High Frequency One-step Gene Replacement in Trichoderma Reesei. II. Effects of Deletions of Individual Cellulase Genes

Overview

Journal Mol Gen Genet

Specialties Genetics
Molecular Biology

Date 1993 Dec 1

PMID 8264527

Citations 34

Authors

P L Suominen

A L Mantyla

T Karhunen

S Hakola

H Nevalainen

Affiliations

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Abstract

Four cellulase genes of Trichoderma reesei, cbh1, cbh2, egl1 and egl2, have been replaced by the amdS marker gene. When linear DNA fragments and flanking regions of the corresponding cellulase locus of more than 1 kb were used, the replacement frequencies were high, ranging from 32 to 52%. Deletion of the major cellobiohydrolase 1 gene led to a 2-fold increase in the production of cellobiohydrolase II; however, replacement of the cbh2 gene did not affect the final cellulase levels and deletion of egl1 or egl2 slightly increased production of both cellobiohydrolases. Based on our results, endoglucanase II accounts for most of the endoglucanase activity produced by the hypercellulolytic host strain. Furthermore, loss of the egl2 gene causes a significant drop in the filter paper-hydrolysing activity, indicating that endoglucanase II has an important role in the total hydrolysis of cellulose.

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