Optimization of culture conditions for a synthetic gene expression in Escherichia coli using response surface methodology: the case of human interferon beta.
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 mg l(-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.
Maldonado
Luz M T Paz
LM
Division of Molecular Biology, Institute for Scientific and Technological Research of San Luis Potosi, Apartado Postal 3-74, Tangamanga, 78231 San Luis Potosi, S.L.P., Mexico.
Hernández
Víctor E Balderas
VE
Rivero
Emilio Medina
EM
Barba de la Rosa
Ana P
AP
Flores
José L Flores
JL
Acevedo
Leandro G Ordoñez
LG
De León Rodríguez
Antonio
A
D019149:Bioreactors / Q000382:microbiology; D018929:Cell Culture Techniques / Q000379:methods*; D003198:Computer Simulation; D004926:Escherichia coli / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D024861:Genetic Enhancement / Q000379:methods; D006801:Humans; D007370:Interferon Type I / Q000096:biosynthesis* / Q000235:genetics; D008954:Models, Biological; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis
D007370:Interferon Type I; D011994:Recombinant Proteins