Solution Chemistry of Dihydroxyacetone and Synthesis of Monomeric Dihydroxyacetone. Chemical research in toxicology Dihydroxyacetone (DHA) is a major byproduct of e-cigarette combustion and is the active ingredient in sunless tanning products. Mounting evidence points to its damaging effects on cellular functions. While developing a simple synthetic route to monomeric [C]DHA for flux metabolic studies that compared DHA and glyceraldehyde (GA) metabolism, we uncovered that solid DHA ages upon storage and differences in the relative abundance of each of its isomer occur when reconstituted in an aqueous solution. While all three of the dimeric forms of DHA ultimately resolve to the ketone and hydrated forms of monomeric DHA once in water at room temperature, these species require hours rather than minutes to reach an equilibrium favoring the monomeric species. Consequently, when used in bolus or flux experiments, the relative abundance of each isomer and its effects at the time of application is dependent on the initial DHA isomeric composition and concentration, and time of equilibration in solution before use. Here, we make recommendations for the more consistent handling of DHA as we report conditions that ensure that DHA is present in its monomeric form while in solutions, conditions used in an isotopic tracing study that specifically compared monomeric DHA and GA metabolism in cells. 10.1021/acs.chemrestox.1c00403

Phosphatidic acid biosynthesis in the model organism yeast Saccharomyces cerevisiae - a survey. Biochimica et biophysica acta. Molecular and cell biology of lipids Phosphatidic acid biosynthesis represents the initial part of de novo formation of all glycerophospholipids (membrane lipids) as well as triacylglycerols (storage lipids), and is thus the centerpiece of glycerolipid metabolism. The universal route of phosphatidic acid biosynthesis starts from the precursor glycerol-3-phosphate and comprises two consecutive acylation reactions which are catalyzed by a glycerol-3-phosphate acyltransferase and a 1-acyl glycerol-3-phosphate acyltransferase. In addition, yeast and mammals harbor a set of enzymes which can synthesize phosphatidic acid from the precursor dihydroxyacetone phosphate. In the present review our current knowledge about enzymes contributing to phosphatidic acid biosynthesis in the invaluable model organism yeast, Saccharomyces cerevisiae, is summarized. A special focus is laid upon the regulation and the localization of these enzymes. Furthermore, research needs for a deeper insight into the high complexity of phosphatidic acid biosynthesis and consequently the entire lipid metabolic network is presented. 10.1016/j.bbalip.2021.158907

Dihydroxyacetone phosphate, DHAP, in the crystalline state: monomeric and dimeric forms. Slepokura Katarzyna,Lis Tadeusz Carbohydrate research It was shown that dihydroxyacetone phosphate may exist in both monomeric DHAP (C(3)H(7)O(6)P) and dimeric DHAP-dimer (C(6)H(14)O(12)P(2)) form. Monomeric DHAP was obtained in the form of four crystalline salts: CaCl(DHAP) x 2.9H(2)O (7a), Ca(2)Cl(3)(DHAP) x 5H(2)O (7b), CaCl(DHAP) x 2H(2)O (7c), and CaBr(DHAP) x 5H(2)O (7d) by crystallization from aqueous solutions containing DHAP acid and CaCl(2) or CaBr(2), or by direct crystallization from a solution containing DHAP precursor and CaCl(2). At least one of the salts is stable and may be stored in the crystalline state at room temperature for several months. The dimeric form was obtained by slow saturation of free DHAP syrup with ammonia at -18 degrees C and isolated in the form of its hydrated diammonium salt (NH(4))(2)(DHAP-dimer) x 4H(2)O (8). The synthesis of the compounds, their crystallization, and crystal structures determined by X-ray crystallography are described. In all 7a-d monomeric DHAP exists in the monoanionic form in an extended (in-plane) cisoid conformation, with both hydroxyl and ester oxygen atoms being synperiplanar to the carbonyl O atom. The crucial structural feature is the coordination manner, in which the terminal phosphate oxygen atoms act as chelating as well as bridging atoms for the calcium cations. Additionally, the DHAP monoanions chelate another Ca(2+) by the alpha-hydroxycarbonyl moiety, in a manner observed previously in dihydroxyacetone (DHA) calcium chloride complexes. In dimeric 8 the anion is a trans isomer with the dioxane ring in a chair conformation with the hydroxyl groups in axial positions and the phosphomethyl group in an equatorial position. 10.1016/j.carres.2009.12.008

Molecular mechanism of GylR-mediated regulation of glycerol metabolism in NRRL 3585. Frontiers in microbiology Glycerol is a readily available and low-cost simple polyol compound, which can be used as a carbon source for microorganisms to produce various value-added products. Understanding the underlying regulatory mechanism in glycerol metabolism is critical for making better use of glycerol for diverse applications. In a few reported strains, the glycerol utilization gene cluster ( operon) was shown to be regulated by the IclR family transcriptional regulator GylR. However, the molecular regulatory mechanism mediated by GylR has not been fully elucidated. In this study, we first analyzed the available genomes in the NCBI Genome database, and found that the operon-like gene clusters are conserved in and several other genera of . By taking NRRL 3585 as a model system, we identified that GylR represses the expressions of operon and by directly binding to their promoter regions. Both glycerol-3-phosphate and dihydroxyacetone phosphate can induce the dissociation of GylR from its binding sequences. Furthermore, we identified a minimal essential operator site (a palindromic 18-bp sequence) of GylR-like regulators in . Our study for the first time reported the binding sequences and effector molecules of GylR-like proteins in . The molecular regulatory mechanism mediated by GylR presumably exists widely in . Our findings would facilitate the design of glycerol utilization pathways for producing valuable products. Moreover, our study provided new basic elements for the development of glycerol-inducible regulatory tools for synthetic biology research in the future. 10.3389/fmicb.2022.1078293

Dissecting carbon metabolism of type strain W29 using genome-scale metabolic modelling. Computational and structural biotechnology journal is a widely-used chassis cell in biotechnological applications. It has recently gained extensive research interest owing to its extraordinary ability of producing industrially valuable biochemicals from a variety of carbon sources. Genome-scale metabolic models (GSMMs) enable analyses of cellular metabolism for engineering various industrial hosts. In the present study, we developed a high-quality GSMM Yli21 for type strain W29 by extensive manual curation with Biolog experimental data. The model showed a high accuracy of 85.7% in predicting nutrient utilization. Transcriptomics data were integrated to delineate cellular metabolism of utilizing six individual metabolites as sole carbon sources. Comparisons showed that 302 reactions were commonly used, including those from TCA cycle, oxidative phosphorylation, and purine metabolism for energy and material supply. Whereas glycolytic reactions were employed only when glucose and glycerol used as sole carbon sources, gluconeogenesis and fatty acid oxidation reactions were specifically employed when fatty acid, alkane and glycerolipid were the sole carbon sources. Further test of 46 substrates for generating 5 products showed that hexanoate outcompeted other compounds in terms of maximum theoretical yield owing to the lowest carbon loss for energy supply. This newly generated model Yli21 will be a valuable tool in dissecting metabolic mechanism and guiding metabolic engineering of this important industrial cell factory. 10.1016/j.csbj.2022.05.018