20 results about "Uridine diphosphate glucose" patented technology

The invention discloses a genetically engineered bacterium for producing uridine diphosphate glucose as well as a construction method and application of the genetically engineered bacterium, and belongs to the technical field of genetic engineering. The following genes are overexpressed in the genetically engineered bacterium: a pyrimidine nucleoside operon gene, a carbamyl phosphate synthetase gene, an aspartate carbamyl transaminase gene, an orotic acid phosphoribose transferase gene, a phosphoribose pyrophosphate synthase gene, a uridylate kinase gene and a glucophosphate mutase gene. A uridine triphosphate glucose-1-phosphate uridine group transferase gene; the genetically engineered bacterium does not express the following genes: a purine nucleotide repressor protein gene, a deoxyribonucleoside triphosphate reductase gene, a nucleoside triphosphate pyrophosphohydrolase gene, a cytidine triphosphate synthase gene, a pyrimidine nucleotidase gene, a nucleotidase gene, an ornithine carbamoyltransferase gene, a UDPG dehydrogenase gene and a lacI repressor protein gene. The genetically engineered bacterium can be used for efficiently synthesizing UDPG by taking glucose as a substrate.

The invention belongs to the field of gene engineering, and particularly relates to application of a willow uridine diphosphate glucose-4-epimerase gene SpUGE1 in plant cell wall component remodeling, organelle cadmium content reduction and cadmium resistance improvement. According to the invention, the SpUGE1 gene in willow is extracted, and is transferred into arabidopsis thaliana by constructing a recombinant plasmid, so that the biological activity, Cd < 2 + > concentration and the like of wild arabidopsis thaliana and transgenic arabidopsis thaliana are compared. The result of the embodiment shows that the transgenic arabidopsis thaliana has certain tolerance to cadmium stress, the cell wall of the transgenic arabidopsis thaliana is obviously thickened, and the content of cadmium in organelles is obviously reduced.

The invention discloses an extraction method of mogroside V, and relates to the technical field of biology. Escherichia coli containing glycosyl transferases UGT-MS1 and UGT-MS2 is cultured, fermented, centrifuged and homogenized to obtain enzyme clear liquid, mogroside IIE, uridine diphosphate glucose, the enzyme clear liquid and salt are added into a phosphate buffer solution for enzyme catalysis to obtain conversion liquid, and the conversion liquid is subjected to enzyme purification to obtain the mogroside V. And finally, carrying out ultrafiltration, column loading and elution, then feeding into alkaline anion exchange resin, and concentrating and drying effluent to obtain mogroside V. The product is high in yield and environment-friendly.

The application relates to the field of biological medicine, and discloses a glucosyltransferase mutant, a coding gene of the glucosyltransferase mutant, a recombination carrier, a recombination strain, an enzyme preparation, and application of the glucosyltransferase mutant. The glucosyltransferase mutant is a point mutation at the R position of the amino acid sequence shown in SEQ ID NO:1, and 4-8 histidines are added at the N end. A method for preparing gastrodin comprises the following steps: contacting the glucosyltransferase mutant with p-hydroxybenzyl alcohol and uridine diphosphate glucose. The glucosyltransferase mutant provided by the application has higher specificity for the substrate p-hydroxybenzyl alcohol, thereby obviously improving the conversion rate of the conversion of p-hydroxybenzyl alcohol into gastrodin, and having a good industrial application prospect.

The present invention discloses a genetically engineered bacterium for the whole-cell catalyzed synthesis of glucoside ferulate and its application, belonging to the technical field of bioengineering. In the present invention, by overexpressing the expression of two key enzymes involved in the synthesis of uridine diphosphate glucose (UDP-glucose) in the host cell, namely phosphoglucomutase and uridine diphosphate glucose pyrophosphorylase genes, the intracellular accumulation level of UDP-glucose is further increased; by transferring the UDP-glucosyltransferase gene into the host cell and achieving its high-efficiency expression, a genetically engineered bacterium capable of biocatalytic synthesis of glucoside ferulate is obtained. By adopting the technical scheme of the present invention, the problem of intracellular UDP-glucose supply can be solved, and glucoside ferulate is prepared by whole-cell catalysis using ferulic acid as a substrate. Finally, the yield of glucoside ferulate reaches 220.47 mg / L, and the molar conversion rate is 60.1%.

Disclosed is a method for efficiently preparing rebaudioside M by using a bio-enzymatic method. The preparation method comprises: adding rebaudioside A, uridine diphosphate glucose, glycosyltransferase, and a salt into a buffer solution, mixing to obtain a mixed solution, performing an enzyme-catalyzed reaction while stirring, and obtaining said rebaudioside M. The amino acid sequence of the glycosyltransferase is one or more of those shown in SEQ ID NO. 1-12. The glycosyltransferase can also be coupled with sucrose synthase for catalysis, so as to use sucrose as a glycosyl donor to, in one step, catalyze rebaudioside A to generate rebaudioside M.

The inventive technology described herein includes systems, methods, and compositions for the upregulation of Uridine diphosphate glucose (UDPG) recycling in a yeast cell, by expressing a heterologous nucleotide sequence, operably linked to a promoter, encoding a phosphoglucomutase enzyme which catalyzes the conversion from glucose-6-phosphate to glucose- 1 -phosphate, and a UDP-glucose pyrophosphorylase enzyme which catalyzes the formation of UDPG from glucose 1 -phosphate and UTP. The increased levels of UDPG substrate produced by the yeast cell may be used as a substrate for a co-expressed UDP -glycosyltransferases enzyme capable of generating cannabinoid glycoside compounds.

The present application relates to a kind of selectively improved glycosyltransferase UGTBL1 mutant and its catalytic synthesis of application of salidroside, belong to genetic engineering and enzyme engineering field.The mutant obtained by single point mutation technology includes the amino acid sequence of 132th amino mutation, the amino acid sequence of the glycosyltransferase is as shown in SEQ ID NO:1.The research results show that the glycosyltransferase mutant S132V, S132G described in the application has good selectivity, can efficiently catalyze tyrosol and uridine diphosphate glucose to synthesize salidroside, and the selectivity is 89%, suitable for the enzyme preparation of salidroside, with wide application prospect.

A use of uridine diphosphate glucose (UDPG) in treatment of abnormal liver glucolipid metabolism. Specifically, a use of UDPG or a pharmaceutical composition comprising same in the preparation of a drug for preventing and / or treating abnormal liver glucolipid metabolism, wherein the abnormal liver glucolipid metabolism is preferably nonalcoholic fatty liver disease, dyslipidemia and related cardiovascular diseases. Treatment using UDPG can reduce lipid synthesis and lipid synthesis related gene expression in the liver.

The application discloses application of a glycosyltransferase Bs-YjiC in Bacillus subtilis 168 and comprises the following steps: catalyzing reaction of ginsenoside Rg2, the glycosyltransferase Bs-YjiC and uridine diphosphate glucose, so as to glycosylate C 20 hydroxyl group of the ginsenoside Rg2 to generate ginsenoside Re; or catalyzing reaction of notoginsenoside R2, the glycosyltransferase Bs-YjiC and uridine diphosphate glucose, so as to glycosylate C 20 hydroxyl group of the notoginsenoside R2 to generate notoginsenoside R1; the application utilizes the glycosyltransferase Bs-YjiC of microbial origin to catalyze C 20 hydroxyl group glycosylation of the ginsenoside Rg2 or the notoginsenoside R2, and since the ginsenoside Re and the notoginsenoside R1 have important pharmacological effects, the glycosyltransferase Bs-YjiC of microbial origin can be utilized to catalyze synthesis of the ginsenoside Re and the notoginsenoside R1 beneficial to human health.

The application discloses application of uridine diphosphate glucose in promoting absorption of heavy metal cadmium by hyperaccumulating plants, and is characterized in that: a water solution of uridine diphosphate glucose is sprayed on leaf surfaces of hyperaccumulating plants in a growth stage, cadmium is enriched by using the growth and heavy metal absorption promoting effect of the uridine diphosphate glucose on the hyperaccumulating plants, and after the hyperaccumulating plants grow, the hyperaccumulating plants are cut or uprooted, and the application is obtained.The application sprays the water solution of the uridine diphosphate glucose on the leaf surfaces of the hyperaccumulating plants, uses the growth and heavy metal absorption promoting effect of the uridine diphosphate glucose on the plants to improve the cadmium accumulation amount of the hyperaccumulating plants, further improves the heavy metal enrichment efficiency, and effectively promotes efficient repair of heavy metal cadmium contaminated soil or water bodies by the hyperaccumulating plants.

The invention belongs to the technical field of genetic engineering, and particularly relates to a genetically engineered bacterium for efficiently synthesizing betacyanin as well as a construction method and application of the genetically engineered bacterium. According to the invention, a gene editing technology is used for jointly enhancing the expression levels of phosphoglucomutase pgm and UDP-glucose pyrophosphorylase galU on a BL21 (DE3) genome, the supply level of uridine diphosphate glucose which is a precursor substance for synthesizing betacyanin can be effectively improved, and a genetically engineered bacterium is constructed by combining a double-plasmid multienzyme expression system and overexpressing hpaBC, dodA, CYP76AD1 and 5GT. The engineering bacterium realizes whole-cell catalytic synthesis of betacyanin by taking tyrosine and glucose as direct substrates for the first time through collaborative optimization of genome editing and a plasmid expression system, and the product yield and the process economy are remarkably improved.

The application discloses a kind of serum-free medium for efficiently enriching tumor stem cells and application thereof, belong to the technical field of cell biology.The serum-free medium includes basic medium, basic additive factor, functional additive component;Among them, functional additive component includes: 1 μM-1mM of alpha-ketoglutaric acid dimethyl ester, 10nM-10 μM of butyl benzyl phthalate, 0.01 μM-1 μM of endothelin-1, 0.01mM-1mM of uridine diphosphate glucose, 1nM-100nM of tetrabromobisphenol A.The application provides stable stemness maintenance conditions for tumor stem cells by adding functional additive component to serum-free medium;At the same time, the synergistic effect between functional additive component and other components significantly improves the enrichment efficiency of tumor stem cells, and has the advantage of low cost;Therefore, it has good application prospect in efficient enrichment of tumor stem cells.

The present invention discloses a method for enzymatically synthesizing nucleotide sugars, which belongs to the field of biotechnology. The present invention uses sucrose as a substrate, and generates uridine diphosphate glucose and fructose by sucrose synthase catalysis, fructose is catalyzed by mannose isomerase or lyxose isomerase to generate mannose, mannose is catalyzed by N-acetyhexose aminophosphokinase to generate mannose-1-phosphate, and mannose-1-phosphate is catalyzed by GDP-Man pyrophosphorylase to obtain guanosine diphosphate mannose. The present invention cascades modules for synthesizing UDP-Glc and synthesizing GDP-Man, uses cheap sucrose as a raw material, and simultaneously obtains UDP-Glc and GDP-Man in the shortest reaction route, thereby reducing the cost of synthesizing high-value-added oligosaccharides and glycosides.

This invention discloses a method for extracting mogroside V, relating to the field of biotechnology. The method involves culturing, fermenting, centrifuging, and homogenizing *E. coli* containing glycosyltransferases UGT-MS1 and UGT-MS2 to obtain an enzyme supernatant. Then, mogroside IIE, uridine diphosphate glucose, the enzyme supernatant, and salt are added to a phosphate buffer solution for enzymatic catalysis to obtain a conversion solution. Finally, the solution is ultrafiltered, loaded onto a column, eluted, and then passed through an alkaline anion exchange resin. The effluent is concentrated and dried to obtain mogroside V. The product has a high yield and is environmentally friendly.

The invention relates to the technical field of medicine purification, and discloses a uridine diphosphate glucose separation and purification method, which comprises the following steps: S1, loading: filtering a uridine diphosphate glucose crude product, loading the filtered uridine diphosphate glucose crude product into a chromatographic column, and taking polystyrene-divinyl benzene monodisperse microspheres as a matrix as a filler of the chromatographic column; s2, elution: eluting the uridine diphosphate glucose adsorbed in the chromatographic column by using an eluent; and S3, collecting and gathering: carrying out segmented collection on the eluted uridine diphosphate glucose solution, and gathering the component liquid meeting the target peak value requirement. The separation and purification method of uridine diphosphate glucose is simple and convenient, the purification period is short, the requirement for high purity of uridine diphosphate glucose can be met only through one-step chromatographic purification, the purity reaches 99% or above, the purification yield is high and stable, and better use prospects are brought.

The invention relates to a method for preparing a C-7-position glycoside compound by using an isoflavone aglycone compound, and belongs to the technical field of biology. According to the method, an isoflavone aglycone compound is taken as a substrate, a belamcanda chinensis glycosyl transferase gene BcUGT009 gene encoding protein, a belamcanda chinensis glycosyl transferase BcUGT032 gene encoding protein or a belamcanda chinensis glycosyl transferase BcUGT124 gene encoding protein is taken as a catalyst, uridine diphosphate glucose is taken as a glycosyl donor, and C-7-site glycosylation reaction is performed to generate the corresponding glycoside compound. According to the application, the functions of the three glycosyl transferase genes BcUGT009, BcUGT032 and BcUGT124 in the blackberry lily are identified and verified for the first time, the effects of the three glycosyl transferase genes in catalyzing flavonoid aglycones to generate C-7 glucoside are defined, the blank of research on a specific flavonoid glycoside biosynthesis pathway of the blackberry lily is filled, and important information is provided for research on a biosynthesis mechanism of active ingredients of medicinal plants.

The invention discloses a recombinant uridine diphosphate glucose dehydrogenase mutant and a preparation method thereof, and relates to the technical field of biology, the mutant is derived from pyogenic streptococcus and is obtained through site-specific mutagenesis (mutation site L102V-H129K-I270G), double tags of His-tag and SUMO-tag are fused at the N terminal, the corresponding protein sequence is SEQIDNO.1, and the coding gene sequence is SEQIDNO.2. Compared with a recombinant wild type UGD, the mutant has the advantages that the mutagenesis efficiency is high, and the mutagenesis efficiency is high. The performance of the mutant is remarkably improved, the catalytic activity reaches 1.8 times of that of a wild type, the residual activity is 69.4% after the mutant is stored at room temperature for 5 days, the soluble expression quantity is improved by 5 times, UDPG can be efficiently promoted to be oxidized to generate UDPGA, the commercial production cost of UDPGA is effectively reduced, and the mutant has a wide application prospect. The invention is suitable for the fields of scientific research reagents, biological medicines, cosmetics and the like.

The utility model discloses an extraction device of mogroside V, and relates to the technical field of biology. An inlet of a reaction tank is respectively communicated with a fermentation liquid tank, a mogroside IIE tank, a uridine diphosphate glucose tank and a salt tank through pipelines, and an outlet of the reaction tank is communicated with a conversion liquid tank through a pipeline; an outlet of the conversion liquid tank is communicated with a macroporous resin column through a pipeline, an inlet of the macroporous resin column is communicated with an eluent tank through a pipeline, an outlet of the macroporous resin column is communicated with an eluent tank, and an outlet of the eluent tank is communicated with an anion exchange resin column through a pipeline. An outlet of the anion exchange resin column is communicated with a concentration tank, an outlet of the concentration tank is communicated with a dryer, and an outlet of the dryer is communicated with a mogroside V tank. The yield and the purity of the product are improved, and the production steps and the production period are shortened.

The invention discloses a method for quantitatively measuring pyrophosphoric acid (hereinafter referred to as PPi) in human plasma. According to the method, a sample and radioactive isotope labeled uridine diphosphate glucose are subjected to an enzymatic reaction in a reaction system, and the radioactive intensity before and after the enzymatic reaction is measured so as to calculate the content of pyrophosphate in the sample. According to the method, unreacted uridine diphosphate glucose is adsorbed by activated carbon before determination, the use concentration of the activated carbon is 3-3.5% (w / v), and the pH value is 7.2-8.5. The method can be used for detecting the content of PPi in plasma and is not influenced by the matrix effect.