Cloning of Transglutaminase Gene from Streptomyces fradiae and its Enhanced Expression in the Original Strain |
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Authors: | Xiaoqiu Liu Xiuqing Yang Fuhong Xie Shijun Qian |
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Institution: | (1) Institute of Microbiology, Chinese Academy of Sciences, Beijing , 100080, China;(2) Graduate School of Chinese Academy of Sciences, Beijing , 100039, China |
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Abstract: | The transglutaminase (TGase) gene of Streptomyces fradiae was cloned. It had an ORF of 1242 bp, encoding a presumed prepro-region of 82 amino acids and a mature TGase of 331 amino acids. Enhanced expression of the TGase was achieved by introducing another copy of TGase gene into the original host genome which was driven by the strong constitutive promoter, “ermE up”, and shown to be expressed at the mRNA and protein levels. TGase activity in the recombinant strain (3.2 U/ml) was improved 1.3-fold when compared to that normally expressed in the original strain (2.4 U/ml). The specific enzyme activity in the recombinant strain (3.8 U/mg) was double that of the original strain (1.9 U/mg). |
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Keywords: | Cloning Enhanced expression Streptomyces fradiae Transglutaminase |
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