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A novel Application Potential of GH6 Cellobiohydrolase Ctcel6 from Thermophilic Chaetomium Thermophilum for Gene Cloning, Heterologous Expression and Biological Characterization.
Author(s):
1. Qinzheng Zhou: Department of Mycology, Shandong Agricultural University, Taian, Shandong 271018, China
2. Jianchao Jia: Department of Mycology, Shandong Agricultural University, Taian, Shandong 271018, China
3. Peng Ji: Department of Mycology, Shandong Agricultural University, Taian, Shandong 271018, China
4. Chao Han: Department of Mycology, Shandong Agricultural University, Taian, Shandong 271018, China
Abstract:
Chaetomium thermophilum is a thermophilic fungus expressed a series of glycoside hydrolases. Genome sequence analysis of C. thermophilium revealed that ctcel6 gene encoded a putative cellobiohydrolase which composed of 397 amino acid residues including a predicted signal peptide sequence. Ctcel6 gene was cloned, heterologously expressed in Pichia pastoris and purified by Ni2+ affinity chromatography. Sequence alignment indicated that CtCel6 enzyme belonged to glycoside hydrolase family 6 (GH6) and the molecular mass of purified recombinant enzyme CtCel6 was 42 kDa by SDS-PAGE analysis. Characterization of recombinant CtCel6 exhibited high hydrolysis activity and excellent thermostability. The optimum reaction temperature and pH was 70oC and pH 5, respectively. The bivalent metallic cations Mg2+ and Ca2+ significantly enhanced the activity of CtCel6. The specific activity of CtCel6 enzyme was 1.27 U/mg and Km value was 0.38 mM on ß-Dglucan. The substrate specificity and hydrolysis products insisted that CtCel6 was an exo-/endo-type cellobiohydrolase. The biochemical properties of recombinant CtCel6 made it potentially effective for bioconversion of biomass and had tremendous potential in industrial applications such as enzyme preparation industry and feed processing industry. 
Page(s): 355-362
Published: Journal: International Journal of Agriculture and Biology, Volume: 19, Issue: 2, Year: 2017
Keywords:
Keywords are not available for this article.
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