Koivula, A., Ruohonen, L., Wohlfahrt, G., Reinikainen, T., Teeri, T.T., Piens, K., Claeyssens, M., Weber, B., Vasella, A., Becker, D., Sinnott, M.L., Zou, J., Kleywegt, G., Szardenings, M., Stahlberg, J. and Jones, T.A. (2002) The Active Site of the Cellobiohydrolase Cel6A from Trichoderma reesei: The Roles of the Aspartic Acids D221 and D175. Journal of the American Chemical Society, 124, 10015-10024. https://doi.org/10.1021/ja012659q

has been cited by the following article:

• TITLE: Study on Cellulase Gene Expressed in Pichia pastoris and Analyses of Its Biochemical Characters

  AUTHORS: Min Lu, Juan Chang, Ping Wang, Qingqiang Yin, Xiaowei Dang, Tianzeng Gao, Fushan Lu

  KEYWORDS: Cellulase Gene, Cloning and Expression, Biochemical Characters, Pichia pastoris

  JOURNAL NAME: Journal of Materials Science and Chemical Engineering, Vol.6 No.7, July 4, 2018

  ABSTRACT: Objectives: In order to increase cellulose degradation, cellulase was expressed in this study. Literature Review: Cellulose is the most abundant organic carbon source on Earth; its enzymatic hydrolysis will be very useful for bioenergy production and resource recycling. Methods: Cellobiohydrlase I (CBH I) gene was amplified from genomic DNA of Trichoderma koningii and inserted into pGAPZα A plasmid to construct the vector of pGAPZαA-CBH I. It was linearized and transformed into Pichia pastoris by electroporation. The recombinant Pichia pastoris was selected and incubated with YPD medium for cellulase secretion. Results: The result showed that CMCase and avicelase activity in the supernatant was 1.1798 U/mL and 0.1276 U/mL, the molecular weight of the expressed protein was 53 kDa determined with SDS-PAGE analyses, and the optimal temperature and pH of the expressed cellulase were 45?C - 50?C and 4.5 - 5.0, respectively. Conclusion: Cellulase gene from T. koningii has been successfully cloned and expressed in Pichia pastoris.