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Effect of fadR gene knockout on the metabolism of Escherichia coli based on analyses of protein expressions, enzyme activities and intracellular metabolite concentrations

Paper ID Volume ID Publish Year Pages File Format Full-Text
18796 42743 2006 9 PDF Available
Title
Effect of fadR gene knockout on the metabolism of Escherichia coli based on analyses of protein expressions, enzyme activities and intracellular metabolite concentrations
Abstract

The metabolic effect of fadR gene knockout on Escherichia coli was investigated based on comparative analyses of protein expressions, enzyme activities and intracellular metabolite concentrations between the parent strain Escherichia coli BW25113 and its fadR knockout mutant JWK1176. The E. coli strains were grown in a minimal medium using glucose as the carbon source. Two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry were used to detect and identify the proteins expressed. Enzyme activities and intracellular metabolite concentrations were also measured. Results showed that the fadR knockout E. coli reduced acetate excretion by 27.3%, accompanied by 8.5% enhancement in biomass yield. Expressions of proteins involved in glucose transport, energy metabolism and some of amino acid biosynthesis pathways were upregulated, whereas those for fatty acids biosynthesis, such as AccB and FabD were downregulated. Besides, UspA and DnaK, the universal stress and heat shock proteins, were induced in response to fadR knockout. There was 3.7- and 1.9-fold increase in the enzyme activities of isocitrate lyase and malate synthase, respectively, in the fadR mutant compared to the parent. Intracellular concentrations of acetyl coenzyme A, pyruvate and phosphoenolpyruvate deceased, whereas the concentrations of isocitrate, α-ketoglutarate, malate, oxaloacetate and aspartate increased. These results suggest that the fadR knockout E. coli activates the glyoxylate shunt and enhances the capacity of energy metabolism and biosynthesis, which leads to the reduction of acetate excretion and improvement of the biomass yield.

Keywords
Escherichia coli; fadR knockout; Metabolic regulation; Protein expression; Enzyme activity; Intracellular metabolite
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Effect of fadR gene knockout on the metabolism of Escherichia coli based on analyses of protein expressions, enzyme activities and intracellular metabolite concentrations
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 38, Issues 3–4, 1 February 2006, Pages 512–520
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