Expression of Trichoderma reesei cellulases CBHI and EGI in Ashbya gossypii

Research output: Contribution to journalArticle

Researchers

  • Orquidea Ribeiro
  • Marilyn Wiebe
  • Marja Ilmen
  • Lucilia Domingues
  • Merja Penttila

Research units

  • Universidade do Minho
  • VTT Technical Research Centre of Finland

Abstract

To explore the potential of Ashbya gossypii as a host for the expression of recombinant proteins and to assess whether protein secretion would be more similar to the closely related Saccharomyces cerevisiae or to other filamentous fungi, endoglucanase I (EGI) and cellobiohydrolase I (CBHI) from the fungus Trichoderma reesei were successfully expressed in A. gossypii from plasmids containing the two micron sequences from S. cerevisiae, under the S. cerevisiae PGK1 promoter. The native signal sequences of EGI and CBHI were able to direct the secretion of EGI and CBHI into the culture medium in A. gossypii. Although CBHI activity was not detected using 4-methylumbelliferyl-beta-d-lactoside as substrate, the protein was detected by Western blot using monoclonal antibodies. EGI activity was detectable, the specific activity being comparable to that produced by a similar EGI producing S. cerevisiae construct. More EGI was secreted than CBHI, or more active protein was produced. Partial characterization of CBHI and EGI expressed in A. gossypii revealed overglycosylation when compared with the native T. reesei proteins, but the glycosylation was less extensive than on cellulases expressed in S. cerevisiae.

Details

Original languageEnglish
Pages (from-to)1437-1446
Number of pages10
JournalApplied Microbiology and Biotechnology
Volume87
Issue number4
Publication statusPublished - Jul 2010
MoE publication typeA1 Journal article-refereed

    Research areas

  • Ashbya gossypii, Recombinant protein production, Trichoderma reesei endoglucanase I, Trichoderma reesei cellobiohydrolase I, Cellulases heterologous expression, HETEROLOGOUS PROTEIN-PRODUCTION, CELLULOSE-BINDING DOMAINS, SACCHAROMYCES-CEREVISIAE, CELLOBIOHYDROLASE-I, PICHIA-PASTORIS, ASPERGILLUS-NIGER, FILAMENTOUS FUNGI, ENDOGLUCANASE-I, RIBOFLAVIN PRODUCTION, GENE-EXPRESSION

ID: 9229117