Production of sterigmatocystin by Aspergillus versicolor and Bipolaris sorokiniana on semisynthetic liquid and solid media.

Higher yields of sterigmatocystin were obtained with Aspergillus versicolor than with Bipolaris sorokiniana both in liquid and on solid media. The optimum temperature for sterigmatocystin production by A. versicolor was 27 to 29 degrees C and 23 degrees C for B. sorokiniana. In liquid shake cultures, production of sterigmatocystin by B. sorokiniana was negligible, whereas maximal production by A. versicolor was 210 mg/liter. On solid substrates, the highest yields (8 g/kg) were obtained with A. versicolor on still cultures of whole corn supplemented with Soytone.     

Combined submerged and solid substrate fermentation for the bioconversion of lignocellulose

A novel two-stage bioreactor has been designed for a combined submerged (SF) and solid substrate fermentation (SSF) of wheat straw. The straw was pretreated with steam, and cellulases from the culture fluid of Trichoderma reesei were adsorbed on it for increased bioconvertibility. SSF was conducted in the top part of the bioreactor by inoculating the straw with a 36-h mycelial culture of T. reesei, or Coriolus versicolor. In the bottom part of the fermenter, Endomycopsis fibuliger was grown in SF. The SF liquor was recirculated through the SSF stage at 24 h intervals to remove glucose and other metabolites that may inhibit growth, and to maintain optimum moisture level and temperature. The removed glucose and other metabolites provided nutrients for the yeast in the SF stage. The combined fermentation resulted in overall higher biomass yield, increased bioconversion, increased cellulase production, and increased digestibility compared with single SSF or SF.     

Selection of white-rot basidiomycetes for bioconversion of mustard (Brassica compestris) straw under solid-state fermentation into energy substrate for rumen micro-organism

Aims: Selection of white‐rot fungi of bio‐conversion of mustard straw (MS) into feed for ruminants. Methods and Results: Mustard straw was cultured with Ganoderma applanatum, Coriolus versicolor and Phanerochaete chrysosporium for solid‐state fermentation at 35°C from 7 to 63 days for dilignification and for 21 days to study dry matter digestibility and protein enrichment. Lignin loss in fungus cultured straw varied between 100 and 470 g kg^?1 lignin. Dilignification was higher between 7 and 28 days fermentation with C. versicolor. Among the three fungi P. chrysosporium was the most effective in degrading lignin for longer fermentation. In‐vitro dry matter digestibility (IVDMD) and crude protein content was higher in C. versicolor cultured straw. Large quantity of straw was cultured by C. versicolor for 21 days, for in vivo evaluation. Mean pH and metabolites of rumen fermentation were not different while, pH and volatile fatty acid increased at 6 h postfermentation on cultured straw feeding. Cultured straw fermentation increased (P = 0·001) small holotricks and reduced (P = 0·005) large holotricks population. Fungus cultures straw did not improve microbial enzyme concentration. Conclusions: Coriolus versicolor and P. chrysosporium were the promising fungus for MS bio‐dilignification. Significance and Impact of the Study: Coriolus versicolor treated MS improved dry matter digestibility and protein content.     

Rice straw-decomposing fungi and their cellulolytic and xylanolytic enzymes

Filamentous fungi colonizing rice straw were collected from 11 different sites in Korea and were identified based on characterization of their morphology and molecular properties. The fungi were divided into 25 species belonging to 16 genera, including 14 ascomycetes, one zygomycete, and one basidiomycete. Fungal cellulolytic and xylanolytic enzymes were assessed through a two-step process, wherein highly active cellulase- and/or hemicellulaseproducing fungi were selected in a first screening step followed by a second step to isolate the best enzymeproducer. Twenty-five fungal species were first screened for the production of total cellulase (TC), endo-beta-1,4 glucanase (EG), and endo-beta-1,4 xylanase (XYL) using solid-state fermentation with rice straw as substrate. From this screening, six species, namely, Aspergillus niger KUC5183, A. ochraceus KUC5204, A. versicolor KUC5201, Mucor circinelloides KUC6014, Trichoderma harzianum 1 KUC5182, and an unknown basidiomycete species, KUC8721, were selected. These six species were then incubated in liquid Mandels' media containing cellulose, glucose, rice straw, or xylan as the sole carbon source and the activities of six different enzymes were measured. Enzyme production was highly influenced by media conditions and in some cases significantly increased. Through this screening process, Trichoderma harzianum 1 KUC5182 was selected as the best enzyme producer. Rice straw and xylan were good carbon sources for the screening of cellulolytic and xylanolytic enzymes.     

Vibrio vulnificus may produce a metalloprotease causing an edematous skin lesion in vivo

Abstract Oxalate decarboxylase was detected both intra- and extracellularly in liquid cultures of Coriolus versicolor . Induction of the enzyme by addition of oxalic acid to the medium on day 6 of growth resulted in a 50-fold increase in specific activity in the mycelia and a 30-fold increase in the extracellular specific activity in the media. The protein was isolated and purified from mycelia, and characterised by polyacrylamide gel electrophoresis and Western blotting against a polyclonal antibody raised to oxalate decarboxylase from Collybia velutipes (Basidiomycete). A major protein band of M _r 59000 cross-reacted with the antibody. Immunogold-cytochemical labelling of ultra-thin sections of beechwood infected with C. versicolor showed that the enzyme was localised close to the plasma membrane and in intracellular vesicles.     

Protein enrichment and digestibility of soft rush (<Emphasis Type="Italic">Juncus effusus</Emphasis>) and rice residues using edible mushrooms <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> and <Emphasis Type="Italic">Pleurotus sajor-caju</Emphasis>

Pleurotus species are found to be among the most efficient lignocellulolytic types of white-rot fungi. Rice is the main grain cultivated in the extreme south of Brazil. Defatted rice bran and straw are by-products of low aggregate value. Soft rush (Juncus effusus) is a common native plant also very abundant in the region. In the present work, we evaluated changes in substrate composition after growth of two white-rot fungal species: Pleurotus ostreatus and Pleurotus sajor-caju, aiming to increase protein content and digestibility from substrates through solid fermentations and obtain edible mushrooms of high aggregate value. For that, defatted rice bran, defatted rice straw and soft rush were utilized as substrate. The influence of the variables thermal treatment temperature of substrate, substrate moisture and concentration were evaluated on the protein content, digestibility and biological efficiency. The highest protein enrichment of rice bran in P. sajor-caju-fermented medium was due the fact that there was no fructification in these media, while for the P. ostreatus-fermented medium, part of the synthesized protein was converted into mushrooms. The highest protein enrichments were verified in medium with 80% moisture and 25% soft rush (47.1% using P. ostreatus and 49.0% using P. sajor-caju). A higher digestible protein increase was obtained for both species in media with 70% moisture and 25% soft rush.     

培养液及其浓度和pH值对草履虫种群增长的影响

采用稻草、小麦、奶粉、酵母、玉米粒、荷叶等6种培养液培养草履虫,结果表明,稻草、玉米粒和奶粉培养液为草履虫的理想培养液。另外,对上述3种理想培养液的不同浓度和稻草培养液的不同pH值影响草履虫的生长繁殖进行了实验研究,统计分析表明影响较为明显。稻草培养液的最适浓度为1%~2%;奶粉培养液的最适浓度为0.1%~0.2%;玉米粒培养液的最适浓度为1%~2%;稻草培养液最适pH值为7.0。     

Comparison of phenanthrene and pyrene degradation by different wood-decaying fungi.

The degradation of phenanthrene and pyrene was investigated by using five different wood-decaying fungi. After 63 days of incubation in liquid culture, 13.8 and 4.3% of the [ring U-14C]phenantherene and 2.4 and 1.4% of the [4,5,9,10-14C]pyrene were mineralized by Trametes versicolor and Kuehneromyces mutabilis, respectively. No 14CO2 evolution was detected in either [14C]phenanthrene or [14C]pyrene liquid cultures of Flammulina velutipes, Laetiporus sulphureus, and Agrocybe aegerita. Cultivation in straw cultures demonstrated that, in addition to T. versicolor (15.5%) and K. mutabilis (5.0%), L. sulphureus (10.7%) and A. aegerita (3.7%) were also capable of mineralizing phenanthrene in a period of 63 days. Additionally, K. mutabilis (6.7%), L. sulphureus (4.3%), and A. aegerita (3.3%) mineralized [14C]pyrene in straw cultures. The highest mineralization of [14C] pyrene was detected in straw cultures of T. versicolor (34.1%), which suggested that mineralization of both compounds by fungi may be independent of the number of aromatic rings. Phenanthrene and pyrene metabolites were purified by high-performance liquid chromatography and identified by UV absorption, mass, and 1H nuclear magnetic resonance spectrometry. Fungi capable of mineralizing phenanthrene and pyrene in liquid culture produced enriched metabolites substituted in the K region (C-9,10 position of phenanthrene and C-4,5 position of pyrene), whereas all other fungi investigated produced metabolites substituted in the C-1,2, C-3,4, and C-9,10 positions of phenanthrene and the C-1 position of pyrene.     

Extracellular oxidative enzyme production and PAH removal in soil by exploratory mycelium of white rot fungi

Selected strains of three species of white rot fungi, Pleurotus ostreatus, Phanerochaete chrysosporium and Trametes versicolor, were grown in sterilized soil from straw inocula. The respective colonization rates and mycelium density values decreased in the above mentioned order. Three- and four-ringed PAHs at 50 ppm inhibited growth of fungi in soil to some extent. The activities of fungal MnP and laccase (units per g dry weight of straw or soil), extracted with 50 mM succinate-lactate buffer (pH 4.5), were 5 to 20-fold higher in straw compared to soil. The enzyme activities per g dry soil in P. ostreatus and T. versicolor were similar, in contrast to P. chrysosporium, where they were extremely low. Compared to the aerated controls, P. ostreatus strains reduced the levels of anthracene, pyrene and phenanthrene by 81–87%, 84–93% and 41–64% within 2 months, respectively. During degradation of anthracene, all P. ostreatus strains accumulated anthraquinone. PAH removal rates in P. chrysosporium and T. versicolor soil cultures were much lower.     

Necrotrophic mycoparasitism of Botrytis cinerea by cellulolytic and ligninocellulolytic Basidiomycetes

Twenty-six isolates representing 17 species of aphyllophoraceous, wood-decaying Basidiomycetes and five species of agaricoid, turf-borne, thatch-decaying Basidiomycetes were screened for their abilities to degrade cellulose, lignin, and melanin by using colorimetric degradation assays on agar media. Selected ligninocellulolytic Basidiomycetes capable of degrading melanin were screened for antagonism of Botrytis cinerea Per.:Fr. The greatest inhibition of Botrytis colony and hyphal growth in vitro was observed in confrontations with Irpex lacteus (Fr.) Fr., Trametes versicolor (L.:Fr.) Pilat, and Chondrostereum purpureum (Pers.:Fr.) Pouzar. Hyphal interference and necrotrophic mycoparasitism by these ligninocellulolytic Basidiomycetes were recognized microscopically as coagulation and degeneration of Botrytis cytoplasm and as coiling and invasion of hyphae, conidiophores, and conidia, respectively. Sclerotia of B. cinerea were killed and parasitized in agar media, straw mulch, or moist sand infested separately with these three mycoparasites.     

Radiation and fermentation treatment of cellulosic wastes

The effect of radiation pasteurization of sugar cane bagasse and rice straw and fermentation using various strains of fungi were studied for upgrading of cellulosic wastes. The initial contamination by fungi and aerobic bacteria both in bagasse and straw was high. The doses of 30 kGy for sterilization and 8 kGy for elimination of fungi were required. Irradiation effect showed that rice straw contained comparatively radioresistant microorganisms. It was observed that all the fungi (Hericium erinacium, Pleurotus djamor, Ganoderma lucidum, Auricularia auricula, Lentinus sajor-caju, Coriolus versicolor, Polyporus arcularius, Coprinus cinereus) grow extending over the entire substrates during one month after inoculation in irradiated bagasse and rice straw with 3% rice bran and 65% moisture content incubated at 30°C. Initially, sugar cane bagasse and rice straw substrates contained 39.4% and 25.9% of cellulose, 22.9% and 26.9% of hemicellulose, and 19.6% and 13.9% of lignin + cutin, respectively. Neutral detergent fibre (NDF) values decreased significantly in sugar cane bagasse fermented byG. lucidum, A. auricula andP. arcularius, and in rice straw fermented by all the 8 strains of fungi. Acid detergent fibre (ADF) values also decreased in bagasse and rice straw fermented by all the fungi.P. arcularius, H. erinacium, G. lucidum andC. cinereus were found to be the most effective strains for delignification of sugar cane bagasse.     

Improvement of xylanase production in solid-state fermentation by alkali tolerant Aspergillus versicolor MKU3

AIMS: To optimize the media components for xylanase production by Aspergillus versicolor MKU3 in solid-state fermentation (SSF). METHODS AND RESULTS: Medium optimization was carried out using De Moe's fractional factorial design with seven components. Maximum production of xylanase 3249.9 U g(-1) was obtained in SSF with an optimized medium containing (g l(-1)): NaNO(3), 20; K(2)HPO(4), 20; MgSO(4), 10; FeSO(4), 0.001; KCl, 1; peptone, 10 and yeast extract, 10. Four components namely NaNO(3), MgSO(4), peptone and K(2)HPO(4) significantly increased the xylanase production by A. versicolor MKU3. CONCLUSIONS: Fractional factorial design was used to optimize the seven components in the fermentation medium for SSF. The optimized media increased xylanase production by 3.4-fold. SIGNIFICANCE AND IMPACT OF THE STUDY: Aspergillus versicolor MKU3 produced maximum xylanase after two steps of media optimization under alkaline condition. This medium will be significant value for xylanase production in SSF.     

Growth of Pleurotus ostreatus on wheat straw and wheat-grain-based media: biochemical aspects and preparation of mushroom inoculum

Mycelial growth, intracellular activity of proteases, laccases and β-1,3-glucanases, and cytoplasmic protein were evaluated in the vegetative phase of Pleurotus ostreatus grown on wheat straw and in wheat-grain-based media in Petri dishes and in bottles. The productivity of the wheat straw and wheat-grain-based spawn in cylindrical polyethylene bags containing 5 kg of chopped straw was also determined. We observed high activity of proteases and high content of intracellular protein in cultures grown on wheat straw. This suggests that the proteases are not secreted into the medium and that the protein is an important cellular reserve. On the contrary, cultures grown on wheat straw secreted laccases into the medium, which could be induced by this substrate. P. ostreatus grown on media prepared with a combination of wheat straw and wheat grain showed a high radial growth rate in Petri dishes and a high level of mycelial growth in bottles. The productivities of wheat straw and wheat-grain-based spawn were similar. Our results show that cheaper and more productive mushroom spawn can be prepared by developing the mycelium on wheat straw and wheat-grain-based substrates.     

Production of bioactive polysaccharides by Inonotus obliquus under submerged fermentation supplemented with lignocellulosic biomass and their antioxidant activity

The effect of lignocellulose degradation in wheat straw, rice straw, and sugarcane bagasse on the accumulation and antioxidant activity of extra- (EPS) and intracellular polysaccharides (IPS) of Inonotus obliquus under submerged fermentation were first evaluated. The wheat straw, rice straw, and sugarcane bagasse increased the EPS accumulation by 91.4, 78.6, and 74.3 % compared with control, respectively. The EPS and IPS extracts from the three lignocellulose media had significantly higher hydroxyl radical- and 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity than those from the control medium. Of the three materials, wheat straw was the most effective lignocellulose in enhancing the mycelia growth, accumulation and antioxidant activity of I. obliquus polysaccharides (PS). The carbohydrate and protein content, as well as the monosaccharide compositions of the EPS and IPS extracts, were correlated with sugar compositions and dynamic contents during fermentation of individual lignocellulosic materials. The enhanced accumulation of bioactive PS of cultured I. obliquus supplemented with rice straw, wheat straw, and bagasse was evident.     

Effect of steam explosion on biodegradation of lignin in wheat straw

The effect of steam explosion pretreatment on biodegradation of lignin in wheat straw was studied in this paper. Through experiments and analysis, 0.8MPa operation pressure and 1:20 wheat straw to water ratio are optimum for destroying lignin and the maximum of lignin loss rate is 19.94%. After steam explosion pretreatment, the wheat straw was retted by Trametes versicolor for 40 days. Biodegradation rate of lignin was tested and the maximum of 55.40% lignin loss rate was found on day 30. During the whole process of both steam explosion pretreatment and biodegradation, 75.34% lignin was degraded, without steam explosion the biodegradation of raw material the degradation rate of lignin was 31.23% only. FT-IR spectroscopy, TGA and SEM were used for further validating the results of biodegradation.     

微生物处理对秸秆结构的影响

使用红外光谱、X射线衍射及扫描电镜对4种经固态培养的微生物降解稻草秸秆的效果进行了研究,并与原稻草粉进行结构对比分析。结果表明：经过高碳低氮培养,秸秆木质素均有明显降解,其中黄孢原毛平革菌达到37.17%。扫描电镜发现秸秆的表面形态和内部结构发生了明显改变,红外光谱中芳香环与醚键吸收明显减少,X射线衍射得到的结晶度最大下降了5.60%。     

Evaluation of support materials for the surface immobilization and decoloration of amaranth by Trametes versicolor

The ability of Trametes versicolor ATCC 20869 to colonize several natural and synthetic materials (wheat straw, jute, hemp, maple woodchips, and nylon and polyethylene teraphthalate fibers) and to subsequently decolorize amaranth was evaluated. Jute was found to be the best support material as T. versicolor grew well on it without color leaching from the support and without loss of the jute's integrity over a 4 week period. The fungus immobilized on jute, straw and hemp decolorized amaranth (50 mg l(-1)) at a rate of about 5 mg l(-1) h(-1) without glucose being added. When 1 g l(-1) glucose was added, the dye was degraded more quickly (about 8 mg l(-1) h(-1)). Decoloration did not occur in a suspension culture without glucose. As the number of decoloration cycles increased, the rate of decoloration decreased. This rate was restored to its original level after the biomass was incubated in fresh growth medium for 5 days. With all immobilization supports, the toxicity of the medium before and after decoloration was the same or lower.     

Purification and characterisation of a calcium-independent lectin (PjLec) from the haemolymph of the shrimp Penaeus japonicus

A natural lectin (nominated PjLec) was isolated from haemolymph of the shrimp Penaeus japonicus by affinity chromatography with fetuin-Sepharose. The result of SDS-PAGE showed that the purified PjLec protein consisted of 37kDa subunits. The native PjLec behaved as a 452kDa protein in gel filtration chromatography. Those data suggest that PjLec is composed of 12 subunits of similar molecular weight. PjLec has a broad spectrum of bacterial-agglutination activities against both Gram-positive and Gram-negative bacteria, including two Vibrio species and two other strains pathogenic for shrimp. In addition, PjLec could agglutinate all the vertebrate erythrocytes tested, and the haemagglutination was calcium-independent. The haemagglutination of PjLec was inhibited by ManNAc, Neu5A and lipopolysaccharide. Bovine submaxillary mucin, which contains mainly Neu5A, was the most potent inhibitor of PjLec (MIC of 0.0006mgml(-1)). The haemagglutination activity of PjLec was stable between pH 6 and pH 8, and was temperature-dependent. Our results suggested that PjLec may be an important humoral defence factor against bacterial infection in P. japonicus.     

Pollution control of swine manure and straw by conversion to Chaetomium cellulolyticum SCP feed

Swine manure has a very high pollution potential and obnoxious odor. Large farms particularly are confronted with a manure disposal problem since environmentally acceptable solutions are now required by government regulations. Swine manure was found to be a good source of supplementary nutrients to ferment wheat straw into single-cell protein (SCP) with Chaetomium cellulolyticum when 0.13g (NH₄)₂SO₄/g solid was used as an additional source of N. In batch fermentations, inhibitory effects, possibly due to soluble released from the straw during alkali or acid pretreatment, were overcome by starting the fermentation at about pH 7.0 and then reducing it to 5.0 during growth. An overall protein productivity of up to 66 mg/L h was obtained from a slurry mixture of 1% w/v solids of manure and straw. This compares favorably with 99 mg/L h when manure was fermented with glucose instead of straw as the main carbon source. A high protein productivity of 200 mg/L h was obtained from a slurry mixture containing anaerobically prefermented swine manure liquor and 1.5% w/v solids from straw. The final products of the manure and straw fermentations contained 25–30% DW crude protein and 6–20% DW cellulose and the materials were free of the original obnoxious odor and undesirable microbial contamination.     

Degradation of myelin proteins by cathepsin B and inhibition by E-64 analogue

Purified human brain myelin was isolated, heat-treated to inactivate the endogenous proteolytic activity and incubated with cathepsin B purified from rat liver, at pH 6.0. Incubation resulted in a marked reduction of myelin basic protein (BP) and partial breakdown of proteolipid protein or Wolfgram protein. Degradation of myelin proteins was inhibited by E-64 analogue (E-64-a). E-64 is a specific thiol protease inhibitor isolated from a solid culture of Aspergillus japonicus. The present study suggests that cathepsin B may play some role in demyelination.