restriction analysis of the ITS1-5.8S rRNA gene-ITS2. When this technique did not permit us to obtain positive results and confirm the PCR-RFLP results, we used the sequence of the D1/D2 domain of the 26S rRNA gene instead for most representative isolates. With these techniques, we identified the most representative yeast species of the three classes of “chicha”: Candida tropicalis, Pichia kluyveri, Pichia guilliermondii, Hanseniapora guilliermondii, Pichia fermentans, Saccharomyces cerevisiae, Candida maltosa, Rhodotorula glutinis, Torulaspora delbrueckii, Hanseniaspora uvarum, Kazachstania exigua

  3. Diversidad de levaduras asociadas a chichas tradicionales de Colombia

    Directory of Open Access Journals (Sweden)

    William Andrés López-Arboleda

    2011-01-01

    , Saccharomyces martiniae, Dekkera anomala, Aureobasidium pullulans y Candida pseudointermedia. La caracterización preliminar de los aislados obtenidos, basada en pruebas de tolerancia a etanol y halo-tolerancia, permitió identificar levaduras nativas con posible utilización biotecnológica en el sector industrial. Palabras clave: Levaduras; diversidad; chicha; identificación molecular; biotecnología Abstract In Colombia, knowledge about yeast communities has been limited because most reports have focused on yeast species with clinical relevance. The spontaneous fermentation of different substrates creates important habitats for analyzing wild yeast populations; for this reason, in this study we isolated and identified yeasts associated with the “chichas” of corn, pineapple, and “arracacha,” which are traditional fermented Colombian beverages. The most representative yeasts were isolated from “chicha” during its three phases of fermentation: initial, tumultuous and final. Initially, we made a partial characterization of isolated yeasts, including macroscopic and microscopic descriptions, physiological tests, and measurement of capacity for producing spores and filaments. However, because these techniques were not sufficient for identification of isolated yeasts to the level of genus and species, the study was complemented by using molecular techniques based on restriction analysis of the ITS1-5.8S rRNA gene-ITS2. When this technique did not permit us to obtain positive results and confirm the PCR-RFLP results, we used the sequence of the D1/D2 domain of the 26S rRNA gene instead for most representative isolates. With these techniques, we identified the most representative yeast species of the three classes of “chicha”: Candida tropicalis, Pichia kluyveri, Pichia guilliermondii, Hanseniapora guilliermondii, Pichia fermentans, Saccharomyces cerevisiae, Candida maltosa, Rhodotorula glutinis, Torulaspora delbrueckii, Hanseniaspora uvarum, Kazachstania exigua

  4. Searching for eukaryotic life preserved in Antarctic permafrost

    DEFF Research Database (Denmark)

    Zucconi, L.; Selbmann, L.; Buzzini, P.

    2012-01-01

    Fungi and yeasts isolated in pure culture from Antarctic permafrost collected at different depths in the McMurdo Dry Valleys were identified with cultural, physiological and molecular methods. Fungi belonged to the genera Penicillium, Eurotium, Cladosporium, Alternaria, Engyodonthium, Aureobasidium...

  5. Enhanced conjugation stability and blood circulation time of macromolecular gadolinium-DTPA contrast agent.

    Science.gov (United States)

    Jenjob, Ratchapol; Kun, Na; Ghee, Jung Yeon; Shen, Zheyu; Wu, Xiaoxia; Cho, Steve K; Lee, Don Haeng; Yang, Su-Geun

    2016-04-01

    In this study, we prepared macromolecular MR T1 contrast agent: pullulan-conjugated Gd diethylene triamine pentaacetate (Gd-DTPA-Pullulan) and estimated residual free Gd(3+), chelation stability in competition with metal ions, plasma and tissue pharmacokinetics, and abdominal MR contrast on rats. Residual free Gd(3+) in Gd-DTPA-Pullulan was measured using colorimetric spectroscopy. The transmetalation of Gd(3+) incubated with Ca(2+) was performed by using a dialysis membrane (MWCO 100-500 Da) and investigated by ICP-OES. The plasma concentration profiles of Gd-DTPA-Pullulan were estimated after intravenous injection at a dose 0.1 mmol/kg of Gd. The coronal-plane abdominal images of normal rats were observed by MR imaging. The content of free Gd(3+), the toxic residual form, was less than 0.01%. Chelation stability of Gd-DTPA-Pullulan was estimated, and only 0.2% and 0.00045% of Gd(3+) were released from Gd-DTPA-Pullulan after 2h incubation with Ca(2+) and Fe(2+), respectively. Gd-DTPA-Pullulan displayed the extended plasma half-life (t1/2,α=0.43 h, t1/2,β=2.32 h), much longer than 0.11h and 0.79 h of Gd-EOB-DTPA. Abdominal MR imaging showed Gd-DTPA-Pullulan maintained initial MR contrast for 30 min. The extended plasma half-life of Gd-DTPA-Pullulan probably allows the prolonged MR acquisition time in clinic with enhanced MR contrast. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. VISUALIZATION OF BIOCHEMICAL ACTIVITY OF MICROORGANISMS GENUS AEROCOCCUS SPECIES AEROCOCCUS VIRIDANS

    Directory of Open Access Journals (Sweden)

    Stepansky DO

    2016-03-01

    Full Text Available Introduction. Aerococci are catalase-negative Gram-positive microorganisms, widespread in the environment, in the cold-blooded microbiocenoses [1, 2] and warm-blooded organisms [3]. Currently, there are seven species of the Aerococcus genus [4]. Type A. viridans got its name as a result of greening blood agar around the growing colonies. It was found that one of the products of A. viridans growth is hydrogen peroxide, has an antagonistic effect on various kinds of bacteria "in vitro" and "in vivo" [5]. It was subsequently found that hydrogen peroxide and superoxide radical are produced as a result of NAD-independent lactatoxidase [6,7] and piruvatoxidase functioning[8]. The oxidative ability of A. viridans is a distinctive feature and allows to visualize these properties using a specific indicator medium [9]. We have developed an indicator that enables to visualize oxidation and reduction properties of aerococci. Material & methods. Auto-symbiotic cultures of A. viridans was used for studying of oxidase - reductase activity, inoculated from human body, an industrial strain of A. viridans 167, used for the preparation of "A-bacterin", culture of Aureobasidium pullulans B5, isolated from the soil and having glucose oxidase activity [10]. Designed indicating medium were also tested in the inoculation of aerococci crops, having lactatoxidase activity of biomaterials from birds and mammals. As a basis for the indicating media (IM IM1 media was selected with following composition (g per 1 liter of water Iodide / potassium 26.0 Soluble starch 10.0 Nutrient agar 30.0 For visualization of oxidase-reductase activity of aerococci acellular components IM4 was developed based on IM3, in which distilled water after double purification was used instead of tap water and highly purified agar-agar was used instead of standard nutrient agar. Results & discussion. IM1 is used to test the oxidative properties of aerococci crops. IM manifests the appearance of a dark

  7. Enhanced conjugation stability and blood circulation time of macromolecular gadolinium-DTPA contrast agent

    Energy Technology Data Exchange (ETDEWEB)

    Jenjob, Ratchapol [Department of New Drug Development, School of Medicine, Inha University, 2F A-dong, Jeongseok Bldg., Sinheung-dong 3-ga, Jung-gu, Incheon 400-712 (Korea, Republic of); Kun, Na [Department of Biotechnology, The Catholic University of Korea, 43 Jibong-ro, Wonmi-gu, Bucheon-si, Gyeonggi-do 420-743 (Korea, Republic of); Ghee, Jung Yeon [Utah-Inha DDS and Advanced Therapeutics, B-403 Meet-You-All Tower, SongdoTechnopark, 7–50, Songdo-dong, Yeonsu-gu, Incheon 406-840 (Korea, Republic of); Shen, Zheyu; Wu, Xiaoxia [Division of Functional Materials and Nano-Devices, Ningbo Institute of Materials Technology & Engineering (NIMTE), Chinese Academy of Sciences, 519 Zhuangshi Street, Zhenhai District, Ningbo, Zhejiang 315201 (China); Cho, Steve K., E-mail: scho@gist.ac.kr [Division of Liberal Arts and Science, GIST College, Gwangju Institute of Science and Technology, Gwangju 500-712 (Korea, Republic of); Lee, Don Haeng [Utah-Inha DDS and Advanced Therapeutics, B-403 Meet-You-All Tower, SongdoTechnopark, 7–50, Songdo-dong, Yeonsu-gu, Incheon 406-840 (Korea, Republic of); Department of Internal Medicine, School of Medicine, Inha University Hospital, Incheon 420-751 (Korea, Republic of); Yang, Su-Geun, E-mail: Sugeun.Yang@Inha.ac.kr [Department of New Drug Development, School of Medicine, Inha University, 2F A-dong, Jeongseok Bldg., Sinheung-dong 3-ga, Jung-gu, Incheon 400-712 (Korea, Republic of)

    2016-04-01

    In this study, we prepared macromolecular MR T1 contrast agent: pullulan-conjugated Gd diethylene triamine pentaacetate (Gd-DTPA-Pullulan) and estimated residual free Gd{sup 3+}, chelation stability in competition with metal ions, plasma and tissue pharmacokinetics, and abdominal MR contrast on rats. Residual free Gd{sup 3+} in Gd-DTPA-Pullulan was measured using colorimetric spectroscopy. The transmetalation of Gd{sup 3+} incubated with Ca{sup 2+} was performed by using a dialysis membrane (MWCO 100–500 Da) and investigated by ICP-OES. The plasma concentration profiles of Gd-DTPA-Pullulan were estimated after intravenous injection at a dose 0.1 mmol/kg of Gd. The coronal-plane abdominal images of normal rats were observed by MR imaging. The content of free Gd{sup 3+}, the toxic residual form, was less than 0.01%. Chelation stability of Gd-DTPA-Pullulan was estimated, and only 0.2% and 0.00045% of Gd{sup 3+} were released from Gd-DTPA-Pullulan after 2 h incubation with Ca{sup 2+} and Fe{sup 2+}, respectively. Gd-DTPA-Pullulan displayed the extended plasma half-life (t{sub 1/2,α} = 0.43 h, t{sub 1/2,β} = 2.32 h), much longer than 0.11 h and 0.79 h of Gd-EOB-DTPA. Abdominal MR imaging showed Gd-DTPA-Pullulan maintained initial MR contrast for 30 min. The extended plasma half-life of Gd-DTPA-Pullulan probably allows the prolonged MR acquisition time in clinic with enhanced MR contrast. - Highlights: • Macromolecule (pullulan) conjugated Gd contrast agent (Gd-DTPA-Pullulan) showed the extended plasma half-life (t{sub 1/2,α} = 0.43 h, t{sub 1/2,β} = 2.32 h) in comparison with Gd-EOB-DTPA • Gd-DTPA-pullulan T1 contrast agent exhibited strong chelation stability against Gd. • The extended blood circulation attributed the enhanced and prolonged MR contrast on abdominal region of rats. • The extended blood circulation may provide prolonged MR acquisition time window in clinics.

  8. Oxidative Stability of Nano-Microstructures containing fish oil

    DEFF Research Database (Denmark)

    García Moreno, Pedro Jesús; Özdemir, N.; Boutrup Stephansen, Karen

    investigated. For that purpose, three different biopolymers namely pullulan, dextran and whey protein concentrate (WPC) were evaluated as encapsulating materials. First, the influence of biopolymer concentration on the physical properties (e.g. viscosity, conductivity and surface tension) of the biopolymer...... solutions and on the morphology of NMS was assayed. Secondly, the oxidative stability of the biopolymer solutions containing emulsified fish oil during storage (14 days at 40 °C) and of NMS loaded with fish oil (e.g. pullulan fibers and dextran and WPC capsules) was determined. Finally, to improve...... the oxidative status of the NMS, pullulan fibers, dextran capsules and WPC capsules were produced by adding neat fish oil instead of emulsified fish oil to the biopolymer solutions. These latter NMS presented a higher oxidative stability, which may be due to a better entrapment of the fish oil into biopolymer...

  9. Interactions between yeasts, fungicides and apple fruit russeting

    NARCIS (Netherlands)

    Gildemacher, P.R.; Heijne, B.; Silvestri, M.; Houbraken, J.; Hoekstra, E.; Theelen, B.; Boekhout, T.

    2006-01-01

    The effect of inoculations with yeasts occurring on apple surfaces and fungicide treatments on the russeting of Elstar apples was studied. Captan, dithianon and a water treatment were implemented to study the interaction between the fungicides, the inoculated yeast species and Aureobasidium

  10. American Peptide Symposium Proceedings (13th) Held in Edmonton, Alberta, Canada on June 20-25, 1993. Peptides: Chemistry, Structure and Biology

    Science.gov (United States)

    1993-06-20

    protein . rceloMEIS w-mdeimidobenzol UtMA I,-Nfoer N-hydroxy- SONaY succinimide ester iniraioa units 14CM melanin concentrating luM keyhole limpet MCPSA...cyclic depsipeptide aureobasidin A (1), isolated as a major component from the culture medium of the black yeast Aureobasidium puUukans R 106, exhibits

  11. Torsion Induced Traumatic Optic Neuropathy (TITON): Animal Model for Diagnostics, Drugs Delivery, and Therapeutics for Injuries to the Cental Nervous System

    Science.gov (United States)

    2016-06-01

    optimization of gel mechanical and swelling properties to mechanically assist the recovery process. 15. SUBJECT TERMS 16. SECURITY CLASSIFICATION OF: 17...polysaccharides including gellan, three varieties of alginate, iota -carrageenan, kappa-carrageenan, xanthan gum, cellulose, carboxymethylcelluose...and pullulan. The most promising candidates were collagen gels alone or in combination with alginate, iota -carrageenan, or gellan gum. Studies are

  12. A novel fermentor system optimized for continuous production of ...

    African Journals Online (AJOL)

    Jane

    2011-08-29

    Aug 29, 2011 ... polyurethane foam with pore size of 1000 Ǻ. The system has ... enhanced solubility in water (Leathers, 2002). ... 8, pressure gauge ; 9, exhaust ; 9a, exhaust ; 10, extra port for .... pullulan production on the outer walls of unit 'B' into the fermentor. ..... retention period of the fermented broth depending on the.

  13. Grewia Gum 1: Some Mechanical and Swelling Properties of ...

    African Journals Online (AJOL)

    Erah

    Methods: Compacts (500 mg) of both freeze-dried and air-dried grewia gum were separately ... grewia gum films were compared with films of pullulan and guar gum which were similarly prepared. .... Freeze-drying was carried out using an.

  14. Development of carbohydrate-based nano-microstructures loaded with fish oil by using electrohydrodynamic processing

    DEFF Research Database (Denmark)

    García Moreno, Pedro Jesús; Özdemir, N.; Boutrup Stephansen, Karen

    2017-01-01

    The encapsulation of fish oil in carbohydrate-based nanomicrostructures obtained by electrohydrodynamic processing was investigated. Solutions of pullulan 200 kDa (15 wt%) and dextran 70 kDa (25 wt%) presented appropriate properties (viscosity, surface tension and conductivity) to allow the forma......The encapsulation of fish oil in carbohydrate-based nanomicrostructures obtained by electrohydrodynamic processing was investigated. Solutions of pullulan 200 kDa (15 wt%) and dextran 70 kDa (25 wt%) presented appropriate properties (viscosity, surface tension and conductivity) to allow...... the formation of nano-microfibers and nano-microcapsules, respectively. Although dextran 70 kDa exhibited antioxidant properties in solution, their capsules produced at lab and pilot-plant scales showed a low oxidative stability both with emulsified and neat oil. Phase separation of solution and opened capsules...... indicated a poor interaction between dextran and fish oil, which suggested that further optimization of the electrospraying solution is necessary. On the contrary, pullulan solutions were optimized to work even at pilot-plant scale. In this case, in spite of the prooxidant effect of pullulan in solution...

  15. Polysaccharide charge density regulating protein adsorption to air/water interfaces by protein/polysaccharide complex formation

    NARCIS (Netherlands)

    Ganzevles, R.A.; Kosters, H.; Vliet, T. van; Stuart, M.A.C.; Jongh, H.H.J. de

    2007-01-01

    Because the formation of protein/polysaccharide complexes is dominated by electrostatic interaction, polysaccharide charge density is expected to play a major role in the adsorption behavior of the complexes. In this study, pullulan (a non-charged polysaccharide) carboxylated to four different

  16. Interactions between fungi colonizing the stem base of winter wheat

    Directory of Open Access Journals (Sweden)

    Urszula Wachowska

    2014-08-01

    Full Text Available In vitro conditions, the interactions betecen the fungi most frequently isolated from the stem base of winter wheat were determined. These were the species from genus Fusarium (F. culmorum, F. avenaceum and F. poae and Rhizoctonia cerealis, Pseudocercosporella herpotrichoides, Alternaria alternata and Aureobasidium bolleyi. Some saprotrophes showed stimulating effect on R. cerealis, P. herpotrichoides and F. poae. Certain species in combined cultures showed exceptionally favourable relationships.

  17. Highly versatile nanohydrogel platform based on riboflavin-polysaccharide derivatives useful in the development of intrinsically fluorescent and cytocompatible drug carriers.

    Science.gov (United States)

    Di Meo, Chiara; Montanari, Elita; Manzi, Lucio; Villani, Claudio; Coviello, Tommasina; Matricardi, Pietro

    2015-01-22

    In this work we describe a new nanohydrogel platform, based on polysaccharides modified with the hydrophobic and fluorescent molecule riboflavin tetrabutyrate, which leads to innovative structures useful for drug delivery applications. Hyaluronic acid and pullulan were chosen as representative of anionic and neutral polysaccharides, respectively, and the bromohexyl derivative of riboflavin tetrabutyrate was chemically linked to these polymer chains. Because of such derivatization, polymer chains were able to self-assemble in aqueous environment thus forming nanohydrogels, with mean diameters of about 312 and 210 nm, for hyaluronan and pullulan, respectively. These new nanohydrogels showed low polydispersity index, and negative ζ-potential. Moreover, the nanohydrogels, which can be easily loaded with model drugs, showed long-term stability in water and physiological conditions and excellent cytocompatibility. All these properties allow to consider these intrinsically fluorescent nanohydrogels suitable for the formulation of innovative drug dosage forms. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. FILMES BIODEGRADÁVEIS DE AMIDO DE MANDIOCA, PULULANA E CELULOSE BACTERIANA

    Directory of Open Access Journals (Sweden)

    Ana Claudia Sueiro

    Full Text Available The largest consumption of plastics in the world is referred to the synthetic polymers, which are not biodegradable and have a non-renewable source, generating a large environmental impact, especially in urban centers. As a result, in the last two decades several polymers obtained from renewable sources (biopolymers have been studied as potential raw materials for the production of new biodegradable materials with different applications. The objectives of this study were to produce biodegradable films based on cassava starch, pullulan and bacterial cellulose, and also to characterize these films according to their microstructure, barrier, thermal and mechanical properties. The addition of bacterial cellulose and pullulan to the starch films resulted in films with more homogeneous surfaces, and also decreased solubility and water vapor permeability, and increased elongation and thermal stability.

  19. Structural Characterization and Bacterial Degradation of Marine Carbohydrates

    Science.gov (United States)

    1993-06-01

    of glycerol, and a mono- or oligosaccharide glycosydically linked to the #3 carbon. The NMR 3 spectra of F3, however, are not consistent with a major... hydrolase activity has been demonstrated in a number of diatoms (Myldestad et al., 1982) and has been detected in beach sands collected from beneath...Four different pullulanases and pullulan hydrolases , with different specificities, have been I found in a range of bacteria. The enzymes in the

  20. Nanogel-quantum dot hybrid nanoparticles for live cell imaging

    International Nuclear Information System (INIS)

    Hasegawa, Urara; Nomura, Shin-ichiro M.; Kaul, Sunil C.; Hirano, Takashi; Akiyoshi, Kazunari

    2005-01-01

    We report here a novel carrier of quantum dots (QDs) for intracellular labeling. Monodisperse hybrid nanoparticles (38 nm in diameter) of QDs were prepared by simple mixing with nanogels of cholesterol-bearing pullulan (CHP) modified with amino groups (CHPNH 2 ). The CHPNH 2 -QD nanoparticles were effectively internalized into the various human cells examined. The efficiency of cellular uptake was much higher than that of a conventional carrier, cationic liposome. These hybrid nanoparticles could be a promising fluorescent probe for bioimaging

  1. Electrospinning of food proteins and polysaccharides

    DEFF Research Database (Denmark)

    Mendes, Ana Carina Loureiro; Boutrup Stephansen, Karen; Chronakis, Ioannis S.

    2017-01-01

    Nano-microfibrous structures of biopolymers with a wide range of compositions, morphologies, mechanical properties and bioactivities could be developed using electrospinning technology. This review focuses on the processing, properties, functionalization and potential applications of electrospun ...... biopolymers. Biopolymers include proteins (gelatin, collagen, elastin, silk, soy zein, gliadin, hordein, amaranth, casein, wheat, whey, marine sources proteins), and polysaccharides (chitosan, starch, alginate, cellulose and cellulose derivatives, pullulan, dextran, cyclodextrins)....

  2. Insulin Inclusion into a Tragacanth Hydrogel: An Oral Delivery System for Insulin

    OpenAIRE

    Mokhamad Nur; Todor Vasiljevic

    2018-01-01

    Nanoparticles or microparticles created by physical complexation between two polyelectrolytes may have a prospective use as an excipient for oral insulin administration. Natural polymers such as tragacanth, alginate, dextran, pullulan, hyaluronic acid, gelatin and chitosan can be potential candidates for this purpose. In this research, insulin particles were prepared by the inclusion of insulin into a tragacanth hydrogel. The effect of the pH and concentration relationship involving polyelect...

  3. Mold attack on frescoes and stone walls of Gradac monastery

    Directory of Open Access Journals (Sweden)

    Stupar Miloš Č.

    2011-01-01

    Full Text Available Microfungi can colonize stone surfaces and form sub-aerial biofilms which can lead to biodeterioration of historic monuments. In this investigation samples for mycological analyses were collected from stone material with visible alteration on stone walls of Gradac monastery exterior. The prevailing fungi found on stone walls were dematiaceous hyphomycetes with melanized hyphae and reproductive structures (Alternaria, Aureobasidium, Cladosporium and Epicoccum species. The frescoes inside the monastery building were also analyzed for the presence of mycobiota. The predominant fungi found on frescoes were osmophilic species from genera Aspergillus and Penicillium. The significant result is identification of human pathogen species Aspergillus fumigatus on frescoes.

  4. High-throughput sequencing of microbial community diversity in soil, grapes, leaves, grape juice and wine of grapevine from China.

    Science.gov (United States)

    Wei, Yu-Jie; Wu, Yun; Yan, Yin-Zhuo; Zou, Wan; Xue, Jie; Ma, Wen-Rui; Wang, Wei; Tian, Ge; Wang, Li-Ye

    2018-01-01

    In this study Illumina MiSeq was performed to investigate microbial diversity in soil, leaves, grape, grape juice and wine. A total of 1,043,102 fungal Internal Transcribed Spacer (ITS) reads and 2,422,188 high quality bacterial 16S rDNA sequences were used for taxonomic classification, revealed five fungal and eight bacterial phyla. At the genus level, the dominant fungi were Ascomycota, Sordariales, Tetracladium and Geomyces in soil, Aureobasidium and Pleosporaceae in grapes leaves, Aureobasidium in grape and grape juice. The dominant bacteria were Kaistobacter, Arthrobacter, Skermanella and Sphingomonas in soil, Pseudomonas, Acinetobacter and Kaistobacter in grape and grapes leaves, and Oenococcus in grape juice and wine. Principal coordinate analysis showed structural separation between the composition of fungi and bacteria in all samples. This is the first study to understand microbiome population in soil, grape, grapes leaves, grape juice and wine in Xinjiang through High-throughput Sequencing and identify microorganisms like Saccharomyces cerevisiae and Oenococcus spp. that may contribute to the quality and flavor of wine.

  5. High-throughput sequencing of microbial community diversity in soil, grapes, leaves, grape juice and wine of grapevine from China

    Science.gov (United States)

    Yan, Yin-zhuo; Zou, Wan; Ma, Wen-rui; Wang, Wei; Tian, Ge; Wang, Li-ye

    2018-01-01

    In this study Illumina MiSeq was performed to investigate microbial diversity in soil, leaves, grape, grape juice and wine. A total of 1,043,102 fungal Internal Transcribed Spacer (ITS) reads and 2,422,188 high quality bacterial 16S rDNA sequences were used for taxonomic classification, revealed five fungal and eight bacterial phyla. At the genus level, the dominant fungi were Ascomycota, Sordariales, Tetracladium and Geomyces in soil, Aureobasidium and Pleosporaceae in grapes leaves, Aureobasidium in grape and grape juice. The dominant bacteria were Kaistobacter, Arthrobacter, Skermanella and Sphingomonas in soil, Pseudomonas, Acinetobacter and Kaistobacter in grape and grapes leaves, and Oenococcus in grape juice and wine. Principal coordinate analysis showed structural separation between the composition of fungi and bacteria in all samples. This is the first study to understand microbiome population in soil, grape, grapes leaves, grape juice and wine in Xinjiang through High-throughput Sequencing and identify microorganisms like Saccharomyces cerevisiae and Oenococcus spp. that may contribute to the quality and flavor of wine. PMID:29565999

  6. A High Molecular-Mass Anoxybacillus sp. SK3-4 Amylopullulanase: Characterization and Its Relationship in Carbohydrate Utilization

    Directory of Open Access Journals (Sweden)

    Kian Mau Goh

    2013-05-01

    Full Text Available An amylopullulanase of the thermophilic Anoxybacillus sp. SK3-4 (ApuASK was purified to homogeneity and characterized. Though amylopullulanases larger than 200 kDa are rare, the molecular mass of purified ApuASK appears to be approximately 225 kDa, on both SDS-PAGE analyses and native-PAGE analyses. ApuASK was stable between pH 6.0 and pH 8.0 and exhibited optimal activity at pH 7.5. The optimal temperature for ApuASK enzyme activity was 60 °C, and it retained 54% of its total activity for 240 min at 65 °C. ApuASK reacts with pullulan, starch, glycogen, and dextrin, yielding glucose, maltose, and maltotriose. Interestingly, most of the previously described amylopullulanases are unable to produce glucose and maltose from these substrates. Thus, ApuASK is a novel, high molecular-mass amylopullulanase able to produce glucose, maltose, and maltotriose from pullulan and starch. Based on whole genome sequencing data, ApuASK appeared to be the largest protein present in Anoxybacillus sp. SK3-4. The α-amylase catalytic domain present in all of the amylase superfamily members is present in ApuASK, located between the cyclodextrin (CD-pullulan-degrading N-terminus and the α-amylase catalytic C-terminus (amyC domains. In addition, the existence of a S-layer homology (SLH domain indicates that ApuASK might function as a cell-anchoring enzyme and be important for carbohydrate utilization in a streaming hot spring.

  7. A screening method for β-glucan hydrolase employing Trypan Blue-coupled β-glucan agar plate and β-glucan zymography.

    Science.gov (United States)

    Park, Chang-Su; Yang, Hee-Jong; Kim, Dong-Ho; Kang, Dae-Ook; Kim, Min-Soo; Choi, Nack-Shick

    2012-06-01

    A new screening method for β-(1,3-1,6) glucan hydrolase was developed using a pure β-glucan from Aureobaisidum pullulans by zymography and an LB-agar plate. Paenibacillus sp. was screened as a producer a β-glucan hydrolase on the Trypan Blue-coupled β-glucan LB-agar plate and the activity of the enzyme was analyzed by SDS-β-glucan zymography. The β-glucan was not hydrolyzed by Bacillus spp. strains, which exhibit cellulolytic activity on CMC zymography. The gene, obtaining by shotgun cloning and encoding the β-glucan hydrolase of Paenibacillus sp. was sequenced.

  8. The application of HP-GFC chromatographic method for the analysis of oligosaccharides in bioactive complexes

    Directory of Open Access Journals (Sweden)

    Savić Ivan

    2009-01-01

    Full Text Available The aim of this work was to optimize a GFC method for the analysis of bioactive metal (Cu, Co and Fe complexes with olygosaccharides (dextran and pullulan. Bioactive metal complexes with olygosaccharides were synthesized by original procedure. GFC was used to study the molecular weight distribution, polymerization degree of oligosaccharides and bioactive metal complexes. The metal bounding in complexes depends on the ligand polymerization degree and the presence of OH groups in coordinative sphere of the central metal ion. The interaction between oligosaccharide and metal ions are very important in veterinary medicine, agriculture, pharmacy and medicine.

  9. Effect of Glycerol, as Cryoprotectant in the Encapsulation and Freeze Drying of Microspheres Containing Probiotic Cells

    Directory of Open Access Journals (Sweden)

    Oana Lelia Pop

    2015-05-01

    Full Text Available It is reported that probiotics provide several health benefits as they help in maintaining a good balance and composition of intestinal flora, and increase the resistance against invasion of pathogens. Ensuring adequate dosages of probiotics at the time of consumption is a challenge, because several factors during processing and storage affect the viability of probiotic organisms. Major emphasis has been given to protect the microorganisms with the help of encapsulation technique, by addition of different protectants. In this study, probiotic cells (Bifidobacterium lactis 300B were entrapped in alginate/pullulan microspheres. In the encapsulation formula glycerol was used as cryoprotectant in the freeze drying process for long time storage. It was observed that the survival of Bifidobacterium lactis 300B when encapsulated without cryoprotectant was higher than the formula with glycerol in the fresh obtained microspheres. The addition of glycerol was in order to reduce the deep freezing and freeze drying damages. In the chosen formulations, glycerol did not proved protection for the entrapped probiotic cells in the freeze drying process, for which the use of glycerol as cryoprotectant for alginate/pullulan Bifidobacterium lactis 300B entrapment is not recommended.

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

    Science.gov (United States)

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

    2011-08-01

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

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

    Science.gov (United States)

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

    2012-03-01

    Maple sap processing and microbial contamination are significant aspects that affect maple syrup quality. In this study, two sample sets from 2005 and 2008 were used to assess the maple syrup quality variation and its relationship to microbial populations, with respect to processing, production site and harvesting period. The abundance of maple sap predominant bacteria (Pseudomonas fluorescens group and two subgroups, Rahnella spp., Janthinobacterium spp., Leuconostoc mesenteroides) and yeast (Mrakia spp., Mrakiella spp.,Guehomyces pullulans) was assessed by quantitative PCR. Maple syrup properties were analyzed by physicochemical and sensorial methods. Results indicate that P. fluorescens, Mrakia spp., Mrakiella spp. G. pullulans and Rahnella spp. are stable contaminants of maple sap, as they were found for every production site throughout the flow period. Multiple factor analysis reports a link between the relative abundance of P. fluorescens group and Mrakia spp. in maple sap with maple and vanilla odor as well as flavor of maple syrup. This evidence supports the contribution of these microorganisms or a consortium of predominant microbial contaminants to the characteristic properties of maple syrup. Copyright © 2011 Elsevier B.V. All rights reserved.

  12. Graphene Oxide Bionanocomposite Coatings with High Oxygen Barrier Properties

    Directory of Open Access Journals (Sweden)

    Ilke Uysal Unalan

    2016-12-01

    Full Text Available In this work, we present the development of bionanocomposite coatings on poly(ethylene terephthalate (PET with outstanding oxygen barrier properties. Pullulan and graphene oxide (GO were used as main polymer phase and nanobuilding block (NBB, respectively. The oxygen barrier performance was investigated at different filler volume fractions (ϕ and as a function of different relative humidity (RH values. Noticeably, the impermeable nature of GO was reflected under dry conditions, in which an oxygen transmission rate (OTR, mL·m−2·24 h−1 value below the detection limit of the instrument (0.01 mL·m−2·24 h−1 was recorded, even for ϕ as low as 0.0004. A dramatic increase of the OTR values occurred in humid conditions, such that the barrier performance was totally lost at 90% RH (the OTR of coated PET films was equal to the OTR of bare PET films. Modelling of the experimental OTR data by Cussler’s model suggested that the spatial ordering of GO sheets within the main pullulan phase was perturbed because of RH fluctuations. In spite of the presence of the filler, all the formulations allowed the obtainment of final materials with haze values below 3%, the only exception being the formulation with the highest loading of GO (ϕ ≈ 0.03. The mechanisms underlying the experimental observations are discussed.

  13. Purification and properties of a thermostable pullulanase from Clostridium thermosulfurgenes EM1 which hydrolyses both. alpha. -1,6 and. alpha. -1,4-glycosidic linkages

    Energy Technology Data Exchange (ETDEWEB)

    Spreinat, A [Goettingen Univ. (Germany, F.R.). Inst. fuer Mikrobiologie; Antranikian, G [Technische Univ. Hamburg-Harburg, Hamburg (Germany, F. R.). Arbeitsbereich Biotechnologie 1

    1990-08-01

    A novel thermostable pullulanase, secreted by the thermophilic anaerobic bacterium Clostridium thermosulfurogenes EM1, was purified and characterized. Applying anion exchange chromatography and gel filtration the enzyme was purified 47-fold and had a specific activity of 200 units/mg. The molecular mass of this thermostable enzyme was determined to be 102 000 daltons and consisted of a single subunit. The enzyme was able to attack specifically the {alpha}-1,6-glycosidic linkages in pullulan and caused its complete hydrolysis to maltotriose. Surprisingly and unlike the enzyme from Klebsiella pneumoniae, the purified enzyme from this anaerobic thermophile exhibited, in addition to its debranching and pullulanase activity, an {alpha}-1,4 hydrolysing activity as well. By the action of this single polypeptide chain various branched and linear polysaccharides were completely converted to two major products, namely maltose and maltotriose. The K{sub m} values of this enzyme for pullulan and amylose were determined to be 1.33 mg/ml and 0.38 mg/ml, respectively. This debranching enzyme displays a temperature optimum at 60deg-65deg C and a pH optimum at 5.5-6.0. The application of this new class of pullulanase (pullulanase type II) in industry will significantly enhance the starch saccharification process. (orig.).

  14. Holographic diffraction gratings as laser radiation protection filters

    International Nuclear Information System (INIS)

    Pantelic, D.; Pantelic, G.

    2006-01-01

    Holographic volume diffraction gratings are used as attenuation filters, due to their selective spectral transmission. They can be tailored to reflect or transmit narrow spectral ranges by adjusting spatial frequency of Bragg grating in carefully chosen photosensitive materials, like silver-halide emulsion or di-chromated gelatin layers. If properly recorded and chemically processed, resulting gratings can significantly attenuate light at wavelengths corresponding to various laser spectral lines. Thus, they can be used as filters in laser protection goggles. We analyze the characteristics of Bragg gratings necessary to obtain high attenuation coefficients. Also, their angular selectivity is taken into account and corresponding experimental conditions are investigated. Although di-chromated gelatin seems to be almost ideal material, due to its almost 100% diffraction efficiency, environmental stability is poor (degradation under humid environment), thus making its practical usage difficult. Thus, we have analyzed alternative materials like di-chromated pullulan, which is stable under normal environmental conditions (without drop in diffraction efficiency after prolonged exposure to humidity). Pullulan is polymer (polysaccharide) of biologic origin produced by certain bacteria. If doped with chromium ions it becomes photosensitive, enabling recording of diffraction gratings with spatial frequency of more than 3000 lines/mm. Material is chemically processed by mixture of isopropyl alcohol and water. Both thick and thin layers can be produced by gravity settling. Spectral properties of resulting gratings are analyzed, showing that they can significantly attenuate laser light of particular wavelength, depending of grating period and its slant angle. (authors)

  15. Physicochemical characterization and oxidative stability of fish oil-loaded electrosprayed capsules: Combined use of whey protein and carbohydrates as wall materials

    DEFF Research Database (Denmark)

    García Moreno, Pedro Jesús; Pelayo, Andres; Yu, Sen

    2018-01-01

    The encapsulation of fish oil in electrosprayed capsules using whey protein and carbohydrates (pullulan and dextran or glucose syrup) mixtures as glassy wall materials was studied. Capsules with fish oil emulsified by using only a rotor-stator emulsification exhibited higher oxidative stability...... than capsules where the oil was emulsified by high-pressure homogenization. Moreover, glucose syrup capsules (with a peroxide value, PV, of 19.7 ± 4.4 meq/kg oil and a content of 1-penten-3-ol of 751.0 ± 69.8 ng/g oil) were less oxidized than dextran capsules after 21 days of storage at 20 °C (PV of 24.......9 ± 0.4 meq/kg oil and 1-penten-3-ol of 1161.0 ± 222.0 ng/g oil). This finding may be attributed to differences in oxygen permeability between both types of capsules. These results indicated the potential of both combinations of whey protein, pullulan, and dextran or glucose syrup as shell materials...

  16. α--AMYLASES OF Aspergillus flavus var. oryzae AND Bacillus subtilis: THE SUBSTRATE SPECIFICITY AND RESISTANCE TO A NUMBER OF CHEMICALLY ACTIVE SUBSTANCES

    Directory of Open Access Journals (Sweden)

    K. V. Avdiyuk

    2013-06-01

    Full Text Available The ability of Aspergillus flavus var. oryzae 80428 and Bacillus subtilis 147 α-amylases to split different carbohydrate-containing substrates, such as maltose, sucrose, trehalose, dextrin, α- and β-cyclodextrin, amylose, amylopectin, glycogen, pullulan, soluble starch, insoluble starch, corn starch, wheat starch, dextran 500 has been studied. It was shown that investigated enzymes differ by substrate specificity. α-Amylase of A. flavus var. oryzae 80428 rapidly hydrolysed soluble potato and wheat starch, while the α-amylase of B. subtilis 147 — only wheat starch. Both enzymes don’t cleave maltose, α-cyclodextrin and dextran 500. A. flavus var. oryzae 80428 α-amylase display very small ability to hydrolyze pullulan, while α-amylase of B. subtilis 147 it does not act in general. The lowest values of Michaelis constant for both enzymes at splitting of glycogen have been obtained, indicating that enzymes have the greatest affinity to this substrate. The studies of influence of chemically active substances on activity of A. flavus var. oryzae 80428 and B. subtilis 147 ?-amylases show there are resistant to urea, deoxycholic acid, Tween-80, Triton X-100 and hydrogen peroxide. It’s indicate the enzymes tested may be competitive in compare with earlier described in literature enzymes. The obtained results give a possibility to propose in future usage these enzymes in different fields of industry, foremost in detergent industry.

  17. A Metabarcoding Survey on the Fungal Microbiota Associated to the Olive Fruit Fly.

    Science.gov (United States)

    Malacrinò, Antonino; Schena, Leonardo; Campolo, Orlando; Laudani, Francesca; Mosca, Saveria; Giunti, Giulia; Strano, Cinzia Patricia; Palmeri, Vincenzo

    2017-04-01

    The occurrence of interaction between insects and fungi is interesting from an ecological point of view, particularly when these interactions involve insect pests and plant pathogens within an agroecosystem. In this study, we aimed to perform an accurate analysis on the fungal microbiota associated to Bactrocera oleae (Rossi) through a metabarcoding approach based on 454 pyrosequencing. From this analysis, we retrieved 43,549 reads that clustered into 128 operational taxonomic units (OTUs), of which 29 resulted in the "core" associate fungi of B. oleae. This fungal community was mainly represented by sooty mould fungi, such as Cladosporium spp., Alternaria spp. and Aureobasidium spp., by plant pathogens like Colletotrichum spp. and Pseudocercospora spp., along with several other less abundant taxa whose ecology is unclear in most of the cases. Our findings lead to new insights into the microbial ecology of this specific ecological niche, enabling the understanding of a complex network of interactions within the olive agroecosystem.

  18. Fungal growth during malting of barley

    Directory of Open Access Journals (Sweden)

    Kocić-Tanackov Sunčica D.

    2005-01-01

    Full Text Available Fungi were isolated and identified in two samples of winter two-row barley (SSK3 and SSK6 harvested in 2003, Kragujevac location, during micromalting. Fungi were isolated and identified in barley before the micromalting, after the 1st, 2nd and 3rd day of steeping, the first day and after the germination after kilning and after malt degermination. The total fungi count was followed in both barley samples, during the mentioned phases. The total count of fungi was also determined in the steeping water, and the isolation and identification was performed after the steeping process. Change of the total count of fungi during barley micromalting was exponentional. During barley micromalting nine fungi genera were isolated: Phoma, Alternaria, Fusarium aspergillus, Cladosporium, Geotrichum, Scopulariopsis, Aureobasidium and Mucor. The most frequent genera were: Phoma, Alternaria and Fusarium. In water for steeping, five genera were identified: Geotrichum, Fusarium, Phoma Cladosporium and Mucor. The most frequent genera was Phoma.

  19. Influence of roadside pollution on the phylloplane microbial community of Alnus nepalensis (Betulaceae)

    International Nuclear Information System (INIS)

    Joshi, S. R.

    2008-01-01

    The North Eastern region of India is undergoing industrial development at a faster rate than expected. Roads form the main system of transportation and communication owing to the hilly topography of the region. Automobiles discharge a number of gaseous and trace metal contaminants. Human activities like stone grinding, road construction and sand milling also increase the atmospheric dust and heavy metal contaminant level. These contaminants get settled on leaf surfaces at roadsides and enter in contact with phylloplane microorganisms. This study compares microorganisms on leaf surfaces of alder (Alnus nepalensis (Betulaceae)) on roadside and non-roadside environments. Two sites dominated by alder were selected. One at a busy road intersection on the National Highway no. 44 in Shillong with high traffic density (8000-9000 heavy vehicles/day), taken as the polluted site and the other one in a forest approximately 500 m away from the roadside considered as the unpolluted site. Analysis of phylloplane microorganisms, lead, zinc, copper, cadmium and sulphur was carried out from leaves. The bacterial population was higher at the unpolluted site. Bacterial population showed a significant negative correlation with lead, zinc, copper, cadmium and sulphur. Similarly, fungal population was higher at the unpolluted site. A total of 29 fungal species were isolated from the phylloplane of A. nepalensis (polluted site 16 species; unpolluted site 28 species). Some fungal forms like Mortierella sp., Fusarium oxysporum and Aureobasidium pollulans were dominant in the polluted site. Numbers of phylloplane fungi and bacteria were significantly reduced in the polluted site. The correlation coefficient indicated a detrimental effect of metals like lead, zinc, copper, cadmium and sulphur on the microbial community of leaf surfaces. The specificity of certain fungi to the unpolluted site may be attributed to their sensitivity to pollution. The predominance of Aureobasidium pollulans

  20. Nanogel-crosslinked nanoparticles increase the inhibitory effects of W9 synthetic peptide on bone loss in a murine bone resorption model

    Directory of Open Access Journals (Sweden)

    Sato T

    2015-05-01

    Full Text Available Toshimi Sato,1 Neil Alles,1,2 Masud Khan,1,3 Kenichi Nagano,1,4 Mariko Takahashi,1 Yukihiko Tamura,1 Asako Shimoda,5,6 Keiichi Ohya,1 Kazunari Akiyoshi,5,6 Kazuhiro Aoki1 1Department of Bio-Matrix (Pharmacology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan; 2Department of Biochemistry, Faculty of Medicine, University of Peradeniya, Peradeniya, Sri Lanka; 3Department of Dental Pharmacology, City Dental College and Hospital, Dhaka, Bangladesh; 4Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, Boston, MA, USA; 5Department of Polymer Chemistry, Graduate School of Engineering, Kyoto University, Katsura, 6ERATO Akiyoshi Bio-Nanotransporter Project, Japan Science and Technology Agency, Katsura Int’tech Center Kyotodaigaku-Katsura, Nishikyo-ku, Kyoto, Japan Abstract: We investigated the biological activity of W9, a bone resorption inhibitor peptide, using NanoClik nanoparticles as an injectable carrier, where acryloyl group-modified cholesterol-bearing pullulan (CHPOA nanogels were crosslinked by pentaerythritol tetra (mercaptoethyl polyoxyethylene. Thirty 5-week-old male C57BL/6J mice were fed a low calcium diet and received once-daily subcutaneous injections of the carrier alone, W9 24 mg/kg/day alone, W9 24 mg/kg/day incorporated in cholesterol bearing pullulan (CHP nanogels, or W9 (8 and 24 mg/kg/day incorporated in NanoClik nanoparticles for 4 days (n=5. Mice that received a normal calcium diet with NanoClik nanoparticle injections without W9 were used as a control group. Radiological analyses showed that administration of W9 24 mg/kg/day significantly prevented low calcium-induced reduction of bone mineral density in the long bones and lumbar vertebrae, but only when the NanoClik nanoparticles were used as a carrier. Histomorphometric analyses of the proximal tibiae revealed that W9 24 mg/kg/day incorporated in NanoClik nanoparticles prevented the increase in bone resorption indices

  1. Effects of frequency and a radical scavenger on ultrasonic degradation of water-soluble polymers.

    Science.gov (United States)

    Koda, Shinobu; Taguchi, Kimihiko; Futamura, Kazunori

    2011-01-01

    Ultrasonic degradation of methyl cellulose, pullulan, dextran and poly(ethylene oxide) in aqueous solutions was investigated at the frequencies of 20 and 500 kHz, where the ultrasonic power delivered into solutions was kept constant (22 W). The number average molecular mass and the polydispersity were obtained as a function of sonication time. The degradation under sonication at the 500 kHz frequency proceeded faster in comparison with the 20 kHz sonication for four polymers. The addition of a radical scavenger, t-BuOH, resulted in suppression of degradation of water-soluble polymers. The degradation rate constants were estimated from the plot of molecular weight against sonication time. The degradation rate of methyl cellulose was the largest one among the investigated polymers. The difference in the degradation rates was discussed in terms of the flexibility and the hydrodynamic radius of polymer chains in aqueous solutions. Copyright © 2010 Elsevier B.V. All rights reserved.

  2. Efficient secretory expression of functional barley limit dextrinase inhibitor by high cell-density fermentation of Pichia pastoris

    DEFF Research Database (Denmark)

    Jensen, Johanne Mørch; Vester-Christensen, Malene Bech; Møller, Marie Sofie

    2011-01-01

    The limit dextrinase inhibitor (LDI) from barley seeds acts specifically on limit dextrinase (LD), an endogenous starch debranching enzyme. LDI is a 14kDa hydrophobic protein containing four disulfide bonds and one unpaired thiol group previously found to be either glutathionylated or cysteinylated...... the identity of the produced glutathionylated LDI-His6. At a 1:1M ratio the recombinant LDI completely inhibited hydrolysis of pullulan catalyzed by 5–10nM LD. LDI retained stability in the pH 2–12 range and at pH 6.5 displayed a half-life of 53 and 33min at 90 and 93°C, respectively. The efficient...

  3. Recent trends on gellan gum blends with natural and synthetic polymers: A review.

    Science.gov (United States)

    Zia, Khalid Mahmood; Tabasum, Shazia; Khan, Muhammad Faris; Akram, Nadia; Akhter, Naheed; Noreen, Aqdas; Zuber, Mohammad

    2018-04-01

    Gellan gum (GG), a linear negatively charged exopolysaccharide,is biodegradable and non-toxic in nature. It produces hard and translucent gel in the presence of metallic ions which is stable at low pH. However, GG has poor mechanical strength, poor stability in physiological conditions, high gelling temperature and small temperature window.Therefore,it is blended with different polymers such as agar, chitosan, cellulose, sodium alginate, starch, pectin, polyanaline, pullulan, polyvinyl chloride, and xanthan gum. In this article, a comprehensive overview of combination of GG with natural and synthetic polymers/compounds and their applications in biomedical field involving drug delivery system, insulin delivery, wound healing and gene therapy, is presented. It also describes the utilization of GG based materials in food and petroleum industry. All the technical scientific issues have been addressed; highlighting the recent advancement. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Iron complexes of pharmaceutical interest: Antianemics

    Directory of Open Access Journals (Sweden)

    Cakić Milorad D.

    2003-01-01

    Full Text Available Preparations based on different compounds of bi- and trivalent iron are curently used for the prevention and therapy of sideropenic anemia in human and veterinary medicine. The application of preparations based on dextran started about 1950. Up to now, synthesis and production of preparations were performed with the purpose of improving pharmacological performance by using dextran oligosaccharides with different weight - average molar masses and their oxidized and hydrogenated derivatives. Synthesis of polynuclear iron(lll complexs with other oligosaccharides (inulin and pullulan and their derivatives was developed, with potential or valid pharmacological activity for sideropenic anemia treatment.A Review of iron(lll complexes with different oligosaccharides, their physico-chemical characterization pharmaco-biological performance, global structure, further research and possible applications of then complexes, are presented in this paper.

  5. High activity and low temperature optima of extracellular enzymes in Arctic sediments: implications for carbon cycling by heterotrophic microbial communities

    DEFF Research Database (Denmark)

    Arnosti, C.; Jørgensen, BB

    2003-01-01

    (chondroitin sulfate, fucoidan, xylan and pullulan) to determine the temperature-activity responses of hydrolysis of a related class of compounds. All 4 enzyme activities showed similarly low temperature optima in the range of 15 to 18degreesC. These temperature optima are considerably lower than most previous......The rate of the initial step in microbial remineralization of organic carbon, extracellular enzymatic hydrolysis, was investigated as a function of temperature in permanently cold sediments from 2 fjords on the west coast of Svalbard (Arctic Ocean). We used 4 structurally distinct polysaccharides...... reports of temperature optima for enzyme activities in marine sediments. At 0degreesC, close to the in situ temperature, these enzyme activities achieved 13 to 38% of their rates at optimum temperatures. In one experiment, sulfate reduction rates were measured in parallel with extracellular enzymatic...

  6. Glycoproteins functionalized natural and synthetic polymers for prospective biomedical applications: A review.

    Science.gov (United States)

    Tabasum, Shazia; Noreen, Aqdas; Kanwal, Arooj; Zuber, Mohammad; Anjum, Muhammad Naveed; Zia, Khalid Mahmood

    2017-05-01

    Glycoproteins have multidimensional properties such as biodegradability, biocompatibility, non-toxicity, antimicrobial and adsorption properties; therefore, they have wide range of applications. They are blended with different polymers such as chitosan, carboxymethyl cellulose (CMC), polyvinyl pyrrolidone (PVP), polycaprolactone (PCL), heparin, polystyrene fluorescent nanoparticles (PS-NPs) and carboxyl pullulan (PC) to improve their properties like thermal stability, mechanical properties, resistance to pH, chemical stability and toughness. Considering the versatile charateristics of glycoprotein based polymers, this review sheds light on synthesis and characterization of blends and composites of glycoproteins, with natural and synthetic polymers and their potential applications in biomedical field such as drug delivery system, insulin delivery, antimicrobial wound dressing uses, targeting of cancer cells, development of anticancer vaccines, development of new biopolymers, glycoproteome research, food product and detection of dengue glycoproteins. All the technical scientific issues have been addressed; highlighting the recent advancement. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Action of amylolytic and pullulytic enzymes from various anaerobic thermophiles on linear and branched glucose polymers

    Energy Technology Data Exchange (ETDEWEB)

    Koch, R [Goettingen Univ. (Germany, F.R.). Inst. fuer Mikrobiologie; Antranikian, G [Technische Univ. Hamburg-Harburg, Hamburg (Germany, F.R.). Arbeitsbereich Biotechnologie 1

    1990-10-01

    A detailed study has been conducted on the action of starch hydrolyzing enzymes from thermophilic anaerobic bacteria belonging to the genera Clostridium, Thermoanaerobacter and Thermobacteroides. The appearance of multiple bands on polyacrylamide gels with amylolytic as well as pullulytic activities was shown to be a general feature of bacteria investigated. Analysis of the hydrolysis products of each protein band clearly demonstrated the capability of these organisms to hydrolyze {alpha}-1,4-glycosidic bonds in linear oligosaccharides and {alpha}-1,6-glycosidic linkages in pullulan. Furthermore, the enzyme system of thermophilic bacteria investigated was also capable of attacking in the {alpha}-1,6-linkages in branched oligosaccharides. Due to the action of these thermoactive enzymes with multiple specificity an almost complete hydrolysis of raw starch and maltodextrin could be achieved under the same conditions and in one step. (orig.).

  8. Structure and function of α-glucan debranching enzymes

    DEFF Research Database (Denmark)

    Møller, Marie Sofie; Henriksen, Anette; Svensson, Birte

    2016-01-01

    α-Glucan debranching enzymes hydrolyse α-1,6-linkages in starch/glycogen, thereby, playing a central role in energy metabolism in all living organisms. They belong to glycoside hydrolase families GH13 and GH57 and several of these enzymes are industrially important. Nine GH13 subfamilies include α......-glucan debranching enzymes; isoamylase and glycogen debranching enzymes (GH13_11); pullulanase type I/limit dextrinase (GH13_12–14); pullulan hydrolase (GH13_20); bifunctional glycogen debranching enzyme (GH13_25); oligo-1 and glucan-1,6-α-glucosidases (GH13_31); pullulanase type II (GH13_39); and α-amylase domains......_39 enzymes could represent a “missing link” between the strictly α-1,6-specific debranching enzymes and the enzymes with dual specificity and α-1,4-linkage preference....

  9. A test for measuring gustatory function.

    Science.gov (United States)

    Smutzer, Gregory; Lam, Si; Hastings, Lloyd; Desai, Hetvi; Abarintos, Ray A; Sobel, Marc; Sayed, Nabil

    2008-08-01

    The purpose of this study was to determine the usefulness of edible taste strips for measuring human gustatory function. The physical properties of edible taste strips were examined to determine their potential for delivering threshold and suprathreshold amounts of taste stimuli to the oral cavity. Taste strips were then assayed by fluorescence to analyze the uniformity and distribution of bitter tastant in the strips. Finally, taste recognition thresholds for sweet taste were examined to determine whether or not taste strips could detect recognition thresholds that were equal to or better than those obtained from aqueous tests. Edible strips were prepared from pullulan-hydroxypropyl methylcellulose solutions that were dried to a thin film. The maximal amount of a tastant that could be incorporated in a 2.54 cm2 taste strip was identified by including representative taste stimuli for each class of tastant (sweet, sour, salty, bitter, and umami) during strip formation. Distribution of the bitter tastant quinine hydrochloride in taste strips was assayed by fluorescence emission spectroscopy. The efficacy of taste strips for evaluating human gustatory function was examined by using a single series ascending method of limits protocol. Sucrose taste recognition threshold data from edible strips was then compared with results that were obtained from a standard "sip and spit" recognition threshold test. Edible films that formed from a pullulan-hydroxypropyl methylcellulose polymer mixture can be used to prepare clear, thin strips that have essentially no background taste and leave no physical presence after release of tastant. Edible taste strips could uniformly incorporate up to 5% of their composition as tastant. Taste recognition thresholds for sweet taste were over one order of magnitude lower with edible taste strips when compared with an aqueous taste test. Edible taste strips are a highly sensitive method for examining taste recognition thresholds in humans. This

  10. Size and shape of soil humic acids estimated by viscosity and molecular weight.

    Science.gov (United States)

    Kawahigashi, Masayuki; Sumida, Hiroaki; Yamamoto, Kazuhiko

    2005-04-15

    Ultrafiltration fractions of three soil humic acids were characterized by viscometry and high performance size-exclusion chromatography (HPSEC) in order to estimate shapes and hydrodynamic sizes. Intrinsic viscosities under given solute/solvent/temperature conditions were obtained by extrapolating the concentration dependence of reduced viscosities to zero concentration. Molecular mass (weight average molecular weight (M (w)) and number average molecular weight (M (n))) and hydrodynamic radius (R(H)) were determined by HPSEC using pullulan as calibrant. Values of M (w) and M (n) ranged from 15 to 118 x 10(3) and from 9 to 50 x 10(3) (g mol(-1)), respectively. Polydispersity, as indicated by M (w)/M (n), increased with increasing filter size from 1.5 to 2.4. The hydrodynamic radii (R(H)) ranged between 2.2 and 6.4 nm. For each humic acid, M (w) and [eta] were related. Mark-Houwink coefficients calculated on the basis of the M (w)-[eta] relationships suggested restricted flexible chains for two of the humic acids and a branched structure for the third humic acid. Those structures probably behave as hydrated sphere colloids in a good solvent. Hydrodynamic radii of fractions calculated from [eta] using Einstein's equation, which is applicable to hydrated sphere colloids, ranged from 2.2 to 7.1 nm. These dimensions are fit to the size of nanospaces on and between clay minerals and micropores in soil particle aggregates. On the other hand, the good agreement of R(H) values obtained by applying Einstein's equation with those directly determined by HPSEC suggests that pullulan is a suitable calibrant for estimation of molecular mass and size of humic acids by HPSEC.

  11. Application of exopolysaccharides to improve the performance of ceramic bodies in the unidirectional dry pressing process

    Science.gov (United States)

    Caneira, Inês; Machado-Moreira, Bernardino; Dionísio, Amélia; Godinho, Vasco; Neves, Orquídia; Dias, Diamantino; Saiz-Jimenez, Cesareo; Miller, Ana Z.

    2015-04-01

    Ceramic industry represents an important sector of economic activity in the European countries and involves complex and numerous manufacturing processes. The unidirectional dry pressing process includes milling and stirring of raw materials (mainly clay and talc minerals) in aqueous suspensions, followed by spray drying to remove excess water obtaining spray-dried powders further subjected to dry pressing process (conformation). However, spray-dried ceramic powders exhibit an important variability in their performance when subjected to the dry pressing process, particularly in the adhesion to the mold and mechanical strength, affecting the quality of the final conformed ceramic products. Therefore, several synthetic additives (deflocculants, antifoams, binders, lubricants and plasticizers) are introduced in the ceramic slips to achieve uniform and homogeneous pastes, conditioning their rheological properties. However, an important variability associated with the performance of the conformed products is still reported. Exopolysaccharides or Extracellular Polymeric Substances (EPS) are polymers excreted by living organisms, such as bacteria, fungi and algae, which may confer unique and potentially interesting properties with potential industrial uses, such as viscosity control, gelation, and flocculation. Polysaccharides, such as pullulan, gellan, carrageenan and xanthan have found a wide range of applications in food, pharmaceutical, petroleum, and in other industries. The aim of this study was the assessment of exopolysaccharides as natural additives to optimize the performance of spray-dried ceramic powders during the unidirectional dry pressing process, replacing the synthetic additives used in the ceramic production process. Six exopolysaccharides, namely pullulan, gellan, xanthan gum, κappa- and iota-carrageenan, and guar gum were tested in steatite-based spray-dried ceramic powders at different concentrations. Subsequently, these ceramic powders were

  12. Fatty acids from oleaginous yeasts and yeast-like fungi and their potential applications.

    Science.gov (United States)

    Xue, Si-Jia; Chi, Zhe; Zhang, Yu; Li, Yan-Feng; Liu, Guang-Lei; Jiang, Hong; Hu, Zhong; Chi, Zhen-Ming

    2018-02-01

    Oleaginous yeasts, fatty acids biosynthesis and regulation in the oleaginous yeasts and the fatty acids from the oleaginous yeasts and their applications are reviewed in this article. Oleaginous yeasts such as Rhodosporidium toruloides, Yarrowia lipolytica, Rhodotorula mucilaginosa, and Aureobasidium melanogenum, which can accumulate over 50% lipid of their cell dry weight, have many advantages over other oleaginous microorganisms. The fatty acids from the oleaginous yeasts have many potential applications. Many oleaginous yeasts have now been genetically modified to over-produce fatty acids and their derivatives. The most important features of the oleaginous yeasts are that they have special enzymatic systems for enhanced biosynthesis and regulation of fatty acids in their lipid particles. Recently, some oleaginous yeasts such as R. toruloides have been found to have a unique fatty acids synthetase and other oleaginous yeasts such as A. melanogenum have a unique highly reducing polyketide synthase (HR-PKS) involved in the biosynthesis of hydroxyl fatty acids. It is necessary to further enhance lipid biosynthesis using metabolic engineering and explore new applications of fatty acids in biotechnology.

  13. Diverse deep-sea fungi from the South China Sea and their antimicrobial activity.

    Science.gov (United States)

    Zhang, Xiao-Yong; Zhang, Yun; Xu, Xin-Ya; Qi, Shu-Hua

    2013-11-01

    We investigated the diversity of fungal communities in nine different deep-sea sediment samples of the South China Sea by culture-dependent methods followed by analysis of fungal internal transcribed spacer (ITS) sequences. Although 14 out of 27 identified species were reported in a previous study, 13 species were isolated from sediments of deep-sea environments for the first report. Moreover, these ITS sequences of six isolates shared 84-92 % similarity with their closest matches in GenBank, which suggested that they might be novel phylotypes of genera Ajellomyces, Podosordaria, Torula, and Xylaria. The antimicrobial activities of these fungal isolates were explored using a double-layer technique. A relatively high proportion (56 %) of fungal isolates exhibited antimicrobial activity against at least one pathogenic bacterium or fungus among four marine pathogenic microbes (Micrococcus luteus, Pseudoaltermonas piscida, Aspergerillus versicolor, and A. sydowii). Out of these antimicrobial fungi, the genera Arthrinium, Aspergillus, and Penicillium exhibited antibacterial and antifungal activities, while genus Aureobasidium displayed only antibacterial activity, and genera Acremonium, Cladosporium, Geomyces, and Phaeosphaeriopsis displayed only antifungal activity. To our knowledge, this is the first report to investigate the diversity and antimicrobial activity of culturable deep-sea-derived fungi in the South China Sea. These results suggest that diverse deep-sea fungi from the South China Sea are a potential source for antibiotics' discovery and further increase the pool of fungi available for natural bioactive product screening.

  14. Influence of roadside pollution on the phylloplane microbial community of Alnus nepalensis (Betulaceae

    Directory of Open Access Journals (Sweden)

    S.R Joshi

    2008-09-01

    Full Text Available The North Eastern region of India is undergoing industrial development at a faster rate than expected. Roads form the main system of transportation and communication owing to the hilly topography of the region. Automobiles discharge a number of gaseous and trace metal contaminants. Human activities like stone grinding, road construction and sand milling also increase the atmospheric dust and heavy metal contaminant level. These contaminants get settled on leaf surfaces at roadsides and enter in contact with phylloplane microorganisms. This study compares microorganisms on leaf surfaces of alder (Alnus nepalensis (Betulaceae on roadside and non-roadside environments. Two sites dominated by alder were selected. One at a busy road intersection on the National Highway no. 44 in Shillong with high traffic density (8 000-9 000 heavy vehicles/day, taken as the polluted site and the other one in a forest approximately 500 m away from the roadside considered as the unpolluted site. Analysis of phylloplane microorganisms, lead, zinc, copper, cadmium and sulphur was carried out from leaves. The bacterial population was higher at the unpolluted site. Bacterial population showed a significant negative correlation with lead, zinc, copper, cadmium and sulphur. Similarly, fungal population was higher at the unpolluted site. A total of 29 fungal species were isolated from the phylloplane of A. nepalensis (polluted site 16 species; unpolluted site 28 species. Some fungal forms like Mortierella sp., Fusarium oxysporum and Aureobasidium pollulans were dominant in the polluted site. Numbers of phylloplane fungi and bacteria were significantly reduced in the polluted site. The correlation coefficient indicated a detrimental effect of metals like lead, zinc, copper, cadmium and sulphur on the microbial community of leaf surfaces. The specificity of certain fungi to the unpolluted site may be attributed to their sensitivity to pollution. The predominance of

  15. Yeast: An Overlooked Component of Bactrocera tryoni (Diptera: Tephritidae) Larval Gut Microbiota.

    Science.gov (United States)

    Deutscher, Ania T; Reynolds, Olivia L; Chapman, Toni A

    2017-02-01

    Yeasts, often in hydrolyzed form, are key ingredients in the larval and adult diets of tephritid fruit fly colonies. However, very little is known about the presence or role of yeasts in the diets of tephritid fruit flies in nature. Previous studies have identified bacteria but not detected yeasts in the gut of Queensland fruit fly, Bactrocera tryoni (Froggatt), one of Australia's most economically damaging insect pests of horticultural crops and of significant biosecurity concern domestically and internationally. Here we demonstrate that cultivable yeasts are commonly found in the gut of B. tryoni larvae from fruit hosts. Analysis of the ITS1, 5.8S rRNA gene, and ITS2 sequences of randomly selected isolates identified yeasts and yeast-like fungi of the genera Aureobasidium, Candida, Cryptococcus, Hanseniaspora, Pichia, and Starmerella. The prevalence of these yeasts in fruits suggests that larvae consume the yeasts as part of their diet. This work highlights that yeasts should be considered in future tephritid larval gut microbiota studies. Understanding tephritid-microbial symbiont interactions will lead to improvements in artificial diets and the quality of mass-reared tephritids for the sterile insect technique. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  16. Gradual Changes of Gut Microbiota in Weaned Miniature Piglets

    Directory of Open Access Journals (Sweden)

    Xianghua Yan

    2016-11-01

    Full Text Available Colonization of gut microbiota in mammals during the early life is vital to host health. The miniature piglet has recently been considered as an optimal infant model. However, less is known about the development of gut microbiota in miniature piglets. Here, this study was conducted to explore how the gut microbiota develops in weaned Congjiang miniature piglets. In contrast to the relatively stabilized gut fungal community, gut bacterial community showed a marked drop in alpha diversity, accompanied by significant alterations in taxonomic compositions. The relative abundances of 24 bacterial genera significantly declined, whereas the relative abundances of 7 bacterial genera (Fibrobacter, Collinsella, Roseburia, Prevotella, Dorea, Howardella, and Blautia significantly increased with the age of weaned piglets. Fungal taxonomic analysis showed that the relative abundances of 2 genera (Kazachstania and Aureobasidium significantly decreased, whereas the relative abundances of 4 genera (Aspergillus, Cladosporium, Simplicillium, and Candida significantly increased as the piglets aged. Kazachstania telluris was the signature species predominated in gut fungal communities of weaned miniature piglets. The functional maturation of the gut bacterial community was characterized by the significantly increased digestive system, glycan biosynthesis and metabolism, and vitamin B biosynthesis as the piglets aged. These findings suggest that marked gut microbial changes in Congjiang miniature piglets may contribute to understand the potential gut microbiota development of weaned infants.

  17. Screening of wheat endophytes as biological control agents against Fusarium head blight using two different in vitro tests.

    Science.gov (United States)

    Comby, Morgane; Gacoin, Marie; Robineau, Mathilde; Rabenoelina, Fanja; Ptas, Sébastien; Dupont, Joëlle; Profizi, Camille; Baillieul, Fabienne

    2017-09-01

    In order to find biological control agents (BCAs) for the management of Fusarium head blight (FHB), a major disease on wheat crops worldwide, 86 microorganisms isolated from inner tissues of wheat plants were discriminated for their ability to inhibit the growth of Fusarium graminearum and Fusarium culmorum by in vitro dual culture assays. A group of 22 strains appeared very effective to inhibit F. graminearum (inhibition of 30-51%) and they were also globally effective in controlling F. culmorum (inhibition of 15-53%). Further evaluation of a subselection of strains by screening on detached spikelets in vitro confirmed three species, namely Phoma glomerata, Aureobasidium proteae and Sarocladium kiliense, that have not yet been reported for their efficacy against Fusarium spp., indicating that looking for BCAs toward FHB among wheat endophytes proved to be promising. The efficacy of some strains turned out different between both in vitro screening approaches, raising the importance of finding the most appropriate screening approach for the search of BCAs. This study pointed out the interest of the test on detached wheat spikelets that provided information about a potential pathogenicity, the growth capacity and efficacy of the endophyte strains on the targeted plant, before testing them on whole plants. Copyright © 2017 Elsevier GmbH. All rights reserved.

  18. ISOLASI DAN KARAKTERISASI JAMUR LIGNINOLITIK SERTA PERBANDINGAN KEMAMPUANNYA DALAM BIODELIGNIFIKASI

    Directory of Open Access Journals (Sweden)

    Putri Elvira Valencia

    2017-09-01

    Full Text Available Fungi can survive in various environments on different media including wood. Lignin in timber is hard to degrade efficiently because of its polymer form, and only a few of it can be hydrolyzed because of its composite and complex structure. Ligninolytic fungi produce an extracellular enzyme to withstand to toxic or mutagenic chemicals exposure and known to degrade different types of pollutant compounds. Lignin decomposers were also known to play a significant role in the pulping process of paper mills, used in waste treatment such as textile and hydrocarbon wastes. This study was conducted to obtain fungal isolates that have de-lignification capability and to compare the ability of fungal isolates to degrade lignin. The research isolated samples from rotten wood and soil using selective lignin medium with tannic acid as sole C source. This study characterized the isolates by its morphology and identified using the Morphology and Taxonomy of Fungi book by Bessey (1950 as a reference. This study compared the ligninolytic capability by measuring the transparent zone formed on selective lignin media. This research found 14 isolates of fungi and all of them had a ligninolytic capability. Aspergillus niger isolate has the highest ligninolytic capability by producing 6.45 cm clear zone diameter on the 7th day. Aureobasidium sp has the smallest clear zone diameter of 1.9 cm within the same period.

  19. Fungicide contamination reduces beneficial fungi in bee bread based on an area-wide field study in honey bee, Apis mellifera, colonies.

    Science.gov (United States)

    Yoder, Jay A; Jajack, Andrew J; Rosselot, Andrew E; Smith, Terrance J; Yerke, Mary Clare; Sammataro, Diana

    2013-01-01

    Fermentation by fungi converts stored pollen into bee bread that is fed to honey bee larvae, Apis mellifera, so the diversity of fungi in bee bread may be related to its food value. To explore the relationship between fungicide exposure and bee bread fungi, samples of bee bread collected from bee colonies pollinating orchards from 7 locations over 2 years were analyzed for fungicide residues and fungus composition. There were detectable levels of fungicides from regions that were sprayed before bloom. An organic orchard had the highest quantity and variety of fungicides, likely due to the presence of treated orchards within bees' flight range. Aspergillus, Penicillium, Rhizopus, and Cladosporium (beneficial fungi) were the primary fungal isolates found, regardless of habitat differences. There was some variation in fungal components amongst colonies, even within the same apiary. The variable components were Absidia, Alternaria, Aureobasidium, Bipolaris, Fusarium, Geotrichum, Mucor, Nigrospora, Paecilomyces, Scopulariopsis, and Trichoderma. The number of fungal isolates was reduced as an effect of fungicide contamination. Aspergillus abundance was particularly affected by increased fungicide levels, as indicated by Simpson's diversity index. Bee bread showing fungicide contamination originated from colonies, many of which showed chalkbrood symptoms.

  20. FT-IR spectroscopy: A powerful tool for studying the inter- and intraspecific biodiversity of cultivable non-Saccharomyces yeasts isolated from grape must.

    Science.gov (United States)

    Grangeteau, Cédric; Gerhards, Daniel; Terrat, Sebastien; Dequiedt, Samuel; Alexandre, Hervé; Guilloux-Benatier, Michèle; von Wallbrunn, Christian; Rousseaux, Sandrine

    2016-02-01

    The efficiency of the FT-IR technique for studying the inter- and intra biodiversity of cultivable non-Saccharomyces yeasts (NS) present in different must samples was examined. In first, the capacity of the technique FT-IR to study the global diversity of a given sample was compared to the pyrosequencing method, used as a reference technique. Seven different genera (Aureobasidium, Candida, Cryptococcus, Hanseniaspora, Issatchenkia, Metschnikowia and Pichia) were identified by FT-IR and also by pyrosequencing. Thirty-eight other genera were identified by pyrosequencing, but together they represented less than 6% of the average total population of 6 musts. Among the species identified, some of them present organoleptic potentials in winemaking, particularly Starmerella bacillaris (synonym Candidazemplinina). So in a second time, we evaluated the capacity of the FT-IR technique to discriminate the isolates of this species because few techniques were able to study intraspecific NS yeast biodiversity. The results obtained were validated by using a classic method as ITS sequencing. Biodiversity at strain level was high: 19 different strains were identified from 58 isolates. So, FT-IR spectroscopy seems to be an accurate and reliable method for identifying major genera present in the musts. The two biggest advantages of the FT-IR are the capacity to characterize intraspecific biodiversity of non-Saccharomyces yeasts and the possibility to discriminate a lot of strains. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Antifungal activity of the pygidial gland secretion of Laemostenus punctatus (Coleoptera: Carabidae) against cave-dwelling micromycetes

    Science.gov (United States)

    Nenadić, Marija; Ljaljević-Grbić, Milica; Stupar, Miloš; Vukojević, Jelena; Ćirić, Ana; Tešević, Vele; Vujisić, Ljubodrag; Todosijević, Marina; Vesović, Nikola; Živković, Nemanja; Ćurčić, Srećko

    2017-06-01

    The antifungal potential of the pygidial gland secretion of the troglophilic ground beetle Laemostenus punctatus from a cave in Southeastern Serbia against cave-dwelling micromycetes, isolated from the same habitat, has been investigated. Eleven collected samples were analyzed and 32 isolates of cave-dwelling fungi were documented. A total of 14 fungal species were identified as members of the genera Aspergillus, Penicillium, Alternaria, Cladosporium, Rhizopus, Trichoderma, Arthrinium, Aureobasidium, Epicoccum, Talaromyces, and Fusarium. Five isolates were selected for testing the antifungal activity of the pygidial gland secretion: Talaromyces duclauxi, Aspergillus brunneouniseriatus, Penicillium sp., Rhizopus stolonifer, and Trichoderma viride. The microdilution method has been applied to detect minimal inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs). The most sensitive isolate was Penicillium sp., while the other isolates demonstrated a high level of resistance to the tested agent. L. punctatus has developed a special mechanism of producing specific compounds that act synergistically within the secretion mixture, which are responsible for the antifungal action against pathogens from the cave. The results open opportunities for further research in the field of ground beetle defense against pathogens, which could have an important application in human medicine, in addition to the environmental impact, primarily.

  2. Mixed submerged fermentation with two filamentous fungi for cellulolytic and xylanolytic enzyme production.

    Science.gov (United States)

    Garcia-Kirchner, O; Muñoz-Aguilar, M; Pérez-Villalva, R; Huitrón-Vargas, C

    2002-01-01

    The efficient saccharification of lignocellulosic materials requires the cooperative actions of different cellulase enzyme activities: exoglucanase, endoglucanase, beta-glucosidase, and xylanase. Previous studies with the fungi strains Aureobasidium sp. CHTE-18, Penicillium sp. CH-TE-001, and Aspergillus terreus CH-TE-013, selected mainly because of their different cellulolytic and xylanolytic activities, have demonstrated the capacity of culture filtrates of cross-synergistic action in the saccharification of native sugarcane bagasse pith. In an attempt to improve the enzymatic hydrolysis of different cellulosic materials, we investigated a coculture fermentation with two of these strains to enhance the production of cellulases and xylanases. The 48-h batch experimental results showed that the mixed culture of Penicillium sp. CH-TE-001 and A. terreus CH-TE-013 produced culture filtrates with high protein content, cellulase (mainly beta-glucosidase), and xylanase activities compared with the individual culture of each strain. The same culture conditions were used in a simple medium with mineral salts, corn syrup liquor, and sugarcane bagasse pith as the sole carbon source with moderate shaking at 29 degrees C. Finally, we compared the effect of the cell-free culture filtrates obtained from the mixed and single fermentations on the saccharification of different kinds of cellulosic materials.

  3. Distribution of microorganisms in medical devices and their inactivation effects by gamma-irradiation

    International Nuclear Information System (INIS)

    Ito, Hitoshi; Ishigaki, Isao

    1986-01-01

    Radiation treatment is getting important position for sterilizing medical devices and for packaging films of foods. Recently, survey of bioburden is an important technique for evaluation of sterility doses for medical devices. However, many studies have been done mainly on the irradiation effects of spore-forming bacteria in medical devices. In this study, radiation sensitivity of spore-forming bacteria and fungi were examined after the survey on distribution of microorganisms in several kinds of medical devices. The main contaminant in disposable syringes, needles and conical flasks were consisted of Bacillus, with lesser amount of Micrococcus, Pseudomonas, Peptococcaceae and fungi as positive of 5 - 60 % in the medical devices which cultivated in thioglycolate broth. Bacillus group were identified as B. pumilus, B. sphaericus, B. coagulans, B. megaterium and etc. Fungi were isolated a lesser amounts compared with spore-forming bacteria and identified as Blastomyces, Penicillium, Haplosporangium, Euricoa and Audeobasidium. Peptococaceae were not isolated after irradiation with a dose of 0.1 Mrad even the samples were contaminated with high percentage. The D 10 values of dryed endospores of Bacillus-isolates which attached to the filter paper with pepton-glycerin were obtained to be 0.11 - 0.19 Mrad. The D 10 values of many isolates of fungi in dry condition were obtained below 0.08 Mrad. However, the isolate of Aureobasidium is radiation-resistant, and it's D 10 values was obtained as 0.28 Mrad under aerobic and anaerobic dry condition. (author)

  4. Study of Geophilic Keratinophilic Fungi in Public Squares of Jaboticabal-SP

    Directory of Open Access Journals (Sweden)

    Adilson César Abreu Bernardi

    2015-12-01

    Full Text Available Considering the use of public parks as a source of leisure for the population, who maintain an intense and direct contact with the ground, which is a potential source of contamination by different causative agents of mycosis, the purpose of the research was to identify genera of keratinophilic geophilic fungi in the soil of squares in Jaboticabal-SP. 60 soil samples from 15 public squares, and 4 samples of each square were selected and collected. To isolate the soil fungi, we used the method of Vanbreuseghem (1952, modified by Machado (1977. Later the fungi were cultivated in Petri dishes containing Sabouraud Agar added to chloramphenicol, and were incubated at room temperature for about ten days. The grown colonies were isolated in test tubes to obtain pure culture. Macroscopic and microscopic analyses of isolated colonies were made and they were identified at a gender level with the aid of an identification guide. From 60 soil samples collected, 39 were positive for keratinophilic fungi, of which 90 fungic colonies were isolated, being identified the genera Penicillium spp. Fonsecaea spp., Rhizopus spp., Microsporum spp., Fusarium spp., Phialophora spp., Aspergillus spp., Acremonium spp., Nigrospora spp., Trichoderma spp., Bipolaris spp., Aureobasidium spp., Curvularia spp., Mucor spp. and Mycelia sterilia. The results allowed us to conclude that these soils represent a diverse microflora capable of degrading keratin substrates, allowing an assessment of the epidemiological potential represented by the soil in the squares of the city of Jaboticabal-SP.

  5. Aislamiento e identificación de microorganismos en biopelículas provenientes del Castillo de Chapultepec, Ciudad de México.

    Directory of Open Access Journals (Sweden)

    E de la Cruz

    2011-12-01

    Full Text Available Today microbiology is based on pure cultures and separate microorganisms, pure cultures really do not exist in nature, because that microorganism are combined into large colonies slimy (biofilms in which the various individuals establish relationships and dependencies. Only in the United State of America is estimated that biofilms cause billions of dollars in energy losses, equipment damage, product contamination and medical infections. This paper presents the results of the isolation of microorganisms by microbiological and molecular methods, from biofilms located in the Castle of Chapultepec in Mexico City. In the biofilm coexist different genres of filamentous fungi such as Cladosporium, Mucor, Alternaria, Aspergillus, Aureobasidium, Rhodotorula and others in those biofilms; besides bacterial genera as Bacillus, Pantoea, Kokuria, etc. Many of these microbial genera have been widely reported as a participating in biodeterioration processes to monuments and others are reported as contributing to its restoration, so this work opens the door to future researches that allow biological restoration of monuments, conservation and development of new biotechnological processes.   Keywords: Microbiology; pure cultures; biofilms; isolation of microorganism; microbiological methods; biodeterioration processes. 

  6. Corrosion of aluminum alloy 2024 by microorganisms isolated from aircraft fuel tanks.

    Science.gov (United States)

    McNamara, Christopher J; Perry, Thomas D; Leard, Ryan; Bearce, Ktisten; Dante, James; Mitchell, Ralph

    2005-01-01

    Microorganisms frequently contaminate jet fuel and cause corrosion of fuel tank metals. In the past, jet fuel contaminants included a diverse group of bacteria and fungi. The most common contaminant was the fungus Hormoconis resinae. However, the jet fuel community has been altered by changes in the composition of the fuel and is now dominated by bacterial contaminants. The purpose of this research was to determine the composition of the microbial community found in fuel tanks containing jet propellant-8 (JP-8) and to determine the potential of this community to cause corrosion of aluminum alloy 2024 (AA2024). Isolates cultured from fuel tanks containing JP-8 were closely related to the genus Bacillus and the fungi Aureobasidium and Penicillium. Biocidal activity of the fuel system icing inhibitor diethylene glycol monomethyl ether is the most likely cause of the prevalence of endospore forming bacteria. Electrochemical impedance spectroscopy and metallographic analysis of AA2024 exposed to the fuel tank environment indicated that the isolates caused corrosion of AA2024. Despite the limited taxonomic diversity of microorganisms recovered from jet fuel, the community has the potential to corrode fuel tanks.

  7. Filamentous fungi associated with natural infection of noble rot on withered grapes.

    Science.gov (United States)

    Lorenzini, M; Simonato, B; Favati, F; Bernardi, P; Sbarbati, A; Zapparoli, G

    2018-05-02

    The effects of noble rot infection of grapes on the characteristics of different types of wine, including Italian passito wine, are well known. Nevertheless, there is still little information on filamentous fungi associated with noble-rotten grapes. In this study, withered Garganega grapes for passito wine production, naturally infected by noble rot, were analyzed and compared to sound grapes. Skin morphology and fungal population on berry surfaces were analyzed. Scanning electron microscopy analysis revealed microcracks, germination conidia and branched hyphae on noble-rotten berries. Penicillium, Aureobasidium and Cladosporium were the most frequent genera present. Analysis of single berries displayed higher heterogeneity of epiphytic fungi in those infected by noble-rot than in sound berries. Penicillium adametzoides, Cladosporium cladospoirioides and Coniochaeta polymorpha were recovered. These, to the best of our knowledge, had never been previously isolated from withered grapes and, for C. polymorpha, from grapevine. This study provided novel data on noble rot mycobiota and suggests that fungi that co-habit with B. cinerea could have an important role on grape and wine quality. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. Potential contribution of microbial communities in technical ceramics for the improvement of rheological properties

    Science.gov (United States)

    Moreira, Bernardino; Miller, Ana Z.; Santos, Ricardo; Monteiro, Sílvia; Dias, Diamantino; Neves, Orquídia; Dionísio, Amélia; Saiz-Jimenez, Cesareo

    2014-05-01

    Several bacterial and fungal species naturally occurring in ceramic raw materials used in construction, such as Aspergillus, Penicillium and Aureobasidium, are known to produce exopolysaccharides (EPS). These polymers excreted by the cells are of widespread occurrence and may confer unique and potentially interesting properties with potential industrial uses, such as viscosity control, gelation, and flocculation, during ceramic manufacturing. In this study, the microbial communities present in clay raw materials were identified by both cultural methods and DNA-based molecular techniques in order to appraise their potential contribution to enhance the performance of technical ceramics through the use of EPS. Mineralogical identification by X- Ray Diffraction (XRD) and Fourier Transform Infrared (FTIR) spectroscopy of the clay raw materials, as well as characterization of rheological properties of ceramic slips were also performed. Microbial EPS production and its introduction into ceramic slips will be then carried out in order to evaluate their effects on the rheological properties of the ceramic slips, powders and conformed bodies. Some positive aspects related to the use of EPS are: reduction of the environmental impact caused by synthetic organic additives, reduction of production costs, as well as the costs related with operator protection systems, gaseous effluent treatments, complex landfill, among others.

  9. MINYAK KELAPA BERPOTENSI SEBAGAI PENGAWET BUAH DAN SAYURAN

    Directory of Open Access Journals (Sweden)

    Nunik Siti Aminah

    2012-07-01

    Full Text Available Abstract. Lack of information about coconut oil for healing some diseases, makes people ignore to use coconut oil. after the Vico (Virgin Coconut Oil recognized, now people pay attention to coconut oil. Coconut oil contain laurat acid in a large amount and this thing with other acids in coconut oil is very potential as fruit and vegetables preservative, because it is cheap, easy to apply, has litlle side effect, safe for non target and environment friendly. This Coconut oil to prolong the time of fruit saving. Pear has the followed by apple, Nephelium litchi, mango, grape and Salacca edulis. For Vegetables : potatotollamed, cucumber,tomato, carrot and paprica. The microba that can be prevented to grow are : Eschericia coli, Penicillium sp, Aspergillus niger and Scopulariopsi sp. The microba that can contaminate the fruits and vegetables are : Klebsiella, Fusariuim, Geotrichum, Scopulariopsis, Penicillium, Aspergillus, Mucor, Rhizopus, Chrysomillia, Botrytis, Synchepalastrum, Aureobasidium, Neosartorya and Monascus. Among all the fungi that produce toxin, which is called micotoxin is very harmful to human and animal health. Using coconut oil as preservatives is very easy if fruits vegetables are in small amount, they only need a paint-brush that already dipin to coconut oil then rub it to clean fruits vegetables. But if the fruit vegetables that will be preserved are in a large container filled with coconut oil is needed to soak the fruits large vegetables for 10 seconds.Key words : Coconut oil, Natural preservative, Fruit and Vegetable.

  10. Characterization of Grewia Gum, a Potential Pharmaceutical Excipient

    Directory of Open Access Journals (Sweden)

    Elijah.I.Nep

    2010-03-01

    Full Text Available Grewia gum was extracted from the inner stem bark of Grewia mollis and characterized by several techniques such as gas chromatography (GC, gel permeation chromatography (GPC, scanning electron microscopy (SEM, differential scanning calorimetry (DSC and thermogravimetric analysis of the extracted sample. Spectroscopic techniques such as x-ray photoelectron spectroscopy (XPS, fourier-transformed infrared (FT-IR, solid-state nuclear magnetic resonance (NMR, and 1H and 13C NMR techniques were also used to characterize the gum. The results showed that grewia gum is a typically amorphous polysaccharide gum containing glucose, rhamnose, galactose, arabinose and xylose as neutral sugars. It has an average molecular weight of 5925 kDa expressed as the pullulan equivalent. The gum slowly hydrated in water, dispersing and swelling to form a highly viscous dispersion exhibiting pseudoplastic flow behaviour. The polysaccharide gum is thermally stable and may have application as stabilizer or suspending agent in foods, cosmetics and in pharmaceuticals. It may have application as a binder or sustained-release polymer matrix in tablets or granulations.

  11. Biopolymer-Based Nanoparticles for Drug/Gene Delivery and Tissue Engineering

    Science.gov (United States)

    Nitta, Sachiko Kaihara; Numata, Keiji

    2013-01-01

    There has been a great interest in application of nanoparticles as biomaterials for delivery of therapeutic molecules such as drugs and genes, and for tissue engineering. In particular, biopolymers are suitable materials as nanoparticles for clinical application due to their versatile traits, including biocompatibility, biodegradability and low immunogenicity. Biopolymers are polymers that are produced from living organisms, which are classified in three groups: polysaccharides, proteins and nucleic acids. It is important to control particle size, charge, morphology of surface and release rate of loaded molecules to use biopolymer-based nanoparticles as drug/gene delivery carriers. To obtain a nano-carrier for therapeutic purposes, a variety of materials and preparation process has been attempted. This review focuses on fabrication of biocompatible nanoparticles consisting of biopolymers such as protein (silk, collagen, gelatin, β-casein, zein and albumin), protein-mimicked polypeptides and polysaccharides (chitosan, alginate, pullulan, starch and heparin). The effects of the nature of the materials and the fabrication process on the characteristics of the nanoparticles are described. In addition, their application as delivery carriers of therapeutic drugs and genes and biomaterials for tissue engineering are also reviewed. PMID:23344060

  12. Biopolymer-Based Nanoparticles for Drug/Gene Delivery and Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Keiji Numata

    2013-01-01

    Full Text Available There has been a great interest in application of nanoparticles as biomaterials for delivery of therapeutic molecules such as drugs and genes, and for tissue engineering. In particular, biopolymers are suitable materials as nanoparticles for clinical application due to their versatile traits, including biocompatibility, biodegradability and low immunogenicity. Biopolymers are polymers that are produced from living organisms, which are classified in three groups: polysaccharides, proteins and nucleic acids. It is important to control particle size, charge, morphology of surface and release rate of loaded molecules to use biopolymer-based nanoparticles as drug/gene delivery carriers. To obtain a nano-carrier for therapeutic purposes, a variety of materials and preparation process has been attempted. This review focuses on fabrication of biocompatible nanoparticles consisting of biopolymers such as protein (silk, collagen, gelatin, β-casein, zein and albumin, protein-mimicked polypeptides and polysaccharides (chitosan, alginate, pullulan, starch and heparin. The effects of the nature of the materials and the fabrication process on the characteristics of the nanoparticles are described. In addition, their application as delivery carriers of therapeutic drugs and genes and biomaterials for tissue engineering are also reviewed.

  13. "Real-time" disintegration analysis and D-optimal experimental design for the optimization of diclofenac sodium fast-dissolving films.

    Science.gov (United States)

    El-Malah, Yasser; Nazzal, Sami

    2013-01-01

    The objective of this work was to study the dissolution and mechanical properties of fast-dissolving films prepared from a tertiary mixture of pullulan, polyvinylpyrrolidone and hypromellose. Disintegration studies were performed in real-time by probe spectroscopy to detect the onset of film disintegration. Tensile strength and elastic modulus of the films were measured by texture analysis. Disintegration time of the films ranged from 21 to 105 seconds whereas their mechanical properties ranged from approximately 2 to 49 MPa for tensile strength and 1 to 21 MPa% for young's modulus. After generating polynomial models correlating the variables using a D-Optimal mixture design, an optimal formulation with desired responses was proposed by the statistical package. For validation, a new film formulation loaded with diclofenac sodium based on the optimized composition was prepared and tested for dissolution and tensile strength. Dissolution of the optimized film was found to commence almost immediately with 50% of the drug released within one minute. Tensile strength and young's modulus of the film were 11.21 MPa and 6, 78 MPa%, respectively. Real-time spectroscopy in conjunction with statistical design were shown to be very efficient for the optimization and development of non-conventional intraoral delivery system such as fast dissolving films.

  14. Alternansucrase acceptor reactions with D-tagatose and L-glucose.

    Science.gov (United States)

    Côté, Gregory L; Dunlap, Christopher A; Appell, Michael; Momany, Frank A

    2005-02-07

    Alternansucrase (EC 2.4.1.140) is a d-glucansucrase that synthesizes an alternating alpha-(1-->3), (1-->6)-linked d-glucan from sucrose. It also synthesizes oligosaccharides via d-glucopyranosyl transfer to various acceptor sugars. Two of the more efficient monosaccharide acceptors are D-tagatose and L-glucose. In the presence of d-tagatose, alternansucrase produced the disaccharide alpha-d-glucopyranosyl-(1-->1)-beta-D-tagatopyranose via glucosyl transfer. This disaccharide is analogous to trehalulose. We were unable to isolate a disaccharide product from L-glucose, but the trisaccharide alpha-D-glucopyranosyl-(1-->6)-alpha-d-glucopyranosyl-(1-->4)-l-glucose was isolated and identified. This is analogous to panose, one of the structural units of pullulan, in which the reducing-end D-glucose residue has been replaced by its L-enantiomer. The putative L-glucose disaccharide product, produced by glucoamylase hydrolysis of the trisaccharide, was found to be an acceptor for alternansucrase. The disaccharide, alpha-D-glucopyranosyl-(1-->4)-L-glucose, was a better acceptor than maltose, previously the best known acceptor for alternansucrase. A structure comparison of alpha-D-glucopyranosyl-(1-->4)-L-glucose and maltose was performed through computer modeling to identify common features, which may be important in acceptor affinity by alternansucrase.

  15. Formation of polyelectrolyte complexes with diethylaminoethyl dextran: charge ratio and molar mass effect.

    Science.gov (United States)

    Le Cerf, Didier; Pepin, Anne Sophie; Niang, Pape Momar; Cristea, Mariana; Karakasyan-Dia, Carole; Picton, Luc

    2014-11-26

    The formation of polyelectrolyte complexes (PECs) between carboxymethyl pullulan and DEAE Dextran, was investigated, in dilute solution, with emphasis on the effect of charge density (molar ratio or pH) and molar masses. Electrophoretic mobility measurements have evidenced that insoluble PECs (neutral electrophoretic mobility) occurs for charge ratio between 0.6 (excess of polycation) and 1 (stoichiometry usual value) according to the pH. This atypical result is explained by the inaccessibility of some permanent cationic charge when screened by pH dependant cationic ones (due to the Hoffman alkylation). Isothermal titration calorimetry (ITC) indicates an endothermic formation of PEC with a binding constant around 10(5) L mol(-1). Finally asymmetrical flow field flow fractionation coupled on line with static multi angle light scattering (AF4/MALS) evidences soluble PECs with very large average molar masses and size around 100 nm, in agreement with scrambled eggs multi-association between various polyelectrolyte chains. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Analysis of the hydrolysis of inulin using real time 1H NMR spectroscopy

    Science.gov (United States)

    Barclay, Thomas; Ginic-Markovic, Milena; Johnston, Martin R.; Cooper, Peter D.; Petrovsky, Nikolai

    2012-01-01

    The hydrolysis of various carbohydrates was investigated under acidic conditions in real time by 1H NMR spectroscopy, with a focus on the polysaccharide inulin. Sucrose was used as a model compound to illustrate the applicability of this technique. The hydrolysis of sucrose was shown to follow pseudo first order kinetics and have an activation energy of 107.0 kJ.mol−1 (s.d. 1.7 kJ.mol−1). Inulin, pullulan and glycogen also all followed pseudo first order kinetics, but had an initiation phase at least partially generated by the protonation of the glycosidic bonds. It was also demonstrated that polysaccharide chain length has an effect on the hydrolysis of inulin. For short chain inulin (DPn 18, s.d. 0.70) the activation energy calculated for the hydrolytic cleavage of glucose was similar to sucrose at 108.5 kJ.mol−1 (std. dev. 0.60). For long chain inulin (DPn 30, s.d. 1.3) the activation energy for the hydrolytic cleavage of glucose was reduced to 80.5 kJ.mol−1 (s.d. 2.3 kJ.mol−1). This anomaly has been attributed to varied conformations for the two different lengths of inulin chain in solution. PMID:22464225

  17. Insulin Inclusion into a Tragacanth Hydrogel: An Oral Delivery System for Insulin

    Directory of Open Access Journals (Sweden)

    Mokhamad Nur

    2018-01-01

    Full Text Available Nanoparticles or microparticles created by physical complexation between two polyelectrolytes may have a prospective use as an excipient for oral insulin administration. Natural polymers such as tragacanth, alginate, dextran, pullulan, hyaluronic acid, gelatin and chitosan can be potential candidates for this purpose. In this research, insulin particles were prepared by the inclusion of insulin into a tragacanth hydrogel. The effect of the pH and concentration relationship involving polyelectrolytes offering individual particle size and zeta potential was assessed by zetasizer and scanning electron microscopy (SEM. Insulin–tragacanth interactions at varying pH (3.7, 4.3, 4.6, or 6, and concentration (0.1%, 0.5%, or 1% w/w were evaluated by differential scanning calorimetry (DSC and ATR Fourier transform infrared (ATR-FTIR analysis. Individual and smaller particles, approximately 800 nm, were acquired at pH 4.6 with 0.5% of tragacanth. The acid gelation test indicated that insulin could be entrapped in the physical hydrogel of tragacanth. DSC thermograms of insulin–tragacanth showed shifts on the same unloaded tragacanth peaks and suggested polyelectrolyte–protein interactions at a pH close to 4.3–4.6. FTIR spectra of tragacanth–insulin complexes exhibited amide absorption bands featuring in the protein spectra and revealed the creation of a new chemical substance.

  18. Mikrobiyel ekzopolisakkaritler

    Directory of Open Access Journals (Sweden)

    Erdi Ergene

    2016-08-01

    Full Text Available Mikrobiyel ekzopolisakkaritler (EPS birçok mikroorganizma tarafından üretilerek dış ortama salgılanan karbonhidrat yapısındaki biyopolimerlerdir. EPS, mikroorganizmayı kurumadan, fagositozdan, faj etkisinden korur, ısı, ışık, ses gibi stres ortamında bariyer görevi görür. Endüstriyel amaçla üretilen EPS’ler genellikle gıda, kozmetik, petrol, kimya gibi alanlarda kullanılmaktadır. Mikrobiyel ekzopolisakkaritler gıdalarda stabilizatör, emülgatör, jelleştirme ajanı, nem tutucu, kıvam verici gibi amaçlarla kullanılmaktadır. Bu amaç için en çok kullanılan mikrobiyel ekzopolisakkaritlerin başında ksantan ve jellan gelmektedir. Bunların yanında, dekstran, kurdlan, levan, pullulan, bakteriyel aljinatlar gibi önemli özelliklere sahip EPS’lerin kısıtlı da olsa kullanımları mevcuttur. Bu derlemede, mikrobiyel EPS’lerin özellikleri ve önemi üzerinde durulmuştur.

  19. Smart stimuli sensitive nanogels in cancer drug delivery and imaging: a review.

    Science.gov (United States)

    Maya, S; Sarmento, Bruno; Nair, Amrita; Rejinold, N Sanoj; Nair, Shantikumar V; Jayakumar, R

    2013-01-01

    Nanogels are nanosized hydrogel particles formed by physical or chemical cross-linked polymer networks. The advantageous properties of nanogels related to the ability of retaining considerable amount of water, the biocompatibility of the polymers used, the ability to encapsulate and protect a large quantity of payload drugs within the nanogel matrix, the high stability in aqueous media, their stimuli responsively behavior potential, and the versatility in release drugs in a controlled manner make them very attractive for use in the area of drug delivery. The materials used for the preparation of nanogels ranged from natural polymers like ovalbumin, pullulan, hyaluronic acid, methacrylated chondroitin sulfate and chitosan, to synthetic polymers like poly (N-isopropylacrylamide), poly (Nisopropylacrylamide- co-acrylic acid) and poly (ethylene glycol)-b-poly (methacrylic acid). The porous nanogels have been finding application as anti-cancer drug and imaging agent reservoirs. Smart nanogels responding to external stimuli such as temperature, pH etc can be designed for diverse therapeutic and diagnostic applications. The nanogels have also been surface functionalized with specific ligands aiding in targeted drug delivery. This review focus on stimuli-sensitive, multi-responsive, magnetic and targeted nanogels providing a brief insight on the application of nanogels in cancer drug delivery and imaging in detail.

  20. Thermo-controlled rheology of electro-assembled polyanionic polysaccharide (alginate) and polycationic thermo-sensitive polymers.

    Science.gov (United States)

    Niang, Pape Momar; Huang, Zhiwei; Dulong, Virginie; Souguir, Zied; Le Cerf, Didier; Picton, Luc

    2016-03-30

    Several thermo-sensitive polyelectrolyte complexes were prepared by ionic self-association between an anionic polysaccharide (alginate) and a monocationic copolymer (polyether amine, Jeffamine®-M2005) with a 'Low Critical Solubility Temperature' (LCST). We show that electro-association must be established below the aggregation temperature of the free Jeffamine®, after which the organization of the system is controlled by the thermo-association of Jeffamine® that was previously electro-associated with the alginate. Evidence for this comes primarily from the rheology in the semi-dilute region. Electro- and thermo-associative behaviours are optimal at a pH corresponding to maximum ionization of both compounds (around pH 7). High ionic strength could prevent the electro-association. The reversibility of the transition is possible only at temperatures lower than the LCST of Jeffamine®. Similar behaviour has been obtained with carboxymethyl cellulose (CMC), which suggests that this behaviour can be observed using a range of anionic polyelectrolytes. In contrast, no specific properties have been found for pullulan, which is a neutral polysaccharide. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Antifungal Activities of Extracts from Selected Lebanese Wild Plants against Plant Pathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Y. Abou-Jawdah

    2004-12-01

    Full Text Available Extracts of nine plant species growing wild in Lebanon were tested for their efficacy against seven plant pathogenic fungi: Botrytis cinerea, Alternaria solani, Penicillium sp., Cladosporium sp., Fusarium oxysporum f. sp. melonis, Rhizoctonia solani and Sphaerotheca cucurbitae. Extracts of three of the plants, Origanum syriacum, Micromeria nervosa and Plumbago maritima, showed the highest levels of in vitro activity against spore germination and mycelial growth of the fungi tested. Inula viscosa showed high activity against spore germination but only moderate activity against mycelial growth. The other five plant species tested Calamintha origanifolia, Micromeria juliana, Ruta sp., Sideritis pullulans and Urginea maritima showed only moderate to low activity against these fungi. Preventive sprays with extracts of O. syriacum, M. nervosa, P. maritima and I. viscosa, applied at concentrations ranging between 4 and 8% to squash and cucumber seedlings, gave efficient protection against gray mold caused by B. cinerea and powdery mildew caused by S. cucurbitae. However, these extracts did not control green mold of citrus fruits caused by Penicillium sp. Thin layer chromatography revealed three inhibitory bands in extracts of O. syriacum, two in I. viscosa and only one in each of the other plants tested: M. nervosa, P. maritima, C. origanifolia and Ruta sp.

  2. Transposon mutagenesis in Bifidobacterium breve: construction and characterization of a Tn5 transposon mutant library for Bifidobacterium breve UCC2003.

    Science.gov (United States)

    Ruiz, Lorena; Motherway, Mary O'Connell; Lanigan, Noreen; van Sinderen, Douwe

    2013-01-01

    Bifidobacteria are claimed to contribute positively to human health through a range of beneficial or probiotic activities, including amelioration of gastrointestinal and metabolic disorders, and therefore this particular group of gastrointestinal commensals has enjoyed increasing industrial and scientific attention in recent years. However, the molecular mechanisms underlying these probiotic mechanisms are still largely unknown, mainly due to the fact that molecular tools for bifidobacteria are rather poorly developed, with many strains lacking genetic accessibility. In this work, we describe the generation of transposon insertion mutants in two bifidobacterial strains, B. breve UCC2003 and B. breve NCFB2258. We also report the creation of the first transposon mutant library in a bifidobacterial strain, employing B. breve UCC2003 and a Tn5-based transposome strategy. The library was found to be composed of clones containing single transposon insertions which appear to be randomly distributed along the genome. The usefulness of the library to perform phenotypic screenings was confirmed through identification and analysis of mutants defective in D-galactose, D-lactose or pullulan utilization abilities.

  3. Transposon mutagenesis in Bifidobacterium breve: construction and characterization of a Tn5 transposon mutant library for Bifidobacterium breve UCC2003.

    Directory of Open Access Journals (Sweden)

    Lorena Ruiz

    Full Text Available Bifidobacteria are claimed to contribute positively to human health through a range of beneficial or probiotic activities, including amelioration of gastrointestinal and metabolic disorders, and therefore this particular group of gastrointestinal commensals has enjoyed increasing industrial and scientific attention in recent years. However, the molecular mechanisms underlying these probiotic mechanisms are still largely unknown, mainly due to the fact that molecular tools for bifidobacteria are rather poorly developed, with many strains lacking genetic accessibility. In this work, we describe the generation of transposon insertion mutants in two bifidobacterial strains, B. breve UCC2003 and B. breve NCFB2258. We also report the creation of the first transposon mutant library in a bifidobacterial strain, employing B. breve UCC2003 and a Tn5-based transposome strategy. The library was found to be composed of clones containing single transposon insertions which appear to be randomly distributed along the genome. The usefulness of the library to perform phenotypic screenings was confirmed through identification and analysis of mutants defective in D-galactose, D-lactose or pullulan utilization abilities.

  4. Status, Antimicrobial Mechanism, and Regulation of Natural Preservatives in Livestock Food Systems.

    Science.gov (United States)

    Lee, Na-Kyoung; Paik, Hyun-Dong

    2016-01-01

    This review discusses the status, antimicrobial mechanisms, application, and regulation of natural preservatives in livestock food systems. Conventional preservatives are synthetic chemical substances including nitrates/nitrites, sulfites, sodium benzoate, propyl gallate, and potassium sorbate. The use of artificial preservatives is being reconsidered because of concerns relating to headache, allergies, and cancer. As the demand for biopreservation in food systems has increased, new natural antimicrobial compounds of various origins are being developed, including plant-derived products (polyphenolics, essential oils, plant antimicrobial peptides (pAMPs)), animal-derived products (lysozymes, lactoperoxidase, lactoferrin, ovotransferrin, antimicrobial peptide (AMP), chitosan and others), and microbial metabolites (nisin, natamycin, pullulan, ε-polylysine, organic acid, and others). These natural preservatives act by inhibiting microbial cell walls/membranes, DNA/RNA replication and transcription, protein synthesis, and metabolism. Natural preservatives have been recognized for their safety; however, these substances can influence color, smell, and toxicity in large amounts while being effective as a food preservative. Therefore, to evaluate the safety and toxicity of natural preservatives, various trials including combinations of other substances or different food preservation systems, and capsulation have been performed. Natamycin and nisin are currently the only natural preservatives being regulated, and other natural preservatives will have to be legally regulated before their widespread use.

  5. Insulin Inclusion into a Tragacanth Hydrogel: An Oral Delivery System for Insulin

    Science.gov (United States)

    Vasiljevic, Todor

    2018-01-01

    Nanoparticles or microparticles created by physical complexation between two polyelectrolytes may have a prospective use as an excipient for oral insulin administration. Natural polymers such as tragacanth, alginate, dextran, pullulan, hyaluronic acid, gelatin and chitosan can be potential candidates for this purpose. In this research, insulin particles were prepared by the inclusion of insulin into a tragacanth hydrogel. The effect of the pH and concentration relationship involving polyelectrolytes offering individual particle size and zeta potential was assessed by zetasizer and scanning electron microscopy (SEM). Insulin–tragacanth interactions at varying pH (3.7, 4.3, 4.6, or 6), and concentration (0.1%, 0.5%, or 1% w/w) were evaluated by differential scanning calorimetry (DSC) and ATR Fourier transform infrared (ATR-FTIR) analysis. Individual and smaller particles, approximately 800 nm, were acquired at pH 4.6 with 0.5% of tragacanth. The acid gelation test indicated that insulin could be entrapped in the physical hydrogel of tragacanth. DSC thermograms of insulin–tragacanth showed shifts on the same unloaded tragacanth peaks and suggested polyelectrolyte–protein interactions at a pH close to 4.3–4.6. FTIR spectra of tragacanth–insulin complexes exhibited amide absorption bands featuring in the protein spectra and revealed the creation of a new chemical substance. PMID:29304023

  6. Effects of Polysaccharide-Based Edible Coatings on Quality and Antioxidant Enzyme System of Strawberry during Cold Storage

    Directory of Open Access Journals (Sweden)

    Li Li

    2017-01-01

    Full Text Available Strawberry is a nutritious, but highly perishable fruit. Three polysaccharide-based edible coatings (alginate, chitosan, and pullulan were applied to postharvest strawberry fruit during cold storage (4°C, and their effects on fruit quality and antioxidant enzyme system were investigated in the present study. The results showed that polysaccharide coatings showed a significant delay in fruit softening and rot and reduced changes in total soluble solid and titratable acidity content during 16 d storage. Polysaccharide coatings also maintained higher ascorbic acid and total phenolic contents than control from day 2 and significantly inhibited fruit decay and respiration after 12 d storage (p<0.05. Polysaccharide treatments enhanced the activities of antioxidant enzymes (peroxidase, catalase, superoxide dismutase, and ascorbate peroxidase so as to prevent lipid peroxidation and reduce membrane damage. Additionally, chitosan coating had the most positive effects on fruit quality amongst three polysaccharide-based edible coatings and presented the highest relative activities of antioxidant enzymes. These results indicated that polysaccharide-based edible coatings were helpful in postharvest quality maintenance of strawberry fruit.

  7. Design of magnetic gene complexes as effective and serum resistant gene delivery systems for mesenchymal stem cells.

    Science.gov (United States)

    Zhang, Tian-Yuan; Wu, Jia-He; Xu, Qian-Hao; Wang, Xia-Rong; Lu, Jingxiong; Hu, Ying; Jo, Jun-Ichiro; Yamamoto, Masaya; Ling, Daishun; Tabata, Yasuhiko; Gao, Jian-Qing

    2017-03-30

    Gene engineered mesenchymal stem cells (MSCs) have been proposed as promising tools for their various applications in biomedicine. Nevertheless, the lack of an effective and safe way to genetically modify these stem cells is still a major obstacle in the current studies. Herein, we designed novel magnetic complexes by assembling cationized pullulan derivatives with magnetic iron oxide nanoparticles for delivering target genes to MSCs. Results showed that this complexes achieved effective gene expression with the assistance of external magnetic field, and resisted the adverse effect induced by serum proteins on the gene delivery. Moreover, neither significant cytotoxicity nor the interference on the osteogenic differentiation to MSCs were observed after magnetofection. Further studies revealed that this effective and serum resistant gene transfection was partly due to the accelerated and enhanced intracellular uptake process driven by external magnetic field. To conclude, the current study presented a novel option for genetic modification of MSCs in an effective, relatively safe and serum compatible way. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Characterization of the oral fungal microbiome (mycobiome in healthy individuals.

    Directory of Open Access Journals (Sweden)

    Mahmoud A Ghannoum

    2010-01-01

    Full Text Available The oral microbiome-organisms residing in the oral cavity and their collective genome-are critical components of health and disease. The fungal component of the oral microbiota has not been characterized. In this study, we used a novel multitag pyrosequencing approach to characterize fungi present in the oral cavity of 20 healthy individuals, using the pan-fungal internal transcribed spacer (ITS primers. Our results revealed the "basal" oral mycobiome profile of the enrolled individuals, and showed that across all the samples studied, the oral cavity contained 74 culturable and 11 non-culturable fungal genera. Among these genera, 39 were present in only one person, 16 genera were present in two participants, and 5 genera were present in three people, while 15 genera (including non-culturable organisms were present in >/=4 (20% participants. Candida species were the most frequent (isolated from 75% of participants, followed by Cladosporium (65%, Aureobasidium, Saccharomycetales (50% for both, Aspergillus (35%, Fusarium (30%, and Cryptococcus (20%. Four of these predominant genera are known to be pathogenic in humans. The low-abundance genera may represent environmental fungi present in the oral cavity and could simply be spores inhaled from the air or material ingested with food. Among the culturable genera, 61 were represented by one species each, while 13 genera comprised between 2 and 6 different species; the total number of species identified were 101. The number of species in the oral cavity of each individual ranged between 9 and 23. Principal component (PCO analysis of the obtained data set followed by sample clustering and UniFrac analysis revealed that White males and Asian males clustered differently from each other, whereas both Asian and White females clustered together. This is the first study that identified the "basal mycobiome" of healthy individuals, and provides the basis for a detailed characterization of the oral mycobiome in

  9. Production of xylanases by mangrove fungi from the Philippines and their application in enzymatic pretreatment of recycled paper pulps.

    Science.gov (United States)

    Torres, Jeremy Martin O; Dela Cruz, Thomas Edison E

    2013-04-01

    Mangrove fungi are vastly unexplored for enzymes with industrial application. This study aimed to assess the biocatalytic activity of mangrove fungal xylanases on recycled paper pulp. Forty-four mangrove fungal (MF) isolates were initially screened for xylanolytic activity in minimal medium with corn cob xylan as the sole carbon source. Eight MF were further cultivated under submerged fermentation for the production of crude xylanases. These crude enzymes were then characterized and tested for the pretreatment of recycled paper pulps. Results showed that 93 % of the tested MF isolates exhibited xylanolytic activity in solid medium. In submerged fermentation, salinity improved the growth of the fungal isolates but did not influence xylanase production. The crude xylanases were mostly optimally active at 50 °C and pH 7. Changes in pH had a greater effect on xylanase stability than temperature. More than half of the activity was lost at pH 9 for majority of the crude enzymes. However, two thermophilic xylanases from Fusarium sp. KAWIT-A and Aureobasidium sp. 2LIPA-M and one alkaliphilic xylanase from Phomopsis sp. MACA-J were also produced. All crude enzymes exhibited cellulase activities ranging from 4 to 21 U/ml. Enzymatic pretreatment of recycled paper pulps with 5 % consistency produced 70-650 mg of reducing sugars per gram of pulp at 50 °C after 60 min. The release of high amounts of reducing sugars showed the potential of mangrove fungal crude xylanases in the local paper and pulp industry. The diverse properties shown by the tested crude enzymes also indicate its potential applications to other enzyme-requiring industries.

  10. Metabolomic and high-throughput sequencing analysis—modern approach for the assessment of biodeterioration of materials from historic buildings

    Science.gov (United States)

    Gutarowska, Beata; Celikkol-Aydin, Sukriye; Bonifay, Vincent; Otlewska, Anna; Aydin, Egemen; Oldham, Athenia L.; Brauer, Jonathan I.; Duncan, Kathleen E.; Adamiak, Justyna; Sunner, Jan A.; Beech, Iwona B.

    2015-01-01

    Preservation of cultural heritage is of paramount importance worldwide. Microbial colonization of construction materials, such as wood, brick, mortar, and stone in historic buildings can lead to severe deterioration. The aim of the present study was to give modern insight into the phylogenetic diversity and activated metabolic pathways of microbial communities colonized historic objects located in the former Auschwitz II–Birkenau concentration and extermination camp in Oświecim, Poland. For this purpose we combined molecular, microscopic and chemical methods. Selected specimens were examined using Field Emission Scanning Electron Microscopy (FESEM), metabolomic analysis and high-throughput Illumina sequencing. FESEM imaging revealed the presence of complex microbial communities comprising diatoms, fungi and bacteria, mainly cyanobacteria and actinobacteria, on sample surfaces. Microbial diversity of brick specimens appeared higher than that of the wood and was dominated by algae and cyanobacteria, while wood was mainly colonized by fungi. DNA sequences documented the presence of 15 bacterial phyla representing 99 genera including Halomonas, Halorhodospira, Salinisphaera, Salinibacterium, Rubrobacter, Streptomyces, Arthrobacter and nine fungal classes represented by 113 genera including Cladosporium, Acremonium, Alternaria, Engyodontium, Penicillium, Rhizopus, and Aureobasidium. Most of the identified sequences were characteristic of organisms implicated in deterioration of wood and brick. Metabolomic data indicated the activation of numerous metabolic pathways, including those regulating the production of primary and secondary metabolites, for example, metabolites associated with the production of antibiotics, organic acids and deterioration of organic compounds. The study demonstrated that a combination of electron microscopy imaging with metabolomic and genomic techniques allows to link the phylogenetic information and metabolic profiles of microbial

  11. Metabolomic and high-throughput sequencing analysis – modern approach for the assessment of biodeterioration of materials from historic buildings

    Directory of Open Access Journals (Sweden)

    Beata eGutarowska

    2015-09-01

    Full Text Available Preservation of cultural heritage is of paramount importance worldwide. Microbial colonization of construction materials, such as wood, brick, mortar and stone in historic buildings can lead to severe deterioration. The aim of the present study was to give modern insight into the phylogenetic diversity and activated metabolic pathways of microbial communities colonized historic objects located in the former Auschwitz II-Birkenau concentration and extermination camp in Oświęcim, Poland. For this purpose we combined molecular, microscopic and chemical methods. Selected specimens were examined using Field Emission Scanning Electron Microscopy (FESEM, metabolomic analysis and high-throughput Illumina sequencing. FESEM imaging revealed the presence of complex microbial communities comprising diatoms, fungi and bacteria, mainly cyanobacteria and actinobacteria, on sample surfaces. Microbial diversity of brick specimens appeared higher than that of the wood and was dominated by algae and cyanobacteria, while wood was mainly colonized by fungi. DNA sequences documented the presence of 15 bacterial phyla representing 99 genera including Halomonas, Halorhodospira, Salinisphaera, Salinibacterium, Rubrobacter, Streptomyces, Arthrobacter and 9 fungal classes represented by 113 genera including Cladosporium, Acremonium, Alternaria, Engyodontium, Penicillium, Rhizopus and Aureobasidium. Most of the identified sequences were characteristic of organisms implicated in deterioration of wood and brick. Metabolomic data indicated the activation of numerous metabolic pathways, including those regulating the production of primary and secondary metabolites, for example, metabolites associated with the production of antibiotics, organic acids and deterioration of organic compounds. The study demonstrated that a combination of electron microscopy imaging with metabolomic and genomic techniques allows to link the phylogenetic information and metabolic profiles of

  12. Metabolomic and high-throughput sequencing analysis-modern approach for the assessment of biodeterioration of materials from historic buildings.

    Science.gov (United States)

    Gutarowska, Beata; Celikkol-Aydin, Sukriye; Bonifay, Vincent; Otlewska, Anna; Aydin, Egemen; Oldham, Athenia L; Brauer, Jonathan I; Duncan, Kathleen E; Adamiak, Justyna; Sunner, Jan A; Beech, Iwona B

    2015-01-01

    Preservation of cultural heritage is of paramount importance worldwide. Microbial colonization of construction materials, such as wood, brick, mortar, and stone in historic buildings can lead to severe deterioration. The aim of the present study was to give modern insight into the phylogenetic diversity and activated metabolic pathways of microbial communities colonized historic objects located in the former Auschwitz II-Birkenau concentration and extermination camp in Oświecim, Poland. For this purpose we combined molecular, microscopic and chemical methods. Selected specimens were examined using Field Emission Scanning Electron Microscopy (FESEM), metabolomic analysis and high-throughput Illumina sequencing. FESEM imaging revealed the presence of complex microbial communities comprising diatoms, fungi and bacteria, mainly cyanobacteria and actinobacteria, on sample surfaces. Microbial diversity of brick specimens appeared higher than that of the wood and was dominated by algae and cyanobacteria, while wood was mainly colonized by fungi. DNA sequences documented the presence of 15 bacterial phyla representing 99 genera including Halomonas, Halorhodospira, Salinisphaera, Salinibacterium, Rubrobacter, Streptomyces, Arthrobacter and nine fungal classes represented by 113 genera including Cladosporium, Acremonium, Alternaria, Engyodontium, Penicillium, Rhizopus, and Aureobasidium. Most of the identified sequences were characteristic of organisms implicated in deterioration of wood and brick. Metabolomic data indicated the activation of numerous metabolic pathways, including those regulating the production of primary and secondary metabolites, for example, metabolites associated with the production of antibiotics, organic acids and deterioration of organic compounds. The study demonstrated that a combination of electron microscopy imaging with metabolomic and genomic techniques allows to link the phylogenetic information and metabolic profiles of microbial communities

  13. Development and functioning of microorganisms in concentration cycles of sulfide copper-nickel and non-sulfide apatite-nepheline ores

    Directory of Open Access Journals (Sweden)

    Fokina N. V.

    2017-03-01

    Full Text Available The number and trophic diversity of bacteria in flotation samples of apatite-nepheline and sulfide copper-nickel ores at the concentration plants of JSC "Apatite" and Kola Mining and Metallurgical Company have been determined. The study of the size and diversity of the microbiota has been conducted by culture on selective nutrient media. The total number and biomass of bacteria have been considered by fluorescence microscopy using Cyclopore polycarbonate membrane filters. Bacteria have been identified by molecular genetic methods. The least amount of both saprotrophic and other trophic groups of bacteria has been observed in the samples of ore and recycled water as at the concentrating factory of Apatit JSC, and also at the plant "Pechenganikel". It has been found out that the bacteria contained in the ore and recycling water flowing from the tailings increased their number during the flotation process due to coming of the nutrients with the flotation reagents, aeration and increased temperature. Strains which occurrence is more than 60 % have been extracted from recycled water and basic flotation products and classified as Pseudomonas. Two strains with occurrence of more than 60 % have been discovered at Apatit JSC and classified as Stenotrophomonas and Acinetobacter. The number of fungi in the cycle of apatite-nepheline ore enrichment at the factories is very low (1 to 24 CFU / 1 ml or 1 g of ore. Fungi of the genus Penicillium have been dominated, fungi of the genera Acremonium, Aureobasidium, Alternaria, Chaetomium have also been detected. At the plant "Pechenganikel" species Aspergillus fumigatus, Penicillium aurantiogriseum and P. glabrum have been extracted. It has been shown that the bacteria deteriorate the apatite flotation as a result of their interaction with active centers of calcium-containing minerals and intensive flocculation decreasing the floatation selectivity. Also some trend of copper and nickel recovery change has been

  14. Fungal levels in the home and allergic rhinitis by 5 years of age.

    Science.gov (United States)

    Stark, Paul C; Celedón, Juan C; Chew, Ginger L; Ryan, Louise M; Burge, Harriet A; Muilenberg, Michael L; Gold, Diane R

    2005-10-01

    Studies have repeatedly demonstrated that sensitization to fungi, such as Alternaria, is strongly associated with allergic rhinitis and asthma in children. However, the role of exposure to fungi in the development of childhood allergic rhinitis is poorly understood. In a prospective birth cohort of 405 children of asthmatic/allergic parents from metropolitan Boston, Massachusetts, we examined in-home high fungal concentrations (> 90th percentile) measured once within the first 3 months of life as predictors of doctor-diagnosed allergic rhinitis in the first 5 years of life. In multivariate Cox regression analyses, predictors of allergic rhinitis included high levels of dust-borne Aspergillus [hazard ratio (HR) = 3.27; 95% confidence interval (CI), 1.50-7.14], Aureobasidium (HR = 3.04; 95% CI, 1.33-6.93), and yeasts (HR = 2.67; 95% CI, 1.26-5.66). The factors controlled for in these analyses included water damage or mild or mildew in the building during the first year of the child's life, any lower respiratory tract infection in the first year, male sex, African-American race, fall date of birth, and maternal IgE to Alternaria > 0.35 U/mL. Dust-borne Alternaria and nonsporulating and total fungi were also predictors of allergic rhinitis in models excluding other fungi but adjusting for all of the potential confounders listed above. High measured fungal concentrations and reports of water damage, mold, or mildew in homes may predispose children with a family history of asthma or allergy to the development of allergic rhinitis.

  15. Development of multifunctional mannan nanogel =

    Science.gov (United States)

    Ferreira, Silvia Alexandra Rodrigues Mendes

    Self-assembled nanogels made of hydrophobized mannan or pullulan were obtained using a versatile, simple, reproducible and low-cost method. In a first reaction pullulan or mannan were modified with hydroxyethyl methacrylate or vinyl methacrylate, further modified in the second reaction with 1-hexadecanethiol. The resultant amphiphilic material self-assembles in water via the hydrophobic interaction among alkyl chains, originating the nanogel. Structural features, size, shape, surface charge and stability of the nanogels were studied using hydrogen nuclear magnetic resonance, cryo-field emission scanning electron microscopy and dynamic light scattering. Above the critical micellar concentration (cmc), evaluated by fluorescence spectroscopy with Nile red and pyrene, spherical polydisperse nanogels reveal long-term colloidal stability in aqueous medium up to six months, with a nearly neutral negative surface charge and mean hydrodynamic diameter in the nanoscale range, depending on the polymer degree of substitution. Nanogel based on vinyl methacrylated mannan was selected for further characterization among others because its synthesis is much easier, cheaper and less time consuming, its cmc and size are smaller, it is less polydisperse, and more stable at pH 3-8, in salt or urea solutions being consequently more suitable for biological applications. Proteins (bovine serum albumin or ovalbumin) and hydrophobic drugs (curcumin) are spontaneously incorporated in the mannan nanogel, being stabilized by the hydrophobic domains randomly distributed within the nanogel, opening the possibility for the development of applications as potential delivery systems for therapeutic molecules. No cytotoxicity is detected up to about 0.4 mg/mL of mannan nanogel in mouse embryo fibroblast cell line 3T3 and mouse bone marrow-derived macrophages (BMDM) using cell proliferation, lactate dehydrogenase and Live/Dead assays. Comet assay, under the tested conditions, reveals no DNA damage in

  16. Wetting of biopolymer coatings: contact angle kinetics and image analysis investigation.

    Science.gov (United States)

    Farris, Stefano; Introzzi, Laura; Biagioni, Paolo; Holz, Torsten; Schiraldi, Alberto; Piergiovanni, Luciano

    2011-06-21

    The surface wetting of five biopolymers, used as coating materials for a plastic film, was monitored over a span of 8 min by means of the optical contact angle technique. Because most of the total variation was observed to occur during the first 60 s, we decided to focus on this curtailed temporal window. Initial contact angle values (θ(0)) ranged from ∼91° for chitosan to ∼30° for pullulan. However, the water drop profile began to change immediately following drop deposition for all biocoatings, confirming that the concept of water contact angle equilibrium is not applicable to most biopolymers. First, a three-parameter decay equation [θ(t) = θ(0) exp(kt(n))] was fit to the experimental contact angle data to describe the kinetics of the contact angle change for each biocoating. Interestingly, the k constant correlated well with the contact angle evolution rate and the n exponent seemed to be somehow linked to the physicochemical phenomena underlying the overall kinetics process. Second, to achieve a reliable description of droplet evolution, the contact angle (CA) analysis was coupled with image analysis (IA) through a combined geometric/trigonometric approach. Absorption and spreading were the key factors governing the overall mechanism of surface wetting during the 60 s analysis, although the individual quantification of both phenomena demonstrated that spreading provided the largest contribution for all biopolymers, with the only exception of gelatin, which showed two quasi-equivalent and counterbalancing effects. The possible correlation between these two phenomena and the topography of the biopolymer surfaces are then discussed on the basis of atomic force microscopy analyses. © 2011 American Chemical Society

  17. Improved Bilayer Resist System Using Contrast-Enhanced Lithography With Water-Soluble Photopolymer

    Science.gov (United States)

    Sasago, Masaru; Endo, Masayuki; Hirai, Yoshihiko; Ogawa, Kazufurni; Ishihara, Takeshi

    1986-07-01

    A new water-soluble contract enhanced material, WSP (Water-soluble Photopolymer), has been developed. The WSP is composed of a mainpolymer and a photobleachable reagents. The mainpolymer is a water-soluble polymer mixed with pullulan (refined through biotechnological process) and polyvinyl-pyrolidone (PVP). The photo-bleachable reagent is of a diazonium compound gorup. The introduction of the mainpolymer and photobleach-able reagent mixture has improved filmity, gas transparency, photobleaching characteristics and solubility in alkaline which are essential to the device fabrication. Submicron photoresist patterns are successfully fabricated by a simple sequence of photolithography process. The WSP layer has been applied to the bilayer resist system--deep-UV portable conformable masking (PCM)--that is not affected by VLSI's topography, and is able to fabricate highly accurate pattern. The aqueous developable layer, PMGI, with high organic solvent resistance is used in the bottom layer. Therefore, no interfacial mixing with conventional positive resist top layer is observed. Furthermore, deep-UV exposure method has been used for the KrF excimer laser optical system in order to increase high throughput. From the experiments, it has been confirmed that good resist transfer profile can be realized by the use of WSP, and that the submicron resist patterns with high aspect-ratio can be developed on the nonplaner wafer with steps of up to 41m by the combination of the WSP with the PCM system. By this technology, has been improved the weak point: variation in the line width due to the thickness of contrast-enhanced layer when the CEL technology is applied, and dependency of both the finished resist profile and the line-width accuracy on the thickness of the top layer resist when the PCM system is adopted.

  18. Possible application of brewer’s spent grain in biotechnology

    Directory of Open Access Journals (Sweden)

    Pejin Jelena D.

    2013-01-01

    Full Text Available Brewer’s spent grain is the major by-product in beer production. It is produced in large quantities (20 kg per 100 liters of produced beer throughout the year at a low cost or no cost, and due to its high protein and carbohydrates content it can be used as a raw material in biotechnology. Biotechnological processes based on renewable agro-industrial by-products have ecological (zero CO2 emission, eco-friendly by-products and economical (cheap raw materials and reduction of storage costs advantages. The use of brewer’s spent grain is still limited, being basically used as animal feed. Researchers are trying to improve the application of brewer’s spent grain by finding alternative uses apart from the current general use as an animal feed. Its possible applications are in human nutrition, as a raw material in biotechnology, energy production, charcoal production, paper manufacture, as a brick component, and adsorbent. In biotechnology brewer’s spent grain could be used as a substrate for cultivation of microorganisms and enzyme production, additive of yeast carrier in beer fermentation, raw material in production of lactic acid, bioethanol, biogas, phenolic acids, xylitol, and pullulan. Some possible applications for brewer’s spent grain are described in this article including pre-treatment conditions (different procedures for polysaccharides, hemicelluloses, and cellulose hydrolysis, working microorganisms, fermentation parameters and obtained yields. The chemical composition of brewer’s spent grain varies according to barley variety, harvesting time, malting and mashing conditions, and a quality and type of unmalted raw material used in beer production. Brewer’s spent grain is lignocellulosic material rich in protein and fibre, which account for approximately 20 and 70% of its composition, respectively.

  19. Polysaccharide characterization by hollow-fiber flow field-flow fractionation with on-line multi-angle static light scattering and differential refractometry.

    Science.gov (United States)

    Pitkänen, Leena; Striegel, André M

    2015-02-06

    Accurate characterization of the molar mass and size of polysaccharides is an ongoing challenge, oftentimes due to architectural diversity but also to the broad molar mass (M) range over which a single polysaccharide can exist and to the ultra-high M of many polysaccharides. Because of the latter, many of these biomacromolecules experience on-column, flow-induced degradation during analysis by size-exclusion and, even, hydrodynamic chromatography (SEC and HDC, respectively). The necessity for gentler fractionation methods has, to date, been addressed employing asymmetric flow field-flow fractionation (AF4). Here, we introduce the coupling of hollow-fiber flow field-flow fractionation (HF5) to multi-angle static light scattering (MALS) and differential refractometry (DRI) detection for the analysis of polysaccharides. In HF5, less stresses are placed on the macromolecules during separation than in SEC or HDC, and HF5 can offer a higher sensitivity, with less propensity for system overloading and analyte aggregation, than generally found in AF4. The coupling to MALS and DRI affords the determination of absolute, calibration-curve-independent molar mass averages and dispersities. Results from the present HF5/MALS/DRI experiments with dextrans, pullulans, and larch arabinogalactan were augmented with hydrodynamic radius (RH) measurements from off-line quasi-elastic light scattering (QELS) and by RH distribution calculations and fractogram simulations obtained via a finite element analysis implementation of field-flow fractionation theory by commercially available software. As part of this study, we have investigated analyte recovery in HF5 and also possible reasons for discrepancies between calculated and simulated results vis-à-vis experimentally determined data. Published by Elsevier B.V.

  20. Identification of Volatile Components of Liverwort (Porella cordaeana Extracts Using GC/MS-SPME and Their Antimicrobial Activity

    Directory of Open Access Journals (Sweden)

    Maria Elisabetta Guerzoni

    2012-06-01

    Full Text Available Chemical constituents of liverwort (Porella cordaeana extracts have been identified using solid-phase microextraction-gas chromatography mass spectrometry (SPME-GC/MS. The methanol, ethanol and ethyl acetate extracts were rich in terpenoids such as sesquiterpene hydrocarbons (53.12%, 51.68%, 23.16%, and monoterpene hydrocarbons (22.83%, 18.90%, 23.36%, respectively. The dominant compounds in the extracts were β-phellandrene (15.54%, 13.66%, 12.10% and β-caryophyllene (10.72%, 8.29%, 7.79%, respectively. The antimicrobial activity of the extracts was evaluated against eleven food microorganisms using the microdilution and disc diffusion methods. The minimum inhibitory concentration (MIC varied from 0.50 to 2.00 mg/mL for yeast strains (Saccharomyces cerevisiae 635, Zygosacharomyces bailii 45, Aerobasidium pullulans L6F, Pichia membranaefaciens OC 71, Pichia membranaefaciens OC 70, Pichia anomala CBS 5759, Pichia anomala DBVPG 3003 and Yarrowia lipolytica RO13, and from 1.00 to 3.00 mg/mL for bacterial strains (Salmonella enteritidis 155, Escherichia coli 555 and Listeria monocytogenes 56Ly. Methanol extract showed better activity in comparison with ethanol and ethyl acetate extracts. High percentages of monoterpene and sesquiterpene hydrocarbons could be responsible for the better antimicrobial activity.

  1. Size and molecular weight determination of polysaccharides by means of nano electrospray gas-phase electrophoretic mobility molecular analysis (nES GEMMA).

    Science.gov (United States)

    Weiss, Victor U; Golesne, Monika; Friedbacher, Gernot; Alban, Susanne; Szymanski, Wladyslaw W; Marchetti-Deschmann, Martina; Allmaier, Günter

    2018-02-21

    Size, size distribution and molecular weight (MW) determination of nanoparticles and that are for example large polymers, are of great interest and pose an analytical challenge. In this context, nano electrospray gas-phase electrophoretic mobility molecular analysis (nES GEMMA) is a valuable tool with growing impact. Separation of single-charged analytes according to their electrophoretic mobility diameter (EMD) starting from single-digit EMDs up to several hundred nm diameters is possible. In case of spherical analytes, the EMD corresponds to the dry nanoparticle size. Additionally, the instrument is capable of number-based, single-particle detection following the recommendation of the European Commission for nanoparticle characterization (2011/696/EU). In case an EMD/MW correlation for a particular compound class (based on availability of well-defined standards) exists, a nanoparticle's MW can be determined from its EMD. In the present study, we focused on nES GEMMA of linear and branched, water-soluble polysaccharides forming nanoparticles and were able to obtain spectra for both analyte classes regarding single-charged species. Based on EMDs for corresponding analytes, an excellent EMD/MW correlation could be obtained in case of the branched natural polymer (dextran). This enables the determination of dextran MWs from nES GEMMA spectra despite high analyte polydispersity and in a size/MW range, where classical mass spectrometry is limited. EMD/MW correlations based on linear (pullulans, oat-ß-glucans) polymers were significantly different, possibly indicating challenges in the exact MW determination of these compounds by, for example, chromatographic and light scattering means. Despite these observations, nES GEMMA of linear, monosaccharide-based polymers enabled the determination of size and size-distribution of such dry bionanoparticles. © 2018 The Authors. Electrophoresis published by Wiley-VCH Verlag GmbH & Co. KGaA.

  2. Present and perspective medical applications of microbial exopolysaccharides

    Directory of Open Access Journals (Sweden)

    Misu - Moscovici

    2015-09-01

    Full Text Available Microbial exopolysaccharides (EPS have found outstanding medical applications since the mid-twentieth century, with the first clinical trials on dextran solutions as plasma expanders. Other EPS entered medicine firstly as conventional pharmaceutical excipients (e.g., xanthan - as suspension stabilizer, or pullulan – in capsules and oral care products. Polysaccharides, initially obtained from plant or animal sources, became easily available for a wide range of applications, especially when they were commercially produced by microbial fermentation. Alginates are used as anti-reflux, dental impressions, or as matrix for tablets. Hyaluronic acid and derivatives are used in surgery, arthritis treatment or wound healing. Bacterial cellulose is applied in wound dressings or scaffolds for tissue engineering. The development of drug controlled-release systems and of micro- and nanoparticulated ones, has opened a new era of medical applications for biopolymers. EPS and their derivatives are well suited potentially non-toxic, biodegradable drug carriers. Such systems concern rating and targeting of controlled release. Their large area of applications is explained by the available manifold series of derivatives, whose useful properties can be thereby controlled. From matrix inclusion to conjugates, different systems have been designed to solubilize, and to assure stable transport in the body, target accumulation and variable rate-release of a drug substance. From controlled drug delivery, EPS potential applications expanded to vaccine adjuvants and diagnostic imaging systems. Other potential applications are related to the bioactive (immunomodulator, antitumor, antiviral characteristics of EPS. The numerous potential applications still wait to be developed into commercial pharmaceuticals and medical devices. Based on previous and recent results in important medical-pharmaceutical domains, one can undoubtedly state that EPS medical applications have a

  3. Systems biology of microbial exopolysaccharides production

    Directory of Open Access Journals (Sweden)

    Ozlem eAtes

    2015-12-01

    Full Text Available Exopolysaccharides (EPS produced by diverse group of microbial systems are rapidly emerging as new and industrially important biomaterials. Due to their unique and complex chemical structures and many interesting physicochemical and rheological properties with novel functionality, the microbial EPSs find wide range of commercial applications in various fields of the economy such as food, feed, packaging, chemical, textile, cosmetics and pharmaceutical industry, agriculture and medicine. EPSs are mainly associated with high-value applications and they have received considerable research attention over recent decades with their biocompatibility, biodegradability, and both environmental and human compatibility. However only a few microbial EPSs have achieved to be used commercially due to their high production costs. The emerging need to overcome economic hurdles and the increasing significance of microbial EPSs in industrial and medical biotechnology call for the elucidation of the interrelations between metabolic pathways and EPS biosynthesis mechanism in order to control and hence enhance its microbial productivity. Moreover a better understanding of biosynthesis mechanism is a significant issue for improvement of product quality and properties and also for the design of novel strains. Therefore a systems-based approach constitutes an important step towards understanding the interplay between metabolism and EPS biosynthesis and further enhances its metabolic performance for industrial application. In this review, primarily the microbial EPSs, their biosynthesis mechanism and important factors for their production will be discussed. After this brief introduction, recent literature on the application of omics technologies and systems biology tools for the improvement of production yields will be critically evaluated. Special focus will be given to EPSs with high market value such as xanthan, levan, pullulan and dextran.

  4. Molecular nuclear imaging for targeting and trafficking

    International Nuclear Information System (INIS)

    Bom, Hee Seung; Min, Jung Jun; Jeong, Hwan-Jeong

    2006-01-01

    Noninvasive molecular targeting in living subjects is highly demanded for better understanding of such diverse topics as the efficient delivery of drugs, genes, or radionuclides for the diagnosis or treatment of diseases. Progress in molecular biology, genetic engineering and polymer chemistry provides various tools to target molecules and cells in vivo. We used chitosan as a polymer, and 99m Tc as a radionuclide. We developed 99m Tc-galactosylated chitosan to target asialoglycoprotein receptors for nuclear imaging. We also developed 99m Tc-HYNIC-chitosan-transferrin to target inflammatory cells, which was more effective than 67 Ga-citrate for imaging inflammatory lesions. For an effective delivery of molecules, a longer circulation time is needed. We found that around 10% PEGylation was most effective to prolong the circulation time of liposomes for nuclear imaging of 99m Tc-HMPAO-labeled liposomes in rats. Using various characteristics of molecules, we can deliver drugs into targets more effectively. We found that 99m Tc-labeled biodegradable pullulan-derivatives are retained in tumor tissue in response to extracellular ion-strength. For the trafficking of various cells or bacteria in an intact animal, we used optical imaging techniques or radiolabeled cells. We monitored tumor-targeting bacteria by bioluminescent imaging techniques, dentritic cells by radiolabeling and neuronal stem cells by sodium-iodide symporter reporter gene imaging. In summary, we introduced recent achievements of molecular nuclear imaging technologies in targeting receptors for hepatocyte or inflammatory cells and in trafficking bacterial, immune and stem cells using molecular nuclear imaging techniques

  5. Yeast β-1,6-glucan is a primary target for the Saccharomyces cerevisiae K2 toxin.

    Science.gov (United States)

    Lukša, Juliana; Podoliankaitė, Monika; Vepštaitė, Iglė; Strazdaitė-Žielienė, Živilė; Urbonavičius, Jaunius; Servienė, Elena

    2015-04-01

    Certain Saccharomyces cerevisiae strains secrete different killer proteins of double-stranded-RNA origin. These proteins confer a growth advantage to their host by increasing its survival. K2 toxin affects the target cell by binding to the cell surface, disrupting the plasma membrane integrity, and inducing ion leakage. In this study, we determined that K2 toxin saturates the yeast cell surface receptors in 10 min. The apparent amount of K2 toxin, bound to a single cell of wild type yeast under saturating conditions, was estimated to be 435 to 460 molecules. It was found that an increased level of β-1,6-glucan directly correlates with the number of toxin molecules bound, thereby impacting the morphology and determining the fate of the yeast cell. We observed that the binding of K2 toxin to the yeast surface receptors proceeds in a similar manner as in case of the related K1 killer protein. It was demonstrated that the externally supplied pustulan, a poly-β-1,6-glucan, but not the glucans bearing other linkage types (such as laminarin, chitin, and pullulan) efficiently inhibits the K2 toxin killing activity. In addition, the analysis of toxin binding to the intact cells and spheroplasts confirmed that majority of K2 protein molecules attach to the β-1,6-glucan, rather than the plasma membrane-localized receptors. Taken together, our results reveal that β-1,6-glucan is a primary target of K2 toxin and is important for the execution of its killing property. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  6. Sustained Ocular Delivery of Ciprofloxacin Using Nanospheres and Conventional Contact Lens Materials

    Science.gov (United States)

    Garhwal, Rahul; Shady, Sally F.; Ellis, Edward J.; Ellis, Jeanne Y.; Leahy, Charles D.; McCarthy, Stephen P.; Crawford, Kathryn S.

    2012-01-01

    Purpose. To formulate conventional contact lenses that incorporate nanosphere-encapsulated antibiotic and demonstrate that the lenses provide for sustained antibacterial activity. Methods. A copolymer composed of pullulan and polycaprolactone (PCL) was used to synthesize core-shell nanospheres that encapsulated ciprofloxacin. Bactericidal activity of the nanosphere-encapsulated ciprofloxacin (nanosphere/cipro) was tested by using liquid cultures of either Staphylococcus aureus or Pseudomonas aeruginosa. Nanosphere/cipro was then incorporated into HEMA-based contact lenses that were tested for growth inhibition of S. aureus or P. aeruginosa in liquid cultures inoculated daily with fresh bacteria. Lens designs included thin or thick lenses incorporating nanosphere/cipro and ciprofloxacin-HCl-soaked Acuvue lenses (Acuvue; Johnson & Johnson Vision Care, Inc., Jacksonville, FL). Results. Less than 2 μg/mL of nanosphere/cipro effectively inhibited the proliferation of cultures inoculated with 107 or 108 bacteria/mL of S. aureus and P. aeruginosa, respectively. HEMA-based contact lenses polymerized with nanosphere/cipro were transparent, effectively inhibited the proliferation of greater than 107/mL of bacteria added daily over 3 days of culture, and killed up to 5 × 109 total microbes in a single inoculation. A thicker lens design provided additional inhibition of bacterial growth for up to 96 hours. Conclusions. Core-shell nanospheres loaded with an antibiotic can be incorporated into a conventional, transparent contact lens and provide for sustained and effective bactericidal activity and thereby provide a new drug delivery platform for widespread use in treating ocular disorders. PMID:22266514

  7. Purification and characterization of an extracellular halophilic and organic solvent-tolerant amylopullulanase from a haloarchaeon, Halorubrum sp. strain Ha25.

    Directory of Open Access Journals (Sweden)

    Mostafa Fazeli

    2013-01-01

    Full Text Available Introduction: Halophiles, especially haloarchaea are one of the most important groups of extremophiles. Halophilic hydrolases have been studied worldwide and have been considered for biotechnology and industrial technologies. This study is the first report in amylopullulanase production in halophilic microorganisms.Materials and methods: A halophilic archaeon, Halorubrum sp. strain Ha25, produced extracellular halophilic organic solvent-tolerant amylopullulanase. The enzyme was purified using ethanol precipitation and anion exchange chromatography method. Molecular mass of purified enzyme was determined by SDS–PAGE method. After purification, the enzyme was characterized. To study the effects of organic solvents in the stability of the enzyme, the enzyme solution was incubated in the presence of various organic compounds and then, residual enzyme activity was measured. Mode of action of the enzyme was determined by thin-layer chromatography.Results: Molecular weight of the purified enzyme was estimated to be 140 kDa by SDS–PAGE method. Optimum temperature for amylolitic and pullulytic activities was 50 °C. Optimum pH for amylolitic activity was 7.0 and for pullulytic activity was 7.5. This enzyme was active over a wide range of concentrations (0-4.5 M of NaCl. The effect of organic solvents on the amylolitic and pullulytic activities showed that this enzyme was more stable in the presence of non-polar organic solvents than polar solvents. The enzyme solely hydrolyzed pullulan and soluble starch to glucose.Discussion and conclusion: Halorubrum sp. strain Ha25 produces thermophilic and extremely halophilic amylopullulanase. The catalytic function under multi extreme condition of high temperature, high salinity, and low water activity might possess biotechnological and commercial values such as treatment waste solutions with starch residues, high salt content and solvents.

  8. Systems Biology of Microbial Exopolysaccharides Production.

    Science.gov (United States)

    Ates, Ozlem

    2015-01-01

    Exopolysaccharides (EPSs) produced by diverse group of microbial systems are rapidly emerging as new and industrially important biomaterials. Due to their unique and complex chemical structures and many interesting physicochemical and rheological properties with novel functionality, the microbial EPSs find wide range of commercial applications in various fields of the economy such as food, feed, packaging, chemical, textile, cosmetics and pharmaceutical industry, agriculture, and medicine. EPSs are mainly associated with high-value applications, and they have received considerable research attention over recent decades with their biocompatibility, biodegradability, and both environmental and human compatibility. However, only a few microbial EPSs have achieved to be used commercially due to their high production costs. The emerging need to overcome economic hurdles and the increasing significance of microbial EPSs in industrial and medical biotechnology call for the elucidation of the interrelations between metabolic pathways and EPS biosynthesis mechanism in order to control and hence enhance its microbial productivity. Moreover, a better understanding of biosynthesis mechanism is a significant issue for improvement of product quality and properties and also for the design of novel strains. Therefore, a systems-based approach constitutes an important step toward understanding the interplay between metabolism and EPS biosynthesis and further enhances its metabolic performance for industrial application. In this review, primarily the microbial EPSs, their biosynthesis mechanism, and important factors for their production will be discussed. After this brief introduction, recent literature on the application of omics technologies and systems biology tools for the improvement of production yields will be critically evaluated. Special focus will be given to EPSs with high market value such as xanthan, levan, pullulan, and dextran.

  9. Minimalism in fabrication of self-organized nanogels holding both anti-cancer drug and targeting moiety.

    Science.gov (United States)

    Kim, Sungwon; Park, Kyong Mi; Ko, Jin Young; Kwon, Ick Chan; Cho, Hyeon Geun; Kang, Dongmin; Yu, In Tag; Kim, Kwangmeyung; Na, Kun

    2008-05-01

    Recent researches to develop nano-carrier systems in anti-cancer drug delivery have focused on more complicated design to improve therapeutic efficacy and to reduce side effects. Although such efforts have great impact to biomedical science and engineering, the complexity has been a huddle because of clinical and economic problems. In order to overcome the problems, a simplest strategy to fabricate nano-carriers to deliver doxorubicin (DOX) was proposed in the present study. Two significant subjects (i) formation of nanoparticles loading and releasing DOX and (ii) binding specificity of them to cells, were examined. Folic acid (FA) was directly coupled with pullulan (Pul) backbone by ester linkage (FA/Pul conjugate) and the degree of substitution (DS) was varied, which were confirmed by 1H NMR and UV spectrophotometry. Light scattering results revealed that the nanogels possessed two major size distributions around 70 and 270 nm in an aqueous solution. Their critical aggregation concentrations (CACs) were less than 10 microg/mL, which are lower than general critical micelle concentrations (CMCs) of low-molecular-weight surfactants. Transmission electron microscopy (TEM) images showed well-dispersed nanogel morphology in a dried state. Depending on the DS, the nanogels showed different DOX-loading and releasing profiles. The DOX release rate from FA8/Pul (with the highest DS) for 24h was slower than that from FA4/or FA6/Pul, indicating that the FA worked as a hydrophobic moiety for drug holding. Cellular uptake of the nanogels (KB cells) was also monitored by confocal microscopy. All nanogels were internalized regardless of the DS of FA. Based on the results, the objectives of this study, to suggest a new method overcoming the complications in the drug carrier design, were successfully verified.

  10. Purification and some properties of a novel maltohexaose-producing exo-amylase from Aerobacter aerogenes.

    Science.gov (United States)

    Kainuma, K; Wako, K; Kobayashi, S; Nogami, A; Suzuki, S

    1975-12-18

    Maltohexaose producing amylase (EC 3.2.1.-) is the fourth known exo-amylase, the three previously known being glucoamylase, beta-amylase and Pseudomonas stutzeri maltotetraose producing amylase. The enzyme after release from Aerobacter aerogenes cells by 0.1% sodium lauryl sulfate extraction was purified by ammonium sulfate precipitation, DEAE-Sephadex column chromatography and Sephadex G-100 gel filtration to 80-fold of the original sodium lauryl sulfate extract activity, It gave a single band on disc electrophoresis, and the molecular weight by gel filtration was 54 000. This amylase showed maximal activity at 50 degrees C and pH 6.80. The pH stability range was relatively wide, the enzyme retaining more than 90% of its initial activity in the range of 6.50-9.0. 80% of the activity was retained after 15 min at 50 degrees C. This enzyme produced maltohexaose from starch, amylose and amylopectin by exo-attack, but did not act on alpha- or beta-cyclodextrin, pullulan or maltohexaitol. Also the enzyme acted on beta-limit dextrins of amylopectin and glycogen to form branched oligosaccharides. The unusual reaction of this enzyme on beta-limit dextrin is discussed from the standpoint of the stereochemistry of 1,4-alpha- and 1,6-alpha-glucosidic bonds. This is the anomalous amylase for which it is recognized that 1,6-alpha-glucosidic linkages in the substrates can mimic the effect of 1,4-alpha-bonds, as previously observed in pseudo-priming reactions of E. coli phosphorylase.

  11. Cohnella amylopullulanases: Biochemical characterization of two recombinant thermophilic enzymes.

    Directory of Open Access Journals (Sweden)

    Fatemeh Zebardast Roodi

    Full Text Available Some industries require newer, more efficient recombinant enzymes to accelerate their ongoing biochemical reactions in harsh environments with less replenishment. Thus, the search for native enzymes from extremophiles that are suitable for use under industrial conditions is a permanent challenge for R & D departments. Here and toward such discoveries, two sequences homologous to amylopullulanases (EC 3.2.1.41, GH57 from an endogenous Cohnella sp., [Coh00831 (KP335161; 1998 bp and Coh01133 (KP335160: 3678 bp] were identified. The genes were heterologously expressed in E. coli to both determine their type and further characterize their properties. The isolated DNA was PCR amplified with gene specific primers and cloned in pET28a, and the recombinant proteins were expressed in E. coli BL21 (DE3. The temperatures and pH optima of purified recombinants Coh 01133 and Coh 00831 enzymes were 70°C and 8, and 60°C and 6, respectively. These enzymes are stable more than 90% in 60°C and 50°C for 90 min respectively. The major reactions released sugars which could be fractionated by HPLC analysis, from soluble starch were mainly maltose (G2, maltotriose (G3 and maltotetraose (G4. The enzymes hydrolyzed pullulan to maltotriose (G3 only. Enzyme activities for both proteins were improved in the availability of Mn2+, Ba2+, Ca2+, and Mg2+ and reduced in the presence of Fe2+, Li2+, Na2+, Triton X100 and urea. Moreover, Co2+, K+, and Cu2+ had a negative effect only on Coh 01133 enzyme.

  12. A thermophilic alkalophilic α-amylase from Bacillus sp. AAH-31 shows a novel domain organization among glycoside hydrolase family 13 enzymes.

    Science.gov (United States)

    Saburi, Wataru; Morimoto, Naoki; Mukai, Atsushi; Kim, Dae Hoon; Takehana, Toshihiko; Koike, Seiji; Matsui, Hirokazu; Mori, Haruhide

    2013-01-01

    α-Amylases (EC 3.2.1.1) hydrolyze internal α-1,4-glucosidic linkages of starch and related glucans. Bacillus sp. AAH-31 produces an alkalophilic thermophilic α-amylase (AmyL) of higher molecular mass, 91 kDa, than typical bacterial α-amylases. In this study, the AmyL gene was cloned to determine its primary structure, and the recombinant enzyme, produced in Escherichia coli, was characterized. AmyL shows no hydrolytic activity towards pullulan, but the central region of AmyL (Gly395-Asp684) was similar to neopullulanase-like α-amylases. In contrast to known neopullulanase-like α-amylases, the N-terminal region (Gln29-Phe102) of AmyL was similar to carbohydrate-binding module family 20 (CBM20), which is involved in the binding of enzymes to starch granules. Recombinant AmyL showed more than 95% of its maximum activity in a pH range of 8.2-10.5, and was stable below 65 °C and from pH 6.4 to 11.9. The kcat values for soluble starch, γ-cyclodextrin, and maltotriose were 103 s(-1), 67.6 s(-1), and 5.33 s(-1), respectively, and the Km values were 0.100 mg/mL, 0.348 mM, and 2.06 mM, respectively. Recombinant AmyL did not bind to starch granules. But the substitution of Trp45 and Trp84, conserved in site 1 of CBM20, with Ala reduced affinity to soluble starch, while the mutations did not affect affinity for oligosaccharides. Substitution of Trp61, conserved in site 2 of CBM20, with Ala enhanced hydrolytic activity towards soluble starch, indicating that site 2 of AmyL does not contribute to binding to soluble long-chain substrates.

  13. Cohnella amylopullulanases: Biochemical characterization of two recombinant thermophilic enzymes.

    Science.gov (United States)

    Zebardast Roodi, Fatemeh; Aminzadeh, Saeed; Farrokhi, Naser; Karkhane, AliAsghar; Haghbeen, Kamahldin

    2017-01-01

    Some industries require newer, more efficient recombinant enzymes to accelerate their ongoing biochemical reactions in harsh environments with less replenishment. Thus, the search for native enzymes from extremophiles that are suitable for use under industrial conditions is a permanent challenge for R & D departments. Here and toward such discoveries, two sequences homologous to amylopullulanases (EC 3.2.1.41, GH57) from an endogenous Cohnella sp., [Coh00831 (KP335161; 1998 bp) and Coh01133 (KP335160: 3678 bp)] were identified. The genes were heterologously expressed in E. coli to both determine their type and further characterize their properties. The isolated DNA was PCR amplified with gene specific primers and cloned in pET28a, and the recombinant proteins were expressed in E. coli BL21 (DE3). The temperatures and pH optima of purified recombinants Coh 01133 and Coh 00831 enzymes were 70°C and 8, and 60°C and 6, respectively. These enzymes are stable more than 90% in 60°C and 50°C for 90 min respectively. The major reactions released sugars which could be fractionated by HPLC analysis, from soluble starch were mainly maltose (G2), maltotriose (G3) and maltotetraose (G4). The enzymes hydrolyzed pullulan to maltotriose (G3) only. Enzyme activities for both proteins were improved in the availability of Mn2+, Ba2+, Ca2+, and Mg2+ and reduced in the presence of Fe2+, Li2+, Na2+, Triton X100 and urea. Moreover, Co2+, K+, and Cu2+ had a negative effect only on Coh 01133 enzyme.

  14. DIVERSIDAD DE LEVADURAS ASOCIADAS A INFLORESCENCIAS DE MANGO Y FLORES DE 'LULO ARBÓREO' DIVERSIDADE DE LEVEDURAS ASSOCIADAS A INFLORESCÊNCIAS DE MANGA E FLORES DE 'LULO ARBÓREO' YEAST DIVERSITY ASSOCIATED TO INFLORESCENCES OF MANGO AND FLOWERS OF 'LULO ARBOREO'

    Directory of Open Access Journals (Sweden)

    JACKELINE GAVIRIA V.

    2012-12-01

    Full Text Available En Colombia, el conocimiento de levaduras ha sido limitado a especies de interés clínico. El micro-hábitat aéreo de las flores, presenta un área de gran interés para el aislamiento de levaduras con importancia en el campo industrial, por su potencial uso como controlador biológico. El objetivo de este trabajo es analizar la diversidad de levaduras asociadas a inflorescencia de Mango (Mangífera índica y flores de Lulo arbóreo (Solanum wrightii. Para dicha finalidad se trabajó con 96 aislados, a los cuales se les realizó caracterización fenotípica y posteriores pruebas de MSP-PCR Fingerprinting. Estas pruebas fueron complementadas con amplificación parcial y secuenciación del segmento 26S del ADNr para identificación a nivel de especie. El patrón de diversidad encontrado refleja una dominancia de la especie Candida leandrae en flores de lulo arbóreo, también se presentaron especies como Cryptococcus laurentii, Candida parazyma, Aeurobasidium pullulans y Pseudozyma tsukubaensis. Las inflorescencias de mango presentaron especies como Sydowia eucalypti, Cryptococcus flavescens, C. laurentii, C. nemorosus, C. heveanensis, Pichia kluyveri, Pseudozyma tsukubaensis y Candida asparagi. Se logró identificar hasta el nivel taxonómico de especie el total de los aislados encontrados, además se hallaron especies con potencial uso biotecnológico.Na Colômbia, o conhecimento das leveduras é limitado a espécies de interesse clinico. O micro-habitat aéreo das flores apresenta um interessante campo para o isolamento de leveduras com importância na indústria pelo seu potencial uso em controle biológico. O objetivo deste trabalho foi analisar a diversidade de leveduras associadas a inflorescências de Manga (Mangifera indica e flores de 'lulo' (Solanum wrightii. Para isso, foram analisados 96 isolados, que foram caracterizados com testes fenotípicos e MSP-PCR Fingerprinting complementadas com sequenciamento parcial da região 26S do r

  15. Endophytic Fungi from Frankincense Tree Improves Host Growth and Produces Extracellular Enzymes and Indole Acetic Acid.

    Science.gov (United States)

    Khan, Abdul Latif; Al-Harrasi, Ahmed; Al-Rawahi, Ahmed; Al-Farsi, Zainab; Al-Mamari, Aza; Waqas, Muhammad; Asaf, Sajjad; Elyassi, Ali; Mabood, Fazal; Shin, Jae-Ho; Lee, In-Jung

    2016-01-01

    Boswellia sacra, an economically important frankincense-producing tree found in the desert woodlands of Oman, is least known for its endophytic fungal diversity and the potential of these fungi to produce extracellular enzymes and auxins. We isolated various fungal endophytes belonging to Eurotiales (11.8%), Chaetomiaceae (17.6%), Incertae sadis (29.5%), Aureobasidiaceae (17.6%), Nectriaceae (5.9%) and Sporomiaceae (17.6%) from the phylloplane (leaf) and caulosphere (stem) of the tree. Endophytes were identified using genomic DNA extraction, PCR amplification and sequencing the internal transcribed spacer regions, whereas a detailed phylogenetic analysis of the same gene fragment was made with homologous sequences. The endophytic colonization rate was significantly higher in the leaf (5.33%) than the stem (0.262%). The Shannon-Weiner diversity index was H' 0.8729, while Simpson index was higher in the leaf (0.583) than in the stem (0.416). Regarding the endophytic fungi's potential for extracellular enzyme production, fluorogenic 4-methylumbelliferone standards and substrates were used to determine the presence of cellulases, phosphatases and glucosidases in the pure culture. Among fungal strains, Penicillum citrinum BSL17 showed significantly higher amounts of glucosidases (62.15±1.8 μM-1min-1mL) and cellulases (62.11±1.6 μM-1min-1mL), whereas Preussia sp. BSL10 showed significantly higher secretion of glucosidases (69.4±0.79 μM-1min-1mL) and phosphatases (3.46±0.31μM-1min-1mL) compared to other strains. Aureobasidium sp. BSS6 and Preussia sp. BSL10 showed significantly higher potential for indole acetic acid production (tryptophan-dependent and independent pathways). Preussia sp. BSL10 was applied to the host B. sacra tree saplings, which exhibited significant improvements in plant growth parameters and accumulation of photosynthetic pigments. The current study concluded that endophytic microbial resources producing extracellular enzymes and auxin could

  16. Endophytic Fungi from Frankincense Tree Improves Host Growth and Produces Extracellular Enzymes and Indole Acetic Acid.

    Directory of Open Access Journals (Sweden)

    Abdul Latif Khan

    Full Text Available Boswellia sacra, an economically important frankincense-producing tree found in the desert woodlands of Oman, is least known for its endophytic fungal diversity and the potential of these fungi to produce extracellular enzymes and auxins. We isolated various fungal endophytes belonging to Eurotiales (11.8%, Chaetomiaceae (17.6%, Incertae sadis (29.5%, Aureobasidiaceae (17.6%, Nectriaceae (5.9% and Sporomiaceae (17.6% from the phylloplane (leaf and caulosphere (stem of the tree. Endophytes were identified using genomic DNA extraction, PCR amplification and sequencing the internal transcribed spacer regions, whereas a detailed phylogenetic analysis of the same gene fragment was made with homologous sequences. The endophytic colonization rate was significantly higher in the leaf (5.33% than the stem (0.262%. The Shannon-Weiner diversity index was H' 0.8729, while Simpson index was higher in the leaf (0.583 than in the stem (0.416. Regarding the endophytic fungi's potential for extracellular enzyme production, fluorogenic 4-methylumbelliferone standards and substrates were used to determine the presence of cellulases, phosphatases and glucosidases in the pure culture. Among fungal strains, Penicillum citrinum BSL17 showed significantly higher amounts of glucosidases (62.15±1.8 μM-1min-1mL and cellulases (62.11±1.6 μM-1min-1mL, whereas Preussia sp. BSL10 showed significantly higher secretion of glucosidases (69.4±0.79 μM-1min-1mL and phosphatases (3.46±0.31μM-1min-1mL compared to other strains. Aureobasidium sp. BSS6 and Preussia sp. BSL10 showed significantly higher potential for indole acetic acid production (tryptophan-dependent and independent pathways. Preussia sp. BSL10 was applied to the host B. sacra tree saplings, which exhibited significant improvements in plant growth parameters and accumulation of photosynthetic pigments. The current study concluded that endophytic microbial resources producing extracellular enzymes and auxin

  17. Evolution de certains composés volatils du bois de chêne au cours de son séchage premiers résultats

    Directory of Open Access Journals (Sweden)

    Pascal Chatonnet

    1994-12-01

    Full Text Available Pendant le séchage des merrains sur parc à l'air libre, on assiste à une profonde modification de la composition chimique et du profil aromatique du bois de chêne. La teneur en eugénol, en aldéhydes phénols et en Β-méthyl-y-octalactone augmente régulièrement pendant le vieillissement du bois. Principalement sous forme trans au début du séchage, la Β-méthyl-y-octalactone s'accumule finalement sous la forme cis plus aromatique. Dans le même temps, on observe également une accumulation importante du précurseur inodore de Β-méthyl-y-octalactone. L'intervention des micro-organismes fréquemment isolés sur le bois en cours de séchage naturel sur la formation de composés aromatiques a été étudiée. La culture de diverses moisissures isolées de bois en cours de séchage naturel sur extraits de bois et directement sur sciure de chêne aboutit dans tous les cas à une forte diminution des teneurs en aldéhydes phénols, notamment de la vanilline. En effet, tous les micro-organismes étudiés (Penicilium sp., Trichoderma sp., Aureobasidium sp. sont incapables de dégrader la lignine car ils ne possèdent pas d'activité ligninase. En revanche, ces champignons possèdent une forte activité oxydo-réductase capable de réduire la vanilline en alcool vanillique inodore. En conséquence, les aldéhydes phénols apparaissant au cours du séchage naturel du merrain ne dérivent pas d'une attaque enzymatique mais plus vraisemblablementde l'acidolyse et de l'oxydation chimique de la lignine du bois. Le stockage du bois à l'air libre en présence d'eau et d'oxygène doit permettre une évolution favorable de son potentiel aromatique tant que la vanilline et la cis Β-méthyly-octalactone s'accumulent.

  18. Film forming microbial biopolymers for commercial applications--a review.

    Science.gov (United States)

    Vijayendra, S V N; Shamala, T R

    2014-12-01

    Microorganisms synthesize intracellular, structural and extracellular polymers also referred to as biopolymers for their function and survival. These biopolymers play specific roles as energy reserve materials, protective agents, aid in cell functioning, the establishment of symbiosis, osmotic adaptation and support the microbial genera to function, adapt, multiply and survive efficiently under changing environmental conditions. Viscosifying, gelling and film forming properties of these have been exploited for specific significant applications in food and allied industries. Intensive research activities and recent achievements in relevant and important research fields of global interest regarding film forming microbial biopolymers is the subject of this review. Microbial polymers such as pullulan, kefiran, bacterial cellulose (BC), gellan and levan are placed under the category of exopolysaccharides (EPS) and have several other functional properties including film formation, which can be used for various applications in food and allied industries. In addition to EPS, innumerable bacterial genera are found to synthesis carbon energy reserves in their cells known as polyhydroxyalkanoates (PHAs), microbial polyesters, which can be extruded into films with excellent moisture and oxygen barrier properties. Blow moldable biopolymers like PHA along with polylactic acid (PLA) synthesized chemically in vitro using lactic acid (LA), which is produced by LA bacteria through fermentation, are projected as biodegradable polymers of the future for packaging applications. Designing and creating of new property based on requirements through controlled synthesis can lead to improvement in properties of existing polysaccharides and create novel biopolymers of great commercial interest and value for wider applications. Incorporation of antimicrobials such as bacteriocins or silver and copper nanoparticles can enhance the functionality of polymer films especially in food packaging

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

    Energy Technology Data Exchange (ETDEWEB)

    Eslaminejad, Touba, E-mail: tslaminejad@yahoo.com [Pharmaceutics Research Centre, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman (Iran, Islamic Republic of); Nematollahi-Mahani, Seyed Noureddin, E-mail: nnematollahi@kmu.ac.ir [Department of Anatomy, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman (Iran, Islamic Republic of); Neuroscience Research Centre, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman (Iran, Islamic Republic of); Afzal Research Institute, Kerman (Iran, Islamic Republic of); Ansari, Mehdi, E-mail: mansari@kmu.ac.ir [Pharmaceutics Research Centre, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman (Iran, Islamic Republic of); Pharmaceutics Research Centre, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman (Iran, Islamic Republic of)

    2016-03-15

    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 IC{sub 50} value of 189 µg/µl. - Highlights: • Magnetite nanopowder was produced by thermal decomposition method. • TEM studies showed the average size of

  20. In-vitro and in-vivo design and validation of an injectable polysaccharide-hydroxyapatite composite material for sinus floor augmentation.

    Science.gov (United States)

    Fricain, J C; Aid, R; Lanouar, S; Maurel, D B; Le Nihouannen, D; Delmond, S; Letourneur, D; Amedee Vilamitjana, J; Catros, S

    2018-04-07

    Polysaccharide-based composite matrices consisting of natural polysaccharides, pullulan and dextran supplemented with hydroxyapatite (Matrix-HA) have recently been developed. The principal objective of this study was to evaluate the capacities of this composite material to promote new bone formation in a sinus lift model in the sheep. Secondary objectives were to evaluate in vitro properties of the material regarding cell adhesion and proliferation. In this report, once such composite matrix was prepared as injectable beads after dispersion in a physiological buffer, and evaluated using a large animal model (sheep) for a sinus lift procedure. In vitro studies revealed that these microbeads (250-550μm in diameter) allow vascular cell adhesion and proliferation of Endothelial Cells (EC) after 1 and 7 days of culture. In vivo studies were performed in 12 adult sheep, and newly formed tissue was analyzed by Cone Beam Computed Tomography (CBCT scanning electron microscopy (SEM) and by histology 3 and 6 months post-implantation. CBCT analyses at the implantation time revealed the radiolucent properties of these matrices. Quantitative analysis showed an increase of a dense mineralized tissue in the Matrix-HA group up to 3 months of implantation. The mineralized volume over total volume after 6 months reached comparable values to those obtained for Bio-Oss ® used as positive control. Histological examination confirmed that the Matrix-HA did not induce any long term inflammatory events, and promoted direct contact between the osteoid tissue and lamellar bone structures and beads. After 6 months, we observed a dense network of osteocytes surrounding both biomaterials as well as a newly vascularized formed tissue in close contact to the biomaterials. In conclusion, the absence of animal components in Matrix-HA, the osteoconductive property of Matrix-HA in sheep, resulting in a dense bone and vascularized tissue, and the initial radiolucent property to follow graft

  1. Incorporation of poly-saccharidic derivatives in model biological systems: monolayers, lamellar phases and vesicles

    International Nuclear Information System (INIS)

    Deme, Bruno

    1995-01-01

    Our aim is to introduce a soluble polymer in a lyotropic lamellar phase, and to modify the force balance in the case of a collapsed system where no repulsive contribution overcomes the van der Waals attraction, except at very short distances where hydration forces dominate (i.e. a collapsed stack of membranes). Mixed layers of a synthetic lecithin (DMPC) and a hydrophobically modified polysaccharide (cholesteryl-pullulan, CHP) have been investigated at the air-water interface by surface tension experiments and by specular reflection of neutrons. The DMPC/CHP/water ternary phase diagram has been determined by small angle X-ray scattering (SAXS) and small angle neutron scattering (SANS). CHP derivatives are associative polymers bearing lateral cholesterol groups that interact with a polar phases such as phospholipid monolayers and biological membranes. These derivatives are surface active and self-aggregate in solution leading to the formation of soluble micellar type aggregates. The interaction of CHP derivatives with lipidic structures involves the anchoring of the cholesterol groups that yields to the tethering of the poly-saccharidic backbones at lipid/water interfaces. These poly-saccharidic backbones are flexible chains in good solvent in water. Using these derivatives and a new preparation procedure, we show that it is possible to avoid the depletion of the polysaccharide due to its steric exclusion by the collapsed DMPC lamellar phase. We are able to prepare samples at thermodynamic equilibrium with the polysaccharide solubilized in the lamellar phase, a situation opposed to the well known behavior of mixed polysaccharide/lecithin Systems commonly used in osmotic stress experiments. Here, the osmotic pressure of the chains confined in the lamellar lattice acts as a new long range repulsive contribution in the DMPC lyotropic L_α phase and results in the swelling of the lamellar phase at large membrane separations (570 A). Such bilayer separations allow out of

  2. A solid phospholipid-bile salts-mixed micelles based on the fast dissolving oral films to improve the oral bioavailability of poorly water-soluble drugs

    Science.gov (United States)

    Lv, Qing-yuan; Li, Xian-yi; Shen, Bao-de; Dai, Ling; Xu, He; Shen, Cheng-ying; Yuan, Hai-long; Han, Jin

    2014-06-01

    The phospholipid-bile salts-mixed micelles (PL-BS-MMs) are potent carriers used for oral absorption of drugs that are poorly soluble in water; however, there are many limitations associated with liquid formulations. In the current study, the feasibility of preparing the fast dissolving oral films (FDOFs) containing PL-BS-MMs was examined. FDOFs incorporated with Cucurbitacin B (Cu B)-loaded PL-sodium deoxycholate (SDC)-MMs have been developed and characterized. To prepare the MMs and to serve as the micellar carrier, a weight ratio of 1:0.8 and total concentration of 54 mg/mL was selected for the PL/SDC based on the size, size distribution, zeta potential, encapsulation efficiency, and morphology. The concentration of Cu B was determined to be 5 mg/mL. Results showed that a narrow size distributed nanomicelles with a mean particle size of 86.21 ± 6.11 nm and a zeta potential of -31.21 ± 1.17 mV was obtained in our optimized Cu B-PL/SDC-MMs formulation. FDOFs were produced by solvent casting method and the formulation with 50 mg/mL of pullulan and 40 mg/mL of PEG 400 were deemed based on the physico-mechanical properties. The FDOFs containing Cu B-PL/SDC-MMs were easily reconstituted in a transparent and clear solution giving back a colloidal system with spherical micelles in the submicron range. In the in vitro dissolution test, the FDOFs containing Cu B-PL/SDC-MMs showed an increased dissolution velocity markedly. The pharmacokinetics study showed that the FDOFs containing PL-SDC-MMs not only kept the absorption properties as same as the PL-SDC-MMs, but also significantly increased the oral bioavailability of Cu B compared to the Cu B suspension ( p < 0.05). This study showed that the FDOFs containing Cu B-PL/SDC-MMs could represent a novel platform for the delivery of poorly water-soluble drugs via oral administration. Furthermore, the integration with the FDOFs could also provide a simple and cost-effective manner for the solidification of PL-SDC-MMs.

  3. Comparison of Chain-Length Preferences and Glucan Specificities of Isoamylase-Type α-Glucan Debranching Enzymes from Rice, Cyanobacteria, and Bacteria.

    Directory of Open Access Journals (Sweden)

    Taiki Kobayashi

    Full Text Available It has been believed that isoamylase (ISA-type α-glucan debranching enzymes (DBEs play crucial roles not only in α-glucan degradation but also in the biosynthesis by affecting the structure of glucans, although molecular basis on distinct roles of the individual DBEs has not fully understood. In an attempt to relate the roles of DBEs to their chain-length specificities, we analyzed the chain-length distribution of DBE enzymatic reaction products by using purified DBEs from various sources including rice, cyanobacteria, and bacteria. When DBEs were incubated with phytoglycogen, their chain-length specificities were divided into three groups. First, rice endosperm ISA3 (OsISA3 and Eschericia coli GlgX (EcoGlgX almost exclusively debranched chains having degree of polymerization (DP of 3 and 4. Second, OsISA1, Pseudomonas amyloderamosa ISA (PsaISA, and rice pullulanase (OsPUL could debranch a wide range of chains of DP≧3. Third, both cyanobacteria ISAs, Cyanothece ATCC 51142 ISA (CytISA and Synechococcus elongatus PCC7942 ISA (ScoISA, showed the intermediate chain-length preference, because they removed chains of mainly DP3-4 and DP3-6, respectively, while they could also react to chains of DP5-10 and 7-13 to some extent, respectively. In contrast, all these ISAs were reactive to various chains when incubated with amylopectin. In addition to a great variation in chain-length preferences among various ISAs, their activities greatly differed depending on a variety of glucans. Most strikingly, cyannobacteria ISAs could attack branch points of pullulan to a lesser extent although no such activity was found in OsISA1, OsISA3, EcoGlgX, and PsaISA. Thus, the present study shows the high possibility that varied chain-length specificities of ISA-type DBEs among sources and isozymes are responsible for their distinct functions in glucan metabolism.

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

    International Nuclear Information System (INIS)

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

    2016-01-01

    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 IC 50 value of 189 µg/µl. - Highlights: • Magnetite nanopowder was produced by thermal decomposition method. • TEM studies showed the average size of the

  5. Novel system for pulse radiolysis with multi-angle light scattering detection (PR-MALLS) - concept, construction and first tests

    Science.gov (United States)

    Kadlubowski, S.; Sawicki, P.; Sowinski, S.; Rokita, B.; Bures, K. D.; Rosiak, J. M.; Ulanski, P.

    2018-01-01

    Time-resolved pulse radiolysis, utilizing short pulses of high-energy electrons from accelerators, is an effective method for rapidly generating free radicals and other transient species in solution. Combined with fast time-resolved spectroscopic detection (typically in the ultraviolet/visible/near-infrared), it is invaluable for monitoring the reactivity of species subjected to radiolysis on timescales ranging from picoseconds to seconds. When used for polymer solutions, pulse radiolysis can be coupled with light-scattering detection, creating a powerful tool for kinetic and mechanistic analysis of processes like degradation or cross-linking of macromolecules. Changes in the light scattering intensity (LSI) of polymer solutions are indicative of alterations in the molecular weight and/or in the radius of gyration, i.e., the dimensions and shape of the macromolecules. In addition to other detection methods, LSI technique provides a convenient tool to study radiation-induced alterations in macromolecules as a function of time after the pulse. Pulse radiolysis systems employing this detection mode have been so far constructed to follow light scattered at a single angle (typically the right angle) to the incident light beam. Here we present an advanced pulse radiolysis & multi-angle light-scattering-intensity system (PR-MALLS) that has been built at IARC and is currently in the phase of optimization and testing. Idea of its design and operation is described and preliminary results for radiation-induced degradation of pullulan as well as polymerization and crosslinking of poly(ethylene glycol) diacrylate are presented. Implementation of the proposed system provides a novel research tool, which is expected to contribute to the expansion of knowledge on free-radical reactions in monomer- and polymer solutions, by delivering precise kinetic data on changes in molecular weight and size, and thus allowing to formulate or verify reaction mechanisms. The proposed method is