Purification and characterization of a novel aminoacylase from Streptomyces mobaraensis.
A novel aminoacylase was purified to homogeneity from culture broth of Streptomyces mobaraensis, as evidenced by SDS-polyacrylamide gel electrophoresis (PAGE). The enzyme was a monomer with an approximate molecular mass of 100 kDa. The purified enzyme was inhibited by the presence of 1,10-phenanthroline and activated by the addition of Co2+. It was stable at temperatures of up to 60 degrees C for 1 h at pH 7.2. It showed broad substrate specificity to N-acetylated L-amino acids. It catalyzed the hydrolysis of the amide bonds of various N-acetylated L-amino acids, except for Nepsilon-acetyl-L-lysine and N-acetyl-L-proline. Hydrolysis of N-acetyl-L-methionine and N-acetyl-L-histidine followed Michaelis-Menten kinetics with K(m) values of 1.3+/-0.1 mM and 2.7+/-0.1 mM respectively. The enzyme also catalyzed the deacetylation of 7-aminocephalosporanic acid (7-ACA) and cephalosporin C. Moreover, feruloylamino acids and L-lysine derivatives of ferulic acid derivatives were synthesized in an aqueous buffer using the enzyme.
Koreishi
Mayuko
M
Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, Tsushima-naka, Japan.
Asayama
Fumiaki
F
Imanaka
Hiroyuki
H
Imamura
Koreyoshi
K
Kadota
Megumi
M
Tsuno
Takuo
T
Nakanishi
Kazuhiro
K
D000581:Amidohydrolases / Q000302:isolation & purification / Q000378:metabolism*; D003035:Cobalt / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors; D004795:Enzyme Stability; D007700:Kinetics; D008970:Molecular Weight; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D003035:Cobalt; D000581:Amidohydrolases; C019672:aminoacylase I