Heterologous co-production of Thermobifida fusca Cel9A with other cellulases in Saccharomyces cerevisiae

Niel Van Wyk, Riaan Den Haan, Willem H. Van Zyl

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The processive endoglucanase Cel9A of the moderately thermophilic actinomycete Thermobifida fusca was functionally produced in Saccharomyces cerevisiae. Recombinant Cel9A displayed activity on both soluble (carboxymethylcellulose) and insoluble (Avicel) cellulose substrates confirming its processive endoglucanase activity. High-performance anionic exchange chromatography analyses of soluble sugars released from Avicel revealed a cellobiose/glucose ratio of 2.5∈±∈0.1. Growth by the recombinant strain on amorphous cellulose was possible due to the sufficient amount of glucose cleaved from the cellulose chain. This is the first confirmed report of S. cerevisiae growing on a cellulosic substrate as sole carbohydrate source while only expressing one recombinant gene. To improve the cellulolytic capability of S. cerevisiae and to investigate the level of synergy among cellulases produced by a recombinant host, the cel9A gene was co-expressed with four cellulase-coding genes of Trichoderma reesei: two endoglucanases cel5A (egII) and cel7B (egI), and two cellobiohydrolases cel6A (cbhII) and cel7A (cbhI). Synergy, especially between the Cel9A and the two cellobiohydrolases, resulted in a higher cellulolytic capability of the recombinant host.

Original languageEnglish
Pages (from-to)1813-1820
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume87
Issue number5
DOIs
Publication statusPublished - Aug 2010
Externally publishedYes

Keywords

  • Cellulases
  • Co-expression
  • Consolidated bioprocessing
  • Phosphoric acid swollen cellulose
  • Saccharomyces cerevisiae

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