The aminopeptidase N-encoding pepN gene of Streptomyces lividans 66.
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.
Butler
M J
MJ
Cangene Corporation, Mississauga, Ontario, Canada.
Aphale
J S
JS
Binnie
C
C
DiZonno
M A
MA
Krygsman
P
P
Soltes
G A
GA
Walczyk
E
E
Malek
L T
LT
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
D001426:Bacterial Proteins; D003470:Culture Media; C029772:arginine beta-naphthylamide; C040135:leucine-beta-naphthylamide; D001120:Arginine; D000626:Aminopeptidases; D018826:CD13 Antigens; D007930:Leucine