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Isobutanol production in engineered Saccharomyces cerevisiae by overexpression of 2-ketoisovalerate decarboxylase and valine biosynthetic enzymes

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Abstract

Engineering of Saccharomyces cerevisiae to produce advanced biofuels such as isobutanol has received much attention because this yeast has a natural capacity to produce higher alcohols. In this study, construction of isobutanol production systems was attempted by overexpression of effective 2-keto acid decarboxylase (KDC) and combinatorial overexpression of valine biosynthetic enzymes in S. cerevisiae D452-2. Among the six putative KDC enzymes from various microorganisms, 2-ketoisovalerate decarboxylase (Kivd) from L. lactis subsp. lactis KACC 13877 was identified as the most suitable KDC for isobutanol production in the yeast. Isobutanol production by the engineered S. cerevisiae was assessed in micro-aerobic batch fermentations using glucose as a sole carbon source. 93 mg/L isobutanol was produced in the Kivd overexpressing strain, which corresponds to a fourfold improvement as compared with the control strain. Isobutanol production was further enhanced to 151 mg/L by additional overexpression of acetolactate synthase (Ilv2p), acetohydroxyacid reductoisomerase (Ilv5p), and dihydroxyacid dehydratase (Ilv3p) in the cytosol.

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Acknowledgments

This research was supported by the Advanced Biomass R&D Center (ABC) of Korea Grant funded by the Ministry of Education, Science and Technology (2010-0029799) and the Marine Biotechnology Program funded by the Ministry of Land, Transport and Maritime Affairs of Korean Government.

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Authors and Affiliations

  1. Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Won-Heong Lee, Hong Nan & Yong-Su Jin

  2. Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Won-Heong Lee & Yong-Su Jin

  3. Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Seoul National University, Seoul, 151-921, Korea

    Won-Heong Lee, Seung-Oh Seo, Yi-Hyun Bae & Jin-Ho Seo

Authors
  1. Won-Heong Lee
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  2. Seung-Oh Seo
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  3. Yi-Hyun Bae
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  4. Hong Nan
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  5. Yong-Su Jin
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  6. Jin-Ho Seo
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Corresponding authors

Correspondence to Yong-Su Jin or Jin-Ho Seo.

Additional information

W.-H. Lee and S.-O. Seo contributed equally to this work.

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Lee, WH., Seo, SO., Bae, YH. et al. Isobutanol production in engineered Saccharomyces cerevisiae by overexpression of 2-ketoisovalerate decarboxylase and valine biosynthetic enzymes. Bioprocess Biosyst Eng 35, 1467–1475 (2012). https://doi.org/10.1007/s00449-012-0736-y

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  • DOI: https://doi.org/10.1007/s00449-012-0736-y

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