Development of a fed-batch process for the production of anticancer drug TATm-survivin(T34A) in Escherichia coli

• Published in: Biochemical engineering journal Vol. 43; no. 2; pp. 163 - 168
• Main Authors: Zhang, Haiyi, Zheng, Yu, Liu, Qinghai, Tao, Xinyi, Zheng, Wenyun, Ma, Xingyuan, Wei, Dongzhi
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier 01.02.2009
• Subjects: Biological and medical sciences; Biotechnology; Escherichia coli; Fundamental and applied biological sciences. Psychology; Antineoplastic agent; Drug; Escherichia coli; TAT; Survivin; Optimization; Design; Production; Bacteria; survivin(T34A); Acetic acid; Operating process; Bioprocess design; Fed-batch culture; Culture; Semicontinuous; Enterobacteriaceae
• ISSN: 1369-703X; 1873-295X
• DOI: 10.1016/j.bej.2008.09.013

A fed-batch process was developed for intracellular production of recombinant TATm-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 TATm-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 TATm-survivin(T34A). As a result, the production of TATm-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.