期刊
BIORESOURCE TECHNOLOGY
卷 319, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2020.124119
关键词
Product inhibition; Cellulase; Lignocellulose; Trichoderma reesei
资金
- National Key R&D Program of China [2018YFA0900500]
- National Natural Science Foundation of China [31470201, 31741003, 31902089]
- Shanghai Agriculture Applied Technology Development Program, China [2018-02-08-00-12-F01555]
Introducing product-activated cellulase genes significantly increases enzyme activity and saccharification efficiency. Improving thermostability of cellulases was achieved through mutants and enhanced enzyme activity. Introducing product-activated cellulase genes is an effective approach to alleviate product inhibition of cellulases.
Product inhibition of cellulase is a challenging issue in industrial processes. Here, we introduced a product activated mushroom cellulase, PaCel3A from Polyporus arcularius, into Trichoderma reesei. The filter paper activity, carboxymethyl cellulase activity, and saccharification efficiency (substrate: pretreated rice straw, PRS) of transformants increased significantly with this enzyme (by 18.4-26.8%, 13.8-22.8%, and 17.0%, respectively). A mutant of PaCel3A, PaCel3AM, obtained based on B-factor analysis, saturated mutagenesis, and residual activity assay, showed improved thermostability. The PRS saccharification efficiency using the cellulase complex from T. reesei transformants overexpressing pacel3am increased by 56.4%-63.0%. In addition, the T. reesei cellulase complex obtained by adding the purified recombinant PaCel3AM from T. reesei (rCel3aM-tr) to hydrolyze PRS resulted in increased reducing sugar yields at all sampling points, outperforming the cellulase complexes without rCel3aM-tr. These results suggest that introducing product-activated cellulase genes is a simple and feasible method to alleviate the product inhibition of cellulase.
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