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Overexpression and characterization of recombinant glutamate decarboxylase from Thermococcus kodakaraensis KOD1

Journal of the Korean Society for Applied Biological Chemistry volume 55, pages 213–218 (2012)Cite this article

Abstract

Glutamate decarboxylase (GAD) (EC 4.1.1.15) catalyzes decarboxylation of glutamic acid to produce gammaaminobutyric acid (GABA). A putative gad gene (tk1814) from an archaeon Thermococcus kodakaraensis KOD1 was cloned and transformed into Escherichia coli to produce a bulk amount of recombinant GAD. Activity of the purified GAD was optimal at 90°C and pH 8.0. Optimal concentration of substrate for conversion into gamma-aminobutyric acid by recombinant GAD was 50 mM monosodium glutamate. Recombinant GAD was confirmed to be monomeric, and its activity was greatly inhibited by various salts such as sodium chloride, Tris-HCl, and sodium phosphate. Km, Vmax, and Kcat values were 9.92 mM, 153.8 μmol min−1 mg−1, and 6.613×103 min−1 respectively.

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Authors and Affiliations

  1. School of Applied Biosciences, College of Agriculture and Life Sciences, Kyungpook National University, Daegu, 702-701, Republic of Korea

    Sung-Jun Hong, Ihsan Ullah, Gun-Seok Park & Jae-Ho Shin

  2. School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu, 702-701, Republic of Korea

    Changhee Lee

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  1. Sung-Jun Hong
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  2. Ihsan Ullah
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  3. Gun-Seok Park
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  4. Changhee Lee
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Correspondence to Jae-Ho Shin.

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Hong, SJ., Ullah, I., Park, GS. et al. Overexpression and characterization of recombinant glutamate decarboxylase from Thermococcus kodakaraensis KOD1. J Korean Soc Appl Biol Chem 55, 213–218 (2012). https://doi.org/10.1007/s13765-012-1006-z

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  • DOI: https://doi.org/10.1007/s13765-012-1006-z

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