cDNA Cloning of a Trichoderma Reesei Cellulase and Demonstration of Endoglucanase Activity by Expression in Yeast

Eur J Biochem. 1997 Oct 15;249(2):584-91. doi: 10.1111/j.1432-1033.1997.00584.x.

Abstract

A Trichoderma reesei cDNA encoding a previously unknown protein with a C-terminal cellulose-binding domain was obtained by complementation screening of a T. reesei cDNA library in a sec1 yeast mutant impaired in protein secretion. The T. reesei protein shows amino acid similarity over its entire length to the Agaricus bisporus cellulose-induced protein CEL1 whose function is not known. These two proteins form a new glycosyl hydrolase family, number 61. Expression of the T. reesei cDNA in yeast showed that it encoded a protein with endoglucanase activity and thus the protein was named EGIV and the corresponding gene egl4. Polyclonal antibodies were prepared against EGIV produced in Escherichia coli and detected a 56-kDa protein in the T. reesei culture supernatant. Northern hybridisation revealed that T. reesei egl4 is regulated in the same manner as other cellulase genes of this fungus.

Publication types

  • Comparative Study

MeSH terms

  • Agaricus / enzymology
  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • Cellulase / chemistry
  • Cellulase / genetics
  • Cellulase / metabolism*
  • Cellulose 1,4-beta-Cellobiosidase
  • Cloning, Molecular
  • DNA, Complementary
  • Fungal Proteins*
  • Gene Library
  • Genes, Fungal
  • Models, Structural
  • Molecular Sequence Data
  • Protein Conformation
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Trichoderma / enzymology*
  • Trichoderma / genetics

Substances

  • DNA, Complementary
  • Fungal Proteins
  • Recombinant Proteins
  • Cellulase
  • EGL4 protein, Hypocrea jecorina
  • Cellulose 1,4-beta-Cellobiosidase

Associated data

  • GENBANK/Y11113