Induction strategies in fed-batch cultures for recombinant protein production in Escherichia coli: Application to rhamnulose 1-phosphate aldolase

• Published in: Biochemical engineering journal Vol. 41; no. 2; pp. 181 - 187
• Main Authors: Pinsach, Jaume, de Mas, Carles, López-Santín, Josep
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 01.09.2008; Amsterdam Elsevier Science; New York, NY
• Subjects: Aldolase; Biological and medical sciences; Biotechnology; Continuous inducer dosage; E. coli; Fed-batch cultures; Fundamental and applied biological sciences. Psychology; Induction strategies; Recombinant protein; Continuous inducer dosage; Recombinant protein; Fed-batch cultures; Aldolase; E. coli; Induction strategies; Enzyme; Induction; Escherichia coli; Induction strategies;Recombinant protein;E. coli;Fed-batch cultures;Continuous inducer dosage;Aldolase; Lyases; Aldehyde-lyases; Carbon-carbon lyases; Batchwise; Production; Rhamnulose-1-phosphate aldolase; Bacteria; Application; Culture; Semicontinuous; Enterobacteriaceae
• ISSN: 1369-703X; 1873-295X
• DOI: 10.1016/j.bej.2008.04.013

Different induction strategies for fed-batch recombinant protein production under the control of the strong T5 promoter in Escherichia coli have been investigated. Since the production of recombinant rhamnulose 1-phosphate aldolase is growth-related, the productivity of the process can be strongly reduced due to the negative effect of protein expression on cell growth. IPTG pulse induction as well as inducer dosage have been applied and their advantages and drawbacks highlighted. Both strategies led to high levels of the recombinant protein, 1000 AU g DCW −1. Inducer concentration and inducer to biomass ratio were identified as the parameters influencing the rate of protein production and final enzymatic activity per gram of biomass. In pulse induction, the maximum enzymatic activity was found at inducer concentration of 70 μM. In continuous induction experiments, inducer concentrations between 4 and 12 μM were identified as the working range in which cell growth and recombinant protein accumulation occurred simultaneously. On the other hand, the amount of IPTG per gram of biomass required was 1.6 μmol IPTG gDCW −1 in pulse induction and between 0.3 and 0.5 μmol IPTG g DCW −1 in continuous induction.