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| Cloning, Expression, Purification and Characterization of L-glutamate Oxidase |
| LU Chan, ZHENG Pu, SUN Zhi-hao |
| The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China |
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Abstract In order to improve microbial production of L-glutamate oxidase, the L-glutamate oxidase gene (LGOX) from Streptomyces sp.X-119-6 was successfully expressed by connected to the expression vector pET28a and then transformed into E.coli BL21(DE3). The HisTrapTMFF affinity chromatography column was used for the purification of LGOX. Enzymatic properties of the recombinant LGOX were also investigated. Results showed that the recombinant LGOX activity could reach 1.1 U/mg at the IPTG concentration of 0.4 mmol/L at 30℃ for induction 6 h. The optimum reaction temperature and pH were 37℃ and 5.0, respectively. The Km was 2.12 mmol/L and Vmax was 1.06μmol/min·mg. The LGOX had high substrate specificity of L-glutamic acid with application prospect.
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Received: 04 March 2013
Published: 25 June 2013
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