Butler M J, Bergeron A, Soostmeyer G, Zimny T, Malek L T
Cangene Corporation, Mississauga, Ontario, Canada.
Gene. 1993 Jan 15;123(1):115-9. doi: 10.1016/0378-1119(93)90549-i.
An aminopeptidase P (PepP)-encoding gene has been cloned from Streptomyces lividans 66 by screening for overexpression of activity using the chromogenic substrate Gly-Pro-beta-naphthylamide as a liquid overlayer on colonies growing on agar medium. The pepP gene was localised by deletion mapping, and the nucleotide sequence was determined. The deduced amino acid sequence was found to display significant similarity to Escherichia coli PepP. The partially purified S. lividans enzyme had a 50-kDa subunit and was present as a homodimer. Direct Edman degradation of the purified protein confirmed that pepP encoded the observed intracellular PepP.
通过使用生色底物甘氨酰-脯氨酰-β-萘酰胺作为液体覆盖层,在琼脂培养基上生长的菌落上筛选活性过表达,从变铅青链霉菌66中克隆了一个编码氨肽酶P(PepP)的基因。通过缺失图谱定位了pepP基因,并测定了核苷酸序列。发现推导的氨基酸序列与大肠杆菌PepP有显著相似性。部分纯化的变铅青链霉菌酶有一个50 kDa的亚基,以同型二聚体形式存在。对纯化蛋白进行直接的埃德曼降解证实pepP编码了观察到的细胞内PepP。