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Improvement of acetic acid tolerance of Saccharomyces cerevisiae using a zinc-finger-based artificial transcription factor and identification of novel genes involved in acetic acid tolerance

  • Bioenergy and biofuels
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Abstract

Acetic acid is present in cellulosic hydrolysate as a potent inhibitor, and the superior acetic acid tolerance of Saccharomyces cerevisiae ensures good cell viability and efficient ethanol production when cellulosic raw materials are used as substrates. In this study, a mutant strain of S. cerevisiae ATCC4126 (Sc4126-M01) with improved acetic acid tolerance was obtained through screening strains transformed with an artificial zinc finger protein transcription factor (ZFP-TF) library. Further analysis indicated that improved acetic acid tolerance was associated with improved catalase (CAT) activity. The ZFP coding sequence associated with the improved phenotype was identified, and real-time RT-PCR analysis revealed that three of the possible genes involved in the enhanced acetic acid tolerance regulated by this ZFP-TF, namely YFL040W, QDR3, and IKS1, showed decreased transcription levels in Sc4126-M01 in the presence of acetic acid, compared to those in the control strain. Sc4126-M01 mutants having QDR3 and IKS1 deletion (ΔQDR3 and ΔIKS1) exhibited higher acetic acid tolerance than the wild-type strain under acetic acid treatment. Glucose consumption rate and ethanol productivity in the presence of 5 g/L acetic acid were improved in the ΔQDR3 mutant compared to the wild-type strain. Our studies demonstrated that the synthetic ZFP-TF library can be used to improve acetic acid tolerance of S. cerevisiae and that the employment of an artificial transcription factor can facilitate the exploration of novel functional genes involved in stress tolerance of S. cerevisiae.

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Acknowledgments

This work was supported by financial support from the National Science Foundation of China (No. 21376043), National High Technology Research and Development Program of China (863 Program, No. 2012AA101805, 2012AA021205), and Program for New Century Excellent Talents, Ministry of Education, China (No. NCET-11-0057). We appreciate the kind help of Dr. Jin-Soo Kim in ToolGen, Inc., South Korea for donating the artificial zinc finger protein (ZFP) library. We also thank Dr. Alan K Chang for improving the language of the manuscript.

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Authors and Affiliations

  1. School of Life Science and Biotechnology, Dalian University of Technology, Dalian, 116024, China

    Cui Ma, Xiaowen Wei, Fei Zhang, Jianren Xu, Xinqing Zhao & Fengwu Bai

  2. School of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai, 200240, China

    Xinqing Zhao & Fengwu Bai

  3. Liaoning Academy of Microbiology, Chaoyang, 122000, China

    Cuihuan Sun

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  1. Cui Ma
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  3. Cuihuan Sun
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Correspondence to Xinqing Zhao.

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Ma, C., Wei, X., Sun, C. et al. Improvement of acetic acid tolerance of Saccharomyces cerevisiae using a zinc-finger-based artificial transcription factor and identification of novel genes involved in acetic acid tolerance. Appl Microbiol Biotechnol 99, 2441–2449 (2015). https://doi.org/10.1007/s00253-014-6343-x

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  • DOI: https://doi.org/10.1007/s00253-014-6343-x

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