Optimization of fermentation conditions for P450 BM-3 monooxygenase production by hybrid design methodology.
- Author:
Yan LU
1
;
Le-he MEI
Author Information
1. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China.
- Publication Type:Journal Article
- MeSH:
Bacillus megaterium;
enzymology;
genetics;
Bacterial Proteins;
biosynthesis;
genetics;
Biotechnology;
Cytochrome P-450 Enzyme System;
biosynthesis;
genetics;
Escherichia coli;
enzymology;
genetics;
Fermentation;
Mixed Function Oxygenases;
biosynthesis;
genetics;
NADPH-Ferrihemoprotein Reductase;
Recombinant Proteins;
biosynthesis;
genetics
- From:
Journal of Zhejiang University. Science. B
2007;8(1):27-32
- CountryChina
- Language:English
-
Abstract:
Factorial design and response surface techniques were used to design and optimize increasing P450 BM-3 expression in E. coli. Operational conditions for maximum production were determined with twelve parameters under consideration: the concentration of FeCl(3), induction at OD(578) (optical density measured at 578 nm), induction time and inoculum concentration. Initially, Plackett-Burman (PB) design was used to evaluate the process variables relevant in relation to P450 BM-3 production. Four statistically significant parameters for response were selected and utilized in order to optimize the process. With the 416C model of hybrid design, response surfaces were generated, and P450 BM-3 production was improved to 57.90x10(-3) U/ml by the best combinations of the physicochemical parameters at optimum levels of 0.12 mg/L FeCl(3), inoculum concentration of 2.10%, induction at OD(578) equal to 1.07, and with 6.05 h of induction.
