铜绿假单胞菌PST-01来源的有机溶剂稳定蛋白酶在单相水-有机溶剂体系中催化的肽合成。

Peptide synthesis catalyzed by organic solvent-stable protease from Pseudomonas aeruginosa PST-01 in monophasic aqueous-organic solvent systems.

作者信息

Ogino H, Yamada M, Watanabe F, Ichinose H, Yasuda M, Ishikawa H

机构信息

Department of Chemical Engineering, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan.

出版信息

J Biosci Bioeng. 1999;88(5):513-8. doi: 10.1016/s1389-1723(00)87668-9.

DOI:10.1016/s1389-1723(00)87668-9

PMID:16232654

Abstract

The equilibrium yields of the peptide Cbz-Arg-Leu-NH2 synthesized from Cbz-Arg and Leu-NH2 using the PST-01 protease in the presence of organic solvents were investigated under various conditions. The equilibrium yields depended little on the concentration of the carboxyl component, but significantly on the concentration of the nucleophile. The optimum temperature and pH for a high equilibrium yield were 30 degrees C and greater than 5.0, respectively. Under optimum conditions the equilibrium yields were 71.6% and 87.7% in the presence of 50% (v/v) DMF and 60% (v/v) DMSO, respectively. Furthermore, the PST-01 protease also catalyzed the syntheses of the dipeptides Cbz-Lys-Leu-NH2, Cbz-Ala-Leu-NH2, Cbz-Ala-Phe-NH2, Cbz-Arg-Leu-NH2, and Cbz-Lys-Phe-NH2 with equilibrium yields of more than 60% in the presence of 50% (v/v) DMF and 50 mM sodium phosphate buffer (pH 7.0).

摘要

研究了在有机溶剂存在下，使用PST - 01蛋白酶由Cbz - Arg和Leu - NH₂合成肽Cbz - Arg - Leu - NH₂在各种条件下的平衡产率。平衡产率对羧基组分的浓度依赖性较小，但对亲核试剂的浓度依赖性显著。实现高平衡产率的最佳温度和pH分别为30℃和大于5.0。在最佳条件下，在50%（v/v）DMF和60%（v/v）DMSO存在下，平衡产率分别为71.6%和87.7%。此外，PST - 01蛋白酶还催化了二肽Cbz - Lys - Leu - NH₂、Cbz - Ala - Leu - NH₂、Cbz - Ala - Phe - NH₂、Cbz - Arg - Leu - NH₂和Cbz - Lys - Phe - NH₂的合成，在50%（v/v）DMF和50 mM磷酸钠缓冲液（pH 7.0）存在下，平衡产率超过60%。