Yeast pyruvate metabolism

Most known thiamin diphosphate-dependent reactions (Table 14-2) can be derived from the five halfreactions, a through e, shown in Fig. 14-3. Each halfreaction is an a cleavage which leads to a thiamin- bound enamine (center, Fig. 14-3) The decarboxylation of an a-oxo acid to an aldehyde is represented by step b followed by a in reverse. The most studied enzyme catalyzing a reaction of this type is yeast pyruvate decarboxylase, an enzyme essential to alcoholic fermentation (Fig. 10-3). There are two 250-kDa isoenzyme forms, one an a4 tetramer and one with an ( P)2 quaternary structure. The isolation of ohydroxyethylthiamin diphosphate from reaction mixtures of this enzyme with pyruvate52 provided important verification of the mechanisms of Eqs. 14-14,14-15. Other decarboxylases produce aldehydes in specialized metabolic pathways indolepyruvate decarboxylase126 in the biosynthesis of the plant hormone indoIe-3-acetate and ben-zoylformate decarboxylase in the mandelate pathway of bacterial metabolism (Chapter 25).1243/127... 

Hohmann, S. (1996) Pyruvate decarboxylases. In F.K. Zimmerman K.D. Entian (Eds.), Yeast Sugar Metabolism Biochemistry, Genetics, Biotechnology, and Applications (pp. 187-212). Boca Raton CRC Press. 

Bertrand, 1994 Allen, 1995) decanal and ( )-2-nonenal, on the other hand, are associated with sawdust or plank odour (Chatonnet and Dubourdieu, 1996 1998). The principal carbonyl compound formed in MLF is 2,3-butanedione (diacetyl), whose level can improve, or affect, the wine with its butter-like or fat note (Davis et al., 1985). Diacetyl and 3-hydroxy-2-butanone (acetoin, the reduced form of diacetyl) are produced by pyruvate metabolism of yeasts and lactic bacteria, and their levels may increase two or three fold with MLF depending on the lactic bacteria strain involved (Davis et al., 1985 Martineau and Henick-Kling, 1995 Radler, 1962 Fornachon and Lloyd, 1965 Rankine et al., 1969 Mascarenhas, 1984). For diacetyl in wine sensory thresholds ranging from 0.2mg/L (in Cbardonnay) to 0.9mg/L (Pinot noir), and 2.8 mg/L (Cabernet Sauvignon wine), are reported (Martineau et al., 1995). 

As the study of CoA developed, it became apparent that the coenzyme was involved in reactions whereby acetate was activated by ATP and subsequently transferred to various acetyl acceptors. In pigeon liver extracts it was shown that acetate could be activated by ATP in the presence of CoA to acetylate sulfanilamide, PABA, histamine, glucosamine, to synthesize acetoacetic acid and citrate. Acetyl phosphate, which has been demonstrated to be a product of pyruvate metabolism in several bacteria and could theoretically be considered to be an intermediate in these reactions, was found to be unable to replace acetate and ATP in animal tissues. Eventually it was shown that there is present in certain bacteria an enzyme, phosphotransacetylase, which could convert acetyl phosphate to a reactive product which was thought to be acetyl-CoA.i 194 isolation of acetyl-CoA from yeast extract by Lynen and Reichert confirmed the idea that acetyl-CA is the reactive 2-carbon unit in these reactions. Stadtman has demonstrated that acetyl-CoA is indeed the product of the action of phosphotransacetylase. Lipmann has recently... 

Presecan-Siedel E et al (1999) Cataholite regulation of the pta gene as part of carbon flow pathways in Bacillus subtilis. J Bacteriol 181(22) 6889-97 Pronk JT, Yde Steensma H, Van Dijken JP (1996) Pyruvate metabolism in Saccharomyces cerevisiae. Yeast 12(16) 1607—33... 

Though use of isolated purified enzymes is advantageous in that undesirable byproduct formation mediated by contaminating enzymes is avoided [37], in many industrial biotransformation processes for greater cost effectiveness the biocatalyst used is in the form of whole cells. For this reason baker s yeast, which is readily available, has attracted substantial attention from organic chemists as a catalyst for biotransformation processes. One of the first commercialized microbial biotransformation processes was baker s yeast-mediated production of (R)-phenylacetyl carbinol, where yeast pyruvate decarboxylase catalyzes acyloin formation during metabolism of sugars or pyruvate in the presence of benzaldehyde [38]. 

Fluoroethanol itself is innocuous towards a variety of tissue constituents, a series of enzymes in rat-liver mince, and the respiration and metabolism in liver, kidney, heart and brain slice.3 After a period of incubation in those tissues known to contain alcohol dehydrogenase, e.g. liver and kidney, the respiration and pyruvate oxidation were strongly inhibited. Likewise, following a period of incubation with yeast, acetate oxidation was blocked. These inhibitions were similar to those produced by fluoroacetate, and the facts can best be explained by the oxidation of fluoroethanol to fluoroacetic acid by alcohol dehydrogenase. 

Some catabolic reactions depend upon ADP, but under most conditions its concentration is very low because it is nearly all phosphorylated to ATP. Reactions utilizing ADP may then become the rate-limiting pacemakers in reaction sequences. Depletion of a reactant sometimes has the effect of changing the whole pattern of metabolism. Thus, if oxygen is unavailable to a yeast, the reduced coenzyme NADH accumulates and reduces pyruvate to ethanol plus C02 (Fig. 10-3). The result is a shift from oxidative metabolism to fermentation. 

Why do we need vitamins Early clues came in 1935 when nicotinamide was found in NAD+ by H. von Euler and associates and in NADP+ by Warburg and Christian. Two years later, K. Lohman and P. Schuster isolated pure cocarboxylase, a dialyz-able material required for decarboxylation of pyruvate by an enzyme from yeast. It was shown to be thiamin diphosphate (Fig. 15-3). Most of the water-soluble vitamins are converted into coenzymes or are covalently bound into active sites of enzymes. Some lipid-soluble vitamins have similar functions but others, such as vitamin D and some metabolites of vitamin A, act more like hormones, binding to receptors that control gene expression or other aspects of metabolism. 

This, the final step in alcohol fermentation, is analogous to lactate fermentation. Both reactions regenerate NAD+ and produce low-molecular-weight, water-soluble, metabolic end products that diffuse out of the cells in which they were produced. In the case of alcoholic fermentation, the second reaction is reversible, so that if oxygen becomes available to previously anaerobic yeast cells, the ethanol is oxidized to acetaldehyde. Unlike lactate fermentation, in which the lactate is oxidized to pyruvate, alcoholic fermentation cannot form pyruvate from acetaldehyde. Instead, the acetaldehyde... 

Pyruvate produced by glycolysis can be used by yeasts for several metabolic pathways. However, yeasts must regenerate NAD+ from the NADH to re-establish the oxydoreduction potential of the cell. This can be done by fermentation or respiration. 

The transformation of pyruvate into ethanal or acetyl-coA is therefore a key point for regulating yeast metabolism (Rib6reau-Gayon et al. 2000c). 

Precursors. Precursors for this reaction are compounds exhibiting keto-enol tau-tomerism. These compounds are usually secondary metabolites derived from the glycolysis cycle of yeast metabolism during fermentation. Pyruvic acid is one of the main precursor compounds involved in this type of reaction. During yeast fermentation it is decarboxylated to acetaldehyde and then reduced to ethanol. Acetone, ace-toin (3-hydroxybutan-2-one), oxalacetic acid, acetoacetic acid and diacetyl, among others, are also secondary metabolites likely to participate in this kind of condensation reaction with anthocyanins. 

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