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

Paper ID Volume ID Publish Year Pages File Format Full-Text
4374 223 2008 7 PDF Available
Title
Induction strategies in fed-batch cultures for recombinant protein production in Escherichia coli: Application to rhamnulose 1-phosphate aldolase
Abstract

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.

Keywords
Induction strategies; Recombinant protein; E. coli; Fed-batch cultures; Continuous inducer dosage; Aldolase
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Induction strategies in fed-batch cultures for recombinant protein production in Escherichia coli: Application to rhamnulose 1-phosphate aldolase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 41, Issue 2, 1 September 2008, Pages 181–187
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us