20 results about "Glucose phosphate" patented technology

The application provides a myo-inositol-3-phosphate synthase mutant, which is obtained by mutating a myo-inositol-3-phosphate synthase with an amino acid sequence as shown in SEQ ID NO: 4 at positions 55, 57 and 273 respectively. The application also provides a preparation method of myo-inositol. The inventors have screened a wild-type myo-inositol-3-phosphate synthase and a myo-inositol monophosphatase with good performance, which can be used in cooperation with a glucan phosphorylase and a glucose phosphate mutase to catalyze starch to generate myo-inositol more efficiently. The myo-inositol-3-phosphate synthase mutant is applied to in-vitro non-fermentation biosynthesis of myo-inositol, combined with myo-inositol monophosphatases E1 and E5, and in a way of feeding whole cells, so that starch can be converted into myo-inositol in a one-pot method, without using NAD + coenzymes, and has a good industrial application prospect.

The invention discloses a genetically engineered bacterium for producing heparinogen and a construction method and application thereof, the genetically engineered bacterium takes Escherichia coli Nissle 1917 as a chassis bacterium, and the construction method comprises the following steps: (1) knocking out endonuclease I coding gene endA in a chassis bacterium genome; (2) knocking out a 6-phosphofructokinase I coding gene pfkA in a chassis bacterium genome; (3) carrying out overexpression on a 6-phosphoglucosamine synthetase encoding gene glmS and / or a UDP-glucose-6-dehydrogenase encoding gene kfiD from a chassis bacterium genome; (4) removing the feedback inhibition of the 6-phosphate glucosamine on the GlmS so as to promote the synthesis of UDP-N-acetylglucosamine; and (5) knocking out the 6-phosphate glucosamine deaminase coding gene nagB in the chassis bacterium genome. The engineering strain obtained by the invention can more efficiently synthesize the proheparin, the shake flask titer reaches 397.9 mg / L, which is increased by about 3 times compared with a wild strain, and the engineering strain has certain industrial application value.

The method for catalytically preparing D-mannose provided in the application can effectively reduce the amount of by-products glucose and fructose, and greatly improve the conversion rate of D-mannose. Specifically, the engineering bacteria expressing isoamylase gene and the engineering bacteria expressing alpha-glucan phosphorylase gene, glucose phosphomutase gene, bifunctional enzyme glucose phosphate isomerase / mannose 6-phosphate isomerase gene and mannose 6-phosphate phosphatase gene are used, starch or starch derivatives are used as substrates, phosphate buffer solution and Mg 2+ A preliminary catalytic system is constructed, and after the reaction is completed, there are unreacted substrates and maltodextrin in the reaction system, the saccharifying enzyme can hydrolyze the unreacted substrates and maltodextrin into glucose, the polyphosphate glucose kinase can convert all the glucose into phosphorylated glucose with the assistance of sodium hexametaphosphate, and then D-mannose is generated, thereby improving the yield and removing the by-products in the system.

This application relates to a phenytoin detection kit. Specifically, the glucose-6-phosphate dehydrogenase mutant of this application contains one mutation selected from the following compared to the wild-type glucose-6-phosphate dehydrogenase: D306C, D375C, or G426C. The detection kit prepared using the glucose-6-phosphate dehydrogenase mutant of this application exhibits high specificity, high sensitivity, ease of operation, short detection time, and accurate quantification, making it suitable for high-throughput detection.

This invention discloses a glucose phosphate mutase gene Rk PGM Its nucleotide sequence is shown in SEQ ID NO:1, and the amino acid sequence encoded by this gene is shown in SEQ ID NO:2; this gene was isolated from *Rhodotorula rubrum* (…). Rhodosporidium kratochvilovae YM25235 was used to ligate the gene to a vector and transfected into Rhodotorula rubrum cells. Experimental results showed that overexpression of Rk... PGM The gene will increase the level of extracellular polysaccharide synthesis in this strain; this invention modifies microorganisms through genetic engineering to improve their ability to secrete extracellular polysaccharides, providing a theoretical basis for increasing the yield of extracellular polysaccharides in Rhodotorula rubrum.

Provided in the present application are a glucose 6-phosphate isomerase OsPGIp1 protein and an encoding gene thereof, and use of the protein and a substance for regulating the expression of the encoding gene of the protein in regulating the height, grain weight, effective tiller number, seed-setting rate, or yield of a plant. The rice gene OsPGIp1 can be used for rice genetic breeding, germplasm resource screening, and mechanism research on breeding.

The invention discloses a genetically engineered bacterium for biosynthesis of inositol and application of the genetically engineered bacterium for biosynthesis of inositol, and the genetically engineered bacterium for biosynthesis of inositol is obtained by co-expressing a sweet orange inositol-3-phosphate synthase gene Csino3 and an escherichia coli inositol-1-monophosphate gene suhB in recombinant escherichia coli and then knocking out a glucose phosphate isomerase gene pgi. According to the technical scheme, the yield of inositol produced by the strain reaches 2.04 g / L and is increased by about 6.6 times compared with that before optimization, the inositol yield is greatly increased, the metabolic flux of glycolysis to glucose-6-phosphoric acid is effectively blocked, carbon metabolic flux is forced to be redirected to a pentose phosphate pathway, glucose-6-phosphoric acid is enriched, and a raw material is provided for inositol production. By blocking downstream branches of a glycometabolism pathway, enrichment of glucose-6-phosphate in cells is realized, so that sufficient precursor substances are provided for synthesis of inositol, and finally the yield of inositol is increased. The technical scheme is expected to be applied to construction of microbial cell factories, so that large-scale and low-cost production of inositol is realized, and theoretical support is provided for development of related industries.

This invention relates to the field of biotechnology, providing a novel NAD-dependent glucose-6-phosphate dehydrogenase, the amino acid sequence of which is shown in SEQ ID NO.2 or SEQ ID NO.3. The invention also provides the encoding gene of the above-mentioned gluconate-6-phosphate dehydrogenase, a recombinant expression vector including this gene, and a recombinant engineered bacterium. Furthermore, the invention provides a method for preparing the above-mentioned gluconate-6-phosphate dehydrogenase and its application in starch hydrogen production. This invention obtains a novel NAD-dependent NAD-dependent gluconate-6-phosphate dehydrogenase by mutating wild-type gluconate-6-phosphate dehydrogenase. + A mutant with better enzyme activity; this mutant is an NAD-dependent glucose-6-phosphate dehydrogenase, which can be used for hydrogen production from starch, effectively reducing the cost of hydrogen production and facilitating industrial production.

This application relates to the use of conjugates in the preparation of diagnostic reagents. Specifically, the glucose-6-phosphate dehydrogenase mutant of this application contains one or a combination of mutations selected from the following, compared to wild-type glucose-6-phosphate dehydrogenase: D306C, D375C, and G426C. Diagnostic kits prepared using the glucose-6-phosphate dehydrogenase mutant of this application exhibit high specificity, high sensitivity, ease of operation, short detection time, and accurate quantification, making them suitable for high-throughput detection.

The invention relates to a uridine diphosphate-glucuronyl transferase mutant and an application of the uridine diphosphate-glucuronyl transferase mutant in catalytic synthesis of glucuronide. The amino acid sequence of the uridine diphosphate-glucuronyl transferase mutant is as shown in SEQ ID NO. 2, and the nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. The invention provides a uridine diphosphate-glucuronyl transferase mutant with high activity and high stability. Compared with wild uridine diphosphate-glucuronyl transferase, the uridine diphosphate-glucuronyl transferase mutant has mutation of 10 amino acid sites. According to experimental determination, the melting temperature of the mutant is increased by 24 DEG C compared with that of a wild type, the activity is increased to 1.65 times, the expression quantity is increased to 4.2 times, and a powerful tool is provided for enzymatic synthesis of glucuronide compounds.

The invention discloses a universal enzyme circulation kit for quantitative detection of six substances and an integrated detection method, and belongs to the technical field of biological diagnosis and detection. According to the method, glucose-6-phosphate, glucose-6-phosphate dehydrogenase and beta-thio-nicotinamide adenine dinucleotide are taken as a core reaction system, and the change of five coenzymes, namely NAD (nicotinamide adenine dinucleotide), NADH (nicotinamide adenine dinucleotide), NADP (nicotinamide adenine dinucleotide phosphate), NADPH (nicotinamide adenine dinucleotide phosphate) and Thio-NAD and G6P are all converted into the absorbance signal change of the Thio-NADH at a dominant wavelength through an enzyme cycle amplification reaction. Quantitative detection of the six known single-component substances can be realized by using the same set of liquid double reagents only by adjusting detection parameters of an automatic biochemical analyzer, such as sample quantity and reading point range. The problem that various special reagents are needed in a traditional method is solved, and the detection efficiency and convenience are remarkably improved.

The application discloses a platycodon grandiflorum phosphoglucoisomerase gene PgGPI and an application of a product coded by the platycodon grandiflorum phosphoglucoisomerase gene PgGPI, and belongs to the technical field of platycodon grandiflorum genes. The platycodon grandiflorum phosphoglucoisomerase gene PgGPI is cloned from the platycodon grandiflorum, and a nucleotide sequence of the platycodon grandiflorum phosphoglucoisomerase gene PgGPI is shown as SEQ ID NO. 1. The PgGPI gene is successfully expressed in Escherichia coli, and it is verified through an enzymatic experiment that the PgGPI has catalytic activity and can effectively isomerize glucose-6-phosphate into fructose-6-phosphate, thereby laying a solid foundation for further exploring a biosynthesis mechanism of the platycodon grandiflorum polysaccharide. In addition, by means of the PgGPI gene and gene engineering technology, it is expected to significantly improve the content of the polysaccharide substance in the platycodon grandiflorum.

The purpose of the present invention is to provide a novel method that can produce a methyl compound by efficiently regenerating SAM using a general organic raw material such as glycine, serine, formic acid, methanol, or glucose and promoting a methylation reaction.　The present invention provides a method for producing a methyl compound using cells modified so that the activity or expression of S-adenosylmethionine (SAM)-dependent methyltransferase is enhanced and the activity or expression of phosphoglucose isomerase (Pgi) is reduced.

This application relates to a tacrolimus detection kit. Specifically, the glucose-6-phosphate dehydrogenase mutant of this application contains one or a combination of mutations selected from the following, compared to the wild-type glucose-6-phosphate dehydrogenase: D306C, D375C, and G426C. The detection kit prepared using the glucose-6-phosphate dehydrogenase mutant of this application exhibits high specificity, high sensitivity, ease of operation, short detection time, and accurate quantification, making it suitable for high-throughput detection.

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 discloses a method for preparing L-arabinose, and belongs to the technical field of preparation of L-arabinose. The invention particularly relates to a method for preparing L-arabinose by utilizing in-vitro biotransformation, which comprises the following steps: converting 6-phosphogluconic acid into D-ribulose 5-phosphoric acid by adopting 6-phosphogluconic acid dehydrogenase; the method comprises the following steps: converting D-ribulose 5-phosphoric acid into D-xylulose 5-phosphoric acid by adopting D-ribulose 5-phosphoric acid 3-epimerase; the method comprises the following steps: converting D-xylulose 5-phosphoric acid into L-ribulose 5-phosphoric acid by adopting L-ribulose 5-phosphoric acid 4-epimerase; under the action of isomerase and dephosphorizing enzyme, the L-ribulose 5-phosphoric acid is converted into the L-arabinose. The method is a brand-new biological method for preparing the L-arabinose, the L-arabinose is obtained with higher yield and higher purity, the purification process is simple, the production cost of the L-arabinose can be reduced, and the preparation process is environment-friendly.

The invention discloses application of glucose 6-phosphate isomerase OsPGIp1 in regulation and control of plant yield and plant height, and belongs to the technical field of gene engineering. The technical problem to be solved by the invention is how to regulate and control the yield and / or plant height of plants. Therefore, the invention provides application of the OsPGIp1 protein or a substance for regulating and controlling the expression of the coding gene of the OsPGIp1 protein in regulating and controlling the plant yield and / or the plant height. The OsPGIp1 protein can be a protein with an amino acid sequence as shown in SEQ ID NO: 3. The invention discloses the negative regulation effect of the OsPGIp1 gene in rice yield and / or plant height for the first time. The rice gene OsPGIp1 can be widely applied to the plant fields of rice genetic breeding, germplasm resource screening and the like, and plays an important role in breeding and mechanism research in the aspects of rice yield, grain weight, effective tiller number and / or maturing rate.

The invention provides a glucan phosphorylase mutant with improved thermal stability and application thereof, and relates to the technical field of enzyme engineering. Amino acids at the 40th site, the 271th site and the 707th site of wild-type glucan phosphorylase with an amino acid sequence shown as SEQ ID NO.1 from Solanum tuberosum are mutated to obtain the glucan phosphorylase mutant with improved thermal stability, the enzyme activity is improved by 39% at 37 DEG C, and the enzyme activity can still be kept about 60% after heat preservation is carried out for 1 hour at 60 DEG C; the amylose can be produced through conversion by taking glucose 1-phosphate as a substrate, and the conversion rate can be up to 98%. When the glucan phosphorylase mutant is applied to production of amylose, the reaction efficiency can be improved, the production period can be shortened, the production cost is further reduced, and the glucan phosphorylase mutant has a good application prospect in the field of starch synthesis of an industrial large system.

The invention relates to the technical field of protease mutation, and discloses a D-psicose 6-phosphoric acid-3-epimerase mutant and an application thereof. According to the invention, D-psicose 6-phosphoric acid-3-epimerase from Pellinia sp. Is used as a wild enzyme, and protein engineering molecular modification is carried out, so that mutants obtained by mutation of three specific amino acid sites, namely F36I, A146D and S202G, are obtained. The mutant has no activity on D-glucose 1-phosphoric acid, D-glucose 6-phosphoric acid, D-glucose, D-fructose and D-psicose, can specifically catalyze isomerization of D-fructose 6-phosphoric acid, and avoids side reaction, so that the high selectivity of D-fructose 6-phosphoric acid is maintained, and the yield of the mutant is improved. The activity of catalyzing the conversion of D-fructose 6-phosphoric acid into D-psicose 6-phosphoric acid is obviously improved, and the yield of multi-enzyme catalytic synthesis of D-psicose is effectively improved.

The invention provides a method for synthesizing D-psicose from glucose by modifying escherichia coli. The modification comprises the following steps: (1) introducing an exogenous D-psicose-6-phosphate epimerase gene alsE with an SUMO tag and an exogenous D-psicose-6-phosphate phosphatase gene a6PP with an SUMO tag, and constructing a metabolic pathway for synthesizing D-psicose from glucose; (2) a phosphofructokinase A gene pfkA, a phosphofructokinase B gene pfkB and a UDP-galactose-4-epimerase gene galE are knocked out, so that side metabolic shunt is reduced, accumulation of a D-psicose synthesis precursor is increased, and the synthesis efficiency of D-psicose is improved; according to the present invention, a 6-glucose phosphate isomerase gene pgi and a phosphogluconate dehydratase gene edd are knocked out, and exogenous NADP + dependent glutamate dehydrogenase gene gdh1 and NAD + dependent glutamate dehydrogenase gene gdh2 are introduced so as to regulate intracellular cofactor balance and restore cell growth, such that the bacterial strain can effectively synthesize the target product while the bacterial strain has good growth performance;