54 results about "Glucose phosphate" patented technology

The invention provides application of acidophilous glycosyltransferase in salidroside production, and belongs to the technical field of biological engineering. The problem of producing salidroside under the acidic condition is solved. Comprising an application of acidophilous glycosyl transferase with an amino acid sequence as shown in SEQ ID NO.1 in salidroside production under an acidic condition and an acidophilous escherichia coli engineering strain for producing salidroside. The escherichia coli engineering strain overexpresses a mutant 3-deoxy-D-arabinoheptulose-7-phosphate (DAHP) synthetase gene aroGfbr, a cyclohexadiene dehydrogenase gene tyrC, a glucose phosphate mutant enzyme gene pgm and a UDP-glucose pyrophosphorylase galU, overexpresses a phenylpyruvate decarboxylase gene ARO10 derived from saccharomyces cerevisiae, and can be used for producing a mutant 3-deoxy-D-arabinoheptulose-7-phosphate mutant enzyme. The kit comprises an ethanol dehydrogenase gene ADH6 and a glycosyl transferase gene LrUGT85AF8. The method is mainly used for producing salidroside under an acidic condition.

The invention provides a hydrogen production reaction system taking formaldehyde and water as co-substrates, which comprises a buffer solution, maltodextrin, formaldehyde, magnesium chloride, manganese chloride, NAD (Nicotinamide Adenine Dinucleotide), benzyl viologen, sodium phosphate, alpha-glucan phosphorylase, phosphoglucomutase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, 6-phosphoglucolactonase, diaphorase and hydrogenase. The enzyme is prepared from 1, 6-hexanediol phosphate synthase, hexanediol phosphate isomerase, glucose phosphate isomerase and 2, 6-hexanediol phosphate synthase. The invention also provides a corresponding hydrogen production method. Formaldehyde and water are utilized to generate hydrogen, so that excessive dependence on agricultural grain resources is avoided; meanwhile, high-energy formaldehyde is used for replacing traditional carbohydrates as a substrate, so that the energy barrier of co-substrate water is reduced, and the process of small molecular utilization of carbohydrates and hydrogen production is simplified; in addition, the method has the advantages of being not harsh in water quality condition and low in cost.

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.

The invention provides genetically engineered saccharomycetes capable of simultaneously producing baicalein and baicalin as well as a construction method and application of the genetically engineered saccharomycetes. According to the invention, by weakening or deleting a glucose 5-phosphate dehydrogenase gene (ZWF1) and phenylpyruvate decarboxylase genes ARO10 and PDC5 in a saccharomyces cerevisiae genome and enhancing the expression of a pre-benzoic acid dehydratase gene Pha2, the metabolic flow of a shikimic acid pathway is increased, the aldrin pathway is blocked, and the yield of the shikimic acid pathway is increased. Therefore, the yield of L-phenylalanine serving as a starting material for synthesizing baicalein and baicalin is increased; meanwhile, exogenous genes PAL, 4CL, CHS, CHI, FNSII, F6H, CPR and F7GAT are integrated onto the genome by virtue of a gene recombination technology, so that baicalein and baicalin are produced from the beginning by taking glucose and glycerol as a mixed carbon source without adding precursor substances such as exogenous L-phenylalanine or malonyl CoA and the like.

The application discloses a bifunctional enzyme glucose phosphate isomerase / mannose phosphate isomerase mutant and application thereof, and belongs to the technical field of enzyme engineering and biological catalysis. The application provides a bifunctional enzyme glucose phosphate isomerase / mannose phosphate isomerase mutant, wherein at least one of the glutamic acid at the 28th position, the arginine at the 83rd position, the valine at the 126th position, the isoleucine at the 161st position, the leucine at the 187th position, the phenylalanine at the 249th position, the leucine at the 319th position and the tyrosine at the 343rd position in the amino acid sequence shown in SEQ ID No. 1 is mutated. The application further improves the conversion efficiency of D-mannose, and overcomes the problem of a large amount of by-products.

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 the 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.

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 discloses a difunctional enzyme glucose phosphate isomerase / mannose phosphate isomerase mutant and application thereof, and belongs to the technical field of enzyme engineering and biological catalysis. The invention provides a difunctional enzyme glucose phosphate isomerase / mannose phosphate isomerase mutant and a preparation method of the difunctional enzyme glucose phosphate isomerase / mannose phosphate isomerase mutant. The amino acid sequence of the mutant is characterized in that at least one site of glutamic acid at the 28th site, arginine at the 83rd site, valine at the 126 site, isoleucine at the 161st site, leucine at the 187th site, phenylalanine at the 249th site, leucine at the 319th site and tyrosine at the 343th site of the amino acid sequence shown in SEQ ID No.1 is mutated. According to the method, the conversion efficiency of D-mannose is further improved, and meanwhile, the problem that a large number of by-products exist is solved.

This application relates to a digoxin detection kit. Specifically, the 6-glucose phosphate dehydrogenase mutant of this application comprises one or a combination of the following mutations compared to wild-type 6-glucose phosphate dehydrogenase: D306C, D375C, and G426C. The detection kit prepared using the 6-glucose phosphate dehydrogenase mutant of this application has strong specificity, high sensitivity, convenient operation, short detection time, and accurate quantitative determination, 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 discloses saccharomyces cerevisiae engineering bacteria for efficiently synthesizing mogroside V based on lipid droplet subcellular organelles and a construction method of the saccharomyces cerevisiae engineering bacteria. The construction method comprises the following steps: integrating squalene epoxidase ERG1, cucurbitadienol synthetase SgCDS, cyclic epoxy hydrolase SgEPH3, cytochrome P450 enzyme CYP87D18 and cytochrome P450 enzyme reductase AtCPR1 at a site GAL80 of a saccharomyces cerevisiae genome; the saccharomyces cerevisiae strain is characterized in that phosphoglucose mutase PGM1, alpha-phosphoglucose mutase PGM2, uridine diphosphate glucose pyrophosphorylase UGP1, glycosyl transferase UGTMG1 and glycosyl transferase SgUGT94-289-3 are integrated at a site Exg1 of a saccharomyces cerevisiae genome. The Saccharomyces cerevisiae engineering bacterium constructed by the invention realizes that the yield of MG-V synthesized by shake flask fermentation is 31.4 mg / L. The Saccharomyces cerevisiae engineering bacterium has the characteristics of strong metabolic flux directionality, high yield and wide industrial application prospect.

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.

This application relates to a method for preparing a conjugate. Specifically, the 6-glucose phosphate dehydrogenase mutant of this application comprises one or a combination of the following mutations compared to wild-type 6-glucose phosphate dehydrogenase: D306C, D375C, and G426C. The detection kit prepared using the 6-glucose phosphate dehydrogenase mutant of this application has strong specificity, high sensitivity, convenient operation, short detection time, and accurate quantitative determination, making it suitable for high-throughput detection.

This application relates to a gentamicin detection kit. Specifically, the 6-glucose phosphate dehydrogenase mutant described herein comprises one or a combination of the following mutations compared to wild-type 6-glucose phosphate dehydrogenase: D306C, D375C, and G426C. The gentamicin detection kit prepared using the 6-glucose phosphate dehydrogenase mutant described herein has strong specificity, high sensitivity, ease of use, short detection time, and minimal inter-batch variability, and has promising application prospects.

The invention relates to a recombinant pichia pastoris strain for producing coupled xylitol by NADPH (nicotinamide adenine dinucleotide phosphate) dependent growth and a construction method of the recombinant pichia pastoris strain, and belongs to the technical field of microbial fermentation. The recombinant pichia pastoris strain provided by the invention contains an NADPH dependent xylitol dehydrogenase gene, a sugar phosphatase gene, a D-arabinol-4-dehydrogenase gene, a fructose-1, 3, 4-triazole-1, 3, 4-triazole-1, 3-triazole-1, 3-triazole-1, 3-triazole-1, 3- The gene sequence is as follows: a 1, 6-diphosphatase gene, a phosphoketolase gene, a phosphate transacetylase gene, a glucose-6-phosphate dehydrogenase gene, a 6-phosphogluconolactonase gene, a 6-phosphogluconate dehydrogenase gene and a ribulose-5-phosphate epimerase gene; gene for expressing xylulokinase is knocked out or down-regulated: NADH dependent type xylitol dehydrogenase gene, ribose-5-phosphate isomerase gene, glutamate dehydrogenase gene, phosphofructokinase gene and phosphoglucose isomerase gene. The yield of xylitol produced by fermentation of the strain is far higher than that in the prior art.

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.

This application relates to a method for preparing conjugates. 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.

The invention discloses a saccharomyces cerevisiae engineering bacterium for efficiently synthesizing mogroside V based on peroxisome and lipid droplet double-cell area compartment and a construction method of the saccharomyces cerevisiae engineering bacterium. According to the construction method disclosed by the invention, squalene epoxidase ERG1 is anchored on the surface of a peroxisome membrane and in lipid droplets respectively; the ERG1 and downstream metabolic enzymes such as cucurbitadienol synthetase, cyclic epoxy hydrolase, cytochrome P450 enzyme, cytochrome P450 enzyme reductase, phosphoglucomutase, alpha-phosphoglucomutase, uridine diphosphate glucose pyrophosphorylase and glycosyltransferase are assembled into a multi-enzyme complex by adopting a multi-enzyme complex directional assembly strategy; the method not only enhances the directionality of metabolic flux, but also improves the catalytic efficiency of the enzyme and reduces the generation of byproducts, thereby realizing efficient synthesis of MG-V.

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 the use of conjugates in the preparation of reagent kits. Specifically, the glucose-6-phosphate dehydrogenase mutant of this application contains a mutation selected from one or a combination of the following, compared to the wild-type glucose-6-phosphate dehydrogenase: D306C, D375C, and G426C. Gentamicin detection kits prepared using the glucose-6-phosphate dehydrogenase mutant of this application exhibit high specificity, high sensitivity, ease of operation, short detection time, and small batch-to-batch variation, demonstrating promising application prospects.

This application relates to the use of conjugates in the preparation of phenytoin detection reagents. Specifically, the 6-glucose phosphate dehydrogenase mutant of the present application comprises one of the following mutations, D306C, D375C, and G426C, compared to wild-type 6-glucose phosphate dehydrogenase. The detection kit prepared using the 6-glucose phosphate dehydrogenase mutant of the present application has strong specificity, high sensitivity, convenient operation, short detection time, and accurate quantitative determination, making it suitable for high-throughput detection.

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.