天蓝色链霉菌66的氨肽酶N编码基因pepN

The aminopeptidase N-encoding pepN gene of Streptomyces lividans 66.

作者信息

Butler M J, Aphale J S, Binnie C, DiZonno M A, Krygsman P, Soltes G A, Walczyk E, Malek L T

机构信息

Cangene Corporation, Mississauga, Ontario, Canada.

出版信息

Gene. 1994 Apr 8;141(1):115-9. doi: 10.1016/0378-1119(94)90137-6.

DOI:10.1016/0378-1119(94)90137-6

PMID:7909302

Abstract

The gene (pepN) encoding an aminopeptidase N (PepN) has been cloned from Streptomyces lividans. This was done using either leucine-beta-naphthylamide or arginine-beta-naphthylamide in a liquid overlayer on colonies growing on agar medium to screen for overproduction of the ability to hydrolyse the substrates. The nucleotide sequence of pepN was determined and shown to encode a 95-kDa protein, which displayed significant homology to PepN proteins from other organisms. Analysis of the overproduced proteinase confirmed that this protein was located intracellularly as a monomeric active species. PepN is a metallo-exopeptidase cleaving next to Leu, Arg and Lys in peptide-bond-containing substrates.

摘要

已从淡紫链霉菌中克隆出编码氨肽酶N（PepN）的基因（pepN）。具体做法是，在琼脂培养基上生长的菌落的液体覆盖层中使用亮氨酸-β-萘酰胺或精氨酸-β-萘酰胺，以筛选水解底物能力的过量表达。测定了pepN的核苷酸序列，结果表明其编码一种95 kDa的蛋白质，该蛋白质与来自其他生物体的PepN蛋白具有显著同源性。对过量表达的蛋白酶的分析证实，该蛋白质以单体活性形式位于细胞内。PepN是一种金属外肽酶，可在含肽键的底物中紧邻亮氨酸、精氨酸和赖氨酸处进行切割。