Heterologous co-production of Thermobifida fusca Cel9A with other cellulases in Saccharomyces cerevisiae
Date
2010
Authors
Van Wyk N.
Den Haan R.
Van Zyl W.H.
Journal Title
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Volume Title
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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. © 2010 Springer-Verlag.
Description
Keywords
Amorphous cellulose, Anionic exchange, Carbohydrate sources, Carboxy methylcellulose, Cellobiohydrolases, Cellulose chain, Cellulose substrates, Cellulosic substrates, Co-expression, Co-production, Coding genes, Consolidated bioprocessing, Endoglucanases, Moderately thermophilic, Phosphoric-acid swollen cellulose, Recombinant hosts, Recombinant strains, S.cerevisiae, Saccharomyces cerevisiae, Soluble sugars, Thermobifida fusca, Trichoderma reesei, Acids, Carbohydrates, Cellulose, Glucose, Phosphoric acid, Substrates, Sugar (sucrose), Yeast, carboxymethylcellulose, cellobiose, cellulase, glucose, bacterium, cellulose, chromatography, enzyme activity, gene expression, growth rate, recombination, sugar, yeast, article, bacterial gene, bacterial growth, bacterial strain, cel5a gene, cel6a gene, cel7a gene, Cel9a gene, enzyme activity, gene expression, Hypocrea jecorina, nonhuman, protein cleavage, protein glycosylation, Saccharomyces cerevisiae, soil microflora, Thermobifida fusca, Actinomycetales, Bacterial Proteins, Cellulases, Cellulose, Chromatography, Ion Exchange, Fungal Proteins, Glucose, Recombinant Proteins, Saccharomyces cerevisiae, Trichoderma, Acids, Carbohydrates, Cellulase, Cellulose, Glucose, Phosphoric Acid, Saccharomyces Cerevisiae, Substrates, Sugar, Hypocrea jecorina, Saccharomyces cerevisiae, Thermobifida fusca
Citation
Applied Microbiology and Biotechnology
87
5
87
5