Exploring improved endoglucanase expression in Saccharomyces cerevisiae strains

被引：40

作者：

du Plessis, Lisa ^{[1
]}

Rose, Shaunita H. ^{[1
]}

van Zyl, Willem H. ^{[1
]}

机构：

[1] Univ Stellenbosch, Dept Microbiol, ZA-7600 Stellenbosch, South Africa

来源：

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | 2010年 / 86卷 / 05期

基金：

新加坡国家研究基金会;

关键词：

Cellulose degradation; Bioethanol; S; cerevisiae; Endoglucanase; TRICHODERMA-REESEI; AMORPHOUS CELLULOSE; ETHANOL-PRODUCTION; YEAST; HYDROLYSIS; GENE; FERMENTATION; CELLULASES; SYNERGISM; XYLOSE;

D O I：

10.1007/s00253-009-2403-z

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The endoglucanase I and II genes (egI or Cel7B and egII or Cel5A) of Trichoderma reesei QM6a were successfully cloned and expressed in Saccharomyces cerevisiae under the transcriptional control of the yeast ENO1 promoter and terminator sequences. Random mutagenesis of the egI-bearing plasmid resulted in a twofold increase in extracellular EGI activity. Both endoglucanase genes were co-expressed with the synthetic, codon-optimised cellobiohydrolase gene (s-cbhI) from T. reesei as well as the beta-glucosidase gene (bgl1) from Saccharomycopsis fibuligera in S. cerevisiae. Extracellular endoglucanase activity was lower when co-expressed with s-cbhI or bgl1. Recombinant strains were able to hydrolyse phosphoric acid swollen cellulose, generating mainly cellotriose, cellobiose and glucose. Cellobiose accumulated, suggesting the beta-glucosidase activity to be the rate-limiting factor. As a consequence, the recombinant strains were unable to produce enough glucose for growth on amorphous cellulose. The results of this study provide insight into further optimisation of recombinantly expressed cellulase combinations for saccharification and fermentation of cellulose to ethanol.

引用

页码：1503 / 1511

页数：9

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