摘要
Factorial design and response surface techniques were used to design and optimize increasing P450 BM-3 expression in E, coll. Operational conditions for maximum production were determined with twelve parameters under consideration： the concentration of FeCl3, induction at OD578 （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 57.90×10^-3 U/ml by the best combinations of the physicochemical parameters at optimum levels of 0,12 mg/L FeCl3, inoculum concentration of 2.10%, induction at OD578 equal to 1.07, and with 6.05 h of induction.
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 FeCl3, induction at OD578 (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 57.90×10?3 U/ml by the best combinations of the physicochemical parameters at optimum levels of 0.12 mg/L FeCl3, inoculum concentration of 2.10%, induction at OD578 equal to 1.07, and with 6.05 h of induction.
基金
Project (No. 30570411) supported by the National Natural ScienceFoundation of China
