Cloning of a Cellobiohydrolase Gene (cbh1) from Aspergillus niger and Heterogenous Expression in Pichia pastoris

Abstract:

A gene encoding a cellobiohydrolase (CBH) was isolated from Aspergillus niger-NL-1 and designated as cbh1. The cbh1 gene contains 1,515 nucleotides with three introns and encodes a 452-amino acid protein with a molecular weight of approximately 60 kDa. The amino acid sequence encoded by cbh1 shows high homology with the sequence of glycoside hydrolase fimily 7. The cellobiohydrolase (cbh1) gene was succussfully expressed in Pichia pastoris KM71H. The recombinant CBHⅠshowed an optimal working condition at 60 °C, pH 4.0 with K_m and V_max toward CMC-Na of 13.81 mM and 0.269 μmol/min, respectively. The enzyme retained more than 80 % of its initial activity after 2 h of incubation at 90 °C and was stable in pH range 1.0～10.0. Because of its moderately stable at high temperature and stability through wide range of pH, this enzyme has potential in various industrial applications.

• Periodical: Advanced Materials Research (Volumes 347-353)
• Pages: 2443-2447
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.347-353.2443
• Online since: October 2011
• Authors: Guo Qing Li, Chang Sheng Chai, Song Fan, Lin Guo Zhao
• Keywords: Aspergillus niger, Cellobiohydrolase I, Cloning, Expression, Pichia pastoris

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