Nucleotide and deduced amino acid sequences of a new subtilisin from an alkaliphilic Bacillus isolate.
The gene for a new subtilisin from the alkaliphilic Bacillus sp. KSM-LD1 was cloned and sequenced. The open reading frame of the gene encoded a 97 amino-acid prepro-peptide plus a 307 amino-acid mature enzyme that contained a possible catalytic triad of residues, Asp32, His66, and Ser224. The deduced amino acid sequence of the mature enzyme (LD1) showed approximately 65% identity to those of subtilisins SprC and SprD from alkaliphilic Bacillus sp. LG12. The amino acid sequence identities of LD1 to those of previously reported true subtilisins and high-alkaline proteases were below 60%. LD1 was characteristically stable during incubation with surfactants and chemical oxidants. Interestingly, an oxidizable Met residue is located next to the catalytic Ser224 of the enzyme as in the cases of the oxidation-susceptible subtilisins reported to date.
Saeki
Katsuhisa
K
Tochigi Research Laboratories of Kao Corporation, 2606 Akabane, Ichikai, Haga, Tochigi, 321-3497, Japan.
Magallones
Marietta V
MV
Takimura
Yasushi
Y
Hatada
Yuji
Y
Kobayashi
Tohru
T
Kawai
Shuji
S
Ito
Susumu
S
D017738:Alkanesulfonic Acids / Q000494:pharmacology; D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins; D020134:Catalytic Domain / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004795:Enzyme Stability / Q000187:drug effects; D006861:Hydrogen Peroxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics; D020860:Subtilisin / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature
D017738:Alkanesulfonic Acids; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D021382:Protein Sorting Signals; D013501:Surface-Active Agents; C030641:alkylbenzyl sulfonic acid; D006861:Hydrogen Peroxide; D012697:Serine Endopeptidases; C088701:protease C; C092955:protease D, Streptomyces griseus; D020860:Subtilisin