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Expression and Characterization of a Novel 1,3-Propanediol Dehydrogenase from Lactobacillus brevis

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Abstract

1,3-Propanediol dehydrogenase (PDOR) is important in the biosynthesis of 1,3-propanediol. In the present study, the dhaT gene encoding PDOR was cloned from Lactobacillus brevis 6239 and expressed in Escherichia coli for the first time. Sequence analysis revealed that PDOR containing two Fe2+-binding motifs and a cofactor motif belongs to the type III alcohol dehydrogenase. The purified recombinant PDOR exhibited a single band of 42 kDa according to SDS-PAGE. Optimal temperatures and pH values of this dehydrogenase are 37 °C, 7.5 for reduction and 25 °C, 9.5 for oxidation, respectively. We found that PDOR was more stable in acid buffer than in alkaline condition, and 60 % of its relative activity still remained after a 2-h incubation at 37 °C. The activity of PDOR can be enhanced in the presence of Mn2+ or Fe2+ iron and inhibited by EDTA or PMSF by different degrees. The K m and V max of this dehydrogenase are 1.25 mM, 64.02 μM min−1 mg−1 for propionaldehyde and 2.26 mM, 35.05 μM min−1 mg−1 for 1,3-PD, respectively. Substrate specificity analysis showed that PDOR has a broad range of substrate specificities. The modeling superposition indicated that the structural differences may account for the diversity of PDORs’ properties. Thus, our PDOR is a potential candidate for facilitating the 1,3-PD biosynthesis.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Nos. 31571806 and 21006041), Project of Jiangsu University (No. 08JDG009), and Jiangsu Government Scholarship for Overseas Studies and PAPD.

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Authors and Affiliations

  1. School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China

    Xianghui Qi, Junhua Yun, Huanhuan Zhang, Fei Wang & Qi Guo

  2. College of Science and Technology, Agricultural University of Hebei, Cangzhou, 061100, China

    Yilin Qi

  3. David Geffen School of Medicine, University of California, Los Angeles, LA, 90095, USA

    Xianghui Qi & Zheng Cao

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  1. Xianghui Qi
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  2. Junhua Yun
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  3. Yilin Qi
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Correspondence to Xianghui Qi or Qi Guo.

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Qi, X., Yun, J., Qi, Y. et al. Expression and Characterization of a Novel 1,3-Propanediol Dehydrogenase from Lactobacillus brevis . Appl Biochem Biotechnol 179, 959–972 (2016). https://doi.org/10.1007/s12010-016-2043-6

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  • DOI: https://doi.org/10.1007/s12010-016-2043-6

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