Secretion of recombinant ribonuclease T1 into the periplasmic space of Escherichia coli with the aid of the signal peptide of alkaline phosphatase

FEBS Letters

Morioka, Hiroshi¹  Uesugi, Seiichi¹  Fujimura, Takao¹  Ikehara, Mono¹  Ohara, Kanako¹  Nishikawa, Satoshi¹  Tanaka, Toshiki¹ 

[1] Faculty of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565, Japan

关键词: Ribonuclease T1;    Alkaline phosphatase;    Signal peptide;    Secretion;    Recombinant protein;    RNase;    ribonuclease;    APase;    alkaline phosphatase;    hGH;    human growth hormone;    IAA;    3-indoleacrylic acid;    pGpC;    5 '-phosphoguanylyl (3'-5')cytidine;   

DOI  :  10.1016/0014-5793(90)80886-N

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The ribonuclease T₁ (RNase T₁) gene was ligated to a synthetic gene for the signal peptide of Escherichia coli alkaline phosphatase. When this fusion gene was expressed in E.Coli under the control of the trp promoter, active RNase T₁ having the correct N-terminal sequence was secreted into the periplasmic space, indicating that the heterologous signal peptide had been cleaved off correctly. The enzyme could be readily purified from the periplasmic fraction with a yield of 1.8 mg from 1 liter culture. Adopting the same strategy, it was possible to produce a labile mutant of RNase T₁ (Glu-58 → Ala mutant) in E. coli, the yield of the purified mutant enzyme being 2.0 mg from 1 liter culture.

【 授权许可】

Unknown