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| Characterization of GshF Expressed in Escherichia coli |
| CHEN Yong-lu, WU Mian-bin, LIN Jian-ping, YANG Li-rong, CEN Pei-lin |
| Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China |
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Abstract Glutathione, an important active tripeptide, is the most common non-protein thiol compound involved in cell activity. It has a wide range of applications in the pharmaceutical, food, cosmetics and other industries because of its antioxidant effect. And improving glutathione production via biosynthesis is the main method and has always been a focus. The bifunctional enzyme GshF discorvered recently can catalyze the two-step reaction of glutathione biosynthesis simultaneously and is expected to improve the catalytic efficiency. However, the properties of the enzyme have not been sufficiently studied. A bifunctional enzyme gene gshF from Streptococcus thermophilus was cloned and expressed in E. coli. GshF reached its highest activity of 44 U/L when the recombinant was induced with 0.1mmol/L IPTG at 30℃ after being cultivated for 4 hours at 37℃. The related enzymatic properties of GshF were achieved after such purification steps as cell lysis, centrifugation and affinity chromatography. The optimum pH and temperature was 8.0 and 37℃ respectively. The stabilities of GshF under its optimum pH and temperature were also studied. The final maximum yield of glutathione was 2.11g/L by enzymatic synthesis of recombinant bacteria. It is of important reference on the application of the enzyme in the biosynthesis of glutathione.
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Received: 17 October 2013
Published: 25 December 2013
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