3.8 Article

Optimization of culture conditions for a synthetic gene expression in Escherichia coli using response surface methodology:: The case of human interferon beta

期刊

BIOMOLECULAR ENGINEERING
卷 24, 期 2, 页码 217-222

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ELSEVIER SCIENCE BV
DOI: 10.1016/j.bioeng.2006.10.001

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codon bias; E. coli BL21-SI; multiple sclerosis; response surface methodology (RSM); synthetic gene; yeast extract

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A human interferon beta (hINF-beta) synthetic gene was optimized and expressed in Escherichia coli BL21-SI using a vector with the T7 promoter. To determine the best culture conditions such as culture medium, temperature, cell density and inducer concentration, we used the response surface methodology and a Box-Behnken design to get the highest hINF-beta production. The maximum hINF-beta production of 61 mgl(-1) was attained using minimum medium and the following predicted optimal conditions: temperature of 32.5 degrees C, cell density of 0.64, and inducer concentration of 0.30 M NaCl. This is the first report showing the successful performance of the BL21-SI system in a minimum medium. The response surface methodology is effective for the optimization of recombinant protein production using synthetic genes. (c) 2006 Elsevier B.V. All rights reserved.

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