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A new NAD(H)-dependent meso-2,3-butanediol dehydrogenase from an industrially potential strain Serratia marcescens H30

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Abstract

The budC gene coding for a new meso-2,3-butanediol dehydrogenase (BDH) from Serratia marcescens H30 was cloned and expressed in Escherichia coli BL21(DE3), purified, and characterized for its properties. The recombinant BDH with a molecular weight of 27.4 kDa exhibited a reversible transformation between acetoin and 2,3-butanediol. In the presence of NADH, BDH could catalyze the reduction of diacetyl and (3R)-acetoin to (3S)-acetoin and meso-2,3-butanediol, respectively, while (3S)-acetoin as a substrate could be further transformed into (2S, 3S)-2,3-butanediol at pH 9.0. For diol oxidation reactions, (3R)-acetoin and (3S)-acetoin were obtained when meso-2,3-butanediol and (2S,3S)-2,3-butanediol were used as the substrates with BDH and NAD+. (2R,3R)-2,3-butanediol was not a substrate for the BDH at all. The low K m value (4.1 mM) in meso-2,3-butanediol oxidation reaction and no activity for diacetyl, acetoin, and 2,3-butanediol as the substrates with NADP+/NADPH suggested that the budC gene product belongs to a NAD(H)-dependent meso-2,3-BDH. Maximum activities for diacetyl and (3S/3R)-acetoin reduction were observed at pH 8.0 and pH 5.0 while for meso-2,3-butanediol oxidation it was pH 8.0. However, the optimum temperature for oxidation and reduction reactions was about 40 °C. In addition, the BDH activity for meso-2,3-butanediol oxidation was enhanced in the presence of Fe2+ and for diacetyl and (3S/3R)-acetoin reduction in the presence of Mg2+ and Mn2+, while several metal ions inhibited its activity, particularly Fe3+ for reduction of diacetyl and acetoin. Sequence analysis showed that the BDH from S. marcescens H30 possessed two conserved sequences including the coenzyme binding motif (GxxxGxG) and the active-site motif (YxxxK), which are present in the short-chain dehydrogenase/reductase superfamily.

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Acknowledgments

This work was supported by the Open Funding Project of the State Key Laboratory of Bioreactor Engineering, the Natural Science Foundation of Fujian Province of China (no. 2011J05048) and Science Fund of the Provincial Education Department of Fujian Province of China (no. JA11089).

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Authors and Affiliations

  1. Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian Province, 350002, People’s Republic of China

    Liaoyuan Zhang, Quanming Xu, Senran Zhan, Hui Lin, Shujing Sun, Li Sha, Kaihui Hu & Xiong Guan

  2. State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, People’s Republic of China

    Yaling Shen

  3. College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, Fujian Province, 350002, People’s Republic of China

    Yongyu Li

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  1. Liaoyuan Zhang
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  2. Quanming Xu
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  3. Senran Zhan
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  4. Yongyu Li
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  5. Hui Lin
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  6. Shujing Sun
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  7. Li Sha
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  8. Kaihui Hu
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  9. Xiong Guan
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  10. Yaling Shen
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Correspondence to Xiong Guan or Yaling Shen.

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Zhang, L., Xu, Q., Zhan, S. et al. A new NAD(H)-dependent meso-2,3-butanediol dehydrogenase from an industrially potential strain Serratia marcescens H30. Appl Microbiol Biotechnol 98, 1175–1184 (2014). https://doi.org/10.1007/s00253-013-4959-x

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  • DOI: https://doi.org/10.1007/s00253-013-4959-x

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