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Journal of Zhejiang University SCIENCE B 2007 Vol.8 No.1 P.27-32


Optimization of fermentation conditions for P450 BM-3 monooxygenase production by hybrid design methodology

Author(s):  LU Yan, MEI Le-he

Affiliation(s):  Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China

Corresponding email(s):   meilh@zju.edu.cn

Key Words:  Optimization, P450 BM-3, Plackett-Burman (PB) design, Hybrid design

LU Yan, MEI Le-he. Optimization of fermentation conditions for P450 BM-3 monooxygenase production by hybrid design methodology[J]. Journal of Zhejiang University Science B, 2007, 8(1): 27-32.

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author="LU Yan, MEI Le-he",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Optimization of fermentation conditions for P450 BM-3 monooxygenase production by hybrid design methodology
%A LU Yan
%A MEI Le-he
%J Journal of Zhejiang University SCIENCE B
%V 8
%N 1
%P 27-32
%@ 1673-1581
%D 2007
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2007.B0027

T1 - Optimization of fermentation conditions for P450 BM-3 monooxygenase production by hybrid design methodology
A1 - LU Yan
A1 - MEI Le-he
J0 - Journal of Zhejiang University Science B
VL - 8
IS - 1
SP - 27
EP - 32
%@ 1673-1581
Y1 - 2007
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2007.B0027

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.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1] Dahiya, N., Tewari, R., Tiwari, R.P., 2005. Chitinase production in solid-state fermentation by Enterobacter sp. NRG4 using statistical experimental design. Curr. Microbiol., 51(4):222-228.

[2] de Coninck, J., Bouquelet, S., Dumortier, V., Duyme, F., Denantes, V.I., 2000. Industrial media and fermentation process for improved growth and protease production by Tetrahymena thermophila BIII. J. Ind. Microbiol. Biotechnol., 24(4):285-290.

[3] Deeni, Y.Y., 2001. Expression, purification and biochemical characterization of a human cytochrome P450 CYP2D6-NADPH cytochrome P450 reductase fusion protein. Arch. Biochem. Biophys., 396(1):16-24.

[4] Helvig, C., Capdevila, J.H., 2000. Biochemical characterization of rat P450 2C11 fused to rat or bacterial NADPH-P450 reductase domains. Biochemistry, 39(17):5196-5205.

[5] Kalil, S.J., Maugeri, F., Rodrigus, M.I., 2000. Response surface analysis and simulation as a tool for bioprocess design and optimization. Process Biochem., 35(6):539-550.

[6] Kar, B., Banerjee, R., Bhattacharyya, B.C., 2002. Optimization of physicochemical parameters for gallic acid production by evolutionary operation-factorial design technique. Process Biochem., 37(12):1395-1401.

[7] Li, Q.S., Schwaneberg, U., Fischer, P., Schmid, R.D., 2000. Directed evolution of the fatty-acid hydroxylase P450 BM-3 into an indole-hydroxylating catalyst. Chemistry—A European Journal, 6(9):1531-1536.

[8] Li, H.M., Mei, L.H., Urlacher, V., Schmid, R.D., 2005. Cytochrome P450 BM-3 mutants with improved catalytic properties hydroxylating indole to indigo by error-prone PCR. Prog. Biochem. Biophys., 32(7):1-6 (in Chinese).

[9] Murthy, M.S.R.C., Swaminathan, T., Rakshit, S.K., Kosugi, Y., 2000. Statistical optimization of lipase catalyzed hydrolysis of methyloleate by response surface methodology. Bioprocess Eng., 22(1):35-39.

[10] Nakamura, K., Martin, M.V., G.uengerich, F.P., 2001. Random mutagenesis of human cytochrome P450 2A6 and screening with indole oxidation products. Arch. Biochem. Biophys., 395(1):25-31.

[11] Ortiz de Montellano, P.R., 1995. Cytochrome P450: Structure, Mechanism, and Biochemistry, 2nd Ed. Plenum, New York.

[12] Plackett, R.L., Burman, J.P., 1946. The design of optimum multifactorial experiments. Biometrika, 33(4):305-325.

[13] Roquemore, K.G., 1976. Hybrid design for quadratic response surfaces. Technometrics, 18(4):419-423.

[14] Schwaneberg, U., Schmidt-Dannert, C., Schmitt, J., Schmid, R.D., 1999. A continuous spectrophotometric assay for P450 BM-3, a fatty acid hydroxylating enzyme, and its mutant F87A. Anal. Biochem., 269(2):359-366.

[15] Stowe, R.A., Mayer, R.P., 1966. Efficient screening of process variables. Ind. Eng. Chem., 58(2):36-40.

[16] Uyar, F., Baysal, Z., 2004. Production and optimization of process parameters for alkaline protease production by a newly isolated Bacillus sp. under solid state fermentation. Process Biochem., 39(12):1893-1898.

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