Optimization of cultural condition to improve recombinant bacteriorhodopsin mutant production in E.coli

At present study, a C-D fragment of Bacterirhodopsin (BR) in recombinant E. coli was expressed. The BR mutant gene was synthesized by consideration of E. coli codon usage. The synthesized gene was cloned in pET31a+ expression plasmid at Nde I and Hind III restriction sites and expressed under T7 promoter successfully. The expressed protein was analyzed by SDS-PAGE. The effect of temperature (A), induction time (B), and time of the process after induction (C), on the protein expression yield (the amount of product per unit dry cell mass) were screened by Yates table analysis. Three factors of A, B and C, and also the interaction of B*C were significant and optimized by Taguchi method. Theoptimized condition was as: temperature 370C, induction time at OD600= 0.7 and time of the process after induction 4 h. Also, the predicted yield at optimized condition was 21.54%.

کلیدواژه ها:

Bacteriorhodopsin (BR) ، Optimization ، Fermentation ، Expression