111 results about "Pyruvic Kinase" patented technology

Compounds and compositions comprising compounds that modulate pyruvate kinase M2 (PKM2) are described herein. Also described herein are methods of using the compounds that modulate PKM2 in the treatment of cancer.

Compounds and compositions comprising compounds that modulate pyruvate kinase M2 (PKM2) are described herein. Also described herein are methods of using the compounds that modulate PKM2 in the treatment of cancer.

The invention relates to a method for determining the content of blood ammonia, and also the reagent kit for blood ammonia diagnosis. The reagent kit comprises cushioning solution, adenosine triphosphate, phosphoenolpyruvate phosphatase, deacidized type coenzyme, ammonia kinase, pyruvate kinase, lactate dehydrogenase, and stabilizer. By mixing sample and reagent of a predetermiend volumetric ratio, generating coupling reaction between them, subjecting the final reactant to biochemiscal analyser, the main wavelength absorbancy variance ratio (speed) can be detected, and the blood ammonia content can thus be measured. The method of the invention can be used to obtain the needed measurement result purely through biochemical analytic instruments, and advantages of the method include higher sensibility, better accuracy, less susceptibility to contamination of internal or external materials, and easy application.

The invention relates to a liquid double reagent diagnostic reagent kit for determining the content of potassium ions in serum and blood plasma. By using the reaction principle that phosphoenolpyruvic acid (PEP) can be catalyzed by K+dependency pyruvate kinase (PK) to be converted into pyruvic acid and taking lactate dehydrogenase (LDH) as indicator enzyme by means of NADH, the reagent kit detects the reduction amount of the NADH at 340 nm and obtains the result that the change of the absorbance every minute (delta / min) is directly proportional to the content of K+, thereby establishing an enzyme coupling method for continuously determining the potassium ions. In addition, enzyme and a primer system required by slowly generating reduction type nicotinoyl coenzyme are added in a system andan enzyme-primer-coenzyme slow reaction system is formed in a reagent, so that the coenzyme can be slowly and circularly supplemented and the reagent can be stabilized when the concentration reaches a certain balance. The liquid double reagent diagnostic reagent kit is convenient and concise to use, carries out rapid detection on a common ultraviolet / visible light analyzer or a semiautomatic / fully-automatic chemical analyzer, needs no special or extra instrument and has low cost.

The invention is about a method of measuring the content of blood ammonia, and it also concerns the reagent box of blood ammonia diagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, 2ú¡ketoglutarate, reduced coenzyme, adenosine triphosphate, phosphoenolpyruvate, glutamate dehydrogenase, glutamate kinase, pyruvate kinase, lactate dehydrogenase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get the content of blood ammonia. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.

Compounds and compositions comprising compounds that modulate pyruvate kinase M2 (PKM2) are described herein. Also described herein are methods of using the compounds that modulate PKM2 in the treatment of cancer.

The invention relates to an Aspergillus niger gene engineering strain efficiently producing malic acid. The Aspergillus niger gene engineering strain is an Aspergillus niger gene engineering strain which knocks out a citric acid transporter gene cexA, and overexpresses a glucose transporter gene mstC, a hexokinase gene hxkA, a phosphofructokinase gene pfkA, and a pyruvate kinase gene pkiA which are derived from Aspergillus niger. The efficiency for producing malic acid of the Aspergillus niger gene engineering strain is significantly improved, the output on the eighth day through fed-batch fermentation in a fermentation tank reaches 195.72-210 g / L, a conversion rate of malic acid to glucose reaches 1.59-1.64 mol / mol, and a fermentation cycle is shorter by one day compared with a starting strain, namely the fermentation cycle is shortened from original 9 days to 8 days. An excellent strain for the preparation of malic acid by microbial fermentation methods is provided.

Compounds of general formula I:and compositions comprising compounds of general formula I that modulate pyruvate kinase are described herein. Also described herein are methods of using the compounds that modulate pyruvate kinase in the treatment of diseases.

The invention relates to a method for cell-free expression of signal protein and an expression system. The method includes joining a target gene of the signal protein on a pIX3.0 vector to obtain expression plasmids; extracting escherichia coli Rosetta (DE3) by S30 buffer solution to obtain escherichia coli extract; preparing an energy supply system comprising PEP (phosphoenolpyruvic acid) / PK (pyruvate kinase), CP (phosphocreatine) / CK (creatine kinase) and glucose; preparing an amino acid mixture and a saline solution; preparing lecithin / cholesterol liposome; adding the expression plasmids into a cell-free protein expression system comprising the escherichia coli extract, the energy supply system, the amino acid mixture, the saline solution and the lecithin / cholesterol liposome for expression. The method is capable of solving the problem of small signal protein expression quantity.

The invention described herein features methods, compositions, and kits that utilize activators of pyruvate kinase M2 (PKM2) for the treatment or amelioration of disorders related to PKM2 function and characterized by abnormally low levels of serine.

Compositions comprising compounds of general formula (I) that modulate pyruvate kinase are described herein. Also described herein are methods of using the compounds that modulate pyruvate kinase in the treatment of diseases.

Methods and compositions are provided for determining ADP in the presence of ATP. These comprise including among the assay reagents at least one of the correcting components creatine phosphokinase and phosphocreatine, pyruvate kinase and phosphoenolpyruvate, peroxidase and a non-interfering peroxidase substrate, and catalase. One aspect of the method employs formation of hydrogen peroxide from the ADP by pyruvate kinase, phosphoenolpyruvate and pyruvate oxidase. The hydrogen peroxide is then determined. A combined reagent having all of the reagents may optionally include a peroxidase when the hydrogen peroxide is to be enzymatically determined. A peroxidase substrate is added to the sample in conjunction with the peroxidase substrate reagent, the mixture incubated and depending on whether the peroxidase substrate is a fluorescer or chemiluminescer, the mixture may be illuminated with excitation light and the emitted light determined as a measure of the ADP in the sample.

[PROBLEMS] To provide lipase activity determination reagents which function by an enzymatic method. The reagents are easy to use in an ordinary clinical examination, have excellent handleability, and are excellent in accuracy and reproducibility. [MEANS FOR SOLVING PROBLEMS] Lipase activity is determined with any of reagents which comprise a low-concentration buffer and a diglyceride dissolved therein. The diglyceride is used as a substrate for lipase, whereby the liquid reagents can have long-term storage stability. One of the reagents converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, pyruvate kinase, lactate dehydrogenase, reduced NAD, ATP, and phosphoenol pyruvate. Another reagent converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, glucose, ADP-dependent hexokinase, glucose-6-phosphate dehydrogenase, oxidized NAD or oxidized NADP, and ATP. A further reagent converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, glycerol-3-phosphate oxidase, peroxidase, and a dye which colors in the presence of hydrogen peroxide.

The invention concerns the use of bioluminescence dependent on the reaction (1): luciferin+ATP+O2+Mg2++luciferase→oxyluciferin+photons for detecting and counting living cells of a given species potentially present in a liquid sample, said use being characterized in that it consists in measuring the total free intracellular adenyl nucleotides (AN) content, expressed in ATP form, of living cells of a given non-viral species, taking into account the fact that the sum of free intracellular ATP, ADP and AMP of said family is constant according to the relationship (2): [AN]=[ATP]+[ADP]+[AMP]=Cte after transforming the free intracellular ATP, ADP and AMP by using myokinase and pyruvate kinase, said measurement being performed (i) without adding ATP and (ii) after adding a known amount of ATP. The invention also concerns a method for detecting and counting cells by ATP-metry.

Compounds and compositions comprising compounds that modulate pyruvate kinase M2 (PKM2) are described herein. Also described herein are methods of using the compounds that modulate PKM2 in the treatment of cancer.

A method for producing a plant with increased yield as compared to a corresponding wild type plant whereby the method comprises at least the following step: increasing or generating in a plant or a part thereof one or more activities of a polypeptide selected from the group consisting of 26S proteasome-subunit, 50S ribosomal protein L36, Autophagy-related protein, B0050-protein, Branched-chain amino acid permease, Calmodulin, carbon storage regulator, FK506-binding protein, gamma-glutamyl-gamma-aminobutyrate hydrolase, GM02LC38418-protein, Heat stress transcription factor, Mannan polymerase II complex subunit, mitochondrial precursor of Lon protease homolog, MutS protein homolog, phosphate transporter subunit, Protein EFR3, pyruvate kinase, tellurite resistance protein, Xanthine permease, and YAR047C-protein.

The present invention relates to prevention of congenital deformations. The invention further relates to cancer inhibition and prevention. The invention further relates to methods and compositions to modulate, antagonize, or agonize disparate signaling pathways that may converge to regulate patterning events and gene expression during prenatal development, post-natal development, and during development in the adult organism. The invention also relates to activators or deactivators of pyruvate kinase M2 (PKM2) for the treatment, prevention, or amelioration of diseases related to PKM2 function.

To provide lipase activity determination reagents which function by the enzymatic method. The reagents are easy to use in an ordinary clinical examination, have excellent handleability, and are excellent in accuracy and reproducibility. Lipase activity is determined with any of reagents which comprise a low-concentration buffer and a diglyceride dissolved therein. The diglyceride is used as a substrate for lipase, whereby the liquid reagents can have long-term storage stability. One of the reagents converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, pyruvate kinase, lactate dehydrogenase, reduced NAD, ATP, and phosphoenol pyruvate. Another reagent converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, glucose, ADP-dependent hexokinase, glucose-6-phosphate dehydrogenase, oxidized NAD or oxidized NADP, and ATP. A further reagent converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, glycero-3-phosphate oxidase, peroxidase, and a dye which colors in the presence of hydrogen peroxide.

The invention relates to a kit for diagnosing / measuring potassium (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of the potassium (ions), and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, adenosine diphosphate, a phosphoenolpyruvic acid, coenzyme A, pyruvate kinase, pyruvate oxidase, NADH peroxydase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of potassium (ions).

The invention provides a novel serology biomarker GSK3beta detection method for cognitive impairment of a diabetic. The method comprises an enzyme activity assay method and a relative quantification dot-blot method. According to the enzyme activity assay method, a GSK-3beta enzyme activity assay kit of GENMED is applied; GSK-3beta phosphorylation is performed under the suppression of GSK-3alpha; along with oxidation reaction of reduced form of nicotinamide-adenine dinucleotid (NADH), a pyruvate kinase and lactic dehydrogenase continuous circulation method reaction system adopts the spectrophotometric method to measure the peak value change after oxidization so as to reflect GSK 3beta activity in a sample. The dot-blot method transfers blood platelet protein to an NC film according to the antigen and antibody fixation reaction principle and then utilizes antibody to perform detection. The novel serology biomarker GSK3beta detection method is used for studying GSK-3beta protein and enzyme activity expression in the diabetic blood and is expected to become a serology biomarker for early diagnosis of Alzheimer's disease (AD).

The invention relates to the technical field of microbial fermentation and particularly discloses a recombinant microbe capable of yielding N-acetylneuraminic acid and application of the recombinant microbe. According to the recombinant microbe provided by the invention, compared with an original strain, the recombinant microbe has lowered expression of pyruvate kinase and / or enzymatic activity; the pyruvate kinase is pyruvate kinase I and / or pyruvate kinase II; and the original strain is a microbe capable of synthesizing the N-acetylneuraminic acid. Compared with the original strain capable of synthesizing the N-acetylneuraminic acid, the recombinant microbe provided by the invention has lowered expression of the pyruvate kinase and / or enzymatic activity. The recombinant microbe is especially adapted for generating the N-acetylneuraminic acid through carrying out fermentation culture by using a cheap low-carbon raw material, i.e., glucose. According to the recombinant microbe provided by the invention, the fermentation level of synthesizing the N-acetylneuraminic acid is high, the production cost of the N-acetylneuraminic acid is reduced remarkably, and thus, the recombinant microbe has industrialized potentials.

Disclosed are pyruvate kinase M2 activators, which are bis sulfonamide piperazinyl and piperidinyl compounds of Formula (I), 2,4-disubstituted 4H-thieno[3,2-c]pyrrole-2-(substituted benzyl)pyridazin-3(2H)-ones of Formula (II) and 6-(3,4-dimethylphenylaminosulfonyl)-3,4-dihydro-1H-quinolin-2-one of formula (III), wherein L, R1, R2, R11 to R16, R21 and R22 are as defined herein, that are useful in treating a number of diseases that are treatable by the activation of PKM2, for example, cancer and anemia.

The invention relates to a separation and purification method of a Basidiomycetes antifungal component; the antifungal component is composed of two amide-type substances, and the molecular weight of the two substances is 5732.39 and 7187.01; the separation and purification process comprises the following steps: Basidiomycetes entity authentication, strain separation and purification, strain identification, mycelium liquid culture, antifungal component extraction, antifungal component separation, antifungal component purification. The method can inhibit chrysosperma mycelium growth and spore germination with 100 percent of inhibition rate, and has obvious inhibiting function to Pinus sylvestris germ at the same time. The Basidiomycetes antifungal substance affects the activity of seven enzymes of pathogenicbacteria, reduces the activity of hexokinase, improves the activity of pyruvate kinase, succinic acid, lactate dehydrogenase and ATP enzyme, and speeds up the respiration of germs.

The invention discloses a detection method and a kit for colorectal cancer cells based on quantum dot labeling. The detection method comprises steps of an anti-human M2-type pyruvate kinase monoclonal antibody preparation process, a quantum dot fluorescent probe preparation process and a process that quantum dot fluorescent probes are added in colorectal cancer cells and of fluorescence images of the colorectal cancer cells are observed under a confocal fluorescence microscope. Through anti-human-M2-type-pyruvate-kinase monoclonal antibodies labeled with quantum dots, a liquid chip or an immunofluorescence detection colorectal cancer cell or protein immunologic diagnosis kit can be formed. The kit has advantages of high sensitivity and high specificity, rapid and accurate detection can be achieved, secreta comprising feces and serum can be detected, and detection accuracy of the colorectal cancer can be raised greatly.

Compounds of general formula I:and compositions comprising compounds of general formula I that modulate pyruvate kinase are described herein. Also described herein are methods of using the compounds that modulate pyruvate kinase in the treatment of diseases.

The invention provides a small molecular activator taking human M2 type pyruvate kinase (M2 type pyruvate kinase, PKM2) as a target spot and application of the small molecular activator, and belongs to the technical field of pharmacy. According to the small molecular activator, the PKM2 is taken as the target spot, and the small molecule activator is screened by combining virtual screening based on butt joint and in-vitro biological activity evaluation. In-vitro activity test results show that two screened compounds have good PKM2 activation activity and good cancer cell inhibition activity, and discovered small molecules can provide a basis for research and development of new drugs for treating PKM2 related diseases, such as cervical cancer, breast cancer, ovarian cancer, prostate cancer,liver cancer, lung cancer, pancreatic cancer, colorectal cancer, leukemia, melanoma and multiple myeloma. A lead compound of the small molecular activator can be further optimized in structure, and arelatively good application prospect is achieved.

The present invention provides a method for producing an aromatic compound such as salicylic acid and a derivative thereof with high productivity using a microorganism. The present invention provides: a method for producing a microorganism having a sugar metabolic pathway modified, including suppressing the expression of a gene encoding a phosphotransferase system enzyme of the microorganism, suppressing the expression of a gene encoding pyruvate kinase of the microorganism, and introducing, into the microorganism, one or more genes encoding an enzyme that enables the microorganism to synthesize an aromatic compound from chorismic acid or isochorismic acid; a modified microorganism obtained by the method; and a method for producing an aromatic compound and a derivative thereof, including culturing the microorganism, and recovering an aromatic compound or the like from the culture.

The invention relates to the technical field of serum potassium detection, in particular to a serum-potassium detection reagent. A reagent R1 is prepared from a buffer solution, a cryptate agent, PEP, ADP, formic acid, formate dehydrogenase, alpha-ketoglutaric acid, NADH, GLDH (glutamate dehydrogenase), polyethylene glycol 6000, ethylene glycol, mannitol, trehalose, BSA, PK (pyruvate kinase), alkyl polyglucoside (APG) and preservative; a reagent R2 is prepared from a buffer solution, polyethylene glycol 6000, ethylene glycol, mannitol, trehalose, BSA, LDH (lactic dehydrogenase), alkyl polyglucoside (APG) and preservative. The Tris buffer solutions are adopted, multiple stabilizers are added, an NADH cyclic regeneration system is introduced, and therefore the stability of the reagent is remarkably improved; as the novel nonionic surfactant alkyl polyglucoside (APG) is added, turbidity of the reaction system can be effectively prevented, and the sensitivity and the stability of the reagent are remarkably improved.