46 results about "Fructokinase" patented technology

The invention relates to the use of isoform-specific fructokinase (ketohexokinase) (KHK) inhibitors alone or in combination with various agents to both prevent and treat a wide variety of diseases including, but not limited to, sugar craving, obesity, features of metabolic syndrome (including insulin resistance, hypertriglyceridemia, hypertension, and fatty liver), polyuria, proximal tubular injury, and diabetic kidney disease.

The invention discloses a construction method of an engineering strain for generating allulose and an application thereof. The construction method comprises the following steps: increasing content ofintracellular fructose 6-phosphoric acid by reducing enzymatic activity of fructose 6-phosphokinase and glucose 6-phosphate dehydrogenase in corynebacterium glutamicum and enhancing enzymatic activityof glucokinase and glucose 6-phophate isomerase by regulating glucose intracellular metabolism; constructing a synthetic route of allulose composed of 6-aloxone phosphate 3-epimerase and 6-aloxone phosphate phosphorylase; constructing a metabolic pathway of fructose composed of fructose transmittase and fructokinase; constructing a metabolic pathway of glycerinum composed of glycerol transmittase, glycerol dehydrogenase and dihydroxyacetone kinase, thereby acquiring a corynebacterium glutamicum recombinant strain. The strain is capable of metabolizing glycerinum, glucose, fructose or saccharose for synthesizing allulose. Compared with the present reported method for compounding allulose through bioconversion of fructose, the construction method provided by the invention has the advantagesof high conversion rate, low production cost, and the like, and is suitable for large-scale production of allulose.

The invention discloses an engineered strain for producing tagatose and a construction method and application thereof. According to the construction method, by regulating intracellular glucose metabolism, the enzymatic activity of 6-phosphofructokinase in cells is reduced, the enzymatic activity of glucokinase and the enzymatic activity of glucose 6-phosphate isomerase are improved, therefore, thecontent of fructose-6-phosphate in the cells is increased, a tagatose synthesis path composed of tagatose-6-phosphate 3-epimeriase and tagatose-6-phosphate phosphorylase is constructed, and a fructose metabolism path composed of fructose permease and fructokinase is constructed. The glutamic acid corynebacterium recombination strain is obtained and can synthesize tagatose by metabolizing glucose,fructose or saccharose. Compared with currently reported fructose biotransformation tagatose synthesis methods, the construction method of the engineered strain has the advantages of being easy to operate and facilitating separation and is suitable for large-scale tagatose production.

The invention relates to the use of isoform-specific fructokinase (ketohexokinase) (KHK) inhibitors alone or in combination with various agents to both prevent and treat a wide variety of diseases including, but not limited to, sugar craving, obesity, features of metabolic syndrome (including insulin resistance, hypertriglyceridemia, hypertension, and fatty liver), polyuria, proximal tubular injury, and diabetic kidney disease.

The invention belongs to the technical field of biological gene engineering and discloses a promoter sequence of a fructokinase gene in an apple, and deletion mutants and application of. A promoter isnamed as FH1 and is a 1780-bp nucleotide sequence at the upper stream of the 5'-end of an MdFRK2 gene coding frame in an apple; and the mutants are sequences obtained by deleting fragments with different lengths from the 5'-end of the sequence of FH1 and are named as FH2, FH3 and FH4 respectively. The invention further discloses application of the MdFRK2 promoter and the deletion mutants of the MdFRK2 promoter in research on plant functional genes. The MdFRK2 promoter can regulate and control the specific expression of a target gene in cambium of stem tip growth points, old leaves, functionalleaf margins and other library tissues of arabidopsis thaliana and poplar, and a fragment from a transcription starting site to upstream-600 bp is determined to be a core fragment of a promoter region of the MdFRK2 promoter. The full length of the MdFRK2 promoter is induced by exogenous sugar and drought, which indicates that the promoter has important application value in industrial developmentof plant genetic engineering.

The invention discloses a genetically engineered bacterium for synthesizing D-allulose, and a construction method and application of the genetically engineered bacterium. The invention provides a bacillus subtilis genetically engineered bacterium capable of synthesizing the D-allulose by D-fructose, and a construction method of the bacillus subtilis genetically engineered bacterium. Particularly,bacillus subtilis WB600 is taken as a host bacterium, a fructokinase gene on the genome of the host bacterium and the genes of PTS system fructose specific translocator components IIABC and IID are knocked out to obtain a bacillus subtilis mutant, and a D-allulose-3-epimerase encoding gene from Arthrobacter globiformis M30 is subjected to free expression in the host bacterium to obtain recombinantbacillus subtilis. By use of the genetically engineered bacterium disclosed by the invention, the substrate D-allulose can be fully utilized, the yield of biosynthesis D-allulose is greatly improvedso as to be favorable for industrial production of the D-allulose, and the genetically engineered bacterium has a wide application prospect.

Belonging to the technical field of bioengineering, the invention relates to a genetic engineering strain producing succinic acid and a method for production of succinic acid by fermenting the strain, particularly a recombinant strain efficiently utilizing cane sugar and molasses to grow and produce succinic acid. The genetic engineering strain producing succinic acid is classified and named as Escherichia coli BA501, and has a preservation registration number of CCTCC NO:M2014014. The construction process of the strain mainly includes: taking Escherichia coli AFP111 that lacks lactate dehydrogenase gene and pyruvate formate lyase activity and has the chromosome ptsG gene undergoing spontaneous mutation as the starting strain, expressing exogenous sucrose permease, sucrose hydrolase and fructokinase genes, and then carrying out continuous domestication cultivation to obtain the strain efficiently utilizing cane sugar and molasses to grow and produce succinic acid. Thus, the synthesis efficiency of succinic acid is greatly improved. The fermentation method adopts a two-stage fermentation way, in the aerobic stage the biomass is improved oxygen, and in the anaerobic stage, fermentation and acid production are achieved.

The invention discloses a phosphofructokinase, and a coding gene and application thereof. The protein has one of the following amino acid residue sequences: 1) amino acid residue sequence disclosed as SEQ ID No.1 in the sequence table; and 2) protein with phosphofructokinase derived from SEQ ID No.1, subjected to substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid residue sequence disclosed as SEQ ID No.1 in the sequence table. The transgenic experiment on the protein indicates that the gene has the function of phosphofructokinase; and after the cDNA (complementary deoxyribonucleic acid) of the gene is transfected into a phosphofructokinase deficient strain RL257, the obtained transgenic strain can normally grow in an M63 basal culture medium which uses mannitol as the unique carbon source.

The invention discloses a method for increasing the plant cellulose content by utilizing an apple fructokinase gene. An apple fructokinase gene MdFRK2, a recombinant overexpression vector containing the apple fructokinase gene MdFRK2 and recombinant bacteria containing the apple fructokinase gene MdFRK2 are involved in the method. The apple fructokinase gene MdFRK2 in the method can be used for increasing the cellulose content; and the method plays an important role in cell wall transformation and green energy development.

The invention discloses a compound native microorganism domestication enzyme for efficiently improving an urban black and odorous water body, and a method for treating the urban and odorous water body. The compound native microorganism domestication enzyme comprises acetyl CoA acyltransferase, 1-phosphofructokinase, pyruvate phosphate dikinase, beta-hydroxybutyryl coenzyme dehydrogenase, 3-phosphoglycerate kinase, NAD-dependent beta-hydroxybutyl coenzyme A dehydrogenase, and thiolase. The method for treating the urban black and odorous water body comprises the following steps: preparation andactivation of the compound native microorganism domestication enzyme; putting of the activated compound native microorganism domestication enzyme into the black and odorous water body for a period oftime; and then aquatic plant construction. The compound native microorganism domestication enzyme of the invention has the advantages of efficiently improving the quality of the black and odorous sewage water body. In addition, the treatment method of the invention has the advantages of efficiently purifying the water body, and continuously adjusting and optimizing the water body environment.

The invention discloses fructokinase from a fungal traditional Chinese medicine and a coding gene and use thereof. The fungal traditional Chinese medicine fructokinase has an amino acid sequence as shown in SEQ ID NO: 1 and a nucleotide sequence of the coding gene as shown in SEQ ID NO: 2. The fungal traditional Chinese medicine fructokinase participates in synthesis of main precursor monosaccharide D-fructose of cordyceps polysaccharide from D-fructose-6-phosphoric acid. A fungal traditional Chinese medicine hirsutella sinensis polysaccharide biosynthetic metabolism pathway is studied in details in principle. A cloned DNA containing the nucleotide sequence can be transferred into engineering bacteria by transduction, transformation and conjugal transfer methods. An expression of a D-fructose biosynthetic gene is regulated, the host is endowed with high expressivity of D-fructose, an effective way is provided for increasing the yield of the D-fructose and a derivative thereof, and the fructokinase has a great application prospect.

The invention provides recombinant escherichia coli for synthesizing glucuronic acid by using glucose and a method for producing glucuronic acid by using the recombinant escherichia coli, and belongs to the field of microbial metabolic engineering. The engineering bacterium is obtained by taking escherichia coli as an original strain and knocking out a glucuronic acid isomerase coding gene uxaC, a 6-phosphofructokinase I coding gene pfkA, a 6-phosphofructokinase II coding gene pfkB and a glucose-6-phosphate dehydrogenase coding gene zwf. The engineering bacterium disclosed by the invention has a good application prospect in industrial production of glucuronic acid.

The present disclosure relates to: a fructokinase variant polypeptide; a polynucleotide encoding the variant polypeptide; a microorganism comprising same; a method for producing an L-amino acid, the method comprising a step for culturing the microorganism in a medium; or a use of the microorganism for L-amino acid production.

Belonging to the technical field of bioengineering, the invention relates to a genetic engineering strain producing succinic acid and a method for production of succinic acid by fermenting the strain, particularly a recombinant strain efficiently utilizing cane sugar and molasses to grow and produce succinic acid. The genetic engineering strain producing succinic acid is classified and named as Escherichia coli BA501, and has a preservation registration number of CCTCC NO:M2014014. The construction process of the strain mainly includes: taking Escherichia coli AFP111 that lacks lactate dehydrogenase gene and pyruvate formate lyase activity and has the chromosome ptsG gene undergoing spontaneous mutation as the starting strain, expressing exogenous sucrose permease, sucrose hydrolase and fructokinase genes, and then carrying out continuous domestication cultivation to obtain the strain efficiently utilizing cane sugar and molasses to grow and produce succinic acid. Thus, the synthesis efficiency of succinic acid is greatly improved. The fermentation method adopts a two-stage fermentation way, in the aerobic stage the biomass is improved oxygen, and in the anaerobic stage, fermentation and acid production are achieved.

The application belongs to the technical field of biology and specifically relates to a method for synthesizing D-tagatose through multi-enzyme cascade catalysis, which uses lactose as a substrate and utilizes beta-galactoside enzyme, L-arabinose isomerase, glucose isomerase, fructokinase, tagatose-1,6-diphosphate aldolase, polyphosphate kinase and phosphatase for synchronous cascade catalysis to synthesize D-tagatose. The method improves the conversion rate of D-tagatose by continuously converting the intermediate product D-glucose into D-tagatose, provides a new method for synthesizing D-tagatose from lactose, and provides certain theoretical basis and technical support for realizing the industrialized production of high-value-added D-tagatose.

The invention discloses a phosphofructokinase 2 mutation and / or knockout recombinant filamentous fungus for ethanol production and construction and ethanol production application thereof, which is characterized in that an endogenous phosphofructokinase 2 gene is knocked out and / or a mutated phosphofructokinase 2 gene is expressed in the filamentous fungus; wherein the mutated phosphofructokinase 2 means that only kinase activity is retained and phosphatase activity is lost or reduced after mutation. Compared with the original strain, the obtained genetic engineering strain has the advantages that the ethanol synthesis capability is improved, and the glucose metabolism rate is accelerated.

The invention discloses a genetic engineering strain for synthesizing rebaudioside M as well as a preparation method and application of the genetic engineering strain, and relates to the technical field of bioengineering. The genetic engineering strain disclosed by the invention contains a coding gene of glycosyl transferase YojK, a coding gene of a glycosyl transferase UGT76G1 mutant P84A / P91A with an amino acid sequence as shown in SEQ ID NO.2, a coding gene of sucrose synthase Sus3, a coding gene of sucrose permease CscB and a coding gene of fructokinase Csck. The genetically engineered bacterium disclosed by the invention can be used for synthesizing rebaudioside M by utilizing a cheap raw material RebA, and the yield of rebaudioside M reaches 30.13 g / L. In addition, the RebM synthesis method directly adopts a fermentation method, enzyme purification is not needed, the technological process is simplified, and the production cost is reduced. The established escherichia coli genetic engineering strain capable of utilizing sucrose shows a good industrial application prospect in the synthesis of RebM.

The invention relates to a protein scaffold mediated multienzyme cascade system and application thereof in preparation of D-tagatose. The protein scaffold mediated multi-enzyme cascade system provided by the invention comprises fructokinase, D-tagatose-6-phosphate 4-epimerase and D-tagatose-6-phosphate phosphatase, and the protein scaffold mediated multi-enzyme cascade system comprises fructokinase, D-tagatose-6-phosphate 4-epimerase and D-tagatose-6-phosphate phosphatase, the D-tagatose-6-phosphoric acid 4-epimerase and the D-tagatose-6-phosphoric acid phosphatase are immobilized on the protein support SH3, and the D-tagatose-6-phosphoric acid 4-epimerase is immobilized on the protein support SH3. According to the system, a plurality of key enzyme spaces in a reaction path are positioned on a protein scaffold through a protein-protein interaction structural domain (SH3), so that a substrate channel and a spatial proximity effect are enhanced, and the yield of D-tagatose is increased. According to the method disclosed by the invention, the yield of the D-tagatose is 3.2-7.7 times that of the original D-tagatose by optimizing reaction conditions.

The invention relates to the use of one or more fructokinase (ketohexokinase) (KHK) inhibitors to both prevent and treat a wide variety of diseases including, but not limited to, alcohol craving, alcohol addiction, alcohol induced liver disease including fatty liver and cirrhosis.

The present invention provides: a lactic acid-producing Escherichia coli comprising at least one gene of a sucrose non-PTS gene group, including at least a sucrose hydrolase gene, provided that a combination of a repressor protein (cscR), a sucrose hydrolase (cscA), a fructokinase (cscK) and a sucrose permease (cscB) and a combination of a sucrose hydrolase (cscA), a fructokinase (cscK) and a sucrose permease (cscB) are excluded, wherein the lactic acid-producing Escherichia coli comprises a lactic acid production enhancing system provided by genetic recombination; and a lactic acid production method including producing lactic acid from a plant-derived sucrose-containing raw material by using the lactic acid-producing Escherichia coli.

The application discloses a kind of added Co 2+ The application discloses a method for improving the yield of D-allulose produced by multi-enzyme catalysis, belonging to the field of biotechnology. The application provides a method for producing D-allulose by multi-enzyme catalysis, which comprises adding sucrose phosphorylase from Bifidobacterium adolescentis, fructokinase from Clostridium acetobutylicum, D-allulose-6-phosphate phosphatase from Clostridium thermocellum, and D-allulose-6-phosphate-3-epimerase from Pantoea sp to a reaction system containing sucrose and ATP, and then adding a metal ion solution to the reaction system to produce D-allulose. The application synthesizes an important low-calorie functional rare sugar D-allulose using inexpensive sucrose as a substrate, and experiments show that the addition of cobalt ions can increase the yield of D-allulose by five times.

The present application relates to the field of metabolic engineering, and provide a method for high-efficiency synthesis of D-allulose by Escherichia coli using nutritional sweetener-sucrose, wherein D-allulose is produced by recombinant Escherichia coli using cheap and traditional nutritional sweetener, and wherein Escherichia coli JM109(DE3) is used as a chassis host, wherein a path for producing D-allulose from sucrose is constructed by introducing exogenous proteins sucrose-6-phosphate hydrolase CscA, sucrose permease CscB, fructokinase CscK, D-allulose-6-phosphate epimerase AlsE and D-allulose-6-phosphate phosphatase A6PP into wild-type Escherichia coli to realize co-expression, and wherein the substrate utilization rate is improved by introducing ptsG-F and knocking out ptsG, fruA, ptsI and ptsH to reprogram the sugar transmembrane transport pathway.

High concentration of free cells of heterofermentative lactic acid bacteria (LAB) in a resting or slowly growing state are used to convert fructose into mannitol. Efficient volumetric mannitol productivities and mannitol yields from fructose are achieved in a process applying cell-recycle, continuous stirred tank reactor and / or circulation techniques with native LAB cells or with LAB cells with inactivated fructokinase gene(s). Mannitol is recovered in high yield and purity with the aid of evaporation and cooling crystallization.

The present disclosure relates to fructokinase variant polypeptides; a polynucleotide encoding the variant polypeptide; a microorganism comprising the same; a method for producing an L-amino acid, said method comprising the step of culturing said microorganism in a culture medium; or to the use of said microorganism in the production of L-amino acids.

The invention provides a prognostic prediction method for a non-small cell lung cancer. The prognostic prediction method comprises the step of detecting phosphorylation levels of fructokinase A and anS659 site of acetyl-coenzyme A synthase kinase 2 of a cancer cell. According to the prognostic prediction method, the phosphorylation expression levels of the fructokinase A and / or the S659 site of the acetyl-coenzyme A synthase kinase 2 are detected through an immunohistochemical technique on the basis of phosphorylation of the fructokinase A and / or the S659 site of the acetyl-coenzyme A synthase kinase 2, and the survival time of the non-small cell lung cancer is independently predicted.

The invention discloses a composite native microorganism domestication enzyme for efficiently improving urban black and odorous water bodies and a method for treating urban black and odorous water bodies. The composite native microorganism domestication enzyme includes acetyl CoA acyltransferase, 1-phosphofructokinase and pyruvate phospho-dikinase. , β-hydroxybutyryl-CoA dehydrogenase, 3-phosphoglycerate kinase, NAD-dependent β-hydroxybutyl-CoA dehydrogenase and thiolase. A method for treating urban black and odorous water bodies includes the following steps: preparing and activating compound native microorganism domestication enzymes; putting the activated compound native microorganism domestication enzymes in black and smelly water bodies for a period of time; and then constructing aquatic plants. The composite indigenous microbial domestication enzyme of the present invention has the advantages of efficiently improving the quality of black and odorous sewage; in addition, the treatment method of the present invention has the advantages of efficiently purifying the water body, and sustainably adjusting and optimizing the water body environment.

Methods for identifying chronic hydrocephalus include assaying for an amount in a biological sample of one or more biomarkers selected from pyruvate dehydrogenase lipoamide kinase isozyme 4 (PDK4), phosphofructokinase-muscle (PFKM), low density lipoprotein receptor adaptor protein 1 (LDLRAP1), glucagon-like peptide-1 receptor (GLP-1R), hemoglobin subunit alpha 1 (HBA1), hemoglobin subunit alpha 2 (HBA2), heme oxygenase 1 (HMOX1), and combinations thereof. Methods for screening for a compound useful for treating chronic hydrocephalus are also provided and include contacting a cell with an effective amount of a test compound, and then detecting an expression or activity level of one or more of the biomarkers.

The application discloses a method for producing D-tagatose by a three-enzyme cascade method, and belongs to the field of biotechnology. In the method, fructose is used as a substrate, a reaction system containing metal ions and ATP is used, fructose kinase FRK derived from Clostridium acetobutylicm, D-tagatose 6-phosphate 4-epimerase GatZ derived from Caldilinea aerophila DSM 14535 and D-tagatose-6-phosphate phosphatase MmPase derived from Methanothermobacter marburgensis are used to produce D-tagatose. The yield of D-tagatose is 2.7-9 times of the original yield by optimizing the temperature, the type and the concentration of the metal ions.

The invention relates to application of a cyanobacteria fructokinase inactivating agent in preparation of cyanobacteria with fructose secretion capability and high fructose yield, and further relates to a reagent for knocking out a fructokinase gene corresponding to fructokinase in the cyanobacteria or a fructokinase function antagonist. The invention also relates to the fructokinase-disabled cyanobacteria and application of the fructokinase-disabled cyanobacteria in fructose production. According to the invention, fructokinase in a carbon circulation path of cyanobacteria is knocked out, a mutant strain accidentally obtains the capacity of mass production of fructose and secretion of fructose out of cells, the fructose yield is increased by hundreds of times, and the possibility is provided for subsequent industrial production of fructose by using cyanobacteria; and a foundation is provided for further metabolic engineering modification by taking cyanobacteria as a chassis algal strain.