Growth recovery on glucose under aerobic conditions of an Escherichia coli strain carrying a phosphoenolpyruvate:carbohydrate phosphotransferase system deletion by inactivating arcA and overexpressing the genes coding for glucokinase and galactose permease.
In Escherichia coli the phosphotransferase system (PTS) consumes one molecule of phosphoenolpyruvate (PEP) to phosphorylate each molecule of internalized glucose. PEP bioavailability into the aromatic pathway can be increased by inactivating the PTS. However, the lack of the PTS results in decreased glucose transport and growth rates. To overcome such drawbacks in a PTS(-) strain and reconstitute rapid growth on glucose phenotype (Glc(+)), the glk and galP genes were cloned into a plasmid and the arcA gene was inactivated. Simultaneous overexpression of glk and galP increased the growth rate and regenerated a Glc(+) phenotype. However, the highest growth rate was obtained when glk and galP were overexpressed in the arcA(-) background. These results indicated that the arcA mutation enhanced glycolytic and respiratory capacities of the engineered strain.
Flores
Noemí
N
Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México.
Leal
Lidia
L
Sigala
Juan Carlos
JC
de Anda
Ramón
R
Escalante
Adelfo
A
Martínez
Alfredo
A
Ramírez
Octavio T
OT
Gosset
Guillermo
G
Bolivar
Francisco
F
D000332:Aerobiosis; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005941:Glucokinase / Q000235:genetics* / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D008954:Models, Biological; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics* / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics; D013045:Species Specificity
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D009004:Monosaccharide Transport Proteins; D012097:Repressor Proteins; C041610:arcA protein, E coli; C043910:galactose permease; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; D005941:Glucokinase; D005947:Glucose