Showing papers on "Methylglyoxal published in 1976"

Purification and properties of formaldehyde dehydrogenase and formate dehydrogenase from Candida boidinii.

Abstract: Formaldehyde hydrogenase and formate dehydrogenase were purified 130-fold and 19-fold respectively from Candida boidinii grown on methanol. The final enzyme preparations were homogenous as judged by acrylamide gel electrophoresis and by sedimentation in an ultracentrifuge. The molecular weights of the enzymes were determined by sedimentation equilibrium studies and calculated as 80000 and 74000 respectively. Dissociation into subunits was observed by treatment with sodium dodecylsulfate. The molecular weights of the polypeptide chains were estimated to be 40000 and 36000 respectively. The NAD-linked formaldehyde dehydrogenase specifically requires reduced glutathione for activity. Besides formaldehyde only methylglyoxal served as a substrate but no other aldehyde tested. The Km values were found to be 0.25 mM for formaldehyde, 1.2 mM for methylglyoxal, 0.09 mM for NAD and 0.13 mM for glutathione. Evidence is presented which demonstrates that the reaction product of the formaldehyde-dehydrogenase-catalyzed oxidation of formaldehyde is S-formylglutathione rather than formate. The NAD-linked formate dehydrogenase catalyzes specifically the oxidation of formate to carbon dioxide. The Km values were found to be 13 mM for formate and 0.09 mM for NAD.

The electronic relaxation of methylglyoxal in the vapor phase

Abstract: The time-resolved fluorescence of methylglyoxal was studied as a function of excitation energy and pressure. In all circumstances a dual fluorescence was observed. From these measurements was obtained the parameters describing the singlet-triplet coupling, the disspative leak rates from singlet and triplet states and the fluorescence quenching constants. Also, the pressure induction of the thermalized phosphorescence was studied. As found previously in biacetyl, the excited manifold should be divided into a black note region and an overlap region, the former being more extensive in methylglyoxal. The decal characteristics of methylglyoxal are intermediate between those of glyoxal and biacetyl. The differences in behavior between the three molecules can be readily understood on the basis of the differences in their level densities, due to the different number of atoms in these molecules.

Some observations on the NADP+-linked oxidation of methylglyoxal catalysed by 2-Oxoaldehyde dehydrogenase

Abstract: In the oxidation of methylglyoxal by 2-oxoaldehyde dehydrogenase, the apparent Km value for NADP+ was about 2.5 times lower than the corresponding Km for NAD+; the apparent Km values for methylglyoxal and for the amine activator L-2-aminopropan-1-ol, with NADP+ as cofactor, were also different from those obtained with NAD+. In the presence of NADP+, the enzyme was not activated by P1, in contrast with the activation of the enzyme when NAD+ was used. The significance of the results is discussed.

The Reaction Mechanism of Rat Liver Glyoxalase I and Its Inhibition by MS-3

Abstract: Glyoxalase I from rat liver was purified about 25-fold by acetone fractionation and ion-exchange chromatography on CM-Sephadex and DEAE-cellulose columns. The kinetic study of the enzymatic reaction supported the one-substrate mechanism : the hemimercaptal adduct produced nonenzymatically from methylglyoxal and glutathione is the substrate. The Km value determined was 0.1 mm and similar to that of porcine erythrocytes enzyme but differed significantly from that of yeast enzyme. It was inhibited by free glutathione competitively (Ki 1.2 mm). Kinetic studies on inhibition of glyoxalase I by MS–3 which was obtained from a cultured mushroom, Stereum hirsutum, indicated the inhibition type was competitive with the hemimercaptal adduct (Ki 4.6 × 10−6 m). By the graphical study of the multiple inhibition kinetics free glutathione and MS–3 were shown to bind at the same sites of the enzyme.

Growth inhibition of Proteus mirabilis by cyclic adenosine 3'-5'-monophosphate.

Abstract: Growth of Proteus mirabilis on a synthetic agar medium containing either glycerol, galactose, or trehalose as the sole source is inhibited by 5 mM cyclic adenosine 3',5'-monophosphate (cAMP). Inhibition on an agar medium is evident as loss of viability, but in broth cAMP only slightly inhibits growth rate. Inhibition is associated with the accumulation of methylglyoxal in the medium. A nonswarming mutant of P. mirabilis is not inhibited by cAMP on either of the three carbon sources, but it is sensitive to exogenous methylglyoxal.

Recovery of methylglyoxal acetal by plural stage distillation with intermediate phase separation

Abstract: A process for the recovery of methylglyoxal acetal from a reaction mixture which has been obtained in the reaction of acetone with an alcohol and a nitrosation agent or oxidizing agent in the presence of an acid catalyst followed by neutralization which comprises diluting the neutralized reaction mixture with such an amount of water that a homogeneous solution is formed, subsequent distillation in a first column in which a mixture of acetone, alcohol and solvent is removed overhead and returned direct to the synthesis and an azeotrope of methylglyoxal acetal, water and high-boilers is withdrawn as a side stream, condensation of the azeotrope thus obtained and separation of the phase rich in methylglyoxal acetal, subsequent distillation of the phase rich in methylglyoxal acetal thus obtained in a second column in which purified methylglyoxal acetal is withdrawn as a side stream and the overhead and bottoms products are returned with the feed to the first column, pure methylglyoxal acetal being obtained as the overhead product in a third column. Methylglyoxal acetal is suitable for the production of animal feed supplements.