Properties and Mode of Action of a Bactericidal Compound (= Methylglyoxal) Produced by a Mutant of
            <i>Escherichia coli</i>

Krymkiewicz, Norberto; Diéguez, Eugenia; Rekarte, U. D.; Zwaig, N.

doi:10.1128/jb.108.3.1338-1347.1971

Cited by 32 publications

(11 citation statements)

References 17 publications

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“…Methylglyoxal is extremely toxic to cells, and a balance of production and detoxification must be maintained. The toxicity is still poorly understood at the molecular level, but this electrophile has the capacity to modify DNA and proteins leading to inactivation of cells (Krymkiewicz et al, 1971;Lo et al, 1994;Papoulis et al, 1995). Escherichia coli cells, stimulated to produce MG by deregulation of the transport and metabolism of glucose-6-phosphate, arabinose, gluconate, glycerol and xylose, rapidly lose viability (Freedberg et al, 1971;Rekarte et al, 1973;Ackerman et al, 1974;Kadner et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

From famine to feast: the role of methylglyoxal production in Escherichia coli

Tötemeyer

Booth

Nichols

et al. 1998

Molecular Microbiology

126

114

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SummaryThe enzyme methylglyoxal synthase (MGS) was partially purified from Escherichia coli extracts, and the amino-terminal sequence of candidate proteins was determined, based on the native protein being a tetramer of about 69 kDa. Database analysis identified an open reading frame in the E. coli genome, YccG, corresponding to a protein of 16.9 kDa. When amplified and expressed from a controlled promoter, it yielded extracts that contained high levels of MGS activity. MGS expressed from the trc promoter accumulated to approximately 20% of total cell protein, representing approximately 900-fold enhanced expression. This caused no detriment during growth on glucose, and the level of methylglyoxal (MG) in the medium rose to only 0.08 mM. High-level expression of MGS severely compromised growth on xylose, arabinose and glycerol. A mutant lacking MGS was constructed, and it grew normally on a range of carbon sources and on low-phosphate medium. However, the mutant failed to produce MG during growth on xylose in the presence of cAMP, and growth was inhibited.

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Section: Introductionmentioning

confidence: 99%

From famine to feast: the role of methylglyoxal production in Escherichia coli

Tötemeyer

Booth

Nichols

et al. 1998

Molecular Microbiology

126

114

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“…Methylglyoxal (MG) is a toxic metabolite for Escherichia coli at millimolar concentrations (Ackerman et al, 1974;Freedberg et al, 1971 ;Krymkiewicz et al, 1971). It is synthesized via a non-phosphorylated pathway from dihydroxyacetone phosphate (Cooper & Anderson, 1970).…”

Section: Introductionmentioning

confidence: 99%

Excretion of Glutathione by Methylglyoxal-resistant Escherichia coli

Murata¹,

Tani²,

Chibata³

1980

Microbiology

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A methylglyoxal-resistant mutant of Escherichia coli B excreted glutathione into the growth medium, especially during growth on medium containing methylglyoxal. In the presence of methylglyoxal, the total amount of glutathione excreted was increased about 50-fold over that of the wild-type strain. The resistant mutant had high activities of two enzyme systems: a glutathione-forming enzyme system (consisting of gamma-glutamylcysteine synthetase and glutathione synthetase) and a glyoxalase system (consisting of glyoxalase I and glyoxalase II). Methylglyoxal resistance appeared to be due to the simultaneous increase in the activities of these two enzyme systems.

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“…Glycerol kinase is allosterically inhibited by fructose-1,6-diphosphate which prevents excessive phosphorylation of the substrate. (It was demonstrated in Es(herichia coli that accumulation of G3P results in growth stasis [2] and overabundance of dihydroxyacetone phosphate results in the lethal production of methylglyoxal [6,12].) Metabolism of glycerol through the dim system occurs by two parallel pathways, one serving for glycerol utilization, the other for the disposal of extra hydrogen.…”

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confidence: 99%

Purification and properties of dihydroxyacetone kinase from Klebsiella pneumoniae

Johnson¹,

Burke²,

Forage³

et al. 1984

J Bacteriol

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Dihydroxyacetone (DHA) kinase of Klebsiella pneumoniae, a gene product of the dha regulon responsible for fermentative dissimilation of glycerol and DHA, was purified 120-fold to a final specific activity of 10 ,umol x min-I x mg of protein-' at 30°C. The enzyme, a dimer of a 53,000 + 5,000-dalton polypeptide, is highly specific for DHA (Kn, ca. 4 ,uM). Glycerol is not a substrate at 1 mM and is not an inhibitor even at 100 mM. The enzyme is not inhibited by 5 mM fructose-1,6-diphosphate. Ca2+ gives a higher enzyme activity than Mg2+ as a cationic cofactor. Escherichia coli glycerol kinase acts on both glycerol and DHA and is allosterically inhibited by fructose-1,6-diphosphate. Antibodies raised against E. coli glycerol kinase cross-reacted with K. pneumoniae glycerol kinase but not with K. pneumoniae DHA kinase.Klebsiella pneumoniae has two different systems for the dissimilation of glycerol ( Fig. 1). With molecular oxygen or nitrate available as an exogenous electron acceptor, the glp regulon is used (14). In the absence of exogenous electron acceptors, the dha regulon is used (5,9,17,23,24,28). Metabolism of glycerol through the glp system is initiated by an ATP-dependent kinase. The product, sn-glycerol 3-phosphate (G3P), is oxidized to dihydroxyacetone phosphate by one of the flavin-linked dehydrogenases. Glycerol kinase is allosterically inhibited by fructose-1,6-diphosphate which prevents excessive phosphorylation of the substrate. (It was demonstrated in Es(herichia coli that accumulation of G3P results in growth stasis [2] and overabundance of dihydroxyacetone phosphate results in the lethal production of methylglyoxal [6,12].) Metabolism of glycerol through the dim system occurs by two parallel pathways, one serving for glycerol utilization, the other for the disposal of extra hydrogen. Utilization of glycerol is catalyzed by an NADlinked dehydrogenase (16,19) which converts the substrate to dihydroxyacetone (DHA). This product is then phosphorylated by an ATP-dependent kinase. Disposal of extra hydrogen is achieved by the sequential action of a B12dependent dehydratase and an NADH-linked oxidoreductase. Glycerol is first converted to 3-hydroxypropionaldehyde which then serves as a hydrogen sink for the regeneration of NAD. The resulting 1,3-propanediol (accounting for about 50% of the glycerol consumed) is excreted into the medium (3, 4, 20, 36). E. coli differs from K. pneuimoniae is possessing only the glp system. In this study, DHA kinase was purified from K. pnelumoniae for characterization and comparison with glycerol kinases from this organism and from E. (coli.Waters Associates, Milford, Mass., and was used on a Waters gradient HPLC system. Ultrogel ACA 34 was from LKB Instruments, Inc., Rockville, Md. Blue Sepharose CL-6B and protein standards for gradient polyacrylamide gel electrophoresis were purchased from Pharmacia Fine Chemicals, Piscataway, N.J. Special enzyme-grade ammonium sulfate was obtained from Schwarz/Mann, Orangeburg, N.Y. Rabbit muscle G3P dehydrogenase was obtained from Boehr...

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Properties and Mode of Action of a Bactericidal Compound (= Methylglyoxal) Produced by a Mutant of Escherichia coli

Cited by 32 publications

References 17 publications

From famine to feast: the role of methylglyoxal production in Escherichia coli

From famine to feast: the role of methylglyoxal production in Escherichia coli

Excretion of Glutathione by Methylglyoxal-resistant Escherichia coli

Purification and properties of dihydroxyacetone kinase from Klebsiella pneumoniae

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