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Simultaneous and direct determination of oxalic acid, tartaric acid, malic acid, vitamin C, citric acid, and succinic acid in Fructus mume by reversed-phase high-performance liquid chromatography.

A method for the simultaneous separation and direct determination of oxalic acid (OA), tartaric acid (TA), malic acid (MA), vitamin C (VC), citric acid (CA), and succinic acid (SA) in Fructus mume using reversed-phase high-performance liquid chromatography with a UV detector in an acidic medium is presented in this study. In the experiment, the optimization of chromatographic conditions (i.e., the pH and flow rate of the mobile phase, the absorption wavelength, and temperature of column) that affect the separation degree and peak shape of organic acids has been obtained. The linear ranges are found to be 0.05-4.7 microg for OA (r = 0.9999), 0.11-10.5 microg for TA (r = 0.9999), 0.114-11.4 microg for MA (r = 0.9999), 0.033-3.30 microg for VC (r = 0.9999), 0.155-15.5 microg for CA (r = 0.9998), and 0.194-19.4 microg for SA (r = 0.9996). For OA, TA, MA, VC, CA, and SA, the even recovery (n = 3) of six effective components are 100.9%, 99.97%, 101.2%, 102.1%, 101.1%, and 100.7%, respectively, and the largest relative standard deviation (n = 11) for the six components is less than 1.7%. The detection limits are 0.01 microg for OA, TA, and VC; 0.05 microg for MA; 0.03 microg for CA; and 0.1 microg for SA. In a single chromatographic run, OA, TA, MA, VC, CA, and SA can be determined in less than 7 min. The method can be used for the purpose of routine analysis and the quality control of a botanic (Fructus mume) containing these effective components.

Ascorbic Acid↗

Influence of pH, malic acid and glucose concentrations on malic acid consumption by Saccharomyces cerevisiae.

Malic acid consumption by Saccharomyces cerevisiae was studied in a synthetic medium. The extent of malic acid degradation is affected by its initial concentration, the extent and the rate of deacidification increased with initial malate concentration up to 10 milligrams. For malic acid consumption, an optimal pH range of 3-3.5 was found, confirming that non-dissociated organic acids enter S. cerevisiae cells by simple diffusion. A full factorial design has been employed to describe a statistical model of the effect of sugar and malic acid on the quantity of malate degraded (milligrams) by a given amount of biomass (milligrams). The results indicated that the initial malic acid concentration is very important for the ratio of malate consumption to quantity of biomass. The yeast was found to be most efficient at higher levels of malate.

Biomass↗

Sucrose and malic acid in the tobacco plant induce hrp regulon in a phytopathogen Ralstonia pseudosolanacearum.

Genes encoding a type III secretion system in Ralstonia pseudosolanacearum are regulated by HrpB as an hrp regulon and induced only in plants. This study aimed to identify the plant signals that induce the hrp regulon and confirm the signal recognition mechanism. Signaling molecules that induce hrpB expression were screened using resting cells of the hrpB-lacZ reporter strain. Only the soluble fraction of smashed tobacco seedlings induced hrpB expression. The heated soluble fraction retained its hrpB-inducing activity, indicating that the signaling molecules were not proteins. When the soluble fraction was fractionated into acidic, neutral, and basic components, both the acidic and neutral fractions induced hrpB expression. As neutral compounds, sucrose, glucose, and fructose have been found to induce hrpB expression. Sucrose-induced hrpB expression was greatly reduced in the prhA mutant, indicating that the TonB-dependent receptor PrhA perceives sugars. Among the organic acids found in the acidic fractions, malic acid most efficiently induced hrpB expression, which was reduced by the mutation of a hybrid histidine kinase gene of a two-component system, rsc1598, indicating that Rsc1598 may sense malic acid. We demonstrated direct binding of Rsc1598 to malic acid using isothermal titration calorimetry.IMPORTANCESimilar to other Gram-negative plant pathogens, the type III secretion system (T3SS) is the most important virulence factor in Ralstonia pseudosolanacearum. The genes for the T3SS are regulated as an hrp regulon, activated only when the pathogen encounters the plants, indicating that the pathogen must sense plant signals. For the first time, we identified two signaling compounds, sucrose and malic acid, that are abundantly found in tobacco roots. The hrp operon was induced even in non-host plants, possibly because sucrose and malic acid are common in plants. We also found that R. pseudosolanacearum membrane proteins received sucrose and malic acid independently. As a next step, antagonists of signaling molecules can be screened.

Malates↗

[Studies on the correlation between production of L-malic acid and some cytosolic enzymes in the L-malic acid producing strain Aspergillus sp. N1-14].

The cytosol enzymatic study in the case of high L-malic acid(LMA) production of Aspergillus sp. N1-14' was reported. The activities of 4 kind enzymes that catalyse the CO2 fixation reactions have been detected, which are pyruvate carboxylase(PC), phosphoenolpyruvate carboxlase (PEPC), phosphoenolpyurvate carboxykinase(PCK) and malic enzyme(ME). With the exception of ME, the linear correlation was found between activities of three carboxlases and the production rate of LMA. The activity of malate dehydrogenase(MDH) was at the level of 2-3 exponential higher than that of the other analysed enzymes, while the activity of succinate dehydrogenase(SDH) was much lower, and as a discrepancy, SDH was in a positive correlation to the content of LMA in fermenting slurry(r = 0.9252). It is shown that the accumulated LMA acted as an activator of SDH. Through dynamic study, it is found that, in contrast with the slow and even increase of biomass, the content of cytosol protein(Cp) sharply fluctuated mainly due to the changes of aeration conditions. The data of the linear correlation coefficients(r) of activities of cytosol enzymes to Cp(PC r = 0.9563, PEPC r = 0.7688, PCK r = 0.7300, MDH r = 0.3920, SDH r = -0.2086) exhibited an inner law of protein synthesis. Experiment of increasing the amount of spore inoculum resulted in increase of LMA and decrease of SA. After fermenting 120 h in a 5 L stirred fermentor, with 3-fold of original spore inoculum 105.88 g/L of LMA was achieved, the overall productivity was 0.883 g/(L.h), the converting rate of glucose to LMA was 78.43%. This result supports the exist of a inner law of protein synthesis in the early period of LMA fermentation by Aspergillus sp. N1-14'.

Aspergillus↗

Final report on the safety assessment of Malic Acid and Sodium Malate.

Malic Acid functions in cosmetic formulations as a pH adjuster, and Sodium Malate functions as a skin conditioning agent-humectant. Malic Acid is reportedly used in almost 50 cosmetic formulations across a range of product types at low concentrations, whereas Sodium Malate is used in only one. As a pH adjuster, Malic Acid is used at low concentrations. One commercial method of preparing Malic Acid is hydration of fumaric acid or maleic acid, and then purified to limit the amount of the starting material present. Because Malic Acid is a component of the Kreb's cycle, another method is fermentation. Malic Acid was relatively nontoxic in acute toxicity studies using animals. In a chronic oral study, feeding Malic Acid to rats resulted only in weight gain changes and changes in feed consumption. Malic Acid did not cause reproductive toxicity in mice, rats, or rabbits. Malic Acid was a moderate to strong skin irritatant in animal tests, and was a strong ocular irritant. Malic Acid was not mutagenic across a range of genotoxicity tests. Malic Acid was irritating in clinical tests, with less irritation seen as pH of the applied material increased. Patients patch tested with Malic Acid, placed on a diet that avoided foods containing Malic or citric acid, and then challenged with a diet high in Malic and citric acid had both immediate urticarial and delayed contact dermatitis reactions. These data were considered sufficient to determine that Malic Acid and Sodium Malate would be safe at the low concentrations at which these ingredients would be used to adjust pH (even though Sodium Malate is not currently used for that purpose). The data, however, were insufficient to determine the safety of these ingredients when used in cosmetics as other than pH adjusters and specifically, the data are insufficient to determine the safety of Sodium Malate when used as a skin conditioning agent-humectant. The types of data required for the Expert Panel to determine the safety of Sodium Malate as a skin-conditioning agent are: concentration of use data; dermal irritation and sensitization data; and ocular irritation data, if available. The data needed to assess the safety of Malic Acid or Sodium Malate for some function other than as a skin-conditioning agent cannot be specified without knowing the intended function. Were these ingredients to be used as exfoliants, for example, data similar to that included in the Cosmetic Ingredient Review safety assessment of Glycolic Acid would be needed. Until these data are available, it is concluded that the available data are insufficient to support the safety of these ingredients in cosmetic formulations for functions other than use as a pH adjuster.

Animals↗

Simultaneous determination of oxalic, fumaric, maleic and succinic acids in tartaric and malic acids for pharmaceutical use by ion-suppression reversed-phase high performance liquid chromatography.

A reliable method for the simultaneous determination of oxalic, fumaric, maleic, and succinic acids in tartaric and malic acids for pharmaceutical use by reversed-phase ion-suppression high performance liquid chromatography is presented. HPLC was achieved on a Nova-Pak C18 column by isocratic elution using water adjusted to pH 2.10-2.15 with perchloric acid, and detection was by UV adsorption at a wavelength of 210 nm. This method was found to be superior to previous liquid chromatography as well as other classical assay, and to be an attractive choice for the analysis of these compounds.

Chromatography, High Pressure Liquid↗

Inducible overexpression of the FUM1 gene in Saccharomyces cerevisiae: localization of fumarase and efficient fumaric acid bioconversion to L-malic acid.

Cloning of the Saccharomyces cerevisiae FUM1 gene downstream of the strong GAL10 promoter resulted in inducible overexpression of fumarase in the yeast. The overproducing strain exhibited efficient bioconversion of fumaric acid to L-malic acid with an apparent conversion value of 88% and a conversion rate of 80.4 mmol of fumaric acid/h per g of cell wet weight, both of which are much higher than parameters known for industrial bacterial strains. The only product of the conversion reaction was L-malic acid, which was essentially free of the unwanted by-product succinic acid. The GAL10 promoter situated upstream of a promoterless FUM1 gene led to production and correct distribution of the two fumarase isoenzyme activities between cytosolic and mitochondrial subcellular fractions. The amino-terminal sequence of fumarase contains the mitochondrial signal sequence since (i) 92 of 463 amino acid residues from the amino terminus of fumarase are sufficient to localize fumarase-lacZ fusions to mitochondria and (ii) fumarase and fumarase-lacZ fusions lacking the amino-terminal sequence are localized exclusively in the cytosol. The possibility that both mitochondrial and cytosolic fumarases are derived from the same initial translation product is discussed.

Cloning, Molecular↗

Differential malic acid degradation by selected strains of Saccharomyces during alcoholic fermentation.

To produce a high-quality wine, it is important to obtain a fine balance between the various chemical constituents, especially between the sugar and acid content. The latter is more difficult to achieve in wines that have high acidity due to excess malic acid, since wine yeast in general cannot effectively degrade malic acid during alcoholic fermentation. An indigenous Saccharomyces paradoxus strain RO88 was able to degrade 38% of the malic acid in Chardonnay must and produced a wine of good quality. In comparison, Schizosaccharomyces pombe strain F effectively removed 90% of the malic acid, but did not produce a good-quality wine. Although commercially promoted as a malic-acid-degrading wine yeast strain, only 18% of the malic acid was degraded by Saccharomyces cerevisiae Lalvin strain 71B. Preliminary studies on the transcriptional regulation of the malic enzyme gene from three Saccharomyces strains, i.e. S. paradoxus RO88, S. cerevisiae 71B and Saccharomyces bayanus EC1118, were undertaken to elucidate the differences in their ability to degrade malic acid. Expression of the malic enzyme gene from S. paradoxus RO88 and S. cerevisiae 71B increased towards the end of fermentation once glucose was depleted, whereas no increase in transcription was observed for S. bayanus EC1118 which was also unable to effectively degrade malic acid.

Base Sequence↗

Modelling malic acid accumulation in fruits: relationships with organic acids, potassium, and temperature.

Malic acid production, degradation, and storage during fruit development have been modelled. The model assumes that malic acid content is determined essentially by the conditions of its storage in the mesocarp cells, and provides a simplified representation of the mechanisms involved in the accumulation of malate in the vacuole and their regulation by thermodynamic constraints. Solving the corresponding system of equations made it possible to predict the malic acid content of the fruit as a function of organic acids, potassium concentration, and temperature. The model was applied to peach fruit, and parameters were estimated from the data of fruit development monitored over 2 years. The predictions were in good agreement with experimental data. Simulations were performed to analyse the behaviour of the model in response to variations in composition and temperature.

Carboxylic Acids↗

Determination of malic acid and other C4 dicarboxylic acids in atmospheric aerosol samples.

An ion chromatographic method was developed which is able to separate five unsubstituted and hydroxy C4 dicarboxylic acids, succinic, malic, tartaric, maleic and fumaric acid, besides the other unsubstituted C2-C5 dicarboxylic acids, oxalic, malonic and glutaric acids, as well as inorganic ions in samples extracted from atmospheric particulate matter. By the application of this method it was found for both rural and urban sites and for various types of air masses that in the summer-time malic acid is the most prominent C4 diacid (64 ng m(-3) by average), exceeding succinic acid concentration (28 ng m(-3) by average) considerably. In winter-time considerably less, a factor of 4-15, C4 acids occurred and succinic acid was more concentrated than malic acid. Tartaric, fumaric and maleic acids were less concentrated (5.1, 5.0 and 4.5 ng m(-3) by average, respectively). Tartaric acid was observed for the first time in ambient air. The results indicate that in particular anthropogenic sources are important for the precursors of succinic, maleic and fumaric acids. Biogenic sources seem to influence the occurrence of malic acid significantly.

Aerosols↗