Proteome analysis of recombinant Escherichia coli producing human glucagon-like peptide-1.
The proteomic response of recombinant Escherichia coli producing human glucagon-like peptide-1 was analyzed by two-dimensional gel electrophoresis. Protein spots in two-dimensional gel could be identified by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and their expression profiles were compared with those of nonproducing cells. Thirty-five intracellular proteins exhibited differential expression levels between the production and control strains. These changes reflected physiological responses to heterologous peptide production in recombinant E. coli. Specifically, physiological changes included the down-regulation of proteins involved in the central carbon metabolism, biosynthesis of cellular building blocks and peptides, and up-regulation of cell protection proteins and some sugar transport proteins. This comprehensive analysis would provide useful information for understanding physiological alterations to heterologous peptide production and for designing efficient metabolic engineering strategies for the production of recombinant peptides in E. coli.
Lee
Dae-Hee
DH
Department of Agricultural Biotechnology and The BioMAX Institute, Seoul National University, Seoul 151-921, South Korea.
Kim
Sung-Gun
SG
Park
Yong-Cheol
YC
Nam
Soo-Wan
SW
Lee
Kelvin H
KH
Seo
Jin-Ho
JH
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D052216:Glucagon-Like Peptide 1 / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D006801:Humans; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
D020543:Proteome; D011994:Recombinant Proteins; D052216:Glucagon-Like Peptide 1