Optimization of fermentation conditions for P450 BM-3 monooxygenase production by hybrid design methodology |
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Authors: | Lu Yan Mei Le-he |
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Institution: | (1) Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, 310027, China |
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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 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.
Project (No. 30570411) supported by the National Natural Science Foundation of China |
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Keywords: | Optimization P450 BM-3 Plackett-Burman (PB) design Hybrid design |
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