A novel, small endoglucanase gene, egl5, from Trichoderma reesei isolated by expression in yeast

Anu Saloheimo, Bernard Henrissat, Anna‐Marja ‐M Hoffrén, Olle Teleman, Merja Penttilä

Research output: Contribution to journalArticleScientificpeer-review

147 Citations (Scopus)

Abstract

A method is presented for the isolation of genes encoding hydrolytic enzymes without any knowledge of the corresponding proteins. cDNA made from the organism of interest is cloned into a yeast vector to construct an expression library in the yeast Saccharomyces cerevisiae. Colonies producing hydrolytic enzymes are screened by activity plate assays. In this work, we constructed a yeast expression library from the filamentous fungus Trichoderma reesel and isolated a new β‐1,4‐endoglucanase gene on plates containing β‐glucan. This gene, eg15, codes for a previously unknown small protein of 242 amino acids. Despite its small size, the protein contains two conservative domains found in Trichoderma cellulases, namely the cellulose‐binding domain (CBD) and the iinker region that connects the CBD to the catalytic core domain. Molecular modelling of the EGV CBD revealed some interesting structural differences compared to the CBD of the major celluiase CBHI from T. reesei. The catalytic core of EGV is unusually small for a ceiiulase and represents a new family of ceilulases (Family K) and of glycosyl hydrolases (Famlly 45) together with the endoglucanase B of Pseudomonas fluorescens and the endoglucanase V of Humicola insolens on the basis of hydrophobic ciuster anaiysis.

Original languageEnglish
Pages (from-to)219-228
Number of pages10
JournalMolecular Microbiology
Volume13
Issue number2
DOIs
Publication statusPublished - 1 Jan 1994
MoE publication typeA1 Journal article-refereed

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Trichoderma
Cellulase
Catalytic Domain
Yeasts
Genes
Cellulases
Pseudomonas fluorescens
Proteins
Glucans
Hydrolases
Enzymes
Saccharomyces cerevisiae
Fungi
Complementary DNA
Amino Acids

Cite this

Saloheimo, Anu ; Henrissat, Bernard ; Hoffrén, Anna‐Marja ‐M ; Teleman, Olle ; Penttilä, Merja. / A novel, small endoglucanase gene, egl5, from Trichoderma reesei isolated by expression in yeast. In: Molecular Microbiology. 1994 ; Vol. 13, No. 2. pp. 219-228.
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A novel, small endoglucanase gene, egl5, from Trichoderma reesei isolated by expression in yeast. / Saloheimo, Anu; Henrissat, Bernard; Hoffrén, Anna‐Marja ‐M; Teleman, Olle; Penttilä, Merja.

In: Molecular Microbiology, Vol. 13, No. 2, 01.01.1994, p. 219-228.

Research output: Contribution to journalArticleScientificpeer-review

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