Abstract
Abstract A fed-batch process was developed for intracellular production of recombinant TAT m -survivin(T34A) in Escherichia coli under the control of T7 promoter. The effects of induction mode and nutritional conditions were investigated. Compared to the one-point addition of inducer, the step-wise addition of isopropyl beta- d -thiogalactopyranoside (IPTG) maintained higher plasmid stability and increased the production level by 52%. Insufficient glucose supply after induction was observed to control acetate accumulation effectively and improved the expression of the target gene evidently. Remarkably, the pre-induction supplement of inorganic nitrogen source had a positive influence on the production of TAT m -survivin(T34A). High ammonium concentration of 4.8 g l −1 was the most efficient in enhancing the production level (as the percentage of total cellular protein) and the specific productivity of TAT m -survivin(T34A). As a result, the production of TAT m -survivin(T34A) was optimized from 11.6% to 36.8% of total cellular protein (corresponding to 1.68 g l (1 ). The findings provide valuable information for optimization of recombinant protein expression.
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