Complete nucleotide sequence of the structural gene for alkaline proteinase from Pseudomonas aeruginosa IFO 3455.
The DNA-encoding alkaline proteinase (AP) of Pseudomonas aeruginosa IFO 3455 was cloned, and its complete nucleotide sequence was determined. When the cloned gene was ligated to pUC18, the Escherichia coli expression vector, the gene-incorporated bacteria expressed high levels of both AP activity and AP antigens. The amino acid sequence deduced from the nucleotide sequence revealed that the mature AP consists of 467 amino acids with a relative molecular weight of 49,507. The amino acid composition predicted from the DNA sequence was similar to the chemically determined composition of purified AP reported previously. The amino acid sequence analysis revealed that both the N-terminal side sequence of the purified AP and several internal lysyl peptide fragments were identical to the deduced amino acid sequences. The percent homology of amino acid sequences between AP and Serratia protease was about 55%. The zinc ligands and an active site of the AP were predicted by comparing the structure of the enzyme with of Serratia protease, thermolysin, Bacillus subtilis neutral protease, and Pseudomonas elastase.
Okuda
K
K
Department of Bacteriology, Yokohama City University School of Medicine, Japan.
Morihara
K
K
Atsumi
Y
Y
Takeuchi
H
H
Kawamoto
S
S
Kawasaki
H
H
Suzuki
K
K
Fukushima
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D004269:DNA, Bacterial / Q000032:analysis; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics*
D004269:DNA, Bacterial; D012697:Serine Endopeptidases; C032923:microbial serine proteinases