30 results about "Fructokinase" patented technology

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.

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 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 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.

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 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.

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.

The invention relates to a method for determining ammonia (ammonia ion) content by utilizing an enzymatic colorimetric method and enzyme-linked immunosorbant assay and composition and components of a reagent for determining ammonia (ammonia ion) content. The technical principle of the method is as follow: determination is completed based on serial catalytic reactions of ammonia kinase, fructokinase and fructose dehydrogenase. The invention also relates to an ammonia (ammonia ion) diagnosis / determination kit. The method has the advantages of high sensitivity and few errors; therefore the method and the kit can be widely applied to clinical medicine / food inspection.

The invention relates to transgenic plants exhibiting increased cell wall content. In one embodiment, transgenic plants engineered to over-express fructokinase (FRK) or both FRK and sucrose synthase (SuSy) are provided. The FRK+SuSy double-transgenic plants of the invention consistently exhibit enhanced cell wall polymer deposition.

The invention relates to the technical field of genetic engineering and biological fermentation, and particularly discloses a recombinant microorganism capable of efficiently utilizing methanol and application of the recombinant microorganism. The invention provides a recombinant microorganism which has increased expression and / or enzymatic activity of methanol dehydrogenase and dihydroxyacetone synthetase and reduced expression quantity of a formaldehyde dissimilatory gene frmAB and expression and / or enzymatic activity of 6-phosphofructokinase compared with an original strain, and the recombinant microorganism has the advantages that the expression quantity of the methanol dehydrogenase and the dihydroxyacetone synthetase is increased, and the expression quantity of the formaldehyde dissimilatory gene frmAB and the expression quantity of 6-phosphofructokinase are reduced; the starting strain is escherichia coli. Specifically, after a formaldehyde dissimilatory gene frmAB, a 6-phosphofructokinase gene pfkAB and an alpha-ketoglutarate dehydrogenase gene sucA are knocked out at the same time, and a 3-phosphoglyceraldehyde dehydrogenase gene gapA is inhibited, the utilization rate of methanol can be further improved, and accumulation of intermediate formaldehyde is reduced.

The present invention relates to Corynebacterium sp. that is transformed with an Escherichia sp.-derived fructokinase gene to express fructokinase showing a sufficient activity of converting fructose into fructose-6-phosphate, thereby preventing unnecessary energy consumption, and a method for producing L-amino acids using the strain. The transformed Corynebacterium sp. of the present invention is able to express fructokinase from the Escherichia-derived fructokinase gene to prevent unnecessary energy consumption during fructose metabolism, leading to more cost-effective production of L-amino acids. Therefore, it can be widely used for the effective production of L-amino acids.

The invention relates to an ammonia (ammonia ion) content determination method employing an enzymatic colorimetric method and an enzyme-linked technology. The invention also relates to the composition and components of a reagent. The technological principle of the determination is that: the determination is achieved with a series of catalytic reactions of ammonia kinase, ketohexokinase fructokinase and mannitol 2-dehydrogenase. The invention also relates to an ammonia (ammonia ion) diagnosis / determination kit. The determination method provided by the invention is advantaged in high sensitivity and low error. Therefore, the determination method and the kit provided by the invention can be widely applied in clinical medicine / food examination.

The invention discloses a method for biocatalytically synthesizing D-(+)-glucuronic acid and an application of the method. The method comprises the following steps: by taking cane sugar as a substrate, adding enzyme to perform a catalytic reaction to obtain D-(+)-glucuronic acid, wherein the enzyme comprises sucrose phosphorylase, phosphoglucose mutase, inositol-1-phosphate synthase, inositol monophosphate esterase and inositol oxidase. The enzyme catalysis system can obtain a very high conversion rate. In addition, phosphate, phosphofructokinase and phosphoglucose isomerase are added into thereaction system, the conversion rate of the D-(+)-glucuronic acid can be further improved. The method is high in raw material utilization rate, high in D-(+)-glucuronic acid conversion rate, simple in step, low in production cost, small in pollution, small in environmental influence and capable of achieving large-scale production of D-(+)-glucuronic acid.

The present invention relates to Corynebacterium sp. that is transformed with an Escherichia sp.-derived fructokinase gene to express fructokinase showing a sufficient activity of converting fructose into fructose-6-phosphate, thereby preventing unnecessary energy consumption, and a method for producing L-amino acids using the strain. The transformed Corynebacterium sp. of the present invention is able to express fructokinase from the Escherichia-derived fructokinase gene to prevent unnecessary energy consumption during fructose metabolism, leading to more cost-effective production of L-amino acids. Therefore, it can be widely used for the effective production of L-amino acids.

Disclosed is a lactic-acid-producing Escherichia coli which has at least one gene including at least a sucrose hydrolase gene and selected from sucrose non-PTS genes (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) and is genetically so modified as to have a system for enhancing the production of lactic acid. Also disclosed is a method for producing lactic acid from a plant-derived sucrose-containing raw material by using the lactic-acid-producing Escherichia coli.