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Bioconversion of Phenylpyruvate to Phenyllactate: Gene Cloning, Expression, and Enzymatic Characterization of d- and l1-Lactate Dehydrogenases from Lactobacillus plantarum SK002

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Abstract

Two DNA fragments containing the entire coding sequences of lactate dehydrogenase (LDH; ldhL1 and ldhD), whose enzymes have high activity for bioconversion of phenylpyruvate (PPA) to phenyllactate (PLA), were amplified from Lactobacillus plantarum SK002 using PCR. Sequencing showed open reading frames of 963 bp (ldhL1) and 999 bp (ldhD) encoding putative proteins of 320 and 332 amino acid residues, respectively. The LDH genes were cloned into an expression vector pET-22b(+) and expressed in Escherichia coli BL21(DE3). The purified recombinant l1-LDH and d-LDH had approximate (SDS-PAGE) molecular weights of 35 and 40 kDa, respectively. l1-LDH and d-LDH had PPA bioconversion specific activities of 71.06 and 215.84 U/mg with K m values of 3.96 and 5.4 mM, respectively. The rl1-LDH and rd-LDH showed maximum enzyme activity at 30 and 40 °C while both had optimum activity at pH 6.0. l1-LDH exhibited a higher pH and temperature stability than d-LDH. The results show that the his-tagged L. plantarum SK002 d- and l1-LDHs are efficient catalysts for bioconversion of PPA to PLA.

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References

  1. Ohhira, I. Kuwaki, S. Morita, H. Suzuki, T. Tomita, S. Hisamatsu, S. et al. (2004). Biocontrol Science, 9(3), 77–81.

    CAS  Google Scholar 

  2. Lavermicocca, P. Valerio, F. & Visconti, A. (2003). Applied and Environmental Microbiology, 69(1), 634–640. doi:10.1128/AEM.69.1.634-640.2003.

    Article  CAS  Google Scholar 

  3. Valerio, F. Lavermicocca, P. Pascale, M. & Visconti, A. (2004). FEMS Microbiology Letters, 233(2), 289–295. doi:10.1111/j.1574-6968.2004.tb09494.x.

    Article  CAS  Google Scholar 

  4. Strom, K. Sjogren, J. Broberg, A. & Schnurer, J. (2002). Applied and Environmental Microbiology, 68(9), 4322–4327. doi:10.1128/AEM.68.9.4322-4327.2002.

    Article  CAS  Google Scholar 

  5. Coloretti, F. Carri, S. Armaforte, E. Chiavari, C. Grazia, L. & Zambonelli, C. (2007). FEMS Microbiology Letters, 271(2), 245–250. doi:10.1111/j.1574-6968.2007.00723.x.

    Article  CAS  Google Scholar 

  6. Vermeulen, N. Ganzle, M. G. & Vogel, R. F. (2006). J. Agric. Food Chem, 54(11), 3832–3839. doi:10.1021/jf052733e.

    Article  CAS  Google Scholar 

  7. Li, X. Jiang, B. & Pan, B. (2007). Biotechnology Letters, 29(4), 593–597. doi:10.1007/s10529-006-9275-4.

    Article  CAS  Google Scholar 

  8. Mu, W. Chen, C. Li, X. Zhang, T. & Jiang, B. (2009). Bioresource Technology, 100(3), 1366–1370. doi:10.1016/j.biortech.2008.08.010.

    Article  CAS  Google Scholar 

  9. Li, X. Jiang, B. Pan, B. Mu, W. & Zhang, T. (2008). Journal of Agricultural and Food Chemistry, 56(7), 2392–2399. doi:10.1021/jf0731503.

    Article  CAS  Google Scholar 

  10. Gaspar, P. Neves, A. R. Shearman, C. A. Gasson, M. J. Baptista, A. M. et al. (2006). FEBS Journal, 274(22), 5924–5936. doi:10.1111/j.1742-4658.2007.06115.x.

    Article  Google Scholar 

  11. Okano, K. Zhang, Q. Shinkawa, S. Yoshida, S. Tanaka, T. Fukuda, H. et al. (2009). Applied and Environmental Microbiology, 75(20), 462–467. doi:10.1128/AEM.01514-08.

    Article  CAS  Google Scholar 

  12. Hummel, W. (1999). Trends in Biotechnology, 17(12), 487–492. doi:10.1016/S0167-7799(98)01207-4.

    Article  CAS  Google Scholar 

  13. Razeto, A. Kochhar, S. Hottinger, H. Dauter, M. Wilson, K. S. & Lamzin, V. S. (2002). Journal of Molecular Biology, 318(1), 109–1. doi:10.1016/S00222836(02)00086-4.

    Article  CAS  Google Scholar 

  14. Kleerebezem, M. Boekhorst, J. Kranenburg, R. Molenaar, D. Kuipers, O. P. Leer, R. et al. (2003). Proceedings of the National Academy of Sciences, 100(4), 1990–1995. doi:10.1073/pnas.0337704100.

    Article  CAS  Google Scholar 

  15. Jia, J. Mu, W. Zhang, T. & Jiang, B. (2009). Food and Fermentation Industries, 35(5), 22–25.

    Google Scholar 

  16. Sambrook, J. & Russell David, D. W. (2002). Molecular Cloning: A Labaratory Manual, 3, 1252–1255.

    Google Scholar 

  17. Hummel, W. Schütte, H. & Kula, M. R. (1983). Applied Microbiology and Biotechnology, 18(2), 75–85. doi:10.1007/BF00500828.

    Article  CAS  Google Scholar 

  18. Copeland, R. A. (2000). Enzymes: A Practical Introduction to Structure, Mechanism, and Data Analysis (pp. 124–128). Wiley: New York.

    Google Scholar 

  19. Origin Lab Corporation. (2003). Origin Pro 7.5 SRI. Version 7.5776(B776). Northampton MA. 01060. USA. www.originlab.com

  20. Taguchi, H. & Ohta, T. (1991). Journal of Biological Chemistry, 266(19), 12588–12594.

    CAS  Google Scholar 

  21. Cristescu, M. E. Innes, D. J. Stillman, J. H. & Crease, T. J. (2008). BMC Evolutionary Biology, 8(1), 268. doi:10.1186/1471-2148-8-268.

    Article  Google Scholar 

  22. Savijoki, K. & Palva, A. (1997). Applied and Environmental Microbiology, 63(7), 2850–2856.

    CAS  Google Scholar 

  23. Tokuda, C. Ishikura, Y. Shigematsu, M. Mutoh, H. Tsuzuki, S. Nakahira, Y. et al. (2003). Journal of Bacteriology, 185(16), 5023–5026. doi:10.1128/JB.185.16.5023-5026.2003.

    Article  CAS  Google Scholar 

  24. Arai, K. Kamata, T. Uchikoba, H. Fushinobu, S. Matsuzawa, H. & Taguchi, H. (2001). Journal of Bacteriology, 183, 397–400. doi:10.1128/JB.183.1.397-400.2001.

    Article  CAS  Google Scholar 

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Acknowledgement

This work was supported by the grants from the National High Technology “863” Program of China, project No. 2006AA10Z334, the Natural Science Foundation of Jiangsu Province, project No. BK2008099 and BK2008003, the Research Program of State Key Laboratory of Food Science and Technology, Jiangnan University, project No. SKLF-MB-200804 and SKLF-TS-200805, and the Program for Innovative Research Team of Jiangnan University, project No. 2008CXTD01.

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  1. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu, 214122, China

    Jianghua Jia, Wanmeng Mu, Tao Zhang & Bo Jiang

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  1. Jianghua Jia
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  2. Wanmeng Mu
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  3. Tao Zhang
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  4. Bo Jiang
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Correspondence to Bo Jiang.

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Jia, J., Mu, W., Zhang, T. et al. Bioconversion of Phenylpyruvate to Phenyllactate: Gene Cloning, Expression, and Enzymatic Characterization of d- and l1-Lactate Dehydrogenases from Lactobacillus plantarum SK002. Appl Biochem Biotechnol 162, 242–251 (2010). https://doi.org/10.1007/s12010-009-8767-9

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