31 results about "Pyruvate kinase" 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.

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 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.

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 invention relates to Corynebacterium glutamicum with high indigo yield as well as a construction method and application thereof, and belongs to the technical field of gene recombination fermentation. The recombinant corynebacterium glutamicum can be used for overexpression of the following genes: a D-glucose-6-phosphate dehydrogenase gene zwf, a II type deoxyarabinose type heptulose phosphate synthase gene aroG and a flavin-containing monooxygenase gene CcFMO. The recombinant corynebacterium glutamicum can weaken the following genes: a pyruvate kinase gene pyk and a tryptophan synthase gene trpB. The corynebacterium glutamicum is used for overexpressing a zwf gene, an aroG gene and a CcFMO gene, weakening a pyk gene and a trpB gene and enhancing the expression quantity of an indole synthetic pathway related enzyme gene, so that the yield of the corynebacterium glutamicum at a shake flask stage reaches 619mg / L in a fermentation production process of indigo under the condition of not additionally adding tryptophan. The invention provides the recombinant corynebacterium glutamicum with high indigo yield, which is low in production cost, good in stability and high in synthesis efficiency, and is suitable for practical popularization and application.

Provided herein is the use of particular dose levels or amounts of certain pyruvate kinase activators or pharmaceutically acceptable salts or compositions thereof, for treating anemia associated with low risk MDS, lower risk MDS and / or intermediate risk MDS (collectively, LRMDS) and other conditions.

The invention relates to compounds of formula (Ia) and to their use in treating or preventing an inflammatory disease, a disease associated with an undesirable immune response, cancer, obesity, a diabetic disease or a blood disorder:wherein RA, RB, RC and RD, X, Y1, Y2, Y3, Z1, Z2 and m are as defined herein.

Provided herein are methods for preparing activators of Pyruvate Kinase (PK) or a salt or a hydrate thereof, as well as intermediates useful in the preparation of such compounds or salts or hydrates thereof. Also provided are a number of crystalline forms of select compounds or salts or hydrates disclosed herein.

The invention discloses a method for detecting the concentration of acetic acid in an anaerobic fermentation system by using a bioelectrochemical sensor, acetic acid is detected by using the bioelectrochemical sensor, the bioelectrochemical sensor comprises a screen-printed electrode, and the working surface of the screen-printed electrode is modified with reduced graphene oxide and polymethylene blue. The working surface of the screen printing electrode is connected with acetokinase, pyruvate kinase and lactic dehydrogenase through a cross-linking agent. According to the method, the adopted sensor has better conductivity, higher response speed and higher current intensity, when the sensor is used for detecting the concentration of acetic acid in a fermentation system, the sensor has the advantages of being easy to operate, low in sample consumption, low in price, high in accurate measurement precision, high in detection speed and the like, and rapid detection of the concentration of acetic acid can be achieved; the acetic acid concentration in the anaerobic fermentation process in different periods can be timely and accurately obtained, effective data can be provided for anaerobic fermentation analysis, and the method has important significance for monitoring the process stability of anaerobic fermentation.

The present application provides a Pichia pastoris engineering bacterium and a kit and application thereof, a method for obtaining the Pichia pastoris engineering bacterium, a method for improving glucose oxidase yield of the Pichia pastoris, and a method for producing glucose oxidase, wherein the Pichia pastoris engineering bacterium carries a glucose oxidase gene and a pyruvate kinase gene. The Pichia pastoris engineering bacterium of the present application can realize high glucose oxidase yield and high enzyme activity, and lays a good foundation for large-scale production of glucose oxidase.

The application relates to the technical field of engineering bacteria, and particularly discloses recombinant Escherichia coli for synthesizing fucosyllactose as well as a construction method and application thereof. In the genome of the recombinant Escherichia coli, phosphofructokinase genes or phosphofructokinase genes and pyruvate kinase genes are knocked out, so that glucose carbon flow is reduced to flow to a tricarboxylic acid cycle required for cell growth, and the intake of glucose carbon flow is forced to flow more to the synthesis of products (GDP-L-fucose in the synthesis of FL), and the synthesis efficiency of the products is improved. Meanwhile, the glucose carbon flow flowing to the cell growth is greatly reduced, and the glucose effect can be removed, so that the tolerance of the fermentation process to excessive glucose residues is improved, and the difficulty of downstream production amplification is reduced. The recombinant Escherichia coli uses glycerol and glucose compound carbon sources as raw materials, uses lactose as a substrate, and has a 3-fucosyllactose yield of 73.88 g / L in a 5L tank fermentation, which is the highest level reported at present.

Provided herein are methods of treating sickle cell disease, methods of increasing hemoglobin levels in a subject diagnosed with sickle cell disease, and methods of reducing hemolysis-associated complications in a subject diagnosed with sickle cell disease, together with kits for such treatments, in a subject in need thereof, comprising administering a Gardos channel inhibitor such as 2,2-bis(4-fluorophenyl)-2-phenylacetamide and a pyruvate kinase activator such as N-(4-((4-(Cyclopropylmethyl)-1-piperazinyl)carbonyl)phenyl)-8- quinolinesulfonamide to the subject. Subjects eligible for treatment include subjects as having hemolysis dominant (HD) sickle cell disease on the basis of laboratory markers, clinical history, and / or sequelae.

This renal fibrosis inhibitor contains a substance that interferes with pyruvate kinase M2.

A joint detection kit for detecting intestinal polyp factors, and a preparation method therefor, a detection method therefor and the use thereof. The joint detection kit at least comprises a plurality of test strips for detecting the following indicators: M2-pyruvate kinase, matrix metalloproteinase 9, myeloperoxidase, glutathione S-transferase Pi, cytidine deaminase, retinol binding protein 4, serine protease inhibitor F2, calprotectin and fecal occult blood. Conjugate pads (2) of each reagent strip are coated with detection antibody-colloidal gold conjugates. Detection lines (3) of the reagent strip are coated with specific capture antibodies, and the specific capture antibodies on each test strip are different. The joint detection performed by the joint detection kit can effectively improve the sensitivity and specificity of the detection on intestinal polyps. The detection time is merely 15 min, with the detection rate of more than 91%. Therefore, the accuracy on the detection and diagnosis of colorectal cancer caused by intestinal polyps is improved.

Provided herein is the use of pyruvate kinase (PK) activators or pharmaceutically acceptable salts or compositions thereof, for treating renal fibrosis and other renal diseases.

The invention relates to application of pyruvate kinase (PKM), triosephosphate isomerase (TPI1), alpha-enolase (ENO1) and lactic dehydrogenase (LDHB) in urine in preparation of medicines for exercise function evaluation (exercise intensity, exercise density, exercise duration, exercise function, exercise posture and physiological state), exercise risk prediction, exerciser physique evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise function evaluation, exercise and scientific exercise guidance, exercise scheme screening and other reagents are applied. Research proves that the proteins PKM, TPI1, ENO1 and LDHB in urine are key proteins for exercise training monitoring for the first time. Compared with the prior art, the expression of the four proteins in a urine sample after movement is obviously increased. The sensitivity and the specificity of various purposes such as function evaluation, exercise risk prediction and scientific exercise guidance of exercise training can be improved through independent use of one of the components and combined use of multiple components.

The application specifically relates to a method for preparing glutathione by immobilized enzyme and application. The method for preparing glutathione provided by the application comprises synthesizing glutathione by using saccharomyces cerevisiae haploid ascospores as a fixed enzyme catalyst to catalyze substrate reaction, using an ATP in-situ regeneration reaction system of phosphoenolpyruvate as an energy supply system, the saccharomyces cerevisiae haploid ascospores are knocked out or inactivated in combination of one or more than two of cell wall beta-glucan synthesis genes, and GSH1 genes, GSH2 genes and PYK1 genes are co-expressed. The saccharomyces cerevisiae haploid ascospores for simultaneously producing GSH1, GSH2 and pyruvate kinase are used as a fixed enzyme catalyst to catalyze synthesis of glutathione, three enzymes no longer need to be added externally, raw material cost is greatly reduced, the obtained product is easy to purify, and the prepared glutathione is high in purity.

Disclosed is an in vitro method for assessing whether a human patient suffering from periodontitis has mild periodontitis or advanced periodontitis. The method is based on the insight to determine a selection of two biomarker proteins. Accordingly, in a sample of saliva a patient suffering from periodontitis, the concentrations are measured of the proteins Pyruvate Kinase (PK) and at least one of Matrix metalloproteinase-9 (MMP9), S100 calcium-binding protein A8 (S100A8), and Hemoglobin subunit beta (Hb-beta); or of the proteins Matrix metalloproteinase-9 (MMP9) and at least one of S 100 calcium-binding protein A8 (S100A8) and S100 calcium-binding protein A9 (S100A9). Based on the concentrations as measured, a value is determined reflecting the joint concentrations for said proteins. This value is compared with a threshold value reflecting in the same manner the joint concentrations associated with advanced periodontitis. The comparison allows assessing whether the testing value is indicative of the presence of advanced periodontitis or of mild periodontitis in said patient. Thereby, typically, a testing value reflecting a joint concentration below the joint concentration reflected by the threshold value is indicative for mild periodontitis in said patient, and a testing value reflecting a joint concentration at or above the joint concentration reflected by the threshold value, is indicative for advanced periodontitis in said patient.

The present application relates to the technical field of bioengineering, and particularly relates to a recombinant Saccharomyces cerevisiae and a construction method and application thereof, comprising the following steps: step one, using the whole genome of Enterococcus faecalis as a template, a recombinant strain SC1-1 is constructed; step two, using the recombinant strain SC1-1 as a primary strain, L-lactate dehydrogenase CYB2 gene of Saccharomyces cerevisiae is knocked out to obtain a recombinant strain SC2; step three, using the recombinant strain SC2 as a primary strain, branched-chain-2-oxo acid decarboxylase THI3 gene of Saccharomyces cerevisiae is knocked out to obtain a recombinant strain SC3; step four, using the whole genome of Saccharomyces cerevisiae as a template, a recombinant strain SC4 is constructed. The present application innovatively optimizes the source of L-lactate dehydrogenase L-LDH, knocks out L-lactate dehydrogenase CYB2 and branched-chain-2-oxo acid decarboxylase THI3, overexpresses pyruvate kinase CDC19 and H(+)-ATPase PMA2, and makes the L-lactic acid production of Saccharomyces cerevisiae increase to 33.45 g / L.

Described herein are methods for using compounds that activate pyruvate kinase.

Provided herein are methods for preparing activators of pyruvate kinase (PK), or salts or hydrates thereof.

The present disclosure provides a recombinant yeast host cell for converting a carbohydrate into a fermentation product with an increased fermentation kinetic. The recombinant yeast host cell comprises an upregulated pyruvate kinase, a first engineered metabolic pathway to convert the carbohydrate into acetyl-coA comprising a heterologous polypeptide having phosphoketolase activity, and a second engineered metabolic pathway to convert acetyl-CoA into the fermentation product. The fermentation kinetic is increased when compared to a recombinant yeast host comprising the first and second engineered pathway but lacking the upregulated pyruvate kinase.

The invention provides a kit for detecting pyruvate kinase and application, and belongs to the technical field of pyruvate kinase detection. The kit comprises a cell ATP removal reagent and a pyruvate kinase detection reagent, the cell ATP removal reagent comprises adenylate cyclase; the pyruvate kinase detection reagent comprises a bioluminescent enzyme, a bioluminescent enzyme substrate, phosphoenolpyruvic acid, adenosine diphosphate and a non-PK kinase inhibitor; the non-PK kinase inhibitor is a myokinase inhibitor and / or a nucleoside diphosphate kinase inhibitor. The kit for detecting the pyruvate kinase can be used for qualitatively or quantitatively detecting the PK activity in a sample. When the kit is used for detecting the light-emitting signal generated by the ATP of the PK reaction product, the kit is suitable for long-time detection, and the enzyme activity is in positive correlation with the amount of the ATP. The kit provided by the invention can be used for detecting the enzyme activity of PK in different samples, and has the advantages of high sensitivity, high specificity, good linear relationship and stable fluorescence signal.

The present invention relates to a novel variant of pyruvate kinase 2 and a method of producing L-aromatic amino acids using the same. The pyruvate kinase 2 variant is obtained by substituting one or more amino acids in the amino acid sequence constituting pyruvate kinase 2 to change the activity of the protein, and a recombinant microorganism comprising the variant is capable of efficiently producing L-tryptophan, L-phenylalanine, or L-tyrosine.

The present application relates to the field of microbial technology, in particular to protein mutants, recombinant bacteria and preparation method and application thereof. The pyruvate kinase mutant provided by the present application has reduced enzyme activity, and the phosphoenolpyruvate carboxylase mutant provided by the present application has enhanced enzyme activity. The pyruvate kinase mutant and the phosphoenolpyruvate carboxylase mutant can promote the accumulation of phosphoenolpyruvic acid when used alone, thereby promoting the synthesis of products such as lysine and other products taking aspartic acid as a precursor and ensuring the good growth of the strain. The two mutants can further improve the yield and conversion rate of the above products when used in combination. The lysine yield and conversion rate of the recombinant microorganism expressing the pyruvate kinase mutant and the phosphoenolpyruvate carboxylase mutant provided by the present application are significantly improved compared with the original strain, and the present application provides advantageous gene and strain resources for the breeding of aspartic acid, its derivatives and aspartic acid family amino acid production strains.

Described herein are methods for using compounds that activate pyruvate kinase.

The invention provides a composite soaking agent for promoting synchronous seedling emergence of Chinese yam tubers as well as a preparation method and application of the composite soaking agent. The composite soaking agent is prepared from the following raw materials in parts by weight: 0.05 to 0.2 part of melatonin nano-liposome, 0.5 to 2.0 parts of sodium lignin sulfonate, 2 to 5 parts of garlic extracting solution, 5 to 10 parts of abscisic acid degrading enzyme activating agent, 3 to 8 parts of mitochondria activating agent, 2 to 6 parts of pyruvate kinase activating agent and 800 to 900 parts of purified water. The compound soaking agent solves the heterogeneity problem of tuber germination from multiple aspects through a five-in-one synergistic network of hormone regulation, energy enhancement, metabolism activation, oxidation resistance and bacterium resistance, and promotes tuber full seedling, so that integrated management of water, fertilizer and illumination in the later period is facilitated, the yield is increased, and the economic benefit is increased. A technical solution of a multi-target and multi-component synergistic mechanism is provided for high-quality and high-yield Chinese yam, and the method has remarkable application and popularization values.

The invention relates to an application of a pyruvate kinase M2 inhibitor in preparation of a neuroprotective agent or an anti-glioma drug, the pyruvate kinase M2 inhibitor can target pyruvate kinase M2 so as to target brain tissues, has high blood brain barrier transmittance and efficient neuroprotective activity and anti-glioma activity, and can be used for preparing neuroprotective agents or anti-glioma drugs. The compound is hopeful to provide more efficient and specific targeting protection for treatment of cerebral apoplexy, or is hopeful to become a glioma treatment drug with a better curative effect.

This invention provides the use of a reagent for increasing pyruvate kinase expression levels in the preparation of a drug for treating ocular neovascularization in subjects. The drug provides long-acting treatment with a high safety profile.