Improved production of ( S)-ketoprofen ester hydrolase by a mutant of Trichosporon brassicae CGMCC 0574.
An efficient screening method following UV mutagenesis yielded a high frequency of improved mutants of Trichosporon brassicae CGMCC 0574, a wild-type esterase-producer capable of enantioselectively hydrolyzing the ethyl ester of ketoprofen [2-(3-benzoylphenyl) propionic acid]. The mutant had an activity 1.8-fold higher than the wild type and was stable in its enzyme production for ten serial transfers. As the best single carbon source, isopropanol improved the specific activity of the enzyme 5-fold; and this did not result from the effect of cell permeabilization. An 18-h culture grown on a medium containing 0.5% glucose plus 0.5% isopropanol produced 3-fold as much esterase as a culture grown on 1% glucose.
Wu
Hui-Yuan
HY
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237 Shanghai, P.R. China.
Xu
Jian-He
JH
Shen
Duan
D
Xin
Qi
Q
D019840:2-Propanol / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D004952:Esters / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006867:Hydrolases / Q000096:biosynthesis*; D006868:Hydrolysis; D007218:Industrial Microbiology / Q000379:methods*; D007660:Ketoprofen / Q000378:metabolism*; D008828:Microbiological Techniques; D016296:Mutagenesis; D014250:Trichosporon / Q000201:enzymology* / Q000235:genetics*; D014466:Ultraviolet Rays
D004952:Esters; D002244:Carbon; D007660:Ketoprofen; D006867:Hydrolases; D005947:Glucose; D019840:2-Propanol