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Acetolactate synthase (AlsS) in Bacillus licheniformis WX-02: enzymatic properties and efficient functions for acetoin/butanediol and l-valine biosynthesis

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Abstract

Acetolactate synthase catalyzes two molecules of pyruvates to form α-acetolactate, which is further converted to acetoin and 2,3-butanediol. In this study, by heterologous expression in Escherichia coli, the enzymatic properties of acetolactate synthase (AlsS) from Bacillus licheniformis WX-02 were characterized. Its K m and k cat for pyruvate were 3.96 mM and 514/s, respectively. It has the optimal activity at pH 6.5, 37 °C and was feedback inhibited by l-valine, l-leucine and l-isoleucine. Furthermore, the alsS-deficient strain could not produce acetoin, 2,3-butanediol, and l-valine, while the complementary strain was able to restore these capacities. The alsS overexpressing strain produced higher amounts of acetoin/2,3-butanediol (57.06 g/L) and l-valine (2.68 mM), which were 10.90 and 92.80% higher than those of the control strain, respectively. This is the first report regarding the in-depth understanding of AlsS enzymatic properties and its functions in B. licheniformis, and overexpression of AlsS can effectively improve acetoin/2,3-butanediol and l-valine production in B. licheniformis. We envision that this AlsS can also be applied in the improvement of acetoin/2,3-butanediol and l-valine production in other microbes.

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Acknowledgements

This work was supported by National Program on Key Basic Research Project (973 Program, No. 2015CB150505), the Science and Technology Program of Wuhan (20160201010086), rural areas of the national science and technology plan in the 12th five-year plan of China (No. 2013AA102801-52).

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  1. Yanli Huo and Yangyang Zhan contributed equally to this article.

Authors and Affiliations

  1. Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, People’s Republic of China

    Yangyang Zhan, Qin Wang, Shunyi Li, Shihui Yang, Christopher T. Nomura & Shouwen Chen

  2. School of Chemical and Biological Engineering, Lanzhou Jiaotong University, Lanzhou, 730070, People’s Republic of China

    Yanli Huo

  3. State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Yanli Huo & Shouwen Chen

  4. Department of Chemistry, The State University of New York College of Environmental Science and Forestry (SUNY ESF), Syracuse, NY, 13210, USA

    Christopher T. Nomura

  5. Tobacco Research Institute of Hubei Province, Wuhan, 430062, People’s Republic of China

    Changjun Wang

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  1. Yanli Huo
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Contributions

SC designed and supervised the study. YH and YZ performed the experiments. YH, YZ, QW, SL, SY, CN, CW and SC analyzed the data and wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Changjun Wang or Shouwen Chen.

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Huo, Y., Zhan, Y., Wang, Q. et al. Acetolactate synthase (AlsS) in Bacillus licheniformis WX-02: enzymatic properties and efficient functions for acetoin/butanediol and l-valine biosynthesis. Bioprocess Biosyst Eng 41, 87–96 (2018). https://doi.org/10.1007/s00449-017-1847-2

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