Source:http://linkedlifedata.com/resource/pubmed/id/12009792
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
2002-5-15
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pubmed:abstractText |
We have developed Escherichia coli strains that internalize glucose utilizing the GalP permease instead of the phosphoenolpyruvate:carbohydrate phosphotransferase system. It has been demonstrated that a strain with these modifications (PTS(-)Glc(+)) can direct more carbon flux into the aromatic pathway than the wild-type parental strain (N. Flores et al., 1996, Nat. Biotechnol. 14, 620-623; G. Gosset et al., 1996, J. Ind. Microbiol. 17, 47-52; J. L. Baéz et al., 2001, Biotechnol. Bioeng. 73, 530-535). In this study, we have determined and compared the carbon fluxes of a wild-type strain (JM101), a PTS(-)Glc(-) strain, and two isogenic PTS(-)Glc(+) derivatives named PB12 and PB13 by combining genetic, biochemical, and NMR approaches. It was determined that in these strains a functional glk gene in the chromosome is required for rapid glucose consumption; furthermore, glucokinase-specific activities were higher than in the wild-type strain. (13)C labeling and NMR analysis allowed the determination of differences in vivo which include higher glycolytic fluxes of 93.1 and 89.2% compared with the 76.6% obtained for the wild-type E. coli. In PB12 and PB13 we found a flux through the malic enzymes of 4 and 10%, respectively, compared to zero in the wild-type strain. While flux through the Pck enzyme was absent in PB12 and PB13, in the wild type it was 7.7%. Finally, it was found that in the JM101 and PB12 strains both the oxidative and the nonoxidative branches of the pentose phosphate pathway contributed to ribose 5-phosphate synthesis, whereas in PB13 this pentose was synthesized almost exclusively through the oxidative branch. The determined carbon fluxes correlate with biochemical and genetic characterizations.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Carbon,
http://linkedlifedata.com/resource/pubmed/chemical/Carbon Isotopes,
http://linkedlifedata.com/resource/pubmed/chemical/Glucokinase,
http://linkedlifedata.com/resource/pubmed/chemical/Glucose,
http://linkedlifedata.com/resource/pubmed/chemical/Membrane Transport Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Monosaccharide Transport Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoenolpyruvate Sugar...,
http://linkedlifedata.com/resource/pubmed/chemical/galactose permease
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
1096-7176
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
4
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
124-37
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:12009792-Biomedical Engineering,
pubmed-meshheading:12009792-Carbon,
pubmed-meshheading:12009792-Carbon Isotopes,
pubmed-meshheading:12009792-Escherichia coli,
pubmed-meshheading:12009792-Glucokinase,
pubmed-meshheading:12009792-Glucose,
pubmed-meshheading:12009792-Magnetic Resonance Spectroscopy,
pubmed-meshheading:12009792-Membrane Transport Proteins,
pubmed-meshheading:12009792-Monosaccharide Transport Proteins,
pubmed-meshheading:12009792-Mutation,
pubmed-meshheading:12009792-Pentose Phosphate Pathway,
pubmed-meshheading:12009792-Phosphoenolpyruvate Sugar Phosphotransferase System
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pubmed:year |
2002
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pubmed:articleTitle |
Analysis of carbon metabolism in Escherichia coli strains with an inactive phosphotransferase system by (13)C labeling and NMR spectroscopy.
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pubmed:affiliation |
Departamento de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, 62250 Cuernavaca, México.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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