• Title/Summary/Keyword: cellulases

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Production of Cellulases by Rhizopus stolonifer from Glucose-Containing Media Based on the Regulation of Transcriptional Regulator CRE

  • Zhang, Yingying;Tang, Bin;Du, Guocheng
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.514-523
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    • 2017
  • Carbon catabolite repression is a crucial regulation mechanism in microorganisms, but its characteristic in Rhizopus is still unclear. We extracted a carbon regulation gene, cre, that encoded a carbon catabolite repressor protein (CRE) from Rhizopus stolonifer TP-02, and studied the regulation of CRE by real-time qPCR. CRE responded to glucose in a certain range, where it could significantly regulate part of the cellulase genes (eg, bg, and cbh2) without cbh1. In the comparison of the response of cre and four cellulase genes to carboxymethylcellulose sodium and a simple carbon source (lactose), the effect of CRE was only related to the concentration of reducing sugars. By regulating the reducing sugars to range from 0.4% to 0.6%, a glucose-containing medium with lactose as the inducer could effectively induce cellulases without the repression of CRE. This regulation method could potentially reduce the cost of enzymes produced in industries and provide a possible solution to achieve the largescale synthesis of cellulases.

Comparative Study on the Effects of Combined Treatments of Lactic Acid Bacteria and Cellulases on the Cell Wall Compositions and the Digestibility of Rhodesgrass (Chloris gayana Kunth.) and Italian Ryegrass (Lolium multiflorum Lam.) Silages

  • Ridla, M.;Uchida, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.4
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    • pp.531-536
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    • 1999
  • This study was conducted to compare the effects of lactic acid bacteria (LAB) or LAB+cellulases on the cell wall compositions and the in vitro dry matter digestibility (IVDMD) of Rhodesgrass (RG) and Italian ryegrass (IRG) silages. LAB (Lactobacillus cassei) at a concentration of $10{\times}10^5\;cfu.g^{-1}$ fresh forage was added to all ensiling samples (except the untreated control) of RG and IRG. The cellulases used were Acremoniumcellulase (A), Meicelase (M) or a mixture of both (AM). Each cellulase was applied at levels of 0.005, 0.01 and 0.02 % fresh sample. The samples were incubated at 20, 30 and $40^{\circ}C$ for about 2 months of storage. LAB inoculation did not affect cell wall components or IVDMD of both the RG and IRG silages, but LAB+cellulase treatments did. Increasing the amount of cellulase addition resulted in further decreases of cell wall concentrations. This reduction more markedly occurred with cellulases A and AM than it did with cellulase M. Cell wall components losses were higher in the IRG silages than in the RG silages. LAB+cellulase treatments decreased IVDMD of the RG silages, but had no effect on the IRG silages. The different effect of LAB+cellulase treatments on cell wall degradation and IVDMD of the RG and IRG silages suggested that RG contains more structural carbohydrates, which were difficult to degrade with cellulase, than did IRG.

Cellulolytic Enzymes from Acrophialophora nainiana

  • Punnapayak, Hunsa
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2005.06a
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    • pp.245-247
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    • 2005
  • A cellulolytic fungus isolated from Agave plantation in northeastern Thailand was identified as Acrophialophora nainiana. The fungus was capable of growing at pH between 3 - 7 and 25 - 45 $^{\circ}C$, with the optimum conditions at pH 5.0 and 40 $^{\circ}C$. The wild isolate produced cellulases, comprising of exoglucanase (0.019 U/mg protein), endoglucanase (0.366 U/mg protein), and ${\beta}$-glucosidase (0.001 U/mg protein). Mutations with UV and NTG produced the UV 10-2 mutant with cellulases activities including exoglucanase (0.093 U/mg protein), endoglucanase (0.585 U/mg protein), and ${\beta}$-glucosidase (0.013 U/mg protein). Purification of the enzymes with ultrafiltration, ammonium sulfate precipitation, and ion-exchange chromatography yielded the maximal cellulase specific activities of 2.736 U/mg protein (exoglucanase), 0.235 U/mg protein (endoglucanase), and 0.008 U/mg protein (${\beta}$-glucosidase). The mutant's cellulases were the most active at pH 5.0 and 60 $^{\circ}C$. Ion-exchange chromatography revealed that A. nainiana UV 10-2 cellulases were comprised of two peaks with one peak showing the single endoglucanase activity while the other peak showed a mixture of the three enzyme activities. Production of A. nainiana UV 10-2 cellulases using banana leaf stalk as the sole carbon source gave comparable yields to that of the pure ${\alpha}$-cellulose. The enzymes were used in the simultaneous saccharification and fermentation (SSF) of plant residue (Coix aquatica) along with Kluveromyces marxianus to produce ethanol. Moreover, when the enzymes were used in the bioscouring process of fabric, the desiravle traits of textile processing including immediate water absorbency, increased in whiteness and reduction of yellowness of the treated fabric were observed.

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Cellulase from the fruiting bodies and mycelia of edible mushrooms: A review

  • Wu, Yuanzheng;Shin, Hyun-Jae
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.127-135
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    • 2016
  • Cellulose is the most abundant organic polymer constituent of the cell wall of green plants and of various forms of algae. The complexity of lignocellulosic biomass is a major challenge in industrial research. Most mushroom species that naturally grow on soil or wood possess cellulases and the corresponding enzymatic system and, potential candidates for the direct bioconversion of softwood polysaccharides into fermentable sugars. However, there have been fewer studies on mushroom cellulases than on fungi such as Trichoderma spp., exploit the full potential of mushroom cellulases. This review will focus on the current status ofmushroom cellulase research and applications and will provide insight into promising future prospects.

Isolation, Identification, and Expression of Microbial Cellulases from the Gut of Odontotermes formosanus

  • Duan, Jiwei;Liu, Jun;Ma, Xueling;Zhang, Yue;Wang, Xiaohua;Zhao, Kai
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.122-129
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    • 2017
  • Termites are destructive to agriculture, forestry, and buildings, but they can also promote agro-ecosystem balance through the degradation of lignocellulose. Termite-triggered cellulose digestion may be clarified through microbial metabolism of cellulose products. In the present study, we characterized the activities of cellulase and its three components synthesized by the cellulase-producing fungal strain HDZK-BYTF620 isolated from the gut of Odontotermes formosanus. The protein components of cellulases were synthesized by strain HDZK-BYTF620, which were isolated and characterized using polyacrylamide gel electrophoresis, and the expression of the cellulases was studied at the proteome level.

Comparative Study on the Effects of Combined Treatments of Lactic Acid Bacteria and Cellulases on the Fermentation Characteristic and Chemical Composition of Rhodesgrass (Chloris gayana Kunth.) and Italian Ryegrass (Lolium multiflorum Lam.) Silages

  • Ridla, M.;Uchida, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.4
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    • pp.525-530
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    • 1999
  • Prior to ensiling Rhodesgrass (RG) and Italian ryegrass (lRG) were treated with lactic acid bacteria (LAB) or with LAB+cellulases to compare their fermentation characteristics and chemical compositions. LAB (Lactobacillus casei) was added to all ensiling materials (except the untreated control) of RG and IRG at a concentration of $1.0{\times}10^5\;cfu.g^{-1}$ fresh forage. The enzymes used were Acremoniumcellulase (A), Meicelase (M) or a mixture of both (AM). Each enzyme was applied at levels of 0.005, 0.01 and 0.02 % of fresh forage. The silages with each treatment were incubated at 20, 30 and $40^{\circ}C$ and stored for about 2 months. While no marked differences were found between the RG and IRG silages with various treatments on dry matter (DM), volatile basic nitrogen (VBN) and water soluble carbohydrate (WSC) contents, there were significant differences in pH value, and lactic acid and butyric acid contents. LAB inoculation did not affect the fermentation characteristics of either the RG or IRG silages. The combined treatments of LAB+cellulases improved the fermentation quality of both the RG and IRG silages as evidenced by the decrease in pH value and increase in lactic acid content. Increasing the amount of added cellulase resulted in a decrease in pH value and an increase in lactic acid content in both the RG and IRG silages. Cellulases A and AM had a greater effect than cellulase M on the fermentation quality of the RG and IRG silages. Incubation temperatures of 30 and $40^{\circ}C$ appeared to be more appropriate environments for stimulating good fermentation than $20^{\circ}C$.

Investigation of the Hydrolysis of Polysaccharides by Crude Cellulases prepared from Several Species of Fungi (몇 종류의 곰팡이에서 분리되는 Crude Cellulase의 다당류 분해능력의 조사)

  • 김은수;김영민;이인규;최태주
    • Korean Journal of Microbiology
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    • v.13 no.3
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    • pp.85-90
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    • 1975
  • Crude cellulases freshly prepared from cultures of Aspergillus niger, Prnicillum motatum, Trichoderma vride 16273 and Trichoderma viride 16374 were assayed on 4 different substrates including Na-CMC, cellulose powder, starch and sucrose. Enzyme prepared from A. niger contained highly active hydrolytic enzymes of the 4 substrates assayed. P. notatum [yielded relatively lower amount of cellulase but the extracts were also highly reactive on starch and sucrose. Trichoderma viride 16274 yielded very little cellulase and invertase, but the extracts showed a high degree of amylase activity. Trichoderma viride 16374, however, yielded collulase comparable to that of Penicillium notatum, but lower activities of amylase and invertase were seen. Commercial cellulases prepared from Penicillium notatum (cellulase[K]) and Trichoderma viride(cellulase[J]) indicated enzyme activities closely parallel to the crude enzymes freshly prepared from fungus cultures. The optimum pH's of cellulolytic activities of cellulase[K] and cellulase[J] were 4.0 and 5.0 respectively. The optimum temperatures of the cellulolytic activities of cellulase[K] and cellualse[J] were 4.0 and 5.0 respectively. The optimum temperatures of the cellulolytic activities of cellulase [K] and cellulase [J] were $60{\circ}C$ and $50{\circ}C$ respectively. Assuming the average molecular weight of Na-CMC is about 115,000, the Km values of cellulase [K] and cellulase[J] were found to be $3.3{\times}10^{-5}/nM$ and $3.3{\times}10^{-4}/nM$ respectively.

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Comparative analysis of silage fermentation and in vitro digestibility of tropical grass prepared with Acremonium and Tricoderma species producing cellulases

  • Khota, Waroon;Pholsen, Suradej;Higgs, David;Cai, Yimin
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.12
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    • pp.1913-1922
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    • 2018
  • Objective: To find out ways of improving fermentation quality of silage, the comparative analysis of fermentation characteristics and in vitro digestibility of tropical grasses silage applied with cellulases produced from Acremonium or Tricoderma species were studied in Thailand. Methods: Fresh and wilted Guinea grass and Napier grass silages were prepared with cellulases from Acremonium (AC) or Trichoderma (TC) at 0.0025%, 0.005%, and 0.01% on a fresh matter (FM), and their fermentation quality, chemical composition and in vitro digestibility were analyzed. Results: All silages of fresh Napier grass were good quality with lower pH, butyric acid, and ammonia nitrogen, but higher lactic acid content than wilted Napier grass and Guinea grass silage. Silages treated with AC 0.01% had the best result in terms of fermentation quality. They also had higher in vitro dry matter digestibility and in vitro organic matter digestibility at 6 and 48 h after incubation than other silages. Silages treated with lower levels at 0.005% or 0.0025% of AC and all levels of TC did not improve silage fermentation. Conclusion: The AC could improve silage fermentation and in vitro degradation of Guinea grass and Napier grass silages, and the suitable addition ration is 0.01% (73.5 U) of FM for tropical silage preparation.

Chitosanolytic Characteristics of Cellulases from Trichoderma viride and Trichoderma reesei (Trichoderma viride 및 Trichoderma reesei 유래 Cellulase의 키토산 분해 특성)

  • Hong, Sang-Pill;Kim, Dong-Soo
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.245-252
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    • 1998
  • The chitosanolytic capabilities of cellulases, glucosidases, proteases and commercial enzymes were evaluated, and effective chitosanolytic cellulases from T. viride, T. reesei and Celluclast, a commercial enzyme from T. reesei were characterized. The reaction of cellulase from T. viride, T. reesei and Celluclast was optimal at pH 5. 0 and $45{\sim}55^{\circ}C$. Max. chitosanolytic activities of cellulases from both T. viride and T. reesei were observed at the enzyme/chitosan ratio=0.1 and chitosan concentration=3.0%. For the possible application of commercial Celluclast to chitosan oligosaccharides production, 3%(w/v) chitosan was reacted with 1%(v/v) Celluclast at pH 5.0 and $55^{\circ}C$. The apparent viscosity decreased by 98% within 30 minutes reaction and Max. contents of 50% EtOH solubles were 70% at 15 hrs reaction. Total reducing sugars were also increased with reaction time and maintained approx. 13.5% after 2hrs reaction. In 15 hrs treated chitosan hydrolyzates, various kinds of chitosan oligosaccharides were produced and contents of chitosan hexamer, known for its antitumor activities, were about 8.0%, about 4 times higher values compared with acid hydrolysis method. The results suggested that chitosan oligosaccharides could be produced with low-cost cellulases from T. reesei.

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Some characters of bacterial cellulases in goats' rumen elucidated by metagenomic DNA analysis and the role of fibronectin 3 module for endoglucanase function

  • Nguyen, Khanh Hoang Viet;Dao, Trong Khoa;Nguyen, Hong Duong;Nguyen, Khanh Hai;Nguyen, Thi Quy;Nguyen, Thuy Tien;Nguyen, Thi Mai Phuong;Truong, Nam Hai;Do, Thi Huyen
    • Animal Bioscience
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    • v.34 no.5
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    • pp.867-879
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    • 2021
  • Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.