Characterization of methylglyoxal synthase in Saccharomyces cerevisiae

Murata, Kousaku; Fukuda, Yasuki; Watanabe, Kunihiko; Saikusa, Toshihiko; Shimosaka, Makoto; Kimura, Akira

doi:10.1016/0006-291x(85)91788-7

Cited by 59 publications

(36 citation statements)

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“…Methylglyoxal pathway is found in T. thermosaccrolyticum [23], C. sphenoides [20] and S. cerevisiae [35,36]. These organisms are reported to ferment common sugars such as glucose, fructose, mannose, galactose, xylose, arabinose, lactose and cellobiose [21,23].…”

Section: Biochemical Pathways For the Production Of 12-propanediolmentioning

confidence: 99%

Microbial production and applications of 1,2-propanediol

et al. 2010

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Section: Biochemical Pathways For the Production Of 12-propanediolmentioning

confidence: 99%

Microbial production and applications of 1,2-propanediol

et al. 2010

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“…In most organisms, this toxic compound is formed as a by-product of glycolysis from triose phosphates through the action of triose-phosphate isomerase (2). But the main source for methylglyoxal in yeast and other microorganisms is dihydroxyacetone phosphate which is converted to methylglyoxal by the methylglyoxal synthase (3,4). As shown in Reaction 1 (where MG, HTA, and GSH indicate methylglyoxal, hemithioacetal, and glutathione, respectively), the first step in the conversion of methylglyoxal into D-lactic acid through the glyoxalase system is the spontaneous formation of hemithioacetal from the reaction of methylglyoxal with reduced glutathione.…”

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confidence: 99%

Identification and Phenotypic Analysis of Two Glyoxalase II Encoding Genes from Saccharomyces cerevisiae,GLO2 and GLO4, and Intracellular Localization of the Corresponding Proteins

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Hadler

et al. 1997

Journal of Biological Chemistry

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We have isolated and characterized two genes coding for the glyoxalase II enzyme from Saccharomyces cerevisiae. The coding region of the GLO2 gene corresponds to a protein with 274 amino acids and a molecular mass of 31,306 daltons. The open reading frame of the GLO4 gene could be translated into a protein with 285 amino acids and a molecular mass of 32,325 daltons. The amino acid sequences of the deduced proteins are 59.1% identical and show high similarities to the sequence of the human glyoxalase II. When grown on either glucose or glycerol as a carbon source, a glo2 glo4 double deletion strain contains no glyoxalase II activity at all and shows no obvious phenotype during vegetative growth. However, this strain showed a similar high sensitivity against exogenous methylglyoxal as compared with a glyoxalase I-deficient strain. Whereas the GLO2 gene is expressed on both glucose and glycerol, the GLO4 gene is only active on glycerol. The active Glo2p protein is localized in the cytoplasm and the active Glo4p in the mitochondrial matrix. Heterologous expression of the full-length GLO2 coding region in Escherichia coli resulted in an active protein. However, to get an active Glo4p protein in E. coli, the putative mitochondrial transit peptide at the N-terminal end had to be removed by shortening the 5 end of the GLO4 open reading frame.

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“…Enzyme activities involved in the L-threonine catabolic pathway and the glycolytic bypath of Saccharomyces cerevisiae Cells were grown on ammonium sulfate or L-threonine as nitrogen sources and were harvested at early log (A610 nm = OS), late log (A610 nm = 1.5), early stationary ( A 6 1 0 , , , , , = 3.0) and late stationary ( A s l o ",,, = 3.6) growth. The activities of the enzymes were determined as described (1) threonine deaminase; (2) threonine aldolase; (3) threonine dehydrogenase; (4) aminoacetone synthase; (5) non-enzymatic reaction; (6) (mono)amine oxidase; (7) methylglyoxal synthase; (8) methylglyoxal reductase; (9) lactaldehyde dehydrogenase; (10) glyoxalase I; (1 1) glyoxalase IT; (1 2) lactate dehydrogenase. The activity directly converting methylglyoxal into either pyruvate or I-hydroxy-2-propanone has not been detected in the yeast strains used 1, all of the enzyme activities in the glycolytic bypath, except for a methylglyoxal synthase activity, appreciably increased when the yeast cells were grown on L-threonine as a nitrogen source.…”

Section: Discussionmentioning

confidence: 99%

“…Methylglyoxal synthase was assayed as described previously [3]. Glyoxalase I and glyoxalase I1 were assayed according to the methods described previously [18, 191.…”

Section: Enzyme Assaymentioning

confidence: 99%

“…Previously, we have purified the yeast methylglyoxal synthase and showed that the activity of the enzyme was regulated closely by phosphorus and/or various intermediates of the glycolytic pathway [3]. Following the study, we examined the enzymes in another methylglyoxalsynthesizing pathway begining with the enzyme threonine dehydrogenase and/or threonine aldolase; we now present the evidence showing that the glycolytic bypath in yeast cells is functioning as a detoxification system of methylglyoxal formed from L-threonine.…”

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Metabolism of 2-oxoaldehydes in yeasts. Possible role of glycolytic bypath as a detoxification system in l-threonine catabolism by Saccharomyces cerevisiae

et al. 1986

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L-Threonine catabolism by Saccharomyces cerevisiae was studied to determine the role of glycolytic bypath as a detoxyfication system of 2-oxoaldehyde (methylglyoxal) formed from L-threonine catabolism. During the growth on L-threonine as a sole source of nitrogen, a large amount of aminoacetone was accumulated in the culture. The enzymatic analyses indicated that L-threonine was converted into either acetaldehyde and glycine by threonine aldolase or 2-aminoacetoacetate by NAD-dependent threonine dehydrogenase. Glycine formed was condensed with acetyl-CoA by aminoacetone synthase to form 2-aminoacetoacetate, a labile compound spontaneously decarboxylated into aminoacetone. The enzyme activities of the glycolytic bypath of the cells grown on L-threonine were considerably higher than those of the cells grown on ammonium sulfate as a nitrogen source. The result indicated the possible role of glycolytic bypath as a detoxification system of methylglyoxal formed from L-threonine catabolism.Methylglyoxal, a toxic compound for cell proliferation at a millimolar concentration, is known to be synthesized from dihydroxyacetone phosphate and aminoacetone by the actions of methylglyoxal synthase [l -31 and amine oxidase [4, 51, respectively. To elucidate the function of methylglyoxal in cell proliferation, we have examined the metabolic fate of methylglyoxal in yeast Saccharomyces cerevisiae cells and found an alternative route for methylglyoxal degradation besides the glyoxalase system consisting of glyoxalase I and glyoxalase 11. In this route, methylglyoxal is converted into L-lactaldehyde by NADPH-dependent methylglyoxal reductase [6] and L-lactaldehyde is then converted to L-lactate by NAD-dependent L-lactaldehyde dehydrogenase [7]. The NADPH-dependent methylglyoxal reductase has been found in all the microbial (Escherichia, Bacillus, Pseudomonas species, Actinomycetes and molds) and mammalian (brain, liver, heart, spleen and kidney of rat) cells at a considerably high activity and was thought to have a significant role in methylglyoxal metabolism in combination with the glyoxalase system (unpublished results).The analyses of all the enzymes responsible for methylglyoxal catabolism in yeast cells strongly suggested that the methylglyoxal was functioning as a regulator of the growth of yeast cells [8, 91, as was first suggested by Szent-Gyorgyi [lo]. Dudani et al. also demonstrated that the activity of the glyoxalase system was a good indicator for yeast cell growth [l 11. So, the synthesis and/or degradation of methylglyoxal seems to be regulated to evade an over-accumulation of Correspondence to K. Murata,

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Characterization of methylglyoxal synthase in Saccharomyces cerevisiae

Cited by 59 publications

References 7 publications

Microbial production and applications of 1,2-propanediol

Microbial production and applications of 1,2-propanediol

Identification and Phenotypic Analysis of Two Glyoxalase II Encoding Genes from Saccharomyces cerevisiae,GLO2 and GLO4, and Intracellular Localization of the Corresponding Proteins

Metabolism of 2-oxoaldehydes in yeasts. Possible role of glycolytic bypath as a detoxification system in l-threonine catabolism by Saccharomyces cerevisiae

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