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Characterization of a glutamate decarboxylase (GAD) gene from Lactobacillus zymae

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Abstract

Lactic acid bacteria (LAB) were isolated from Kimchi, a Korean traditional fermented vegetable food. LAB accumulating GABA (γ-aminobutyric acid) in the culture media were screened by TLC analysis. One isolate, GU240, produced the highest amount of GABA among the 3,000 isolates and identified as a Lactobacillus zymae strain. Glutamate decarboxylase (GAD) gene was cloned and over-expressed in E. coli BL21(DE3) using pET26b(+). The recombinant GAD was purified by using a Ni–NTA column. Its size was 53 kDa by SDS-PAGE. Maximum GAD activity was at pH 4.5 and 41 °C and the activity was dependent on pyridoxal 5′-phosphate. Km and Vmax of LzGAD were 1.7 mM and 0.01 mM/min, respectively, when glutamate was used as a substrate.

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Acknowledgments

This work was supported by Geochang HanSsal and Geochang county Regional Agriculture Specialization fund. J. Y. Park and S.-J. Jeong were supported by BK21 Plus from MOE, Republic of Korea.

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Authors and Affiliations

  1. Division of Applied Life Science (BK21 Plus), Graduate School, Institute of Agriculture and Life Science, Gyeongsang National University, Jinju, 660-701, Republic of Korea

    Ji Yeong Park, Seon-Ju Jeong & Jeong Hwan Kim

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  1. Ji Yeong Park
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  2. Seon-Ju Jeong
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  3. Jeong Hwan Kim
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Correspondence to Jeong Hwan Kim.

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Park, J.Y., Jeong, SJ. & Kim, J.H. Characterization of a glutamate decarboxylase (GAD) gene from Lactobacillus zymae . Biotechnol Lett 36, 1791–1799 (2014). https://doi.org/10.1007/s10529-014-1539-9

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  • DOI: https://doi.org/10.1007/s10529-014-1539-9

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