Batch production of deacetyl 7-aminocephalosporanic acid by immobilized cephalosporin-C deacetylase.
Bacillus subtilis SHS0133 cephalosporin-C deacetylase (CAH) overexpressed in Escherichia coli was immobilized on an anion-exchange resin, KA-890, using glutaraldehyde. The activity yield of immobilized enzyme was approximately 55% of the free enzyme. The pH range for stability of the immobilized enzyme (pH 5-10) was broader than that for free enzyme. The K(m)(app) value of immobilized enzyme for 7-aminocephalosporanic acid (7-ACA) was similar to that of the free enzyme. This immobilized enzyme obeyed Michaelis-Menten kinetics similar to those of the free enzyme. A batch-type reactor with a water jacket was employed for deacetylation of 7-ACA using CAH immobilized on KA-890. Ten kilograms of 7-ACA were completely converted to deacetyl 7-ACA at pH 8.0 within 90 min. The reaction kinetics agreed well with a computer simulation model. Moreover, the immobilized enzyme exhibited only a slight loss of the initial activity even after repeated use (52 times ) over a period of 70 days. This reaction will thus be useful for the production of cephalosporin-type antibiotics.
Takimoto
Akio
A
Discovery Research Laboratories, Shionogi & Co. Ltd, 2-1-3, Kuise Terajima 2-chome, 660-0813, Amagasaki, Hyogo, 660-0813, Japan. akio.takimoto@shionogi.co.jp
Takakura
Tomoaki
T
Tani
Hiroyoshi
H
Yagi
Shigeo
S
Mitsushima
Kenji
K
D001412:Bacillus subtilis / Q000201:enzymology*; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002511:Cephalosporins / Q000378:metabolism; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics
D002511:Cephalosporins; D004800:Enzymes, Immobilized; C030735:7-aminocephalosporanic acid; D002265:Carboxylic Ester Hydrolases; C019861:cephalosporin-C deacetylase