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KMID : 1094720080130010007
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Biotechnology and Bioprocess Engineering
2008 Volume.13 No. 1 p.7 ~ p.13
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Optimization of human interferon gamma production in Escherichia coli by response surface methodology
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Victor E. Balderas Hernandez
Luz M. T. Paz Maldonado
Emilio Medina Rivero
Ana P. Barba de la Rosa
Leandro G. Ordonez Acevedo
Antonio De Leon Rodriguez
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Abstract
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The production of human interferon gamma (hIFN-γ) using a synthetic gene in Escherichia coli BL21-SI was optimized by response surface methodology (RSM) and a Box-Behnken design. The process variables studied were temperature, bio-mass concentration at induction time and the NaCl concentration as inducer. According to the Box-Behnken design, a second order response function was developed. The optimal expression conditions were a temperature of 32.6°C, induction biomass of 0.31 g/L and 0.3 M NaCl in minimal medium. The model prediction for the maximum hIFN-γ production was 77.3 mg/L, which corresponded satisfactorily with the experimental data. The hIFN-γ concentration attained under optimized conditions was 13-times higher than that obtained using the non-optimized conditions. We conclude that RSM is an effective method for the optimization of recombinant protein expression using synthetic genes in E. coli.
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KEYWORD
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Escherichia coli BL21-SI, minimum medium, therapeutic protein, synthetic gene
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