Cloning of transglutaminase gene from Streptomyces fradiae and its enhanced expression in the original strain.
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).
Liu
Xiaoqiu
X
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Yang
Xiuqing
X
Xie
Fuhong
F
Qian
Shijun
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis; D013302:Streptomyces / Q000235:genetics*; D011503:Transglutaminases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
D011994:Recombinant Proteins; D011503:Transglutaminases