Methylglyoxal

About: Methylglyoxal is a research topic. Over the lifetime, 2844 publications have been published within this topic receiving 102037 citations. The topic is also known as: acetylformaldehyde & pyruvaldehyde.

Glyoxal oxidase of Phanerochaete chrysosporium: Its characterization and activation by lignin peroxidase (hydrogen peroxide/methylglyoxal/white-rot fungus/wood decay)

Abstract: Glyoxal oxidase (GLOX) is an extracellular H2O2-generating enzyme produced by ligninolytic cultures of Phanerochaete chrysosporium. The production, purification, and partial characterization of GLOX from agitated cultures are described here. High-oxygen levels are critical for GLOX production as for lignin peroxidase. GLOX purified by anion- exchange chromatography appears homogeneous by NaDod- SO4/PAGE (molecular mass = 68 kDa). However, analysis by isoelectric focusing indicates two major bands (pI 4.7 and 4.9) that stain as glycoproteins as well as for H2O2-producing activity in the presence of methylglyoxal. Purified GLOX shows a marked stimulation in activity when incubated with Cu2+; full activation takes more than 1 hr with 1 mM CuSO4 at pH 6. The steady-state kinetic parameters for the GLOX oxidation of methylglyoxal, glyceraldehyde, dihydroxyacetone, glycol- aldehyde, acetaldehyde, glyoxal, glyoxylic acid, and formal- dehyde, were determined by using a lignin peroxidase coupled- assay at pH 4.5. Of these substrates, the best is the extracellular metabolite methylglyoxal with a Km of 0.64 mM and an apparent rate of catalysis, kcat, of 198 s-1 under air-saturated conditions. The Km for oxygen is greater than the concentration of oxygen possible at ambient pressure—i.e., >1.3 mM at 25°C. Importantly, oxygen-uptake experiments show that pu- rified GLOX is inactive unless coupled to the peroxidase reaction. With this coupled reaction, for each mol of methyl- glyoxal, veratryl alcohol (a lignin peroxidase substrate), and oxygen consumed, 1 mol each of pyruvate and veratraldehyde is produced. The importance of these results is discussed in relation to the physiology of lignin biodegradation and possible extracellular regulatory mechanisms for the control of oxidase and peroxidase activities.

Trapping reactions of reactive carbonyl species with tea polyphenols in simulated physiological conditions

Abstract: The carbonyl stress that leads to the formation of advanced glycation end products (AGEs) in diabetes mellitus has drawn much attention recently. Reactive alpha-dicarbonyl compounds, such as glyoxal (GO) and methylglyoxal (MGO), have been shown to be a high potential glycation agent in vitro and in vivo. In this study, epicatechins in green tea and theaflavins in black tea were found to be able to reduce the concentration of MGO in physiological phosphate buffer conditions. Modified MGO derivatization for GC/flame ionization detector (FID) method in quantification was systematically conducted. In molar ratio of 3 (MGO/polyphenol), theaflavin-3,3'-digallate (TF3) in theaflavins and (-)-epigallocatechin (EGC) in epicatechins showed the highest MGO reduction at 66.65 and 45.74%, respectively, after 1 h of incubation. In kinetic study (molar ratio of MGO/polyphenol = 1:1), rapid MGO reduction occurred within 10 min. Identities of primary adducts between (-)-epigallocatechin gallate (EGCG) and MGO were determined. Newly generated stereoisomers at the C8 position of EGCG A-ring were isolated with a chiral column, and structurally confirmed by 2-D NMR analyses.

Measurement of methylglyoxal by stable isotopic dilution analysis LC-MS/MS with corroborative prediction in physiological samples

Abstract: This protocol describes a method for the detection and quantification of methylglyoxal (MG), the major physiological substrate of the cytosolic glyoxalase system. Accumulation of MG, also called dicarbonyl stress, is implicated in tissue damage in aging and disease. Measurement of MG is important in physiological studies, in the development of glyoxalase 1 (Glo1) inducer and inhibitor therapeutics, and in the characterization of medical products, especially dialysis fluids, and of thermally processed foods and beverages. MG can be derivatized with 1,2-diaminobenzene (DB), resulting in an adduct that can be detected using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Quantification is achieved by stable isotopic dilution analysis with [(13)C3]MG. Pre-analytic processing at ambient temperature, under acidic conditions with peroxidase inhibition, avoids artifactual overestimation of MG. Estimates obtained from physiological samples can be validated by kinetic modeling of in situ rates of protein glycation by MG for confirmation of the results. This procedure was developed for the analysis of cultured cells, plasma and animal tissue samples, and it can also be used to analyze plant material. Experimental measurement requires 4.5 h for sample batch pre-analytic processing and 30 min per sample for LC-MS/MS analysis.

Quercetin Inhibits Advanced Glycation End Product Formation by Trapping Methylglyoxal and Glyoxal

Abstract: Methylglyoxal (MGO) and glyoxal (GO) not only are endogenous metabolites but also exist in exogenous resources, such as foods, beverages, urban atmosphere, and cigarette smoke. They have been identified as reactive dicarbonyl precursors of advanced glycation end products (AGEs), which have been associated with diabetes-related long-term complications. In this study, quercetin, a natural flavonol found in fruits, vegetables, leaves, and grains, could effectively inhibit the formation of AGEs in a dose-dependent manner via trapping reactive dicarbonyl compounds. More than 50.5% of GO and 80.1% of MGO were trapped at the same time by quercetin within 1 h under physiological conditions. Quercetin and MGO (or GO) were combined at different ratios, and the products generated from this reaction were analyzed with LC-MS. Both mono-MGO and di-MGO adducts of quercetin were detected in this assay using LC-MS, but only tiny amounts of mono-GO adducts of quercetin were found. Additionally, di-MGO adducts were observed...