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KMID : 0380620170490060610
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Korean Journal of Food Science and Technology
2017 Volume.49 No. 6 p.610 ~ p.616
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Overproduction of a γ-glutamyltranspeptidase from Bacillus amyloliquefaciens in Bacillus subtilis through medium optimization
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Cho Hye-Bin
Jetendra Kumar Roy
Park Wu-Jin
Jeon Byoung-Oon
Kim Young-Wan
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Abstract
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γ-Glutamyltranspeptidase (GGT, EC 2.3.2.2) transfers γ-glutamyl moiety from glutamine to amino acids or peptides and hydrolyzes glutamine to glutamate and ammonia. In order to overproduce γ-glutamyltranspeptidase from Bacillus amyloliquefaciens (BAGGT), the encoding gene was cloned and expressed in Bacillus subtilis. The productivity of BAGGT in Bacillus subtilis was improved by 42-fold by using a dual-promoter system that was generated by combining promoters from B. subtilis α-amylase and BAGGT genes. Through optimization of medium composition by Plackett-Burman design and central composition design, BAGGT was produced at 18.3×107 U/L of culture in the optimized medium. Compared to previously used Luria-Bertani medium, the optimized culture medium (15 g/L molasses, 60 g/L corn steep liquor, 6 g/L yeast extract, 4 g/L NaCl, 6 g/L K2HPO4, and 2 g/L KH2PO4), resulted in a 4.3-fold increase in production of BAGGT.
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KEYWORD
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Bacillus subtilis, dual-promoter system, γ-glutamyltranspeptidase, medium optimization
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