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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
Authors:Flores Noemí  Leal Lidia  Sigala Juan Carlos  de Anda Ramón  Escalante Adelfo  Martínez Alfredo  Ramírez Octavio T  Gosset Guillermo  Bolivar Francisco
Institution:Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México.
Abstract: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.
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