12 results about "Uridine diphosphate glucose" patented technology

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 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.

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 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.