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Cloning, Characterization, and Expression in Saccharomyces Cerevisiae of Endoglucanase I from Trichoderma Reesei

Bio/Technology volume 5, pages 60–64 (1987)Cite this article

Abstract

We report the cloning, partial characterization, and expression in yeast of the endoglucanase I (EGI) gene from Trichoderma reesei. DNA sequencing revealed significant homology at the amino acid level between EGI and exocellobiohydrolase I (CBHI), but there are differences in codon utilization at homologous amino acids and in the intron/exon structure. These possibly reflect a mechanism for preventing recombination between closely related genes of the cellulase family. The coding sequence for the mature protein with its signal peptide was inserted into an expression plasmid containing yeast transcription control sequences. Yeast colonies transformed with this plasmid secrete enzymatically active hyperglycosylated EGI to the culture medium. This novel glycosylation appears to render the enzyme more resistant to thermal inactivation.

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Author notes

  1. Michael A. Innis: Corresponding author

Authors and Affiliations

  1. Department of Microbial Genetics, Cetus Corporation, 1400 Fifty-Third Street, Emeryville, CA, 94608

    Janelle N. Van Arsdell, Shirley Kwok, David H. Gelfand & Michael A. Innis

  2. Department of Molecular Biology, Cetus Corporation, 1400 Fifty-Third Street, Emeryville, CA, 94608

    Vicki L. Schweickart & Martha B. Ladner

Authors
  1. Janelle N. Van Arsdell
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  2. Shirley Kwok
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  3. Vicki L. Schweickart
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  4. Martha B. Ladner
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  5. David H. Gelfand
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  6. Michael A. Innis
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Van Arsdell, J., Kwok, S., Schweickart, V. et al. Cloning, Characterization, and Expression in Saccharomyces Cerevisiae of Endoglucanase I from Trichoderma Reesei. Nat Biotechnol 5, 60–64 (1987). https://doi.org/10.1038/nbt0187-60

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