Cloning and sequence analysis of the gene (eprA1) encoding an extracellular protease from Aeromonas hydrophila.
A gene (eprA1) encoding the extracellular protease of Aeromonas hydrophila AH1 has been cloned and sequenced. Nucleotide sequence analysis of eprA1 predicted a single open reading frame (ORF) of 1038 bp encoding a 346 amino acid (aa) polypeptide, with a potential 21-aa signal peptide. When the eprA1 gene was expressed in minicells, one major band of approx. 37 kDa was identified, while protease activity staining experiments identified a caseinolytic band of approx. 29 kDa determined by SDS-PAGE analysis of the minicells. The deduced C-terminal aa region (Arg-290 to Gly-313) showed sequence homology to partial C-terminal sequences of other zinc metalloproteases including Penicillium citrinum metalloprotease (PlnC), Aspergillus oryzae metalloprotease (NpII), Aspergillus flavus metalloprotease (MepA), and Aspergillus fumigatus metalloprotease (Mep20), particularly with respect to zinc-binding residues.
Chang
T M
TM
Department of Biochemistry, Medical College, National Cheng Kung University, Tainan, Taiwan.
Liu
C C
CC
Chang
M C
MC
D016980:Aeromonas hydrophila / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D002364:Caseins / Q000378:metabolism; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial / Q000235:genetics*; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000737:chemistry; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015032:Zinc
D001426:Bacterial Proteins; D002364:Caseins; D008024:Ligands; D011994:Recombinant Proteins; D010450:Endopeptidases; C108879:eprA1 protease, Aeromonas hydrophila; D015032:Zinc