Structural gene and complete amino acid sequence of Pseudomonas aeruginosa IFO 3455 elastase.
The DNA encoding the elastase 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, bacteria carrying the gene exhibited high levels of both elastase activity and elastase antigens. The amino acid sequence, deduced from the nucleotide sequence, revealed that the mature elastase consisted of 301 amino acids with a relative molecular mass of 32,926 daltons. The amino acid composition predicted from the DNA sequence was quite similar to the chemically determined composition of purified elastase reported previously. We also observed nucleotide sequence encoding a signal peptide and "pro" sequence consisting of 197 amino acids upstream from the mature elastase protein gene. The amino acid sequence analysis revealed that both the N-terminal sequence of the purified elastase and the N-terminal side sequences of the C-terminal tryptic peptide as well as the internal lysyl peptide fragment were completely identical to the deduced amino acid sequences. The pattern of identity of amino acid sequences was quite evident in the regions that include structurally and functionally important residues of Bacillus subtilis thermolysin.
Fukushima
J
J
Department of Bacteriology, Yokohama City University School of Medicine, Japan.
Yamamoto
S
S
Morihara
K
K
Atsumi
Y
Y
Takeuchi
H
H
Kawamoto
S
S
Okuda
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D005796:Genes; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010196:Pancreatic Elastase / Q000235:genetics*; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013820:Thermolysin / Q000235:genetics
D010196:Pancreatic Elastase; D013820:Thermolysin