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Heterologous expression of cellulase genes in natural Saccharomyces cerevisiae strains

  • Bioenergy and biofuels
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Abstract

Enzyme cost is a major impediment to second-generation (2G) cellulosic ethanol production. One strategy to reduce enzyme cost is to engineer enzyme production capacity in a fermentative microorganism to enable consolidated bio-processing (CBP). Ideally, a strain with a high secretory phenotype, high fermentative capacity as well as an innate robustness to bioethanol-specific stressors, including tolerance to products formed during pre-treatment and fermentation of lignocellulosic substrates should be used. Saccharomyces cerevisiae is a robust fermentative yeast but has limitations as a potential CBP host, such as low heterologous protein secretion titers. In this study, we evaluated natural S. cerevisiae isolate strains for superior secretion activity and other industrially relevant characteristics needed during the process of lignocellulosic ethanol production. Individual cellulases namely Saccharomycopsis fibuligera Cel3A (β-glucosidase), Talaromyces emersonii Cel7A (cellobiohydrolase), and Trichoderma reesei Cel5A (endoglucanase) were utilized as reporter proteins. Natural strain YI13 was identified to have a high secretory phenotype, demonstrating a 3.7- and 3.5-fold higher Cel7A and Cel5A activity, respectively, compared to the reference strain S288c. YI13 also demonstrated other industrially relevant characteristics such as growth vigor, high ethanol titer, multi-tolerance to high temperatures (37 and 40 °C), ethanol (10 % w/v), and towards various concentrations of a cocktail of inhibitory compounds commonly found in lignocellulose hydrolysates. This study accentuates the value of natural S. cerevisiae isolate strains to serve as potential robust and highly productive chassis organisms for CBP strain development.

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Acknowledgments

The authors would like to thank the National Research Foundation (NRF) for financial support to the Chair of Energy Research: Biofuels and other clean alternative fuels (grant number UID 86423 awarded to WHvZ). The authors would like to thank Dr. Neil Jolly from ARC Infruitec-Nietvoorbij, Stellenbosch South Africa for making the natural S. cerevisiae strains available for this study.

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Authors and Affiliations

  1. Department of Microbiology, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa

    Steffi A. Davison & Willem Heber van Zyl

  2. Department of Biotechnology, University of the Western Cape, Bellville, 7535, South Africa

    Riaan den Haan

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  1. Steffi A. Davison
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  2. Riaan den Haan
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  3. Willem Heber van Zyl
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Correspondence to Willem Heber van Zyl.

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Davison, S.A., den Haan, R. & van Zyl, W.H. Heterologous expression of cellulase genes in natural Saccharomyces cerevisiae strains. Appl Microbiol Biotechnol 100, 8241–8254 (2016). https://doi.org/10.1007/s00253-016-7735-x

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