259 results about "Oxaloacetic acid" patented technology

A non-naturally occurring microbial organism having a 3-hydroxypropanoic acid (3-HP) pathway includes at least one exogenous nucleic acid encoding 3-HP pathway enzyme expressed in a sufficient amount to produce 3-HP. The 3-HP pathway includes a 2-keto acid decarboxylase, a CoA-dependent oxaloacetate dehydrogenase, or a malate decarboxylase. A method for producing 3-HP includes culturing a non-naturally occurring microbial organism having a 3-HP pathway that includes at least one exogenous nucleic acid encoding a 3-HP pathway enzyme expressed in a sufficient amount to produce 3-HP under conditions and for a sufficient period of time to produce 3-HP. The 3-HP pathway includes a 2-keto acid decarboxylase, a CoA-dependent oxaloacetate dehydrogenase, or a malate decarboxylase.

4-(Indol-3-ylmethyl)-4-hydroxy-2-oxoglutarate, which is useful as an intermediate in the synthesis of monatin, may be synthesized from indole pyruvic acid and pyruvic acid (and / or oxaloacetic acid) by using a novel aldolase derived from the genus Pseudomonas, Erwinia, Flavobacterium, or Xanthomonas.

The present invention relates to a method for producing optically active IHOG, which can in turn be used for the production of monatin. The present invention further relates to a method for producing optically active monatin, and aldolase used for these methods. As such, the present invention enables the synthesis of 4-(Indole-3-ylmethyl)-4-hydroxy-2-oxoglutaric acid with high optical purity, which is useful as an intermediate in the synthesis of optically active monatin, from indole pyruvic acid and pyruvic acid (or oxaloacetic acid).

Methods of obtaining mutant nucleic acid sequences that demonstrate elevated oxaloacetate α-decarboxylase activity are provided. Compositions, such as genetically modified microorganisms that comprise such mutant nucleic acid sequences, are described, as are methods to obtain the same.

The invention provides a gene engineer strain of Aspergillus niger with high L-malic acid yield and a construction method thereof. The method utilizes gene knockout technique to disrupt the Aspergillus niger oxaloacetic acid hydrolase gene (oahA) so as to obtain the malic acid-producing strain S2. After 7 days of shake flask fermentation, 100 g / L glucose is converted into 120.4 + / - 2. 8 g / L of L-malic acid, the conversion of malic acid and malic acid to glucose is 1.62 mol / mol, which is 81% of the highest conversion in theory. A good strain for the industrialized production of malic acid is provided.

A method for constructing an ALA production bacterial strain, the method enhances the activity of related enzymes promoting the synthesis of oxaloacetate and in the 5-aminolevulinic acid (ALA) production bacterial strain, or introducing exogenous related enzymes promoting the synthesis of oxaloacetate, such as phosphoenolpyruvate carboxylase or pyruvate carboxylase, and / or reducing the activity of related enzymes in the downstream metabolic pathway of succinyl coenzyme A in the bacterial strain, such as succinyl coenzyme A synthetase or succinate dehydrogenase, and / or reducing the activity of phosphoenolpyruvate carboxylated kinase and / or malic enzyme. An ALA high-yield bacterial strain constructed by utilizing the method, and method for utilizing the bacterial strain to prepare ALA.

The present invention provides a new binary organic acid production strain, preparation and applications thereof, particular a genetically modified engineering strain for binary organic acid synthesis. According to the present invention, an exogenous binary organic acid synthesis positive regulation gene is introduced into the engineering strain, and / or a binary organic acid synthesis negative regulation gene is down-regulated; compared to the starting strain, the binary organic acid production capacity of the engineering strain is significantly improved, wherein the binary organic acid comprises malic acid, succinic acid, fumaric acid, oxaloacetic acid, glutaric acid, and adipic acid; and the experiment results prove that the genetically modified engineering strain being subjected to the up-regulation of one or a plurality of the positive regulation genes and / or the down-regulation of one or a plurality of the positive regulation genes can resist the high temperature, effectively utilizes monosaccharides, polysaccharides, glycan or mixed saccharides, and particularly utilize the cheap carbon source (such as cellulose and the like) to synthesize the binary organic acid in the high-yield manner.

The present invention relates to methods of producing a polypeptide comprising culturing a mutant of a parent cell, wherein the mutant produces less oxaloacetate hydrolase than the parent cell, wherein the oxaloacetate hydrolase (i) has an amino acid sequence that has at least 90% identity with amino acids 1-341 of SEQ ID NO: 2: (ii) is encoded by a nucleic acid sequence having at least 90% homology with a nucleotide sequence comprising nuclotides 1157-1411, 1504-1651 and 1764-2383 of SEQ ID NO: 1: (iii) is encoded by a nucleic acid sequence which hybridizes under high stringency conditions with the nucleic acid sequence of SEQ ID NO: 1, the cDNA sequence of SEQ ID NO: 1, the complementary strand of SEQ ID NO: 1, and / or the complementary strand of the cDNA sequence of SEQ ID NO: 1: and / or (iv) a subsequence of one or both of (i) and (ii), wherein the subsequence encodes a fragment that has oxaloacetate hydrolase activity.

The present invention describs methods of increasing the production of a fine chemical, e.g., lysine from a microorganism, e.g., Corynebacterium by way of deregulating an enzyme encoding gene, i.e., fructose-l,6-bisphosphatase. In a preferred embodiment, the invention provides methods of increasing the production of lysine in Corynebacterium glutamicum by way of increasing the expression of fructose-1,6-bisphosphatase activity. The invention also provides a novel process for the production of lysine by way of regulating carbon flux towards oxaloacetate (OAA). In a preferred embodiment, the invention provides methods for the production of lysine by way of utilizing fructose or sucrose as a carbon source.

The invention relates to the field of medicaments and discloses an antialcoholismic composition. The raw materials of the antialcoholismic composition comprise: one or a mixture of more than two of S-adenosyl methionine, cysteine and methionine, one of a mixture of D-glyceric acid and D-glycerate, one or a mixture of more than two of succinic acid, fumaric acid, citric acid, oxaloacetic acid and malic acid, one or a mixture of more than two of vitamin B1, vitamin B6 and vitamin B2, one or a mixture of coenzyme Q10 or coenzyme I, L-glutamine, fortune eupatorium herb, artemisia capillaries, white paeony root and root of tall monkshood. The invention also provides a preparation method of the antialcoholismic composition. The antialcoholismic composition disclosed by the invention is taken after wine drinking for reducing absorption of alcohol, accelerating alcohol metabolism, as well as tonifying stomach and spleen and preventing drunkenness and alcoholism. The antialcoholismic composition is quick in effectiveness and has a bright application prospect.

The invention discloses a construction method of a metabolic engineering escherichia coli strain for producing hydracrylic acid from acetic acid. Escherichia coli reformed by metabolic engineering isutilized for producing hydracrylic acid by fermenting acetic acid; and construction ways are as follows: constructing a metabolic way for producing hydracrylic acid from acetyl CoA, and / or over-expressing relevant genes of an acetic acid intake way so as to increase the transmission rate of acetic acid, and / or blocking or decreasing TCA circle so as to increase metabolic flow of acetyl CoA flowingtowards a target metabolite, and / or reducing decarboxylic reaction between malic acid or oxaloacetic acid so as to delete a byproduct generation way, and / or deleting key genes in an alcohol production way so as to regulate metabolic flow of an acetyl CoA node and / or regulating intracellular oxidation reduction balance through coenzyme engineering. According to the construction method, a production way for exogenously expressing hydracrylic acid is constructed, the metabolic ways, the adjustment and control are analyzed, and escherichia coli is reformed by virtue of a metabolic engineering measure, so that the prepared strain can be utilized for producing hydracrylic acid in a culture medium taking acetic acid as a carbon source.

The present invention relates to electrochemical detection technology, and is quick detecting method of aspartate aminotransferase vitality. Electrochemical or optical change is produced corresponding to the vitality of the measured matter. The method includes: A. sucking the measured matter liquid drop into the reaction area between electric poles to generate oxidation-reduction on the surface of the electric poles so as to produce electron transfer and form current; and B. converting the aspartic acid into glutamic acid and oxaloacetic acid under the cooperated action of current and auxiliary factors and measuring the amount of glutamic acid and oxaloacetic acid so as to obtain the corresponding aspartate aminotransferase vitality. The present invention has raised response speed and detection sensitivity of current type enzyme sensor and lowered reaction voltage.

The invention discloses a method for knocking out pckA to promote synthesis of acetylglucosamine through bacillus subtilis and belongs to the field of genetic engineering. The method comprises the steps that BSGNK-PxylA-glmS-P43-GNA1 serves as original strains, phosphoenolpyruvate carboxylase (pckA) genes are knocked out through homologous recombination, the reaction that enolphosphopyruvate is converted into oxaloacetic acid in host bacterium cells is blocked, the carbon flux flowing to a tricarboxylic acid cycle is reduced, and accumulation of acetylglucosamine is promoted. In the process that a composite culture medium is used for fermentation, the acetylglucosamine yield of recombination bacillus subtilis with pckA knocked out reaches 29.27 g / L and is increased by 28.1% compared with that of control strains. The method lays a foundation for transforming bacillus subtilis to produce acetylglucosamine in metabolic engineering.

A recombinantly modified Corynebacterium glutamicum microorganism with an improved 1,4-butanediol (1,4-BDO) productivity relative to an unmodified Corynebacterium glutamicum microorganism, wherein activity of an enzyme catalyzing a conversion reaction between malate and oxaloacetate is inactivated or reduced relative to an unmodified Corynebacterium glutamicum microorganism, as well as a method of making and using same.

The invention relates to a protein-free, hydrolysate-free and serum-free culture medium and a preparation method thereof. The culture medium contains the following substances: manganese gluconate, calcium citrate malate, disodium edetate, progesterone, alanyl-glutamine,glycerophosphate, fructose, vitamin C, vitamin E, sodium selenite, hydroxypropyl-beta-cyclodextrin, cholesterol, malic acid, oxaloacetic acid, vitamin B12, lipoic acid, ferrous sulfate, ferric citrate, uric acid, taurine, reduced glutathione, zinc sulfate, copper sulfate and ferric nitrate. The protein-free, hydrolysate-free and serum-free culture medium has the advantages that the cost is low, the difference among different batches of products is small, the purification step of down-stream products is simplified, and potential safety hazards are reduced.

The invention relates to a preparation for improving the color and sweet of fruits such as apple and pear, the preparation is aqueous solution agent. The red rate and sweetness of the fruit can be improved through establishing the metabolism control technology of synthesis of apple and pear color and synthesis of soluble sugar. The components and the concentration of the preparation are as follows: acetylsalicylic acid is 1.0 to 3.0 mg / L, galactose is 0.5 to 1.5 g / L, malic acid is 0.1 to 3.0 g / L, phenyalanine is 0.1 to 1.0 g / L, oxaloacetic acid is 0.1 to 1.0 g / L, boric acid is 0.1 to 2.0 g / L,monopotassium phosphate is 0.8 to 1.5 g / L, copper sulphate is 0.1 to 2.0 g / L, ammonium molybdate is 50 to 200 mg / L, twee 100 is 1.0 to 4.0 ml / L, little ethanol, and the balance water. The preparationmethod is as follows: 1, weighing up the acetylsalicylic acid, galactose, malic acid, phenyalanine, oxaloacetic acid, boric acid, monopotassium phosphate, copper sulphate, ammonium molybdate and twee100 according to a certain proportion, 2, the acetylsalicylic acid is dissolved by the ethanol, the phenyalanine is dissolved by distilled water at 100 DEG C, the monopotassium phosphate, copper sulphate and ammonium molybdate are dissolved by distilled water at 80 to 90 DEG C, other components are dissolved by distilled water at room temperature, all the components are dissolved and mixed, and then are stirred evenly to make the volume to the required volume, thereby obtaining the preparation. The preparation can improve the coloring rate of Nanguo pear by 15.8 percent. The preparation is convenient and time-saving to use, has no chemical injury and has no toxic side effect to human body and environment.

The invention discloses a composite dechlorination agent for removing high-concentration chlorine-containing wastewater and an application method thereof, which belong to the field of sewage treatment in environment protection. The composite dechlorination agent is prepared by compounding organic acids and inorganic salts, wherein the organic acids include two or more than two of nicotinic acid, citric acid, succinic acid, beta-ketoglutaric acid, and ketosuccinic acid; the inorganic salts include one or two of mercury sulfate, cadmium sulfate and copper sulfate; and the weight percentages of the organic acids and inorganic salts are 15-80% and 20-85% respectively. The application method comprises the steps of: adding the prepared composite dechlorination agent in percentage by weight of 0.05-0.5% into the high-concentration chlorine-containing wastewater and continuously stirring for 10-30min; adding a methylene dichloride solvent; standing for layering, separating, and subjecting obtained wastewater to air flotation treatment; and finally scraping off a layer of residue floating on the surface of the wastewater by a residue scraping machine to obtain the dechlorinated wastewater as the yielding water, wherein the dechlorinated wastewater can be lower than the national emission standard I. The composite dechlorination agent and the application method disclosed by the invention have the characteristics of simple technological process, smaller dosage of chemical agent, lower running expense, no sludge, high removal rate of chlorine ions in the wastewater, and the like; and the removal rate of chlorine ions in the wastewater can reach above 99.9%.

The invention discloses a rice heavy metal absorption blocking agent and a use method thereof. The blocking agent includes one or more effective components of alpha-ketoglutarate, glycerol and oxaloacetic acid. The aim of preventing heavy metal from being absorbed and transferred to rice can be achieved through leaf surface spraying in the rice tillering period, heading period and / or postulation period. The contents of lead, cadmium and other heavy metal polluting rice in rice fields can be remarkably reduced, and rice produced in rice fields slightly or intermediately polluted by heavy metal meets the food safety standard. The effective spraying concentration of the blocking agent is low, the blocking agent can be sprayed after being mixed with pesticides for preventing and treating rice diseases and pests, and cost is low.

Provided are methods for the detection and diagnosis of Hypothyroidism. The methods are based on the discovery that altered levels of selected analytes in sample fluid, typically blood samples, of patients are supportive of a diagnosis of Hypothyroidism. At least twenty-four new biomarkers for hypothyroidism are thus disclosed (singly or in any combination), Thyroid Stimulating Hormone, Interleukin-12p40, Tumor Necrosis Factor Alpha, Tissue Factor, Interleukin-15, Insulin, Immunoglobulin E, Growth Stimulating Hormone, Calcitonin, Prostate-Specific Antigen, Interleukin-4, Granulocyte Macrophage Colony Stimulating Factor, Matrix Metalloproteinase 9, Lymphotactin, Fatty Acid Binding Protein, Alpha Fetoprotein, Alpha-2 Macroglobulin, Serum Glutamic Oxaloacetic Transaminase, Matrix Metalloproteinase 3, Cancer Antigen 125, Mumps Antibody, Double Stranded DNA Antibody, Proliferating Cell Nuclear Antigen Antibody, Smith Antibody, or Herpes Simplex Virus 1 Glycoprotein D Antibody. Altogether the concentrations of one or more of these analytes, as well as Thyroid Stimulating Hormone, or any combination thereof, provide a sensitive and selective picture of the patient's condition, namely, whether the patient is suffering from Hypothyroidism. Kits containing reagents to assist in the analysis of fluid samples are also described.

The invention discloses a kit and a method for content determination of acetyl coenzyme A. The kit comprises a reagent I, a reagent II, a reagent III, a reagent IV and a reagent V, wherein the reagent I is prepared from Tris-HCl, polyvinylpyrrolidone, mercaptoethanol, phenylmethylsulfonyl fluoride and glycerinum; the reagent II is prepared from malic dehydrogenase; the reagent III is prepared from citrate synthase; the reagent IV is prepared from malic acid and an oxidation type coenzyme I; and the reagent V is prepared from Tris-HCl. The malic dehydrogenase can be used for catalyzing malic acid and the oxidation type coenzyme I to generate oxaloacetic acid and a reduced coenzyme I; the citrate synthase is used for catalyzing acetyl coenzyme A and oxaloacetic acid to generate citric acid and a coenzyme A; by virtue of a coupled reaction of the malic dehydrogenase and the citrate synthase, the content of the acetyl coenzyme A and the generation rate of the reduced coenzyme I are in a direct proportion; the climbing speed of the light absorption value at 340nm reflects the content of the acetyl coenzyme A. The kit disclosed by the invention is simple and convenient to operate, high in detection sensitivity and high in recovery rate, and the cost of the reagents is remarkably lowered. The detection steps are further simplified, so that the kit is simpler and more convenient and efficient to test.

The invention relates to a kind of enzyme colorimetric method and a method of the coupling method measuring the density of citric acid and a diagnosis reagent box applying the citrate synthetase and coupling malic dehydrogenase enzymatic reaction continues monitoring method / ratio colorimetric method. The reaction of citrate synthetase enzymolysis citric acid generates oxaloacetic acid and then through the effect of coupling malic dehydrogenase at last oxidizes the reduced coenzyme (there is a absorption peak at the site of 340nm) becoming the coenzyme (there is a absorption peak at the site of 340nm) so we can assay the degree / speed of the fall-way absorbance at the place of 340nm. It can measure and calculate the density of citric acid by measuring the degree / speed of the fall-way absorbance at the place of 340nm. The method has high specificity and it is not polluted by the endogenous and exogenous object and the test result is precise and accurate. The invention can get the array result by the ultraviolet / visible light analytic instrument so it is convenience to extend and apply.

The invention discloses a construction method of a metabolic engineering escherichia coli strain for producing acetone or isopropanol by virtue of acetic acid. Through metabolic engineering transformed escherichia coli, the acetone or isopropanol is produced by conducting fermentation with the acetic acid as a carbon source; and transforming pathways include the follows: constructing a metabolic pathway of producing the acetone or isopropanol through acetyl CoA, and / or promoting over-expression of acetone intake pathway related genes so as to enhance the transport rate of the acetone, and / or blocking off TCA circulation or down-regulating the TCA circulation so as to increase an acetyl CoA metabolic flux flowing to a target metabolic product, and / or relieving a decarboxylic reaction between malic acid and oxaloacetic acid to delete a by-product generating pathway, and / or deleting key genes in an ethanol producing pathway to regulate an acetyl CoA node metabolic flux. According to the construction method provided by the invention, through analysis on the metabolic pathway as well as regulation and control, the escherichia coli is transformed by virtue of genetic engineering means; and with the application of the produced strain, the acetone and the isopropanol can be produced in a culture medium that the acetic acid is taken as the carbon source.

The invention discloses a vegetable heavy metal blocking agent and a use method thereof. The effective component of the blocking agent is one of or several of alpha-ketoglutaric acid, glycerol and oxaloacetic acid; the purposes of blocking heavy metal absorption and transferring to edible parts can be realized through foliage spraying from vegetable cultivation to harvesting. According to the vegetable heavy metal blocking agent disclosed by the invention, the contents of lead, cadmium and other heavy metals at the edible parts can be obviously reduced, so that vegetables produced in vegetable fields mildly and moderately polluted by the heavy metals conform to food safety standards. The blocking agent disclosed by the invention is low in effective spraying concentration and can be sprayed after being mixed with pesticides for preventing and treating vegetable diseases and insect pests, thereby being low in cost.

The present invention relates to a plant nutrient active agent, it is an aqueous solution containing several components. It is characterized by that it contains at least two preservative components, the described preservative component is carboxylic acid or carboxylate containing no benzene ring, and its concentration is not less than 0.001 mole / L. The described preservative component can be two kinds or more than two kinds selected from acetic acid, acetate, diacetic acid, diacetate, dehydroacetic acid, dehydroacetate, oxaloacetic acid, oxaloacetate, propanoic acid, propionate, acrylic, acid, acrylate and others.