Metabolic engineering of Escherichia coli: construction and characterization of a gltA (citrate synthase) knockout mutant.
E. coli is one of the most important host organisms for recombinant protein production. However, growth and recombinant protein production can be limited by acetate accumulation during high-cell-density fermentations. Some of the strategies used to overcome this problem are based on the alteration of the genotype of the host. This paper discusses the construction and characterization of an E. coli gltA- knockout mutant. The knockout of the gene was confirmed by the loss of citrate synthase activity in an enzyme assay. Also the growth rate of the mutant on Luria Broth and Luria Broth + acetate was reduced.
Vandedrinck
S
S
Laboratory of Industrial Microbiology and Biocatalysis, Ghent University, Coupure Links 653, B-9000 Gent, Belgium.
Deschamps
G
G
Sablon
E
E
Vandamme
E J
EJ
D002950:Citrate (si)-Synthase / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D015202:Protein Engineering / Q000379:methods*
D017931:DNA Primers; D029968:Escherichia coli Proteins; D002950:Citrate (si)-Synthase