35 results about "Glucokinase" patented technology

The invention discloses escherichia coli recombinant engineering bacteria for producing citicoline as well as a construction method and application of the escherichia coli recombinant engineering bacteria, and belongs to the technical field of biology. A genome cytidine deaminase coding gene cdd is knocked out on the basis of Escherichia coli K-12 MG1655, a temperature control expression vector PBV220 is used for expressing a SpnCCT-SpnCKI-pRpLpyrG mutant gene cluster, the expression of choline transporter protein BetT is enhanced, the gene locus of Escherichia coli ptsH-ptsI-crr is destroyed, meanwhile, a glucose permease gene glf and a glucokinase gene glk from Zymomonas mobilis are introduced, and the mutant gene cluster of the Escherichia coli K-12 MG1655 is obtained. The escherichia coli recombinant engineering bacterium ZMCB02NCL capable of efficiently biosynthesizing citicoline is obtained, and the preservation number of the escherichia coli recombinant engineering bacterium ZMCB02NCL is CCTCC NO: M 2025483.

The present disclosure relates to a combination therapy comprising a first AAV vector genome comprising an insulin expression cassette; and a second AAV vector genome comprising a glucokinase expression cassette; wherein the first AAV vector genome and the second AAV vector genome are in a ratio selected from the group consisting of 1:0.25-0.75, 1:1.75-2.25, and 1:3.75-4.25, and methods for using the same for treating diabetes.

The present invention relates to lactose-positive, sucrose-negative, Streptococcus thermophilus strains. The strains are carrying one or more mutation in one or more gene of the sucrose regulon and optionally one or more further mutation affecting the glucose porter, such as affecting glcU and / or its expression. The strains of the present invention may further carry mutations in genes encoding a protein of the mannose-glucose-specific PTS and a glucokinase, a protein of the mannose-glucose-specific PTS and the catabolite control protein A (CcpA), and / or a protein of the mannose-glucose-specific PTS, a glucokinase and CcpA. The inventive strains may, when fermenting milk do this without the need or with a need for adding a reduced amount of sucrose, optionally providing a low-lactose fermented milk and / or a fermented milk not undergoing post-acidification when stored at fermentation temperature. The invention also concerns a composition comprising at least one, lactose-positive, sucrose-negative, Streptococcus thermophilus strain of the invention, and the use of this strain or composition to manufacture a fermented dairy product.

The invention relates to the technical field of biology, in particular to a method for increasing the growth rate of a Cupriavidus hookworm strain with carbon dioxide as a carbon source and an engineering strain. Glucokinase genes participating in glucose metabolism in the Cupriavidus hookworm strain are knocked out, alkaline phosphatase genes and haloid acid dehalogenase family (HAD) phosphatase genes are knocked out on the basis, and the growth rate of the Cupriavidus hookworm strain under the condition that carbon dioxide serves as a carbon source is increased through the operation. The bacterial strain with the increased proliferation rate can be used for producing single-cell protein by taking carbon dioxide as a raw material and is beneficial to fixation of greenhouse gas carbon dioxide. Meanwhile, the production efficiency can be remarkably improved by utilizing the chassis cell to construct a metabolic engineering-based product synthesis strain.

The invention relates to a hyaluronic acid high-yield strain based on metabolic flow optimization as well as a construction method and application thereof, and belongs to the technical field of genetic engineering. According to the corynebacterium glutamicum, sources of key rate-limiting enzymes UDP-glucose dehydrogenase and glutamine-fructose-6-phosphate aminotransferase are screened, and heterologous expression of hyaluronic acid synthase and overexpression of endogenous inositol permease and glucokinase are combined, so that the corynebacterium glutamicum capable of highly producing hyaluronic acid is constructed in a form of optimizing metabolic flux; according to the invention, the hyaluronic acid is used as a substrate, polyhydroxybutyrate synthase clusters are further overexpressed in the substrate, the glucose uptake and utilization efficiency of a host are improved, the metabolic coupling of cofactors is realized, the yield of the hyaluronic acid reaches 45g / L, the production intensity is 1.25 g / L / h, a brand new tool is provided for the production of the hyaluronic acid, and the method has an excellent application prospect.

This invention relates to a blackcurrant and orange juice mixed soft drink, belonging to the field of beverage technology. The mixed soft drink contains blackcurrant extract, orange juice, white hyacinth bean seed extract, mogrosides, steviol glycosides, etc. The blackcurrant extract is a product with a lower than 5kDa obtained by enzymatic hydrolysis of blackcurrants with hemicellulase and pectinase, followed by sequential enzymatic hydrolysis with nattokinase, fig protease, papain, and glucokinase. The white hyacinth bean seed extract is a product with a lower than 5kDa obtained by trypsin hydrolysis of white hyacinth bean seeds, followed by co-fermentation with Propionibacterium finigense, Lactobacillus brevis, and Bifidobacterium longum. The blackcurrant extract and white hyacinth bean seed extract have excellent anti-fatigue, antioxidant, and immune-enhancing effects, making the mixed soft drink both delicious and healthy.

Described are compositions and methods for inhibition of glucokinase (GCK) gene expression and protein production. RNA interference (RNAi) agents for inhibiting the expression of GCK gene are described. The GCK RNAi agents disclosed herein may be targeted to cells, such as hepatocytes, for example, by using conjugated targeting ligands. Pharmaceutical compositions comprising one or more GCK RNAi agents optionally with one or more additional therapeutics are also described.

The invention discloses a method for biosynthesizing rebaudioside based on an in-vivo constructed UDP (User Datagram Protocol) recovery system, and belongs to the technical field of biology. A UDP recovery system is constructed in saccharomyces cerevisiae, specifically, the saccharomyces cerevisiae M24 is taken as a host, synthetic pathway coding genes including a sucrose synthase gene SUS3 and a polyphosphate dependent glucokinase gene PPGK are integrally expressed through a CRISPR Cas9 editing technology, and the genetically engineered bacterium is used for fermentation production of rebaudioside. The gene for endogenous regulation and control of UDP glucose synthesis is enhanced, a UDP recovery system is constructed at the same time, supply of UDP glucose is improved so as to improve the catalytic efficiency of enzyme, and therefore the yield of rebaudioside is increased. The breakthrough provides key technical support for industrial fermentation synthesis of rebaudioside, especially rebaudioside M, and has significant economic benefits and environmental protection values.

The invention relates to a composition for producing a fermented milk product comprising: (i) a starter culture comprising a glucose-fermenting Streptococcus thermophilus (St) strain and a glucose-fermenting Lactobacillus delbrueckii subsp. bulgaricus (Lb) strain; and (ii) a glucose-deficient Streptococcus thermophilus (St) strain, wherein said St strain is galactose-fermenting and carries a mutation in the DNA sequence of the glcK gene encoding a glucokinase protein, which mutation inactivates the glucokinase protein or has a negative effect on expression of the gene.

Providing modified insulin and glucokinase nucleic acids for treating diabetes SOLUTION: Reducing hyperglycaemia and maintaining euglycaemia are the goals of all therapeutic approaches to the T1DM and T2DM. Current therapy for most diabetics is based on regular subcutaneous injections of both short-acting and long-acting insulin preparations. The present disclosure relates to modified nucleic acid sequences encoding insulin and glucokinase, expression cassettes and delivery vectors comprising the same, and methods for their delivery to treat diabetes.SELECTED DRAWING: None

The invention discloses an Escherichia coli strain capable of efficiently secreting and expressing alpha-lipoic acid and application of the Escherichia coli strain, and belongs to the technical field of biology. Escherichia coli is used as a chassis strain for synthesizing lipoic acid by a microbiological method, and an efficient production strain is constructed by optimizing a metabolic pathway; the related modification comprises the following steps: overexpressing a lipoic amidase coding gene eflpA, a truncated thioesterase mutant 'TesA coding gene, a lipoyl synthase coding gene lipA, an iron-sulfur cluster regeneration gene cluster coding gene iscSUA, an S-adenosylmethionine synthetase coding gene sam2 and a 5'-methyl adenosine thioate / S-adenosylhomocysteine nucleoside enzyme coding gene mtn, and carrying out gene coding. A fatty acid degrading enzyme coding gene fadE, a thioesterase coding gene tesA, a pyruvate formate lyase gene cluster coding gene pflAB and a glucokinase coding gene glk are knocked out. According to the invention, the Escherichia coli realizes efficient de novo synthesis and secretory expression of lipoic acid, and finally the extracellular yield in the shake flask fermentation process reaches 16.96 mg / L.

The present invention relates to a composition for producing a fermented milk product comprising (i) at least one Streptococcus thermophilus (St) strain, wherein the St strain is galactose-fermenting, wherein the strain carries a mutation in the DNA sequence of the gick gene encoding a glucokinase protein, wherein the mutation inactivates the glucokinase protein or has a negative effect on expression of the gene, and (ii) at least one Lactobacillus delbrueckii subsp. bulgaricus (Lb) strain, wherein the Lb strain is lactose-deficient and capable of metabolizing a non-lactose carbohydrate.

The present invention relates to a microorganism of the genus Corynebacterium with improved L-lysine production capacity and a method for producing L-lysine using the same. The microorganism of the genus Corynebacterium has enhanced activity of glucose-promoting diffusion transporters and glucokinase, thereby improving the production yield of L-lysine compared to the parent strain or cases where glucose-promoting diffusion transporters or glucokinase are enhanced alone.

The present invention relates to a microorganism belonging to the genus Corynebacterium and having improved L-lysine production capacity, and a method for producing L-lysine using the same, the microorganism being capable of producing L-lysine by enhancing the activity of a glucose-promoting diffusion transporter and a glucokinase. Therefore, the production yield of L-lysine can be improved compared with the condition of enhancing the parent strain, the glucose promoting diffusion transporter or the glucokinase alone.

The present invention relates to the field of microbial technology, and specifically discloses a recombinant microorganism and its application in the fermentative production of ectoine. Compared with the starting strain, the recombinant microorganism of the present invention expresses the ectABC gene of Chromohalobacter salexigens, expresses the glucose facilitator gene of Zymomonas mobilis glf while reducing the ptsG gene expression, and also overexpresses glucokinase glk, preferably also overexpresses phosphoenolpyruvate carboxylase ppc, and more preferably replaces the original promoter of the isocitrate dehydrogenase gene icd with a growth-phase-dependent promoter. Without significantly affecting the growth performance of the bacterial cells, the recombinant microorganism of the present invention can significantly improve the yield and productivity of ectoine in fermentative production, thereby enhancing the efficiency of industrial production of ectoine and reducing the production cost.

Described are compositions and methods for inhibition of glucokinase (GCK) gene expression and protein production. RNA interference (RNAi) agents for inhibiting the expression of GCK gene are described. The GCK RNAi agents disclosed herein may be targeted to cells, such as hepatocytes, for example, by using conjugated targeting ligands. Pharmaceutical compositions comprising one or more GCK RNAi agents optionally with one or more additional therapeutics are also described.

The application discloses application of polyphosphate-dependent glucokinase in glycosylation modification of pentacyclic triterpenoid compounds, a sugar group donor of the glycosylation modification is sucrose, the polyphosphate-dependent glucokinase is used for converting byproduct fructose into fructose-6-phosphate, and the polyphosphate-dependent glucokinase is CgPPGK with an amino acid sequence as shown in SEQ ID NO:1 or Asp-PPGK with an amino acid sequence as shown in SEQ ID NO:2. The polyphosphate-dependent glucokinase CgPPGK or Asp-PPGK is coupled to a traditional UDP-glucose regeneration system in the application, a coupled three-enzyme UDP-glucose regeneration system is formed, consumption of fructose is realized, more UDP-glucose is generated in the reaction, glycosylation efficiency and sucrose utilization rate are improved, and the glycosylation modification capacity for a substrate is obviously higher than that of an original traditional UDP-glucose regeneration system.

The invention relates to the technical field of biochemical engineering, and particularly discloses a recombinant microorganism for producing L-histidine and application of the recombinant microorganism. Compared with an original strain, the recombinant microorganism has the advantages that the expression of a ptsG gene is reduced while galactose transporter protein galP and glucokinase glk are overexpressed, and / or one or more of transketolase tktA, transketolase tktB, transaldolase talA or transaldolase talB are overexpressed, and the original strain is escherichia coli capable of producing histidine. The recombinant microorganism provided by the invention can improve the production efficiency of histidine and reduce the cost of industrial production of histidine.

Described are compositions and methods for inhibition of glucokinase (GCK) gene expression and protein production. RNA interference (RNAi) agents for inhibiting the expression of GCK gene are described. The GCK RNAi agents disclosed herein may be targeted to cells, such as hepatocytes, for example, by using conjugated targeting ligands. Pharmaceutical compositions comprising one or more GCK RNAi agents optionally with one or more additional therapeutics are also described.

The invention relates to a construction and fermentation method of recombinant corynebacterium glutamicum for high-yield indigoidine, and the recombinant corynebacterium glutamicum is obtained by taking corynebacterium glutamicum as an original strain and transforming one or more of A-D items; the method comprises the following steps: A, knocking out a gene for coding glucose transporter on a genome, and integrating a glucose permease gene and a glucokinase gene; b, knocking out a gene for coding gamma-glutamyl kinase on a genome, and integrating a glk gene at the same time; c, overexpressing an ornamental blue synthetase gene and a 4 '-phosphopantetheinyl transferase gene; and D: overexpressing a glutamine synthetase gene, an alpha-ketoglutarate dehydrogenase inhibiting enzyme gene, a glutamate dehydrogenase gene and an isocitrate dehydrogenase gene. According to the method, glutamine can be efficiently converted, the yield of the indigoidine is greatly increased, and a foundation is laid for industrial production of the indigoidine.

The invention discloses a glucose metabolism reconstruction-based engineering strain for efficiently synthesizing rhamnolipid and a construction method thereof, the construction method comprises the following steps: taking pseudomonas putida KT2440 or a flag gene cluster knockout strain thereof as an original strain, optimizing the genome of the original strain through gene modification, and introducing recombinant plasmids to obtain the engineering strain for efficiently synthesizing rhamnolipid; the modification comprises at least one of (1) to (6): (1) heterologous expression of rhamnolipid synthesis related genes; (2) replacing the rmlA gene with the gene rmlA *; (3) knocking out related genes for coding Gad; (4) heterologous expression of a galP gene encoding glucose transporter protein; (5) heterologous expression of the glf gene encoding the glucose transporter; and (6) overexpressing and coding the glk gene of glucokinase. The method disclosed by the invention has the beneficial effects that efficient directional distribution of glucose to a rhamnolipid synthesis route is realized, and the synthesis efficiency of the rhamnolipid is remarkably improved.

The present invention relates to specific uses of lactose-positive Streptococcus thermophilus strains in cheese production. The strains used in accordance with the present invention can carry one or more mutations in the glcK gene encoding a glucokinase, which mutations induce a reduction in glucokinase activity, carry mutations in the ccpA gene, and / or carry mutations in at least one of the genes encoding a mannose-glucose specific PTS protein. The invention further relates to compositions comprising a strain of the invention used according to the invention in combination with at least one strain of Lactobacillus and / or one strain of Mesophilic Lactobacillus, methods for making cheese, and cheese obtained by these methods.

The invention discloses an N-acetylglucosamine transferase mutant and application thereof in production of L-tryptophan, and belongs to the technical field of genetic engineering. In order to solve the problems of yield limitation and the like caused by excessive consumption of phosphoenolpyruvic acid in the production of L-tryptophan, the invention weakens a phosphoprotein coding gene crr on escherichia coli; meanwhile, a uridine diphosphate galactose pyranose mutase gene glf and a glucokinase gene glk of the pseudomonas mobilis and an endogenous galactose H (+) transporter enzyme gene galP of escherichia coli are integrated at a pflB site, and a genetically engineered bacterium is obtained; mutagenesis is carried out on the basis to obtain a mutant strain with improved L-tryptophan yield; and the N-acetylglucosamine transferase mutants nagC and nagE are obtained by virtue of sequencing. Fermentation proves that overexpression of the mutant nagC or nagE or simultaneous overexpression of the mutants nagC and nagE in the genetically engineered bacteria can improve the yield of L-tryptophan.

The invention relates to bacillus subtilis for producing D-psicose and application of the bacillus subtilis, and belongs to the technical field of microorganisms. The invention discloses a recombinant bacillus subtilis obtained by performing gene editing by using a CRISPR / Cpf1 system, a D-psicose 6-phosphoric acid 3-epimerase coding gene A6PE, a glucokinase coding gene glck and a glucose transporter coding gene glf are integrally expressed in a starting bacillus subtilis genome, and a 6-phosphofructokinase coding gene pfkA is knocked out, so that the recombinant bacillus subtilis is obtained. A metabolic pathway for producing D-psicose by taking glucose as a substrate is constructed in bacillus subtilis, and the production efficiency is improved in a form of enhancing glucose uptake and conversion capacity, so that the yield of D-psicose of the constructed recombinant bacillus subtilis at a 250 mL shake flask fermentation level reaches 23 g / L. The bacillus subtilis for producing D-psicose provided by the invention has the advantages of directional regulation and control of metabolic flux, high substrate utilization rate, low production cost and the like, and is suitable for industrial production.

The present disclosure relates to modified nucleic acid sequences encoding insulin and glucokinase, expression cassettes and delivery vectors comprising the same, and methods for delivery of the same for treating diabetes.

The present invention discloses a recombinant engineering bacterium of Escherichia coli for producing citicoline and a construction method and application thereof, belonging to the field of biotechnology; Escherichia coli K‑12 MG1655-based knockout of the genomic cytidine deaminase gene cdd The temperature-controlled expression vector PBV220 was used to express SpnCCT‑SpnCKI‑pRpLpyrG Mutating the gene cluster enhances the expression of the choline transporter BetT and disrupts Escherichia coli ptsH‑ptsI‑ crr locus, and simultaneously introduced from Zymomonas mobilis ( Zymomonas mobilis ) glucose permease gene glf and glucokinase genes glk , and obtained the recombinant Escherichia coli ZMCB02NCL capable of efficiently biosynthesizing citicoline, with the deposit number being CCTCC NO: M 2025483.

The present disclosure relates to modified nucleic acid sequences encoding insulin and glucokinase, expression cassettes and delivery vectors comprising the same, and methods of delivering them to treat diabetes. Certain embodiments are directed to a polynucleotide encoding a human insulin protein, the polynucleotide comprising: (i) a nucleotide sequence encoding a signal peptide, where optionally, the signal peptide is not a wild-type preproinsulin signal sequence; and (ii) a nucleotide sequence encoding a proinsulin polypeptide comprising an amino acid modification at a position selected from amino acids B10, B28, and / or B29 of the human insulin B chain, C1 and / or C32 of the human insulin C chain, or any combination thereof, relative to the corresponding amino acid position in wild-type proinsulin; and optionally, the polynucleotide further comprises a cleavage site.

This invention discloses a method for constructing a zebrafish model of fatty liver based on glucokinase overexpression and its application. The method includes the following steps: using cDNA as a template, a zebrafish GCK gene fragment is obtained by PCR amplification; the vector fragment and the GCK gene fragment are homologously recombined using a homologous recombinase to obtain a recombinant plasmid; the recombinant plasmid is transformed into competent cells, and an endotoxin-free plasmid is extracted as a zebrafish GCK expression vector; a microinjection system is used to inject the plasmid into one-cell-stage fertilized eggs; after the fertilized eggs hatch into juveniles, fluorescent juveniles are selected for further culture until sexual maturity; sexually mature males and females are paired to spawn, and a stably heritable GCK-overexpressing zebrafish strain is selected from the offspring. This invention can obtain a zebrafish strain with stable high GCK expression, which can be used as a zebrafish model of fatty liver based on GCK overexpression.

The invention discloses a glucokinase mutant for enhancing the glucose utilization rate of escherichia coli and application of the glucokinase mutant. The invention belongs to the technical field of biology, and particularly relates to a glucokinase mutant for enhancing the glucose utilization rate of escherichia coli and application of the glucokinase mutant. The mutant protein disclosed by the invention is any one of the following proteins: 1) a protein obtained by mutating the 142 A of a sequence 1 into V or L and keeping other amino acid residues unchanged; 2) a protein obtained by mutating R in the 188th sequence of the sequence 1 into H and keeping other amino acid residues unchanged; and 3) a protein obtained by mutating the 142 of the sequence 1 into V or L, mutating R of the 188 of the sequence 1 into H and keeping other amino acid residues unchanged. The encoding gene of glucokinase in receptor escherichia coli is knocked out, and the encoding gene of mutant protein is introduced into the receptor escherichia coli, so that the enzyme activity of the obtained recombinant escherichia coli is 7.3 times that of wild escherichia coli, and the glucose utilization rate of escherichia coli is increased.

The present invention relates to a novel mutant of 5-dehydro-2-deoxyglucono kinase and a method for producing 5'-inosinic acid using the same. In the 5-dehydro-2-deoxyglucono kinase mutant, one or more amino acids in the amino acid sequence constituting 5-dehydro-2-deoxyglucono kinase are substituted, resulting in a change in the activity of the protein, and a recombinant microorganism containing the same can efficiently produce 5'-inosinic acid.