261 results about "Pyruvic acid" patented technology

The invention provides a valine production strain and a construction method and application thereof, a designed acetohydroxy acid synthase mutant is that the 88th basic group of an ilvB gene is changed from a to c, the 382nd basic group is changed from a to g, the 413th basic group is changed from c to t, the gene sequence of a designed artificial operon comprises a promoter, an ilvB (A138V) gene or ilvB (Q30K, S128G, A138V) gene of coding mutated acetohydroxy acid synthase, and an ilvN (G20D, I21D, I21D, I21D, I21D, I21D, I21D, I21D) gene. I22F) gene, a pyk gene for coding pyruvate kinase, and a terminator; by designing a specific acetohydroxyacid synthase mutant and related biological materials and artificial operon, the strain constructed by directional modification of the strain by using a pK18mobsacB system gene editing technology based on allele exchange has the advantages of good genetic stability, high fermentation yield and the like, and valine can be stably produced.

The invention belongs to the technical field of preparation of organic chemicals, and particularly relates to trifluoropyruvic acid and a method for preparing trifluoropyruvic acid from trifluoroacetone. The method comprises the following steps: dissolving trifluoroacetone in a chlorinated solvent, adding a catalyst, introducing chlorine, carrying out a closed reaction, controlling the reaction temperature, cooling to room temperature after the reaction is finished to obtain a 1, 1, 1-trichlorotrifluoroacetone reaction mixture, adding an alkali liquor to carry out a hydrolysis reaction, separating out a water phase after the reaction is finished, and drying to obtain the trifluoropyruvic acid. And continuously applying the chlorination solvent, continuously adding the acid into the water phase to adjust the pH value to 3-4 to obtain a trifluoropyruvic acid water solution, finally extracting and pulping to obtain a wet trifluoropyruvic acid product, and purging with air to obtain a pure trifluoropyruvic acid product. According to the method for preparing trifluoropyruvic acid from trifluoroacetone, the raw materials are easy to obtain, the process is stable, the yield is high, the cost is low, and the invention further provides a trifluoropyruvic acid product.

The present application relates to a mutant gene encoding phosphoenolpyruvate carboxylase containing a mutant 5' untranslated region (5'UTR), a polyhydroxyalkanoate (PHA)-producing microorganism containing the gene, and a PHA production method using the same.

The invention discloses a gelatin microcarrier lysate as well as a preparation method and application thereof. The gelatin microcarrier lysis buffer comprises lyase, a cell protective agent, a cell energy metabolism protective agent, an antioxidant, a buffer solution and a chelating agent. The gelatin microcarrier lysate has extremely strong protectiveness to cells, abandons damaging trypsin and EDTA, and innovatively adds a cell protection combination of a cell protective agent L-arginine, a cell energy metabolism protective agent sodium pyruvate and an antioxidant vitamin C, so that the cell survival rate (gt; 98%) and the retention rate (gt; the method is high in cell yield, good in safety and wide in range.

The present disclosure relates to novel compounds that are capable of inhibiting the mitochondrial pyruvate carrier and promoting hair growth. The disclosure further relates to methods of promoting hair growth or treating conditions or disorders affecting hair growth, such as baldness or alopecia.

The invention discloses a synthesis method of a mono / dibromo pyridine derivative, which comprises the following steps: reacting a pyridine derivative with dimethyl butynedioate and methyl pyruvate at room temperature by using acetonitrile as a solvent to generate a dearomatized heteroaromatic compound; in an oxygen environment, copper bromide is used for brominating the dearomatized heteroaromatic compound to realize dibromination, and if potassium carbonate is additionally added in the reaction, monobromination is realized; and finally, respectively generating mono / dibromopyridine derivatives under hydrochloric acid hydrolysis. A carbon halogen bond of bromopyridine has very high activity, and various reactions such as Suzuki coupling, Sonogashira coupling, Heck reaction, amination reaction and the like can be carried out. On the basis, the polysubstituted bromopyridine can be subjected to selective reaction, and many new ideas can be provided for organic synthesis. The method disclosed by the invention is simple and safe to operate and environment-friendly, the cost is lower by using the copper bromide, and the required reaction conditions are simple.

The invention discloses a construction method of an escherichia coli mutant for producing succinic acid by fermentation of a synthetic medium. The method comprises the following steps: firstly, knocking out a lactic dehydrogenase gene ldhA, a pyruvate formate lyase gene pflB, a ptsG gene responsible for encoding a phosphotransferase system EIIBC protein, an ethanol dehydrogenase gene adhE, an acetokinase-phosphate transacetylase gene ackA-pta, and a ptsG gene responsible for encoding a phosphotransferase system EIIBC protein in escherichia coli; a phosphoenolpyruvate carboxykinase gene pck from bacillus subtilis is integrated at an SS9 safety site of a strain to obtain escherichia coli ESC6 with high succinic acid yield; and mutating one or more loci in one or more genes of a glucose-transcriptional inhibition factor gene mlc, a nitrate response regulatory factor gene narL and a cyclic adenylate receptor protein gene crp to obtain the escherichia coli mutant capable of producing succinic acid by fermentation of a synthetic culture medium, wherein the one or more loci in one or more genes of the glucose-transcriptional inhibition factor gene mlc, the nitrate response regulatory factor gene narL and the cyclic adenylate receptor protein gene crp are mutated. The strain can be fermented in a synthetic medium to produce succinic acid, so that the fermentation cost is greatly reduced, and the strain has a very wide application prospect.

The invention discloses a modified aureobasidium pullulans strain based on reduction of malic acid pathway consumption and application of the modified aureobasidium pullulans strain. Gene for coding NADPH dependent malic enzyme and phosphoenolpyruvate carboxykinase in an aureobasidium pullulans genome is directionally knocked out. The transformation blocks a key consumption branch for synthesis and accumulation of the cytoplasmic malic acid, and effectively guides a central carbon metabolic flow to be more efficiently guided to a target product. Experimental results show that the polymalic acid yield and the saccharic acid conversion rate of the double-knockout engineering strain D3N5-delta3617 / deltapepck in fermentation are remarkably improved compared with those of an original strain, and the biomass is also increased. The invention provides a high-performance strain with important application value for industrial efficient fermentation production of polymalic acid.

The present disclosure relates to a genetically modified microorganism satisfying some of predetermined conditions. The predetermined conditions include: (I) succinate dehydrogenase activity or fumarate reductase activity being reduced or inactivated relative to a wild-type microorganism; (II) lactate dehydrogenase activity being reduced or inactivated relative to the wild-type microorganism; (III) the genetically modified microorganism having modified phosphoenolpyruvate carboxylase activity showing resistance to feedback inhibition by aspartic acid in wild-type phosphoenolpyruvate carboxylase activity, or exogenous phosphoenolpyruvate carboxylase activity having higher resistance to feedback inhibition by aspartic acid than that of the wild-type phosphoenolpyruvate carboxylase activity shown by the wild-type microorganism; and (IV) pyruvate:quinone oxidoreductase being reduced or inactivated relative to the wild-type microorganism.

The microalgae phenol degradation and carbon fixation technology aims to provide a method for improving key enzymes to promote euglena to degrade phenol and enhance carbon fixation. The method comprises the following steps: adjusting the gene expression amount of three key enzymes, H+ transport ATPase, pyruvate decarboxylase and isocitrate dehydrogenase in euglena cells by performing phenol concentration gradient domestication on the euglena cells; performing large-scale culture growth on the euglena domesticated strains, adding appropriate phenol in the culture medium, and continuously feeding the gas containing CO2; calculating the carbon fixation rate of the euglena by measuring the carbon element content in the biomass of the algal liquid during the culture process; after the culture is completed, the euglena polysaccharide content in the unit volume of the algal liquid is measured, and the phenol content in the algal liquid is measured to calculate the phenol degradation rate. The euglena cells are domesticated by using the phenol concentration gradient, the gene expression amount of the key enzymes is improved by several times, the phenol degradation capacity of the euglena cells is enhanced, the carbon fixation rate is improved, and the polysaccharide content in the cells is also improved.

Methods for preparing pyruvate kinase (PK) activators or salts or hydrates thereof are provided herein. Synthetic methods for preparing pyruvate kinase (PK) activators, or salts or hydrates thereof, are provided herein. Such activators may be prepared using compounds having formula (A-1) and / or formula (B-1). Specific PK activators and various intermediates prepared by and / or used by the synthetic methods described herein are also disclosed.

The invention discloses application of eupatolide in preparation of a medicine for preventing and treating inflammatory bowel disease, application of eupatolide in preparation of a medicine for preventing and treating inflammatory bowel disease and application of a medicine preparation containing eupatolide in preparation of a medicine for preventing and treating inflammatory bowel disease, and researches that eupatolide (EPT) can be used for preparing the medicine for preventing and treating inflammatory bowel disease through targeting pyruvate kinase isoenzyme 2 (PKM2, Gene ID: 5135). The inflammatory response, glycolysis metabolism and mitochondrial homeostasis are synergistically regulated and controlled, so that the inhibition effect on the inflammatory bowel disease is exerted. Meanwhile, EPT has the advantages of being low in dosage, feasible in oral administration, free of obvious toxic and side effects and the like, and a solid experimental basis is provided for further developing EPT into candidate drugs for preventing and treating inflammatory bowel diseases.

Methods of treating or preventing conditions in a subject are disclosed, the methods including administering to the subject a therapeutically effective amount of ethyl pyruvate, or a derivative or analog thereof, or a pharmaceutical composition thereof. Methods of treating or preventing conditions in a subject including administering to the subject a therapeutically effective amount of ethyl pyruvate, or a derivative or analog thereof, or a pharmaceutical composition thereof, and one or more additional therapeutic agents, are described.

The invention relates to a pyruvate kinase-targeted compound shown as a formula I, a preparation method and application thereof, and application of the pyruvate kinase-targeted compound serving as a plant antibacterial agent, and part of the pyruvate kinase-targeted compound shows excellent bactericidal activity.

The invention discloses an application of a pyruvate dehydrogenase kinase (PDKS) inhibitor in preparation of a composition for treating Alzheimer's disease (AD), and particularly discloses an application of the PDKs inhibitor in preparation of the composition for treating the AD. According to the invention, the level of oxidative phosphorylation (OXPHOS) of microglial cells is obviously enhanced, the inflammatory response of the microglial cells is effectively reduced, the migration and aggregation capabilities of the microglial cells are obviously improved, the capability of the microglial cells to phagocytize and degrade extracellular beta-amyloid protein (A beta) plaque is obviously enhanced, the cognitive function defect of AD model mice is effectively improved, and the application of the microglial cells in the treatment of the AD model mice is promoted. The preparation is simple, and the application is wide.

The application discloses a chromogenic culture medium for detecting enteropathogenic Escherichia coli, which contains peptone, sodium pyruvate, sodium chloride, D-sorbitol, D-malic acid, bile salt, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium hydroxide, kaolin, bromothymol blue, crystal violet, polyethylene glycol and agar, and the rest is water. The chromogenic culture medium is simple in composition, convenient to prepare and use, and can be used for identifying enteropathogenic Escherichia coli EPEC according to the color of bacterial colonies, and has the advantages of specificity, high efficiency, rapidness, low cost, simple result judgment and the like, and has a wide application prospect in food safety monitoring.

The present invention provides: a culture medium composition which contains a pyruvate dehydrogenase kinase inhibitor and is used for promoting the production of a virus having a target gene mounted thereon when culturing a virus-producing cell into which the target gene has been introduced; a culture medium composition for promoting the production of a virus or a virus-like particle when culturing a cell that produces the virus or the virus-like particle; and the like.

Described herein are compounds that activate pyruvate kinase, pharmaceutical compositions, and methods of using the same. The compounds are represented by Formula (I), wherein R 1 , R 2 , R a , R b , R j , R k , and Q are as defined herein.

The application belongs to the field of biological analysis, and provides a method for precise metabolic flow analysis of TCA cycle based on isotope tracing technology combined with high-resolution mass spectrometry. By optimizing mass spectrometry parameters and liquid chromatography method, one injection within 6 min is realized, and high-throughput, high-sensitivity and high-resolution detection is realized on 11 biomarkers related to TCA cycle metabolic pathways. At the same time, the application innovatively uses LC-MS / MS to decipher 13 C the stepwise position-specific transfer from glucose to subsequent metabolites by glycolysis and TCA cycle, thereby calculating 13 C the integrated steady-state analysis of key metabolic rates of position-specific transfer from sequential precursors to their products (pyruvate dehydrogenase, beta-oxidation, pyruvate carboxylase, isocitrate dehydrogenase and pyruvate cycle). This technology has wide applicability and can potentially characterize mitochondrial metabolism of any tissue or cell.

Provided herein are amorphous and crystalline hemisulfate salt forms of the formula.Also provided are pharmaceutical compositions comprising the amorphous and crystalline hemisulfate salt forms, methods for their manufacture, and uses thereof for treating conditions associated with pyruvate kinase such as e.g., pyruvate kinase deficiency.

The invention discloses a genetically engineered bacterium for efficiently producing D-pantothenic acid as well as a construction method and application of the genetically engineered bacterium, the construction method comprises the following steps: (a) taking corynebacterium glutamicum DPA2 as a chassis bacterium, enhancing the expression of at least one pantothenic acid main pathway gene in a chassis bacterium genome, and introducing plasmid pEC-XK99E-panBCilvC to obtain an engineered strain for modifying the pantothenic acid main pathway gene; (b) knocking out at least one heteroacid pathway gene in the engineering strain obtained in the step (a), carrying out in-situ integration on a panBCC gene at an alaT gene locus, and introducing a plasmid pEC-XK99E-panBCilvC to obtain a heteroacid pathway modified engineering strain; and (c) knocking out the pyruvic acid shunt gene pyc in the heteroacid pathway modified strain obtained in the step (b), enhancing the expression of an odx gene, and introducing a plasmid pEC-XK99E-panBCilvC to obtain the engineering strain for regulating and controlling the pyruvic acid shunt. The genetically engineered bacterium has the beneficial effects that the genetically engineered bacterium for efficiently producing D-pantothenic acid is obtained, and the yield and the conversion rate are higher; the level of the modified strain in the aspect of fermentation production of D-pantothenic acid is obviously improved.

The invention relates to an establishment method and application of a micropterus salmoides muscle cell line, and belongs to the technical field of cell culture. The invention provides a culture solution which takes an M199 culture medium as a basic culture medium and further comprises sodium pyruvate, hydroxyethyl piperazine ethanesulfonic acid, beta-mercaptoethanol, penicillin, streptomycin and fetal calf serum. The culture solution provides optimal conditions for micropterus salmoides muscle cell culture, the culture cost is also reduced, the micropterus salmoides muscle cell line is established by adopting the culture solution, the method is simple and easy to operate and high in repeatability, the established cell line is good in stability, continuous passage and ultralow-temperature cryopreservation can be realized, the cell division speed is high, the passage time is short, the adherence rate is high, and the method is suitable for large-scale popularization and application. In the method, the micropterus salmoides muscle primary cell culture time is short, the number of obtained cells is large, the activity is good, and passage can be carried out in only 8 days.

The invention relates to the technical field of biological medicines, in particular to application of tetramer-form pyruvate kinase type 2 (PKM2) in preparation of a medicine for treating obesity. Pyruvate kinase type 2 is a key enzyme in a glycolytic pathway and has two conformations of dimer and tetramer, and the tetramer-form enzyme has high activity. The invention discovers that the PKM2 tetramer stabilizer TEPP-46 can stabilize PKM2 in a high-activity tetramer state and improve obesity-related metabolic disorder induced by high fat diet. The invention discloses a new way for regulating and controlling energy metabolism by stabilizing the form of the PKM2 tetramer, and provides a new medicine application direction for treating obesity and related metabolic diseases.

The invention relates to the technical field of blood detection, in particular to a formula of an optimized alanine aminotransferase kit, which has the technical scheme that lactic dehydrogenase, NADH (nicotinamide adenine dinucleotide), DCA-Na, glycerol and a Proclin-300 preservative are dissolved in a reagent I Tris buffer solution of the ALT kit; l-alanine, alpha-ketoglutaric acid, glycerol and a Proclin-300 preservative are dissolved in a Tris buffer solution of the reagent II, DCA-Na in the kit is an activator of a pyruvate-lactic dehydrogenase compound and inhibits the kinase activity of pyruvate dehydrogenase, glycerol is an enzyme activity stabilizer, Proclin-300 is a preservative, an instrument is used for detection at the main wavelength of 340 nm and the auxiliary wavelength of 380 nm, and the detection result is accurate. And calculating the ALT activity according to the decreasing rate of NADH (nicotinamide adenine dinucleotide).

The invention relates to the technical field of NK cell culture, and discloses an NK culture medium containing hypotaurine and application of the NK culture medium to improvement of NK cell proliferation and killing activity, and the NK culture medium comprises a basic NK cell culture medium and hypotaurine; the basic NK cell culture medium does not contain hypotaurine and comprises amino acid, vitamins, inorganic salt, trace elements, D-glucose and sodium pyruvate. When the basic NK cell culture medium is an RPMI1640 culture medium, the concentration of the hypotaurine is 0.1-1 mM; when the basic NK cell culture medium is an IMDM culture medium, the concentration of the hypotaurine is 0.4-4 mM. According to the scheme, the hypotaurine is added into the basic NK cell culture medium to obtain the NK culture medium, the NK culture medium can increase the number of NK cells when the NK cells are cultured in vitro, and the killing activity of the cultured NK cells on tumor cells is remarkably improved.

This invention discloses an in vitro maturation culture medium for ferret oocytes and its preparation method, belonging to the field of animal embryo engineering technology. It comprises the following components: TCM199 basal culture medium, 50 mL fetal bovine serum, 25 IU follicle-stimulating hormone, 25 IU luteinizing hormone, 0.5 mg 17β-estradiol, 0.005 mg epidermal growth factor, 12.1 mg sodium pyruvate, 0.2 mM cysteine, 0.1 mM cysteine, 0.030 g penicillin, 0.025 g streptomycin sulfate, as well as an antioxidant complex, a combination of growth factors, and extracellular matrix components. This invention provides a comprehensive and suitable nutritional environment for ferret oocytes by precisely formulating a composite culture medium containing multiple key components. Specifically, the hormone combination at specific concentrations accurately simulates the in vivo physiological environment, effectively regulating the oocyte maturation process; multiple growth factors work synergistically to promote oocyte growth, development, and differentiation; and the addition of the antioxidant complex effectively eliminates free radicals generated during in vitro culture, reducing oxidative stress damage to oocytes.

The invention discloses a genetically engineered bacterium for synthesizing N-acetyl blue as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium is prepared by knocking out a lactic dehydrogenase gene ldh and inserting a phosphoketolase gene xfpK on a knockout site, knocking out a pyruvate oxidase gene poxB and inserting a phosphate transacetylase gene pta on a knockout site of a microorganism for producing N-acetyl blue, and knocking out an aconitase inhibition factor gene acnR, inserting a glutamate dehydrogenase gene gdhA on a knockout site, knocking out a glutamine synthetase adenylation transferase gene glnE, and inserting an alpha-ketoglutarate dehydrogenase inhibition factor gene odhI on a knockout site to obtain the strain. According to the method, the supply of the precursor acetyl coenzyme A is enhanced by introducing a non-oxidative glycolysis pathway, the pH dynamic regulation system balances the metabolic flux, and the double breakthrough of the yield and the carbon recovery rate of the N-acetyl blue is realized by combining a two-stage fermentation process.

This invention belongs to the fields of fermentation and genetic engineering technology, and provides a high-yield hexanoic acid-producing engineered *Escherichia coli* strain, its construction method, and its applications. This engineered strain is *Escherichia coli*... E. coli w3110 was the starting strain, overexpressing the acetyl-CoA transferase gene. act and β-ketothiolase gene bktB Overexpression of trans-enoyl-CoA reductase gene ter 3-Hydroxybutyryl-CoA dehydrogenase gene hbd, and 3-hydroxybutyryl coenzyme A dehydratase gene crt Or, simultaneously not expressing the pyruvate formate lyase gene. pflB or lactate dehydrogenase gene ldhA This engineered bacteria can convert glucose into hexanoic acid, a high-value product, with a yield of 4.968 g / L after 18 hours of fermentation. It adopts aerobic fermentation, resulting in rapid cell growth, a short fermentation cycle, and a high acid production rate. The fermentation process is simple, easy to control, and has low production costs, which is conducive to its promotion and application in industrial production.

This invention discloses a method for the efficient synthesis of gastrodin using recombinant Corynebacterium glutamicum, belonging to the fields of genetic engineering, fermentation engineering, and synthetic biology. This invention uses Corynebacterium glutamicum as the host bacterium and knocks out… pobA The gene was expressed, and the engineered strain WN01 / pB6 was constructed using the expression plasmid pEC-XK99E to express cladoid-pyruvate lyase, carboxylic acid reductase, and glycosyltransferase, achieving heterologous synthesis of gastrodin. Further overexpression was then performed. aroG Genes, overexpression of protein-coding genes that activate carboxylic acid reductase sfp The engineered strain WN05 / pB6 was able to synthesize 7.37 g / L of gastrodin at the shake flask level, and the gastrodin yield reached 17.77 g / L at 72 h of batch fermentation in a 5-L fermenter, with a yield of 0.247 g / L / h. This provides a brand-new biomanufacturing platform for the industrial production of gastrodin.

The present invention relates to a method for producing α-sialyl-β-D-galactoside saccharides, particularly α-sialyl-(2→3)-β-D-galactoside saccharides and α-sialyl-(2→6)-β-D-galactoside saccharides, from a β-D-galactoside saccharide, a sialic acid donor, and an enzyme with β-galactoside α-sialyltransferase activity. The enzymes with β-galactoside α-sialyltransferase activity used herein do not exhibit catalytic activity towards the hydrolysis of cytidine 5'-monophospho-N-acetyl-neuraminic acid to cytidine and N-acetyl-neuraminic acid. The method can be performed in vitro and in vivo using a genetically engineered cell comprising a nucleic acid encoding said enzyme. Further, said process may be adapted to produce the sialic acid donor CMP-Neu5Ac from low-cost substrates N-acetyl-D-glucosamine (GlcNAc), pyruvate, a cytidine phosphate (CMP, CDP or CTP) and polyphosphate in a single reaction mixture with a set of optionally immobilized or optionally co-immobilized enzymes comprising N-acylglucoamine 2-epimerase (AGE), an N-acetylneuraminate lyase (NAL), an N-acylneuraminate cytidylyltransferase (CSS), optional a uridine kinase (UDK), a uridine monophosphate kinase and a polyphosphate kinase 3 (PPK3).