Ozaki E, Sakimae A, Numazawa R
Central Research Laboratories, Mitsubishi Rayon Co., Ltd., Hiroshima, Japan.
Biosci Biotechnol Biochem. 1995 Jul;59(7):1204-7. doi: 10.1271/bbb.59.1204.
The esterase gene (est) of Pseudomonas putida MR-2068 was cloned into Escherichia coli JM109. An 8-kb inserted DNA directed synthesis of an esterase in E. coli. The esterase gene was in a 1.1-kb PstI-ClaI fragment within the insert DNA. The complete nucleotides of the DNA fragment containing the esterase gene were sequenced and found to include a single open reading frame of 828 bp coding for a protein of 276 amino acid residues. The open reading frame was confirmed by N-terminal amino acid sequence analysis of the purified esterase. A potential Shine-Dalgarno sequence is followed by the open reading frame. The esterase activity of the recombinant E. coli was more than 200 times higher than that of parental strain, P. putida MR-2068.
恶臭假单胞菌MR-2068的酯酶基因（est）被克隆到大肠杆菌JM109中。一段8kb的插入DNA在大肠杆菌中指导酯酶的合成。酯酶基因位于插入DNA内一个1.1kb的PstI-ClaI片段中。对包含酯酶基因的DNA片段的完整核苷酸进行了测序，发现其包含一个828bp的单一开放阅读框，编码一个由276个氨基酸残基组成的蛋白质。通过对纯化酯酶的N端氨基酸序列分析证实了该开放阅读框。开放阅读框之前有一个潜在的Shine-Dalgarno序列。重组大肠杆菌的酯酶活性比亲本菌株恶臭假单胞菌MR-2068高200多倍。
