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Research Article

Physiologic Consequences of Glucose Transport and Phosphoenolpyruvate Node Modifications in Bacillus subtilis 168

Cabrera-Valladares N. · Martínez L.M. · Flores N. · Hernández-Chávez G. · Martínez A. · Bolívar F. · Gosset G.

Author affiliations

Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM), Cuernavaca, México

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J Mol Microbiol Biotechnol 2012;22:177–197

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Article / Publication Details

First-Page Preview
Abstract of Research Article

Published online: July 26, 2012
Issue release date: August 2012

Number of Print Pages: 21
Number of Figures: 4
Number of Tables: 6

ISSN: 1464-1801 (Print)
eISSN: 1660-2412 (Online)

For additional information: https://www.karger.com/MMB

Abstract

The phosphoenolpyruvate (PEP) node is an important carbon distribution point in the central metabolic networks; therefore, its modification is a common strategy employed for developing microbial production strains. In this study, mutants of Bacillus subtilis 168 were generated with deletions of pykA (which encodes pyruvate kinase), ptsG (which encodes the glucose-specific IICBAGlc component) or the ptsGHI operon [which encodes IICBAGlc, HPr protein and enzyme I from the PEP:sugar phosphotransferase system (PTS)]. These modifications caused a reduction in the initial rate of [14C]-glucose import, corresponding to 10.99, 2.83 and 0.50% of that found in B. subtilis 168 for strains with inactive pykA, ptsG or ptsGHI genes, respectively. Characterization of derivative strains lacking 3-dehydroquinate synthase activity showed that inactivation of pykA leads to an 8-fold increase in carbon flow to the common aromatic pathway. Quantitative real-time PCR analyses of 76 genes from several functional classes revealed a carbon starvation transcriptional pattern that includes a partial gluconeogenic response and overexpression of genes encoding non-PTS glucose importers in the strains lacking functional pykA, ptsG or ptsGHI genes. A transcriptional response consistent with pyruvate limitation was also detected, which includes upregulation of genes encoding malic enzymes that generate pyruvate from malate.

© 2012 S. Karger AG, Basel


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Article / Publication Details

First-Page Preview
Abstract of Research Article

Published online: July 26, 2012
Issue release date: August 2012

Number of Print Pages: 21
Number of Figures: 4
Number of Tables: 6

ISSN: 1464-1801 (Print)
eISSN: 1660-2412 (Online)

For additional information: https://www.karger.com/MMB


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