黄曲霉毒素生物防控菌的筛选及鉴定

筛选黄曲霉毒素生物防控菌,为黄曲霉毒素的生物防控提供支持。以花生原产地土壤为材料,采用牛津杯法筛选所需菌株。对筛选出的拮抗菌株进行抑制产毒曲霉菌株的生长、产孢、降解黄曲霉毒素实验。筛选出2株黄曲霉毒素生防细菌,编号21-1-2、17-3,经鉴定,拮抗菌21-1-2为枯草芽胞杆菌,拮抗菌17-3为地衣芽胞杆菌。分别对拮抗菌对曲霉孢子萌发的抑制、抑制黄曲霉的生长和菌丝延长以及减少黄曲霉毒素的产生、对黄曲霉毒素的分解作用等几个方面进行研究,结果表明,拮抗菌可以明显抑制产毒曲霉孢子的萌发、生长、菌丝的延长,减少黄曲霉毒素的产生以及分解黄曲霉毒素。     

A thermostable lipase produced by a newly isolated <Emphasis Type="Italic">Geotrichum</Emphasis>-like strain,R59

Growth and production of lipase by a new Geotrichum-like strain, R59, were studied. Production of extracellular lipase was substantially enhanced when the initial pH of the culture medium, types of carbon and nitrogen sources, substances probably stimulating the lipase biosynthesis, the temperature, and time of growth were optimized. Sucrose and triolein were the most effective carbon sources for lipase production. Maximum lipase activity (146 U/ml^–1) was obtained with urea as the nitrogen source. Growth at 30°C, an initial pH of 6.0 and incubation time of 48 h were found as optimum conditions for cell growth and production of lipase by Geotrichum-like strain R59. The enzyme was thermostable and exhibited very high activity after 1 h incubation at 60°C.     

Enzymatic properties of lipase and characteristics production byLactobacillus delbrueckii subsp.bulgaricus

Some properties of an extracellular lipase produced byLactobacillus delbrueckii subsp.bulgaricus were studied. Maximum enzyme activity was found against olive and butter oil as enzyme substrates. Addition of 9% acacia gum, 0.1% Na-deoxycholate and 0.01 M CaCl₂ to the enzyme reaction mixture increased-lipase activity from 5.3 to 14.5 (FFA/mg protein/minute) at pH 6.0 and at 40° C. Maximum lipase production was reached in the presence of glucose as a sole source of carbon, wheat bran as nitrogen source, olive oil as a sole lipid source and butyric acid as fatty acid supporting the growth medium. An initial pH value of the culture medium of 6.0 and a temperature of 35° C gave the highest lipolytic activity.     

Pectinolytic activity of bacteria isolated from soil and two fungal strains during submerged fermentation

Seven different strains were selected for their ability to degrade citrus pectin. Alkaline pectinases were produced by five bacterial soil isolates, whereas two fungal strains produced pectinase in an acidic environment. The bacteria were isolated from soil of a plum orchard in Northern Ireland. These isolates produced significant amounts of pectin lyase (PL) and polygalacturonase (PG) with maximum activities of 30.1 and 29.1 U/ml respectively. Fungal strains Aspergillus sp. and PN-1 produced four different pectinolytic activities; endo-PG, exo-PG, pectin esterase (PE) and PL. The Aspergillus sp. produced higher amounts of pectinase than PN-1. The Aspergillus sp. excreted highly stable pectinases, which may be of importance for industrial applications.     

Production of lipase free of citrinin by Penicillium citrinum

Lipase (Glycerol ester hydrolase E.G. 3.1.1.3) from a Brazilian strain of Penicillium citrinum free of the mycotoxin citrinin has been investigated. Citrinin production was inhibited by using culture medium containing olive oil, soybean oil and corn oil as carbon sources. Potassium concentration and pH play an important role in citrinin production. Potassium concentration lower than 30 mM and pH below 4.5 inhibited the mycotoxin production. P. citrinum produced lipase free of extraneous proteins and citrinin when cultured using, as nitrogen source, ammonium sulphate (lipase activity of 7.88 U/mg) and yeast extract (lipase activity of 4.95 U/mg) with olive oil as carbon source. This data is relevant to the larger scale production of lipases for food technology applications, from Penicillium citrinum.     

Clavatol and patulin formation as the antagonistic principle of <Emphasis Type="Italic">Aspergillus clavatonanicus</Emphasis>, an endophytic fungus of <Emphasis Type="Italic">Taxus mairei</Emphasis>

Many endophytic fungi are known to protect plants from plant pathogens, but the antagonistic mechanism has rarely been revealed. In this study, we wished to learn whether an endophytic Aspergillus sp., isolated from Taxus mairei, would indeed produce bioactive components, and if so whether (a) they would antagonize plant pathogenic fungi; and (b) whether this Aspergillus sp. would produce the compound also under conditions of confrontation with these fungi. The endophytic fungal strain from T. mairei was identified as Aspergillus clavatonanicus by analysis of morphological characteristics and the sequence of the internal transcribed spacers (ITS rDNA) of rDNA. When grown in surface culture, the fungus produced clavatol (2′,4′-dihydroxy-3′,5′-dimethylacetophenone) and patulin (2-hydroxy-3,7-dioxabicyclo [4.3.0]nona-5,9-dien-8-one), as shown by shown by NMR, MS, X-ray, and EI-MS analysis. Both exhibited inhibitory activity in vitro against several plant pathogenic fungi, i.e., Botrytis cinerea, Didymella bryoniae, Fusarium oxysporum f. sp. cucumerinum, Rhizoctonia solani, and Pythium ultimum. During confrontation with P. ultimum, A. clavatonanicus antagonized its growth of P. ultimum, and both clavatol as well as patulin were formed as the only bioactive components, albeit with different kinetics. We conclude that A. clavatonanicus produces clavatol and patulin, and that these two polyketides may be involved in the protection of T. mairei against attack by plant pathogens by this Aspergillus sp.     

Streptomyces sp. TEM 33 possesses high lipolytic activity in solid-state fermentation in comparison with submerged fermentation

Solid-state fermentation (SSF) is a bioprocess that doesn’t need an excess of free water, and it offers potential benefits for microbial cultivation for bioprocesses and product development. In comparing the antibiotic production, few detailed reports could be found with lipolytic enzyme production by Streptomycetes in SSF. Taking this knowledge into consideration, we prefer to purify Actinomycetes species as a new source for lipase production. The lipase-producing strain Streptomyces sp. TEM 33 was isolated from soil and lipase production was managed by solid-state fermentation (SSF) in comparison with submerged fermentation (SmF). Bioprocess-affecting factors like initial moisture content, incubation time, and various carbon and nitrogen additives and the other enzymes secreted into the media were optimized. Lipase activity was measured as 1.74 ± 0.0005 U/g dry substrate (gds) by the p-nitrophenylpalmitate (pNPP) method on day 6 of fermentation with 71.43% final substrate moisture content. In order to understand the metabolic priority in SSF, cellulase and xylanase activity of Streptomyces sp. TEM33 was also measured. The microorganism degrades the wheat bran to its usable form by excreting cellulases and xylanases; then it secretes the lipase that is necessary for degrading the oil in the medium.     

Biotechnological conversion of waste cooking olive oil into lipid-rich biomass using Aspergillus and Penicillium strains

Aims: In this study, we have investigated the biochemical behaviour of Aspergillus sp. (five strains) and Penicillium expansum (one strain) fungi cultivated on waste cooking olive oil. The production of lipid‐rich biomass was the main target of the work. In parallel, the biosynthesis of other extracellular metabolites (organic acids) and enzyme (lipase) and the substrate fatty acid specificity of the strains were studied. Methods and Results: Carbon‐limited cultures were performed on waste oil, added in the growth medium at 15 g l^?1, and high biomass quantities were produced (up to c. 18 g l^?1, conversion yield of c. 1·0 g of dry biomass formed per g of fat consumed or higher). Cellular lipids were accumulated in notable quantities in almost all cultures. Aspergillus sp. ATHUM 3482 accumulated lipid up to 64·0% (w/w) in dry fungal mass. In parallel, extracellular lipase activity was quantified, and it was revealed to be strain and fermentation time dependent, with a maximum quantity of 645 U ml^?1 being obtained by Aspergillus niger NRRL 363. Storage lipid content significantly decreased at the stationary growth phase. Some differences in the fatty acid composition of both cellular and residual lipids when compared with the initial substrate fat used were observed; in various cases, cellular lipids more saturated and enriched with arachidic acid were produced. Aspergillus strains produced oxalic acid up to 5·0 g l^?1. Conclusions: Aspergillus and Penicillium strains are able to convert waste cooking olive oil into high‐added‐value products. Significance and Impact of the Study: Increasing fatty wastes amounts are annually produced. The current study provided an alternative way of biovalourization of these materials, by using them as substrates, to produce added‐value compounds.     

Characterization and optimization of extracellular alkaline lipase production by Alcaligenes sp. Using stearic acid as carbon source

The aim of this study was to improve the production of an extracellular alkaline lipase from Alcaligenes sp. (ATCC 31371) by optimization of the culture medium, for economic production of biodiesel from waste vegetable oil. A number of carbon sources including different types of starch, sugar, sugar alcohol, organic acids, and surfactants were investigated. Polyoxyethylene (20) sorbitan tristearate, whose side chain is stearic acid, was the most effective carbon source for lipase production. Box-Behnken experimental design was used for three factors (soy protein, sodium nitrate, and stearic acid) and the optimal composition for maximum lipase production (1.7-fold enhancement) was established as soy protein 4.07%, sodium nitrate 0.17%, and stearic acid 0.28% at 28°C with an agitation rate of 220 rpm for 24 h. The enzyme was purified to homogeneity and the recovery of the lipase activity was 7.8% with a 30-fold purification. The estimated molecular size of the protein determined by SDS-PAGE was 33 kDa. The optimum pH and temperature of the purified lipase was 8.5 and 40°C, respectively. The purified enzyme was stable in the pH range of 6.0 and 9.5 and in the temperature range of 20 and 50°C.     

Extracellular lipase production by a sapwood-staining fungus,Ophiostoma piceae

The extracellular lipase production of a sapwood-staining fungus, Ophiostoma piceae, grown in liquid media, was optimally active at pH 5.5 and 37°C. Although glucose, fructose, sucrose, starch and dextrin, as carbon sources for growth gave similar mycelial yields, which were higher than those obtained with arabinose, galactose or raffinose, the cells growing on those carbohydrates produced little extracellular lipase. However, both high biomass and lipase activity were obtained when plant oils (olive, soybean, corn, sunflower seed, sesame, cotton seed or peanut) were used as carbon sources. Among the nitrogen sources examined, Casamino acids gave the best growth, whereas (NH₄)₂SO₄ gave the best lipase production. The highest lipase productivity seen was obtained in a medium with olive oil as carbon source and a combination of (NH₄)₂SO₄and peptone as nitrogen source.The authors are with Forest Products Biotechnology, Department of Wood Science, Facully of Forestry, University of British Columbia, Vancouver, BC, V6T 1Z4, Canada     

Removing of Crude Oil From Polluted Areas Using the Isolated Fungi From Tehran Oil Refinery

The pollution of soil and the subsurface environment by crude oil spill and petroleum products spill is a major concern around the world. The aim of this research was to investigate the ability of fungi isolated from Tehran oil refinery area in removing crude oil and to evaluate their enzymatic activities. Plant root samples were collected from the polluted and control areas, and rhizospheral fungi were isolated and determined using the laboratory methods and taxonomic keys. Seven fungal species were isolated and then cultured in potato dextrose agar (PDA) media containing 0–15% (v/v) crude oil. Oil removal was determined after a one-month growth of fungal colonies and then compared with the control media. The results showed that the studied fungi were able to remove crude oil from the media. The highest removal efficiency was observed in Aspergillus sp. Total protein content and enzymatic activity (of peroxidase and catalase) increased with increasing crude oil pollution. The highest enzymatic activity was evaluated in Aspergillus sp. growing in media containing 15% petroleum and the lowest activity was found in non-polluted groups. Results showed that there is a direct correlation between oil-removing potency and enzymatic activity. Aspergillus sp. showed the highest enzyme activity and also the highest petroleum removal efficiency.     

Formation of lipolytic enzymes by Brevibacterium linens

When Brevibacterium linens ATCC 9172 was grown in shake flasks, it produced a cell-associated lipase with a specific activity of 152 to 188 U g^–1 cells depending on the composition of the growth medium. There was no growth in media containing tributyrine as the sole carbon source. The cell-associated lipase had maximum activity at pH 8.0 and 37 °C and was strongly inhibited by 3,4-dichloroisocoumarin, an inhibitor specific for serine esterases. Cell-associated activity was released from the cells by treatment with lysozyme. The kinetics of lipase formation was closely related to the amount of biomass formed during growth.     

Preparation and testing of Sardinella protein hydrolysates as nitrogen source for extracellular lipase production by Rhizopus oryzae

Various fish protein hydrolysates (FPH) from sardinella (Sardinella aurita) were used as nitrogen sources for the production of extracellular lipase by the filamentous fungus Rhizopus oryzae. The best results were obtained with defatted meat–fish protein hydrolysates (DMFPH), indicating the presence in the lipid fraction of some constituents which may repress lipase synthesis. Furthermore, it was found that the extensive hydrolysis of fish proteins resulted in a higher lipase production. The use of 40 g DMFPH l^–1 for the growth of Rhizopus oryzae in medium R1 resulted in a lipase production of 394 U ml^–1, higher than the yield obtained with standard soy peptone as nitrogen source (373 U ml^–1). The most appropriate medium for the growth and the production of lipase is composed only of 24 g DMFPH l^–1 and 10 g glucose l^–1, indicating that the strain can obtain its nitrogen and salts requirements directly from fish substrate.     

Tannase production by Aspergillus fumigatus MA under solid-state fermentation

A tannase yielding fungal culture identified as Aspergillus fumigatus MA was isolated from the effluent collected from a local small scale tannery. The fungal culture produced high yields of extracellular tannase under solid-state fermentation (SSF) using different agro forest residues such as Amla leaves (Phyllanthus emblica), Ber leaves (Zyzyphus mauritiana), Jamun leaves (Syzygium cumini), Jamoa leaves (Syzygium sp.) and Keekar leaves (Acacia nilotica). Among different substrates used, Jamun leaves yielded maximal extra-cellular production of tannase. Various parameters were studied to optimize the extracellular yield of tannase under SSF. The maximum yield of 174.32 U g⁻¹ was obtained at 25°C after 96 h of incubation at pH 5.0. The tap water was used as a moistening agent. A substrate to tap water ratio of 1:1 was found to best for tannase production. Supplementation of the medium with ammonium sulfate as nitrogen source had enhanced tannase production whereas glucose had decreased the enzyme production. This is the first report on production of tannase by Aspergillus fumigatus MA, giving a much higher yield of enzyme under SSF with Jamun leaves as the substrate.     

高寒森林溪流微生物群落结构的季节性变化

高寒森林溪流不仅是区域河流的源头,而且是联系陆地与水域的生态纽带。微生物活动可能成为控制溪流生态系统过程的关键因子,但其结构与动态过程缺乏必要关注。因此,结合同步温度动态监测,采用实时荧光定量PCR和DGGE技术,在2014年到2015年冻融季节和生长季节关键时期对比研究了川西高寒森林溪流和森林林下土壤中微生物群落的动态特征。研究结果发现,高寒森林溪流具有较低的真菌和细菌群落丰度;与森林土壤相同,溪流在冻融季节表现出相对生长季节更高的真菌/细菌比,而且从冻融季节到生长季节,溪流微生物丰度动态也表现出明显的季节性变化特征。与森林土壤不同的是,溪流中细菌和真菌的丰度及其Shannon-Wiener多样性指数的最高值均出现在生长季节而不是冬季冻融季节,并且溪流中细菌丰度在季节性变化的不同时期具有显著差异(P0.05)。此外,森林土壤细菌类群以芽孢杆菌属(Bacillus sp.)比例相对较高,真菌类群则以格孢菌属(Pleosporales sp.)、曲霉属(Aspergillus sp.)和其他一些子囊菌门(Ascomycota)的类群为优势;而溪流细菌类群以红球菌属(Rhodococcus sp.)为主,真菌类群则以曲霉属和空团菌属(Cenococcum sp.)为主。同时,季节性变化中温度、p H、水溶性有机碳和溶解氧等环境因子可显著影响溪流微生物群落结构及其组成,这些环境因子在高寒森林溪流微生物群落的季节性变化过程中具有重要的作用。     

Lipase production byAspergillus andPenicillium species

Forty each of aspergilli and penicillia were screened for extracellular lipase production on agar plates and in liquid medium containing olive oil as substrate. Twenty-nine aspergilli and twenty-six penicillia produced lipase. Out of these, 19 aspergilli and 22 penicillia showed activity both on Nile blue sulfate and glycerol tributyrate agar plates while only 10 aspergilli and 4 penicillia showed a positive response to glycerol tributyrate agar alone. The screening revealed 11Aspergillus spp. and 15Penicillium spp. as new lipase producers. Pig fat as an economic substrate for lipase production was also investigated.     

Studies on a novel carbon source and cosolvent for lipase production by<Emphasis Type="Italic"> Candida rugosa</Emphasis>

Oleic acid esters were shown to be the best carbon source for both cell growth and lipase production by Candida rugosa. Use of a cosolvent, dodecane, in fermentations improved the solubility of solid substrates and increased oxygen solubility. This resulted in the highest lipase activity in batch fermentation with glycerol trioleate and dodecane. Lipase activity reached 77.1 units ml^–1.     

Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) depolymerase from Aspergillus sp. NA-25

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) degrading thermophilic fungus was isolated from soil sample collected from waste disposal site, Islamabad, Pakistan. It was able to grow efficiently on a medium containing PHBV as a sole source of carbon and has been identified as Aspergillus sp. NA-25 by 18S rRNA. Using 9% of inoculum maximum production of PHBV depolymerase was observed at 45°C, pH 7.0 in the presence of 0.2% lactose as an additional carbon source. PHBV depolymerase was purified by precipitation with 80% ammonium sulfate and gel filtration chromatography on Sephadex G-75. The four enzyme forms obtained after gel filtration were analyzed on SDS-PAGE and their molecular weights (36, 68, 72 and 90 kDa) were determined. They were characterized on the basis of effect of different temperatures, pH, metal ions and different reagents on the PHBV activity and stability. It is obvious that the fungal strain Aspergillus sp. NA-25 is capable of degrading PHBV with the help of different types of depolymerases.     

Isolation of a novel lipase producing fungal isolate Aspergillus fumigatus and production optimization of enzyme

Abstract

Fungal lipases occupy a place of prominence among biocatalysts owing to their novel, multifold applications and resistance to high temperature and other operational conditions. In the present study, Aspergillus fumigatus isolated from oil-contaminated soil produced good amount of lipase activity with galactose (1%) as carbon source and peptone (0.1%) as nitrogen source after 72?h of incubation in the production medium at 45?°C and pH 10.0. The isolated enzyme was found to give its optimum reaction temperature at 40?°C and pH 9.0 with the substrate used as p-nitrophenyl benzoate. The activity of lipase was inhibited by the presence of metal ions. A 6.68-fold increase for lipase production was obtained by one variable at a time. Based on the findings of present study, lipase of A. fumigatus is a potential lipase and a candidate for industrial applications such as bioremediation, detergent, leather and pharmaceutical industries.     

High activity xylanase production by Streptomyces olivaceoviridis E-86

The effects of different factors on xylanase production by Streptomyces olivaceoviridis E-86 were studied under shake flask conditions. The best initial pH value of growth medium for xylanase production was pH 6.0. Corn cob xylan and beef peptone were the best C source and N source, respectively. The enzyme activity was doubled by addition of 1.5% (v/v) Tween-80 in the medium. By the combination of the above variables, the highest xylanase activity obtained was 1653 U/ml which is the highest ever reported from Streptomyces sp.