title: The aminopeptidase N-encoding pepN gene of Streptomyces lividans 66.
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.
authors.affiliation: Cangene Corporation, Mississauga, Ontario, Canada.
authors.forename: M J
authors.initials: MJ
authors.lastname: Butler
authors.forename: J S
authors.initials: JS
authors.lastname: Aphale
authors.forename: C
authors.initials: C
authors.lastname: Binnie
authors.forename: M A
authors.initials: MA
authors.lastname: DiZonno
authors.forename: P
authors.initials: P
authors.lastname: Krygsman
authors.forename: G A
authors.initials: GA
authors.lastname: Soltes
authors.forename: E
authors.initials: E
authors.lastname: Walczyk
authors.forename: L T
authors.initials: LT
authors.lastname: Malek
mesh_terms: D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001120:Arginine / Q000031:analogs & derivatives; D001426:Bacterial Proteins / Q000032:analysis; D001483:Base Sequence; D018826:CD13 Antigens; D003001:Cloning, Molecular; D003470:Culture Media; D005798:Genes, Bacterial / Q000235:genetics*; D015698:Genomic Library; D007930:Leucine / Q000031:analogs & derivatives; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; C029772:arginine beta-naphthylamide; C040135:leucine-beta-naphthylamide; D001120:Arginine; D000626:Aminopeptidases; D018826:CD13 Antigens; D007930:Leucine