51 results about "Glycosyltransferase" patented technology

The application discloses a mycobacterium maltose transferase Co GlgE and an encoding gene and application thereof.The maltose transferase Co The amino acid sequence of the GlgE is shown as SEQ ID NO.1.The application discloses a maltose transferase Corallococcus derived from mycobacterium sp.EGB Co GlgE, which can disperse maltoligosaccharide with a polymerization degree (DP) X into products with a DP of X ± 2n (X ≥ 3). Co The GlgE has glycosylation modification effects on alpha-arbutin and beta-arbutin, the biological activity of the modified arbutin is significantly improved, and the application value of the arbutin as an additive of whitening skin care products is improved.

The present disclosure relates to a method for improving the quality of glycosylated antibody drug conjugates (DAR distribution, stability, etc.) using activated carbon. Specifically, the present disclosure employs hydrophobic interaction chromatography (HIC) column, protein A chromatography, activated carbon stirring, and buffer exchange, alone or in series, to remove free glycosyltransferases. As a result, the purity of the glycosylated antibody drug conjugates is improved, and the degradation of the antibody drug conjugates is prevented. Ultimately, the drug substance (DS) of OBI-902 (anti-TROP2 antibody drug conjugate) and OBI-904 (anti-Nectin-4 antibody drug conjugate) is further determined.

This invention discloses a mogroside glycosyltransferase mutant and its application in the production of sympathomimetic I, belonging to the fields of genetic engineering and enzyme engineering. The amino acid sequence of the mogroside glycosyltransferase mutant is shown in SEQ ID NO:2. Compared with the glycosyltransferase SgUGT94-289-3, the mogroside glycosyltransferase mutant of this invention (M92T / H96M / V148W protein) can significantly increase the production of sympathomimetic I. Specifically, it can increase the yield of sympathomimetic I to 81% and the specificity to 100% in enzyme engineering and biosynthesis, meeting the requirements for industrial production and possessing significant application value.

This invention discloses a method for the efficient synthesis of gastrodin using recombinant Corynebacterium glutamicum, belonging to the fields of genetic engineering, fermentation engineering, and synthetic biology. This invention uses Corynebacterium glutamicum as the host bacterium and knocks out… pobA The gene was expressed, and the engineered strain WN01 / pB6 was constructed using the expression plasmid pEC-XK99E to express cladoid-pyruvate lyase, carboxylic acid reductase, and glycosyltransferase, achieving heterologous synthesis of gastrodin. Further overexpression was then performed. aroG Genes, overexpression of protein-coding genes that activate carboxylic acid reductase sfp The engineered strain WN05 / pB6 was able to synthesize 7.37 g / L of gastrodin at the shake flask level, and the gastrodin yield reached 17.77 g / L at 72 h of batch fermentation in a 5-L fermenter, with a yield of 0.247 g / L / h. This provides a brand-new biomanufacturing platform for the industrial production of gastrodin.

The application discloses a key type specificity changed alpha-glucosyltransferase mutant, and belongs to the technical field of enzyme engineering and starch modification. The method is characterized in that, by analyzing the combination of Lf2970GtfB 4,3-alpha-glucosyltransferase in the active center donor / acceptor sub-site and the substrate, part of key amino acids of Lf2970GtfB are mutated into different types of amino acids, the mutant is prepared by changing the enzyme and substrate interaction force, space steric hindrance and other factors affecting the orientation of the substrate in the active center, the transglycosylation is changed, and the bond type composition of the product is changed. The application changes the bond type specificity of Lf2970GtfB 4,3-alpha-glucosyltransferase by mutation, the obtained mutant has novel 4,3 / 6-alpha-glucosyltransferase activity, and is helpful to synthesizing unique structure and more valuable alpha-glucan by using starch substrates.

This invention discloses a method for reducing the viscosity of potato starch, belonging to the fields of enzyme engineering and biomodified starch technology. This invention provides a method for preparing macrocyclodextrin by reacting starch with 4-α-glycosyltransferase, which is then added to potato starch to provide a suitable substrate for subsequent starch processing. This application in starch processing offers a new approach and has great potential and application prospects.

The application provides a mutant of acidophilic glycosyltransferase at the 400th position and application thereof in production of salidroside, and belongs to the technical field of bioengineering. The application solves the problem of low synthesis efficiency of salidroside under acidic conditions. The application comprises an acidophilic glycosyltransferase mutant, wherein the glycosyltransferase mutant is obtained by mutating the 400th position of the amino acid shown in SEQ ID NO. 2. The mutant is mainly used for high-yield production of salidroside under acidic conditions.

The utility model discloses a kind of multi-enzyme step-by-step preparation glucosyl steviol glycoside's device, it is related to glucose steviol glycoside production technical field, the inlet of first reaction tank is respectively communicated with raw material tank, dextrin tank, cyclodextrin glucoside transferase tank and first sodium hydroxide solution tank, the outlet of first reaction tank is communicated with hydrolysis tank, the outlet of hydrolysis tank is communicated with second reaction tank, the inlet of second reaction tank is respectively communicated with sucrose tank, sucrose synthase tank, uridine diphosphate tank, UGT glycosyltransferase tank and lye tank, the outlet of second reaction tank is communicated with concentration tank by pipeline, the lower outlet of concentration tank is communicated with drier, the outlet of drier is communicated with glucosyl steviol glycoside tank. Make residual substrate conversion is relatively more optimal steviol glycoside in taste, improve the taste, quality and yield of product.

The present disclosure provides enzymes and methods of using those enzymes to transfer sugar moieties to substrate steviol glycosides. Specifically, designed beta-1,2-glycosyltransferases and sucrose synthases are used in one-pot reactions to convert stevioside and Reb A to Reb E and Reb D. Additionally, designed beta-1,2-glycosyltransferases can be used in one-pot reactions with sucrose synthases and beta-1,3-glycosyltransferases to produce Reb M from stevioside and / or Reb A.

The application discloses a glycosyltransferase mutant and application thereof, and the mutant is obtained by single mutation or multiple mutations of amino acids at positions 58, 181 or 371 of the amino acid sequence shown in SEQ ID NO. 4 or single mutation or multiple mutations of amino acids at positions 54, 152, 241 or 272 of the amino acid sequence shown in SEQ ID NO. 6. The glycosyltransferase is rationally modified by homologous modeling and sequence homology analysis, and a glycosyltransferase mutant with obviously improved enzyme activity is screened. Compared with the wild type, the relative enzyme activity of the mutants UGTM1-3 and UGTM2-4 is respectively improved by 2.88 times and 3.60 times compared with that before modification, and the mutants have great industrial application potential.

The present invention relates to a method for producing a plant extract comprising rubusoside, to a method for glycosylating rubusoside, to the use of a natural deep eutectic solvent in such a method and to the use of a glycosyltransferase in such a method.

The present application relates to the technical field of genetic engineering, and particularly relates to a salicylic acid glycosyltransferase and use thereof. The present application finds a new gene (nucleotide sequence as shown in SEQ ID NO. 3) in highland barley, the protein (amino acid sequence as shown in SEQ ID NO. 4) expressed by the gene can convert salicylic acid into salicylic acid glucoside, and improve the level of highland barley resistance to powdery mildew. The present application also performs in-vitro expression on the gene fragment, obtains the salicylic acid glycosyltransferase, and successfully prepares salicylic acid glucoside by taking glucose as a glycosyl donor and salicylic acid as a receptor in an in-vitro reaction. The present application also transfers the gene into tobacco, so that the salicylic acid glycosyltransferase is also expressed in the tobacco plant, further produces salicylic acid glucoside, and improves the value. The new gene, the recombinant vector, the recombinant bacteria, the protein and the transgenic plant provided by the present application all have good application prospects.

This invention provides recombinant polynucleotides comprising nucleic acids encoding CYP76AD6 or related genes, and their use for the production of L-DOPA from tyrosine and for the treatment of dopamine-responsive disorders such as Parkinson's disease. This invention also provides recombinant polynucleotides comprising nucleic acids encoding CYP76AD1 and / or CYP76AD6, nucleic acids encoding DOPA 4,5-dioxygenase (DOD) such as beet DODA1, and in some cases, nucleic acids encoding betaine-related glucosyltransferases such as the four o'clock gene cyclic-DOPA 5-O-glucosyltransferase (cDOPA5GT), and their use for the production of betaine. Finally, this invention provides chimeric polypeptides, expression vectors, cells, compositions, and organisms (including plants), and their use in the various methods of this invention.

The application discloses a connecting peptide and application thereof in promoting glucose methylation modification of a compound. Glycosyltransferase-methyltransferase fusion protein genes connected by connecting peptides, genes LtLasS1 and LtLasS2 encoding benzenediol lactone synthetase are co-expressed, and it is determined through liquid phase and mass spectrum analysis of fermentation products that the fusion protein can realize glucose methylation modification on benzenediol lactone. The conversion efficiency of the fusion protein using DL-4 connecting peptide on benzenediol lactone is the highest, and the relative conversion rate is 65.54+ / -3.75%. In addition, the glucose methylation conversion efficiency of the artificial fusion protein constructed by feeding the substrate kaempferol is tested, and the conversion efficiency of the fusion protein using DL-4 connecting peptide on the substrate is 6 times that of the conversion efficiency of the substrate by using a single glycosyltransferase and methyltransferase respectively, and the relative conversion rate is increased to 60.12+ / -4.79%.

The present disclosure relates, in part, to addition of glucosyltransferase type enzymes for producing and / or modifying alpha-glucan type structures in dough such as bread dough. Such enzymes encompass various glucosyltransferases where these enzymes can transfer glucose from sucrose for production and / or modification of alpha-glucan type polymers in bread dough. Further disclosed are methods of preparing a baked food product by baking a dough, comprising incorporating into the dough one or more of the foregoing enzymes, wherein the enzyme can transfer a glucose-based moiety from sucrose to the non-reducing end of a saccharide acceptor, thereby improving texture, specific volume, dough stability, dough viscosity, anti-staling attributes and / or moisture content, for example.

This invention discloses a method for in vitro synthesis of natural H11 glycopeptides from *Haemaphysalis contortus*, belonging to the field of biomedicine. The method includes the following steps: preparing a glycosyl donor compound using sialic acid glycopeptide SGP as a raw material; activating the glycosyl donor to a glycooxazoline; transglycosylation of the glycooxazoline with an H11 glycopeptide chain carrying acetylglucosamine at the N site to obtain the glycopeptide compound; catalyzing the glycopeptide compound with core fucosyltransferase 8, *Haemaphysalis contortus*-derived galactosyltransferase 1, β-1,4-galactosyltransferase, and fucosyltransferase to obtain corresponding glycosyl modifications from the donor, yielding a natural *Haemaphysalis contortus* H11 glycopeptide containing an LDNF structure and a *Gal(β1,4)Fuc* nematode-specific epitope. This invention can accurately mimic the nematode-specific glycosylation modification of natural H11, with high product uniformity, and can provide an antigen source for vaccine development for *Haemaphysalis contortus* disease.

The application discloses a glycosyltransferase UGT91C1 mutant and a method for catalyzing synthesis of rebaudioside D, and belongs to biological catalytic synthesis. The glycosyltransferase UGT91C1 mutant is any one of the following (A)-(C): A) a protein obtained by making any one or more mutations in the amino acid sequence shown in SEQ ID NO. 1: the 89th amino acid is mutated from N to Y; the 155th amino acid is mutated from M to L; the 274th amino acid is mutated from S to T; and the 361st amino acid is mutated from N to S; (B) a protein having 95% or 98% or above identity with the amino acid sequence defined in (A) and having the same function; and (C) a fusion protein obtained by connecting a tag to the end of the protein defined in (A) or (B). The glycosyltransferase UGT91C1 mutant is obtained by mutation screening from a wild-type glycosyltransferase UGT91C1, has higher enzyme activity and catalytic rate, and can efficiently synthesize rebaudioside D by taking rebaudioside A as a substrate.

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express recombinant genes encoding UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce steviol glycosides, e.g., Rebaudioside A and / or Rebaudioside D, which can be used as natural sweeteners in food products and dietary supplements.

The application provides a mutant of an acidophilic glycosyltransferase and application thereof in production of salidroside, and belongs to the technical field of bioengineering. The application solves the problem of low synthesis efficiency of salidroside under acidic conditions. The application comprises a mutant of an acidophilic glycosyltransferase, wherein the glycosyltransferase mutant is obtained by mutating any one of positions 87, 210, 293, 356, 399, 400 and 415 of the amino acid sequence shown in SEQ ID NO. 2. The mutant is mainly used for high-yield production of salidroside under acidic conditions.

The invention belongs to the field of gene engineering and enzyme engineering, and particularly relates to an activity-improved UDP glycosyl transferase mutant and application thereof. The glycosyl transferase mutant is a four-combination mutant N2HN1I, and the amino acid sequence of the glycosyl transferase mutant is as shown in SEQ ID NO. 4. The mutant provided by the invention has higher catalytic activity, the conversion rate of a substrate crocetin is high, and the mutant has important industrial application value.

The application provides an improved recombinant bacillus Calmette-Guerin (BCG) and application thereof, and belongs to the technical field of biotechnology. The BCG ManLAM mutant strain is obtained by knocking out the coding gene of a key glycosyltransferase MptC for synthesizing the terminal mannose of ManLAM in the BCG, and is used as the improved BCG. Compared with the wild-type BCG, the improved BCG constructed in the application can promote the functional maturation of dendritic cells and improve the anti-tuberculosis infection immune response, can be used as a new vaccine for preventing tuberculosis or as an active ingredient for preparing a medicine for preventing and treating tuberculosis, and has a wide prospect in the development and clinical application of the new vaccine. In addition, the application provides a new tool and idea for the prevention and treatment of tuberculosis.

This invention belongs to the field of biotechnology, specifically relating to a bifunctional glycosyltransferase protein, LbUGT41, its encoding gene, and its applications. The LbUGT41 gene was cloned from Ningxia wolfberry, with a full-length nucleotide sequence of 1347 bp encoding a 448-amino acid protein. Functional verification showed that LbUGT41 possesses bifunctional catalytic activity: it can catalyze the 3-O-glucosylation of quercetin to generate isoquercitrin using UDP-glucose as a sugar donor; it can also catalyze the hydrolysis of isoquercitrin to generate quercetin in the presence of UDP. This invention provides a novel biocatalyst for the targeted modification and transformation of flavonoids, and also provides key genes and theoretical basis for research on the regulation of plant flavonoid metabolism and molecular breeding of wolfberry quality.

The application discloses a glycosyltransferase coded by Dendrobium huoshanense UGT70 gene and application of the glycosyltransferase in synthesis of antioxidant flavone disglycosides, and belongs to the technical field of biotechnology. The UGT70 gene nucleotide sequence codes an amino acid sequence as shown in SEQ ID NO. 2. The flavone disglycosides are obtained by using the UGT70 protein to perform enzyme catalysis in vitro by a biosynthesis method, and a new method for synthesizing the flavone disglycosides is provided.

The present application relates to an optimized catalytic synthesis method of rare ginsenoside and application thereof. Specifically, the present application provides a reaction system for co-catalyzing rare ginsenoside by glycosyltransferase and sucrose synthase, compares, mutates and screens the enzyme, optimizes the activity of glycosyltransferase and sucrose synthase and co-expression mode, and further improves the production efficiency of rare ginsenoside by optimizing the concentration of crude enzyme solution, improving the solubility of substrate and the like, so that the substrate conversion rate is increased from about 20% to more than 95%. The method of the present application is suitable for the production of different types of rare ginsenoside and has high industrial production and application value.

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 present invention relates to chimeric GT-B type glycosyltransferases, comprising an N-terminal acceptor binding site, a C-terminal donor binding site, and a linker loop linking the N-terminal acceptor binding site and the C-terminal donor binding site, wherein (i) the N-terminal acceptor binding site domain comprises or consists of an amino acid sequence having at least 80 %, preferably at least 90 %, more preferably 100 % sequence identity to the amino acid sequence set forth in SEQ ID NO: 1; and / or (ii) the linker loop comprises or consists of an amino acid sequence having at least 80 %, preferably at least 90 %, more preferably 100 % sequence identity to the amino acid sequence set forth in SEQ ID NO: 4; and / or (iii) the C-terminal donor binding site domain comprises or consists of an amino acid sequence having at least 80 %, preferably at least 90 %, more preferably 100 % sequence identity to the amino acid sequence set forth in SEQ ID NO: 2 or comprises or consists of an amino acid sequence having at least 80 %, preferably at least 90 %, more preferably 100 % sequence identity to the amino acid sequence set forth in SEQ ID NO: 3; and / or (iv) the C-terminal donor binding site domain is a chimeric C-terminal donor binding site domain comprising a first amino acid sequence derived from a glycosyltransferase distinct from the glycosyltransferase from which the N-terminal acceptor binding site domain is derived, and a second amino acid sequence derived from the same glycosyltransferase as the N-terminal acceptor binding site domain, wherein the first amino acid sequence is linking the linker loop and the second amino acid sequence; to nucleic acid molecules encoding such GT-B type glycosyltransferases; to a method of production of such chimeric GT-B type glycosyltransferases; to a method of production of polysaccharides using such chimeric GT-B type glycosyltransferases and the polysaccharides obtainable by such a method; to polysaccharides defined by formula (I); to compositions comprising such polysaccharides; and further to uses of such polysaccharides as thickening and / or gelling agents and / or as rheology modifiers and / or as 3D-printing inks and / or coatings.

The present application relates to a rock white bifunctional glycosyltransferase BpUGT1 Gene and application in preparation of arbutin and gastrodin, belong to the field of biotechnology. The rock white bifunctional glycosyltransferase BpUGT1 The gene nucleotide sequence is as shown in SEQ ID NO.1, and the sequence full length is 1431bp; the amino acid sequence of the encoded protein is as shown in SEQ ID NO.2, and 476 amino acid residues are encoded. The rock white bifunctional glycosyltransferase BpUGT1 The gene can be used as the biosynthesis regulation gene of arbutin and gastrodin, and is applied to the preparation of arbutin and gastrodin, and the application prospect is remarkable, and easy to popularize and apply.