THE AMINOPEPTIDASE N-ENCODING PEPN GENE OF STREPTOMYCES-LIVIDANS-66

被引:17
作者
BUTLER, MJ
APHALE, JS
BINNIE, C
DIZONNO, MA
KRYGSMAN, P
SOLTES, GA
WALCZYK, E
MALEK, LT
机构
[1] Cangene Corporation, Mississauga
关键词
INTRACELLULAR PEPTIDASE; EXPRESSION SCREENING; CHROMOGENIC PEPTIDE SUBSTRATES; PROTEIN OVERPRODUCTION;
D O I
10.1016/0378-1119(94)90137-6
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
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.
引用
收藏
页码:115 / 119
页数:5
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