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Influence of process temperature on recombinant enzyme activity in Escherichia coli fed-batch cultures

Paper ID Volume ID Publish Year Pages File Format Full-Text
17658 42687 2008 6 PDF Available
Title
Influence of process temperature on recombinant enzyme activity in Escherichia coli fed-batch cultures
Abstract

The influence of proteolysis over recombinant protein quality has been studied using rhamnulose 1-phosphate aldolase (RhuA) production as case example. Progressive induction by means of continuous isopropyl-β-d-thiogalactopyranoside (IPTG) dosage in Escherichia coli fed-batch cultures led to high specific levels of recombinant protein. However, the specific activity profile did not correlate to the specific protein content when the process was run at 37 °C and there was a decrease of the enzyme activity along the induction phase. Specific activity loss depending on the presence of an energy source was observed at short term, but protein degradation due to the action of energy-independent metalloproteases occurred after a longer time period. The effects of lowering the temperature were analysed on both mechanisms, and a reduction of the specific activity loss was observed when the process temperature was decreased to 28 °C. Lower plasmid copy number and specific production rates probably alleviated the metabolic load on host cell during recombinant protein overexpression, and a high increase of the enzyme activity was achieved in high cell density fed-batch cultures under these conditions.

Keywords
Recombinant protein production; Temperature effect; Proteolysis; Aldolases; Fed-batch cultures
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Influence of process temperature on recombinant enzyme activity in Escherichia coli fed-batch cultures
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 43, Issue 7, 10 December 2008, Pages 507–512
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