Enhancement of catalytic activity and thermostability of a thermostable cellobiohydrolase from Chaetomium thermophilum by site-directed mutagenesis

International Journal of Biological Macromolecules
Chao HanQunqing Wang

Abstract

Enzymatic saccharification of lignocellulosic biomass is increasingly applied in agricultural and industrial applications. Nevertheless, low performance in the extreme environment severely prevents the utilization of commercial enzyme preparations. To obtain cellobiohydrolases with improved catalytic activity and thermostability, structure-based rational design was performed based on a thermostable cellobiohydrolase CtCel6 from Chaetomium thermophilum. In the present study, four conserved and noncatalytic residue substitutions were generated via site-directed mutagenesis. Mutations were heterologously expressed in yeast Pichia pastoris, purified, and ultimately assayed for enzymatic characteristics. The mutant Y119F increased the catalytic activity 1.82-, 1.65- and 1.43-fold against β-d-glucan, phosphoric acid swollen cellulose (PASC) and carboxymethylcellulose sodium (CMC-Na), respectively. In addition, S131 W effectively enhanced the enzyme's heat resistance to elevated temperatures. The half-life (t1/2) of this mutant enzyme was increased 1.42- and 2.40-fold at 80 °C and 90 °C, respectively, compared to the wild-type. This study offers initial insight into the biological function of the conserved and noncatalytic residues of...Continue Reading

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Citations

Sep 1, 2020·Applied Microbiology and Biotechnology·Xiuyun WuLushan Wang
Mar 1, 2020·International Journal of Molecular Sciences·Francisca ContrerasMehdi D Davari
Dec 28, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jing-Jing ChenPing Zhu
Nov 25, 2020·Applied Microbiology and Biotechnology·Benjarat BunterngsookVerawat Champreda
Dec 15, 2020·International Journal of Biological Macromolecules·Kemin LvGerhard Schenk
Jan 10, 2021·International Journal of Biological Macromolecules·Zhaoxing LiuNing Hao
Feb 9, 2021·Biotechnology and Applied Biochemistry·Isabela Victorino da Silva AmattoHamilton Cabral
Jul 2, 2021·Biotechnology and Bioengineering·Subrata PramanikMehdi D Davari
Aug 10, 2021·International Journal of Biological Macromolecules·Anica DadwalTulasi Satyanarayana

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