• Title/Summary/Keyword: Cellobiohydrolases

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Description of Cellobiohydrolases Ce16A and Ce17A from Trichoderma reesei Using Langmuir-type Models

  • Kim, Dong-Won;Hong, Young-Gwan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.2
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    • pp.89-94
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    • 2001
  • The binding of cellobiohydrolases to cullulose is a crucial initial step in cellulose hydrolysis. In the search for a detailed understanding of the function of cellobiohydrolases, much information concerning how the enzymes and their constituent catalytic and cellulose-binding changes during hydrolysis is still needed. The adsorption of purified two cellobiohydrolases (Ce17A and Ce16A) from Trichoderma reesei cellulase to microcrystalline cellulose has been studied. Cellobiohydrolase II (Ce16A) does not affect the adsorption of cellobiohydrolase I (Ce17A) significantly, and there are specific binding sites for both Ce17A and Ce16A. The adsorption affinity and tightness of the cullulase binding domain (CBD) for Ce17A are larger than those of the CBD for Ce16A. The CBD for Ce17A binds more rapidly and tightly to Avicel than the CBD for Ce16A. The decrease in adsorption observed when the two cellobihydrolases are studied together would appear to be the result of competition for binding sites on the cellulose. Ce17A competes more efficiently for binding sites than Ce16A. Competition for binding sites is the dominating factor when the two enzymes are acting together, furthermore adsorption to sites specific for Ce17A and Ce16A, also contributes to the total adsorption.

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Effect of Metal Ions on the Degradation and Adsorption of Two Cellobiohydrolases on Microcrystalline Cellulose

  • Kim, Dong Won;Jang, Yeong Heon;Kim, Chang Seok;Lee, Nam Su
    • Bulletin of the Korean Chemical Society
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    • v.22 no.7
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    • pp.716-720
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    • 2001
  • To test the metal ion effect, hydrolysis experiments for two cellobiohydrolases (CBHⅠ and CBH Ⅱ) from Trichoderma reesei have been carried out in the presence of 10 mM metal ions, such as Cu++, Mn++, Ca++, Hg++, Ba++, Pb++, and Cd++. The addition of Mn++, Ba++, and Ca++(10 mM) during the hydrolysis of Avicel PH 101 caused an increase in the total reducing sugar (TRS) for CBH Ⅰ by 142, 135, and 114 percent, respectively. Those for CBH Ⅱ increased by 177, 175, and 115 percent, respectively. The Mn++ was the most stimulatory metal ion, whereas Hg++ was the most inhibitory metal ion. The adsorption experiments were performed to investigate how the influence of Mn++ and Hg++ on the hydrolysis is related to the adsorption of cellobiohydrolases on cellulose. The increase in TRS during hydrolysis by adding Mn++ caused an increase in adsorption affinity (Kad) and tightness (ΔHa). While, the decrease of TRS during hydrolysis by adding Hg++ caused a decrease in the adsorption affinity (Kad) and tightness (ΔHa). These results indicate the changes in the tightness and affinity of adsorption by adding metal ions play a crucial role in the degradation of the microcrystalline cellulose.

Bioconversion of Lignocellulose Materials

  • Pothiraj, C.;Kanmani, P.;Balaji, P.
    • Mycobiology
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    • v.34 no.4
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    • pp.159-165
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    • 2006
  • One of the most economically viable processes for the bioconversion of many lignocellulosic waste is represented by white rot fungi. Phanerochaete chrysosporium is one of the important commercially cultivated fungi which exhibit varying abilities to utilize different lignocellulosic as growth substrate. Examination of the lignocellulolytic enzyme profiles of the two organisms Phanerochaete chrysosporium and Rhizopus stolonifer show this diversity to be reflected in qualitative variation in the major enzymatic determinants (ie cellulase, xylanase, ligninase and etc) required for substrate bioconversion. For example P. chrysosporium which is cultivated on highly lignified substrates such as wood (or) sawdust, produces two extracellular enzymes which have associated with lignin deploymerization. (Mn peroxidase and lignin peroxidase). Conversely Rhizopus stolonifer which prefers high cellulose and low lignin containg substrates produce a family of cellulolytic enzymes including at least cellobiohydrolases and ${\beta}-glucosidases$, but very low level of recognized lignin degrading enzymes.

Efficient Constitutive Expression of Cellulolytic Enzymes in Penicillium oxalicum for Improved Efficiency of Lignocellulose Degradation

  • Waghmare, Pankajkumar Ramdas;Waghmare, Pratima Pankajkumar;Gao, Liwei;Sun, Wan;Qin, Yuqi;Liu, Guodong;Qu, Yinbo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.740-746
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    • 2021
  • Efficient cellulolytic enzyme production is important for the development of lignocellulose-degrading enzyme mixtures. However, purification of cellulases from their native hosts is time- and labor-consuming. In this study, a constitutive expression system was developed in Penicillium oxalicum for the secreted production of proteins. Using a constitutive polyubiquitin gene promoter and cultivating with glucose as the sole carbon source, nine cellulolytic enzymes of different origins with relatively high purity were produced within 48 h. When supplemented to a commercial cellulase preparation, cellobiohydrolase I from P. funiculosum and cellobiohydrolase II from Talaromyces verruculosus showed remarkable enhancing effects on the hydrolysis of steam-exploded corn stover. Additionally, a synergistic effect was observed for these two cellobiohydrolases during the hydrolysis. Taken together, the constitutive expression system provides a convenient tool for the production of cellulolytic enzymes, which is expected to be useful in the development of highly efficient lignocellulose-degrading enzyme mixtures.