Induction strategies in fed-batch cultures for recombinant protein production in Escherichia coli: Application to rhamnulose 1-phosphate aldolase
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 r...
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Published in | Biochemical engineering journal Vol. 41; no. 2; pp. 181 - 187 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
Lausanne
Elsevier B.V
01.09.2008
Amsterdam Elsevier Science New York, NY |
Subjects | |
Online Access | Get full text |
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Summary: | 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. |
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ISSN: | 1369-703X 1873-295X |
DOI: | 10.1016/j.bej.2008.04.013 |