title: The thioesterase I of Escherichia coli has arylesterase activity and shows stereospecificity for protease substrates.
abstract: A thioesterase I gene was recloned and sequenced from Escherichia coli strain JM109. The overexpressed, matured enzyme from JM109 was purified to homogeneity. The enzyme showed broad hydrolytic activity toward three kinds of substrates including acyl-CoAs, esters, and amino acid derivatives. The enzyme had a kcat/Km value of 0.363 s-1 microM-1, for a typical thioesterase I substrate, palmitoyl-CoA. The arylesterase activity of the enzyme was observed by its ability to hydrolyze several aromatic esters including alpha-naphthyl acetate, alpha-naphthyl butyrate, phenyl acetate, benzyl acetate, and eight p-nitrophenyl esters. In kinetic studies a chymotrypsin-like substrate (an amino acid derivative), N-carbobenzoxy-L-phenylalanine p-nitrophenyl ester (L-NBPNPE), was the best substrate for the enzyme with a catalytic efficiency (kcat/Km) of 4.00 s-1 microM-1, which was 23 times higher than that of the enantiomer D-NBPNPE (0.171 s-1 microM-1). It was concluded that the thioesterase I of E. coli had arylesterase activity and it possessed stereospecificity for protease substrates.
authors.affiliation: Department of Marine Food Science, National Taiwan Ocean University, Keelung.
authors.forename: Y L
authors.initials: YL
authors.lastname: Lee
authors.forename: J C
authors.initials: JC
authors.lastname: Chen
authors.forename: J F
authors.initials: JF
authors.lastname: Shaw
mesh_terms: D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D002918:Chymotrypsin / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D008049:Lipase / Q000235:genetics; D010170:Palmitoyl-CoA Hydrolase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D002265:Carboxylic Ester Hydrolases; C019734:arylesterase; D008049:Lipase; D010170:Palmitoyl-CoA Hydrolase; D002918:Chymotrypsin