• Title/Summary/Keyword: cellulases

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Efficient Isolation and Characterization of a Cellulase Hyperproducing Mutant Strain of Trichoderma reesei

  • Zou, Zongsheng;Zhao, Yunying;Zhang, Tingzhou;Xu, Jiaxing;He, Aiyong;Deng, Yu
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1473-1481
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    • 2018
  • A cellulase hyperproducing mutant strain, JNDY-13, was obtained using the ARTP mutation system and with Trichoderma reesei RUT-C30 as the parent strain. Whole-genome sequencing of JNDY-13 confirmed that 105 of the 653 SNPs were point mutations, 336 mutations were deletions and 165 were insertions. Moreover, 99 mutations were insertions and duplications. Among all the mutations, the one that occurred in the galactokinase gene might be related to the production of cellulases in T. reesei JNDY-13. Moreover, the up-regulation of cellulase and hemicellulase genes in JNDY-13 might contribute to higher cellulases production. Under optimal conditions, the highest cellulase activity by batch fermentation reached 4.35 U/ml, and the highest activity of fed-batch fermentation achieved was 5.40 U/ml.

Selection of Multienzyme Complex-Producing Bacteria Under Aerobic Cultivation

  • Pason Patthra;Chon Gil-Hyong;Ratanakhanokchai Khanok;Kyu Khin Lay;Jhee Ok-Hwa;Kang Ju-Seop;Kim Won-Ho;Choi Kyung-Min;Park Gil-Soon;Lee Jin-Sang;Park Hyun;Rho Min-Suk;Lee Yun-Sik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1269-1275
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    • 2006
  • The selection of multienzyme complex-producing bacteria under aerobic condition was conducted for improving the degradation of lignocellulosic substances. The criteria for selection were cellulase and xylanase enzyme production, the presence of cellulose-binding domains and/or xylan-binding domains in enzymes to bind to insoluble substances, the adhesion of bacterial cells to insoluble substances, and the production of multiple cellulases and xylanases in a form of a high molecular weight complex. Among the six Bacillus strains, isolated from various sources and deposited in our laboratory, Paenibacillus curdlanolyticus B-6 strain was the best producer of cellulase and xylanase enzymes, which have both cellulose-binding factors (CBFs) and xylan-binding factors (XBFs). Moreover, multiple carboxymethyl cellulases (CMCases) and xylanases were produced by the strain B-6. The zymograms analysis showed at least 9 types of xylanases and 6 types of CMCases associated in a protein band of xylanase and cellulase with high molecular weight. These cells also enabled to adhere to both avicel and insoluble xylan, which were analyzed by scanning electron microscopy. The results indicated that the strain B-6 produced the multienzyme complex, which may be cellulosome or xylanosome. Thus, P. curdlanolyticus B-6 was selected to study the role and interaction between the enzymes and their substrates and the cooperation of multiple enzymes to enhance the hydrolysis due to the complex structure for efficient cellulases and xylanases degradation of insoluble polysaccharides.

Immobilization of Cellulases from Fomitopsis pinicola and Their Changes of Enzymatic Characteristics (흡착법에 의한 Fomitopsis pinicola 유래 cellulase의 고정화와 그에 따른 효소특성 변화)

  • Shin, Keum;Kim, Tae-Jong;Kim, Young-Kyoon;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.3
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    • pp.251-261
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    • 2010
  • Cellulase from Formiptosis pinicola KMJ812 is an efficient cellulose degradation enzyme complex, especially with a high ${\beta}$-glucosidase activity. In this study, the change in enzymatic characteristics by immobilization and the reduction of immobilized enzyme activity by repeated usages were evaluated using cellulases from F. pinicola KMJ812. Among tested four resins, Duolite A568 resin had the best enzyme activity yield with 61.7% cellulase activity and 64.4% ${\beta}$- glucosidase activity during the cellulase immobilization. The best reaction temperature was $55^{\circ}C$ for both cellulase and ${\beta}$-glucosidase activities which were higher than the unimmobilized soluble cellulases. The best reaction pH was 4.0 for cellulase activity which was a little more basic than a soluble form and 4.5 for ${\beta}$-glucosidase activity. The immobilized cellulase activity was remained 98% of the beginning activity after 72 h incubation at $50^{\circ}C$ and 50% of the beginning activity after eight times usage at $50^{\circ}C$.

Effects of Combined Treatment of Lactic Acid Bacteria and Cell Wall Degrading Enzymes on Fermentation and Composition of Rhodesgrass (Chloris gayana Kunth.) Silage

  • Ridla, M.;Uchida, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.5
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    • pp.522-529
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    • 1998
  • This experiment was conducted to study the effects of lactic acid bacteria (LAB) inoculation either alone or in combination with cell wall degrading enzymes on the fermentation characteristics and chemical compositions of Rhodesgrass silage. Over to 1 kg of fresh Rhodesgrass sample a treatment of inoculant LAB with or without addition of an enzyme of Acremoniumcellulase (A) or Meicelase (M) or a mixture of both enzymes (AM) was applied. The treatments were control untreated, LAB-treated (application rate $1.0{\times}10^5cfu/g$ fresh sample), LAB+A 0.005%, LAB+A 0.01%, LAB+A 0.02%, LAB+M 0.005%, LAB+M 0.01%, LAB+M 0.02 %, LAB+AM 0.005%, LAB+AM 0.01%, and LAB+AM 0.02%. The sample was ensiled into 2-L vinyl bottle silo, with 9 silages of each treatment were made. Three silages of each treatment were incubated at 20, 30 and $40^{\circ}C$ for 2-months of storage period. All silages were well preserved with their fermentation quality has low pH values (3.91-4.26) and high lactic acid concentrations (4.11-9.89 %DM). No differences were found in fermentation quality and chemical composition of the control untreated silage as compared to the LAB-treated silage. Combined treatment of LAB+cellulases improved the fermentation quality of silages measured in terms of lower (p < 0.01) pH values and higher (p < 0.05) lactic concentrations than those of LAB-treated silages. Increasing amount of cellulase addition resulted in decrease (p < 0.05) of pH value and increase (p < 0.05) of lactic acid concentration. LAB + cellulase treatments (all cellulase types) reduced (p < 0.01) NDF, ADF and in vitro dry matter digestibility of silages compared with the control untreated silages. The fermentation quality and the rate of cell wall reduction were higher (p < 0.01) in the silages treated with LAB + cellulase A than in the silages treated with either LAB+cellulase M or LAB + cellulase AM. Incubation temperature of $40^{\circ}C$ was likely to be more appropriate environment for stimulating the fermentation of Rhodesgrass silages than those of 20 and $30^{\circ}C$.

Production of Cellulolytic Enzymes by Trichoderma harzianum FJ1 in Solid State Fermentation. (Trichoderma harzianum FJ1의 고체상태배양에 의한 섬유소분해효소의 생산)

  • 유승수;김경철;김성준
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.257-263
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    • 2003
  • The cellulases production in solid state fermentation (SSF) of Trichoderma harzianum FJ1 with high cellulases productivity using cellulosic wastes was investigated. Physical and chemical conditions of the fermentation, such as moisture content, initial pH, and composition of mixed substrate (wine waste, rice straw, and soybean flour) on FPase (Filter paper activity) production were examined. The enzyme production was optimized in the conditions of moisture content of 70%, pH 5.0, 3$0^{\circ}C$, and 1:1:1 composition of mixed substrate containing wine waste, rice straw, and soybean flour. The highest activities of FPA, CMCase, Xylanase, $\beta$-glucosidase, and Avicelase in the optimized culture conditions were 15.2, 69.1, 83.9, 29.2, and 4.2 unit/g-SDW in 5 day cultivation, respectively. Economical and efficient production of cellulolytic enzymes by T harzianum FJ1 using cellulosic wastes in solid state fermentation will contribute to the biological saccharification of cellulosic wastes with enormous potential resource value in future.

Physiological Diversity between Morphological Phenotypes of Botrytis cinerea (잿빛곰팡이병균(Botrytis cinerea) 형태형 간의 생리적 다양성)

  • Kim, Byung-Sup;Park, Eun-Woo;Roh, Seong-Hwan;Cho, Kwang-Yun
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.320-329
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    • 1997
  • Botrytis cinerea isolates obtained from infected plants of cucumber, tomato, and strawberry were divided into three groups (sporing, sclerotial, and mycelial types). Of which sclerotial types were the major group. There were no correlations between morphological phenotypes and responses to benzimidazole and dicarboximide fungicides. External structure of conidia of three phenotypes by scanning electron microscope was the same with verrucose surface. Mycelial type was the most virulent on fruits of eggplants. Comparative tests were carried out to examine correlations between the virulence and production of fungal enzymes such as phenol oxidases, pectin methyl esterases (PME), amylases, cellulases, ureases, ${\beta}-glucosidases$, and proteinases. There was no correlation among the phenotypes in production of phenol oxidases and ${\beta}-glucosidases$. However, there were significantly different from each other in PME, amylase, cellulase, urease, and protease activity.

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Biopolishing of Cotton Fabric using Crude Cellulases from Acanthophysium sp. KMF001 (목재부후균, Acanthophysium sp. KMF001, 유래 섬유소분해효소를 이용한 섬유 개량)

  • Shin, Keum;Yoon, Sae-Min;Kim, Juhea;Kim, Young-Kyoon;Kim, Tae-Jong;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.44 no.3
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    • pp.381-388
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    • 2016
  • Biopolishing using cellulases was introduced in the production of cotton fabric in order to improve the quality of fabric environmental friendly and is commonly used in the textile industry. In this study, the application of a crude cellulase from Acanthophysium sp. KMF001, which was excellent for the saccharification of cellulose, on biopolishing was evaluated. The optimum treatment biopolishing condition was at $50^{\circ}C$ and pH 4.5 for 60 minutes with 10% crude cellulase of fabric weight. After the optimized biopolishing, the crude cellulase of Acanthophysium sp. KMF001 reduced the tensile strength of the tested cotton fabric less than a commercial cellulase. The appearance of the cotton fabric after the treatment of the crude cellulase of Acanthophysium sp. KMF001 was similar to the fabric after a commercial cellulase treatment. All these results support that the crude cellulase of Acanthophysium sp. KMF001 was a good biopolishing cellulase.