J1 acylase, a glutaryl-7-aminocephalosporanic acid acylase from Bacillus laterosporus J1, is a member of the α/β-hydrolase fold superfamily

• Published in: FEBS letters 2006-02, Vol.580 (5), p.1465-1471
• Main Authors: Yau, Ming-Hon, Wang, Jun, Tsang, Paul W.K., Fong, Wing-Ping
• Format: Article
• Language: English
• Subjects: 7-ACA; 7-ADCA; 7-aminocephalosporanic acid; 7-aminodesacetoxycephalosporanic acid; Acetobacter; Acetobacter turbidans α-amino acid ester hydrolase; AEH; Amidohydrolases - chemistry; Amidohydrolases - genetics; Amidohydrolases - metabolism; AtAEH; Bacillus - enzymology; Bacillus laterosporus; Bacterial Proteins - chemistry; Base Sequence; Catalytic triad; Cephalosporins; cocaine esterase; CocE; GCA; GL-7-ACA; GL-7-ADCA; glutaryl-7-aminocephalosporanic acid; Glutaryl-7-aminocephalosporanic acid acylase; glutaryl-7-aminodesacetoxycephalosporanic acid; Homology modeling; Hydrolases - chemistry; J1 acylase; Kinetics; Models, Molecular; Oxyanion-hole; PepX; Protein Conformation; Protein Structure, Secondary; RhCocE; Rhodococcus; Rhodococcus sp. strain MB1 cocaine esterase; SCOP; SDS–PAGE; Sequence Alignment; sodium dodecyl sulfate–polyacrylamide gel electrophoresis; Solibacter usitatus S15 peptidase; structural classification of protein; SuPep; X-prolyl dipeptidyl aminopeptidase; Xanthomonas citri α-amino acid ester hydrolase; XcAEH; α-amino acid ester hydrolase; α/β-Hydrolase
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2006.01.069

J1 acylase, a glutaryl-7-aminocephalosporanic acid acylase (GCA) isolated from Bacillus laterosporus J1, has been conventionally grouped as the only member of class V GCA, although its amino acid sequence shares less than 10% identity with members of other classes of GCA. Instead, it shows higher sequence similarities with Rhodococcus sp. strain MB1 cocaine esterase ( RhCocE) and Acetobacter turbidans α-amino acid ester hydrolase ( AtAEH), members of the α/β-hydrolase fold superfamily. Homology modeling and secondary structure prediction indicate that the N-terminal region of J1 acylase has an α/β-hydrolase folding pattern. The catalytic triads in RhCocE and AtAEH were identified in J1 acylase as S125, D264 and H309. Mutations to alanine at these positions were found to completely inactivate the enzyme. These results suggest that J1 acylase is a member of the α/β-hydrolase fold superfamily with a serine–histidine–aspartate catalytic triad.