1167results about "Glycosyltransferases" patented technology

Aspects of the disclosure relate to complexes comprising a muscle-targeting agent covalently linked to a molecular payload. In some embodiments, the muscle-targeting agent specifically binds to an internalizing cell surface receptor on muscle cells. In some embodiments, the molecular payload inhibits expression or activity of DUX4. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide or RNAi oligonucleotide.

Aspects of the disclosure relate to complexes comprising a muscle-targeting agent covalently linked to a molecular payload. In some embodiments, the muscle-targeting agent specifically binds to an internalizing cell surface receptor on muscle cells. In some embodiments, the molecular payload promotes the expression or activity of a functional dystrophin protein. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide, e.g., an oligonucleotide that causes exon skipping in a mRNA expressed from a mutant DMD allele.

Glycosyltransferase UGT76G1 and UGT91C1 mutants and a method for catalytic synthesis of rebaudioside A, D, and M by using same, pertaining to the technical field of biocatalytic synthesis. The glycosyltransferase UGT76G1 mutant can effectively synthesize rebaudioside A using stevioside STV as a substrate. The glycosyltransferase UGT91C1 mutant is obtained by mutation screening of wild-type glycosyltransferase UGT91C1, has higher enzyme activity and catalytic rate, and can effectively synthesize rebaudioside D using rebaudioside A as a substrate. The glycosyltransferase UGT76G1 mutant can also be obtained by mutation and subsequent screening of glycosyltransferase UGT76G1, has higher catalytic activity, and can effectively synthesize rebaudioside M using rebaudioside D and UDPG as substrates.

Provided herein, inter alia, is an engineered non-plant cell that produces a hyoscyamine alkaloid product, a precursor of a hyoscyamine alkaloid product, or a derivative of a hyoscyamine alkaloid product. Also described is a method for producing a scopolamine alkaloid, a precursor of a scopolamine alkaloid product, or a derivative of a scopolamine alkaloid product using a cell.

The present invention provides a fucosyltransferase polypeptide, which can be applied to the synthesis of human milk oligosaccharides (HMOs) and belongs to the field of enzyme engineering technology. The fucosyltransferase polypeptide described in the present application comprises a polypeptide having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, and 100% identical amino acid sequence to the polypeptide shown in SEQ ID NO:1-25. The transferase polypeptide described in the present application improves the ability of the polypeptide shown in SEQ ID NO:1 to catalyze the synthesis of 2'-FL while also improving the specificity of the polypeptide catalyzing the substrate.

The invention discloses a fusion protein, a fusion protein gene, a recombinant cell, a recombinant antigen, a reagent for detecting an anti-myelin sheath related glycoprotein antibody and a preparation method, a detection method and application of the reagent, the fusion protein is MAG, GlcAT-P and HNK-1ST fusion protein, and the MAG, GlcAT-P and HNK-1ST fusion protein is formed by sequentially connecting MAG, a GGGGS sequence, EGFP, P2A, GlcAT-P, T2A and HNK-1ST; the amino acid sequence of the MAG, GlcAT-P and HNK-1ST fusion protein is as shown in SEQ ID NO. 1; the kit can be used for detecting the MAG autoantibody, has excellent detection sensitivity and specificity, and can ensure the detection accuracy especially for the detection in a low titer range.

The present disclosure relates to enriched engineered glycoprotein populations with a therapeutically optimized heterogeneous glycan profile. The heterogeneous glycan profile comprises sialylated complex type glycans, which are associated with improved efficacy of the glycoprotein. The present disclosure also relates to compositions and uses of the engineered glycoprotein populations and cell-based method of producing them.

The invention discloses a flavonoid glycoside glycosyltransferase LbUGT71BX1 in laportea bulbifera as well as the coding gene and the use thereof. The amino acid sequence of the flavonoid glycoside glycosyltransferase LbUGT71BX1 is shown as SEQ ID NO: 2. The nucleotide sequence of the coding gene of the flavonoid glycoside glycosyltransferase LbUGT71BX1 is shown as SEQ ID NO: 1. According to the invention, on the basis of relevant results of the second-generation transcriptome and the third-generation full-length transcriptome sequencing of the laportea bulbifera, the last-step key enzyme LbUGT71BX1 for the synthesis of the flavonoid glycoside in the laportea bulbifera is screened and identified by using a reverse genetics method, filling the terminal blank of the biosynthesis pathway of the flavonoid glycoside in the laportea bulbifera.

The invention relates to the technical field of gene engineering, in particular to a glycosyl transferase mutant, a sucrose synthase mutant and application of the glycosyl transferase mutant and the sucrose synthase mutant in synthesis of rebaudioside M. Wild glycosyl transferase and wild sucrose synthase are modified according to a directed evolution theory to obtain a glycosyl transferase mutant and a sucrose synthase mutant. Rebaudioside M is obtained by catalyzing rebaudioside D through a glycosyl transferase mutant and sucrose synthase mutant co-expression strain, and the catalytic efficiency is superior to that of a wild enzyme. The rebaudioside M is obtained by completely catalyzing 110 g / L of rebaudioside D through the co-expression strain of the glycosyl transferase mutant and the sucrose synthase mutant, only 20 h is needed, the conversion rate reaches 97%, the yield of the rebaudioside M reaches 122 g / L, the raw material conversion rate and the yield of the rebaudioside M are effectively increased, and meanwhile the synthesis time is shortened.

Provided herein is a method for efficiently capping RNA in vitro. In some embodiments the capping reaction may be done at high temperature using Vaccinia capping enzyme or a variant thereof. In other embodiments, the capping reactions may comprise a capping enzyme from a large virus of amoeba, e.g., Faustovirus, mimivirus or moumouvirus, or a variant thereof. Compositions and kits for practicing the method are also provided.

The invention provides a uracil production strain and a construction method and application thereof.According to the strain, on an E.coli UR14 genome by means of a CRIPSR / Cas9 gene editing technology, firstly, psuG genes, preTA genes, rutA genes and upp genes are knocked out, so that decomposition of uracil is blocked; then, a uridine phosphorylase gene udp and a pyrimidine-5 '-nucleotide nucleotidase gene ppnN are subjected to overexpression, and synthesis and accumulation of uracil are synergistically enhanced; and finally, overexpression of the ribose phosphate mutase gene pgm further enhances the conversion of a by-product ribose phosphate 1-precursor 5-ribose phosphate 1-pyrophosphate and improves the carbon utilization rate, and the obtained strain has good genetic stability and high fermentation yield, can stably produce uracil, and has wide application prospects.

The invention discloses a novel biosynthesis path for producing 3 '-phosphoadenosine-5'-phosphosulfuric acid and application, and belongs to the technical field of bioengineering. According to the invention, through recombinant expression and purification of ribose kinase, ribose phosphate pyrophosphate kinase, ribose phosphate transferase, adenine thiotransferase, adenosine phosphate sulfate kinase and polyphosphate kinase 2, a six-enzyme cascade reaction taking ribose and adenosine as substrates in vitro is constructed, the molar conversion rate reaches 22.85%, and the escherichia coli and bacillus subtilis PAPS cell factory can be used for preparing the recombinant ribose phosphate pyrophosphate kinase, the ribose phosphate transferase, the adenine thiotransferase, the adenosine phosphate sulfate kinase and the polyphosphate kinase 2. The yield of PAPS in a 5-L fermentation tank reaches 7.60 g / L and 5.03 g / L respectively. The PAPS synthesis route provided by the invention provides an extensible, economical and efficient solution for the production of sulfated compounds.

The invention belongs to the technical field of genetic engineering, and particularly relates to a high-chitin-content engineering bacterium as well as a construction method and application thereof. According to the invention, enzyme related to filamentous fungus chitin synthesis is subjected to overexpression through a genetic engineering means to construct engineering bacteria, the obtained engineering bacteria have high chitin content, and a high-strength mycelium material can be obtained through conventional solid state fermentation. The invention provides a feasible solution with controllable product quality for the production of the high-strength mycelium material, and has good application value.

The purpose of the present invention is to provide a method for producing a methyl compound. Provided is a method for producing a methyl compound using formic acid.

The present disclosure relates to cellular compositions that are modified to introduce a nucleic acid or vector expressing the same. Such compositions may be used to deliver nucleic acid to a target cell and treat disease such as cancer.

The present disclosure provides guided microbial remodeling (GMR) methods for the rational improvement of plant-associated microbes to perform plant-beneficial functions. The GMR methods described herein allow for non-intergeneric genetic optimization of key regulatory networks within the microbes, which improve plant-beneficial functions over wild-type microbes but don't have the risks associated with transgenic approaches (e.g., unpredictable gene function, public and regulatory concerns, etc.). The present disclosure also provides remodeled microbes and compositions thereof. The utilization of remodeled microbes and compositions thereof will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived fertilizers.

The invention discloses a method for biosynthesizing epigallocatechin gallate glucoside, and belongs to the technical field of biocatalytic synthesis. According to the method, a sucrose phosphorylase mutant is adopted as a catalyst, the sucrose phosphorylase mutant is subjected to single-point mutation or combined mutation on the basis of an amino acid sequence SEQ ID NO.1, and the enzyme activity of the sucrose phosphorylase is improved through mutation; the mutant is utilized to realize efficient catalytic synthesis of epigallocatechin gallate glucoside, and in the whole catalytic process, the synthesis method is mild in condition, simple to operate, short in reaction time and high in catalytic efficiency, and has a good application prospect.

The invention discloses a method for improving the synthesis level of L-tryptophan in escherichia coli based on ARTP mutagenesis high-throughput screening, and belongs to the technical field of bioengineering. According to the invention, the L-tryptophan biosensor pSensor-trp3 is constructed, and the L-tryptophan biosensor pSensor-trp3 is good in response under the condition that the exogenous addition concentration range of L-tryptophan is 0-300mg. L <-1 >. High-throughput screening is performed by using a flow cytometry, a dominant mutant strain YB-2 is obtained through iterative mutagenesis, and the shake flask L-tryptophan yield and the sugar-acid conversion rate of the dominant mutant strain YB-2 are 5.68 g.L <-1 > and 0.103 g.g <-1 > respectively and are increased by 11.4% and 10.7%. Through second-generation whole genome re-sequencing comparative analysis, it is speculated that the genes ynfB and waaO possibly have a promoting effect on synthesis of L-tryptophan. The L-tryptophan biosensor used in the invention also provides reference for high-throughput screening and transformation of other related L-tryptophan production strains.

The invention relates to a series of fucosyl transferase polypeptides which can be applied to synthesis of human milk oligosaccharides (HMOs), and belongs to the technical field of enzyme engineering and genetic engineering. The invention provides an alpha-1, 3-fucosyltransferase polypeptide, which comprises a polypeptide with an amino acid sequence which is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to that of a polypeptide shown in SEQ ID NO: 1. Or a polypeptide having at least 70%, 75%, 80%, 85%, 90%, 95%, 99%, 100% of the same amino acid sequence as the polypeptide M1-21. The invention provides a polypeptide as shown in SEQ ID NO: 1 and M1-M21 polypeptides. The 3-FL catalytic synthesis capability of the polypeptide as shown in SEQ ID NO: 1 is improved; and the specificity of a catalytic substrate is improved.

The invention relates to the technical field of synthetic biology and microbial engineering, in particular to recombinant escherichia coli for de novo synthesis of forsythiaside A. The invention provides recombinant escherichia coli for de novo synthesis of forsythiaside A. A preparation method of the recombinant escherichia coli comprises a step of improving an escherichia coli starting strain, and the escherichia coli starting strain is escherichia coli MG1655. In the fermentation process of the recombinant escherichia coli, effective accumulation of forsythiaside A in fermentation liquor can be achieved, and a foundation can be laid for subsequent construction of high-yield forsythiaside A engineering bacteria.

The invention discloses a Pseudomonas putida high-yield rhamnolipid fermentation medium based on a step-by-step optimization strategy, an optimization method and application thereof, through the step-by-step strategy of component screening-single-factor optimization-multi-factor interactive optimization, a mixed carbon source and a mixed nitrogen source are taken as cores, and a Plackett-Burman test, a steepest climbing test and Box-Behnken response surface design are combined, so that the high-yield rhamnolipid fermentation medium for the pseudomonas putida high-yield rhamnolipid is obtained. Optimizing to obtain a special fermentation culture medium; two genetic engineering strains estA-2 and estA-3 are constructed by heterologous expression of key genes, and the supply and extracellular transport efficiency of a rhamnolipid synthesis precursor is enhanced; the recombinant strain is applied to the optimized culture medium, and efficient production of a shake flask and a 5L fermentation tank is achieved. The rhamnolipid yield of the estA-3 strain in a 5L fermentation tank reaches 35.59 + / -2.18 g / L, the conversion rate is 0.282 g / g, the productivity is 0.404 g / L / h, the rhamnolipid yield, the conversion rate and the productivity are obviously superior to those in the prior art, and a key technical support is provided for industrial production of rhamnolipid.

The invention discloses a production method of fucosylated oligosaccharide. The core of the method is as follows: the GDP-D-rhamnose is converted into the GDP-L-fucose by using the GDP-D-rhamnose-3, 5-epimerase; the GDP-D-rhamnose is obtained by converting the GDP-D-mannose-4, 6-dehydratase and the GDP-4-keto-6-deoxy D-mannose reductase by taking the GDP-D-mannose as a substrate, and the GDP-D-rhamnose is obtained by taking the GDP-D-mannose as a substrate. The invention relates to a method for producing a fucosylated oligosaccharide, which is characterized in that a GDP-D-mannose-4, 6-dehydratase, a GDP-4-keto-6-deoxyD-mannose reductase, a GDP-D-rhamnose-3, 5-epimerase and a fucosyltransferase are expressed in a genetically modified host cell, and the fucosylated oligosaccharide can be produced by the way mentioned above.

The present disclosure provides engineered bacterial cells having one or more genetic modifications that result in increased production of expression products, such as increased production of plasmids with reduced occurrence of plasmid concatemers or multimers. Also disclosed herein are expression systems for product manufacturing in media having reduced antibiotic concentration, as well as methods of making cells for use in the expression systems. The cells and expression systems increase the growth rate and production yields of biomolecules produced within the host bacterial host cells without the use of antibiotics, and can produce the product with little to no endotoxins present.

The invention discloses recombinant escherichia coli capable of increasing the yield of fucosyllactose and a construction method and application of a fucosyltransferase mutant of the recombinant escherichia coli, and belongs to the field of synthetic biology and metabolic engineering. According to the invention, recombinant escherichia coli B20 and B21 are constructed based on key enzyme engineering transformation. Under the shake flask condition, the 3-FL production capacity of the recombinant strain is obviously improved to 6.20 g / L and 6.34 g / L from the initial 3.45 g / L, and the amplification reaches 79.7% and 83.8% respectively. The yields of 3-FL of the strain B20 and the strain B21 respectively reach 53.88 g / L and 54.64 g / L under the culture condition of a 3L fermentation tank. The recombinant escherichia coli for expressing the 2 '-FL is constructed, and the yield of the 2'-FL reaches 64.86 g / L in a 3L fermentation tank. The invention provides a new technical scheme for efficient microbial synthesis of the 3-FL.

This disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, this disclosure is in the technical field of cultivation or fermentation of metabolically engineered cells. This disclosure describes a cell metabolically engineered for production of a mixture of at least three different oligosaccharides. Furthermore, this disclosure provides a method for the production of a mixture of at least three different oligosaccharides by a cell as well as the purification of at least one of the oligosaccharides from the cultivation.

The invention relates to the technical field of gene engineering, in particular to a glycosyl transferase mutant and application thereof in synthesis of rebaudioside M. Wild type glycosyl transferase is modified through a directed evolution theory to obtain the glycosyl transferase mutant, so that the efficiency of synthesizing rebaudioside D through reaction of rebaudioside A is effectively improved. Besides, recombinant bacteria capable of simultaneously expressing the glycosyl transferase mutant, a second glycosyl transferase gene and a sucrose synthase gene are constructed, a wet cell obtained by induced culture of the recombinant bacteria or a crude enzyme liquid extracted by crushing the wet cell is used as a catalyst, rebaudioside A is used as a substrate, sucrose is used as an auxiliary substrate, and the rebaudioside M is catalytically synthesized by a one-pot method. According to the present invention, the complete catalysis of 100 g / L of rebaudioside A to produce rebaudioside M only needs 24 h, the conversion rate reaches 97.5%, the yield of rebaudioside M reaches 130 g / L, the raw material conversion rate and the yield of rebaudioside M are effectively improved, and the synthesis time is shortened.

The invention belongs to the technical field of gene engineering, and particularly relates to application of a TaMGD3 gene in improvement of plant starch quality and cultivation of a high-starch transgenic plant, including application of the TaMGD3 gene in promotion of wheat grain starch synthesis, application of the TaMGD3 gene in cultivation of the high-starch and / or high-amylopectin transgenic plant, and application of the TaMGD3 gene in improvement of plant starch quality and cultivation of the high-starch and / or high-amylopectin transgenic plant. The invention also discloses an application of the TaMGD3 gene in improvement of plant starch characteristics. A nucleotide sequence of the TaMGD3 gene is shown as SEQ ID NO. 1. The invention also provides a cultivation method of transgenic wheat. In wheat cultivation, wheat amylopectin quality improvement is realized through overexpression of the gene shown in SEQ ID NO.1. The TaMGD3 gene can improve the starch content of wheat grains, influence the particle size distribution of the wheat grain starch, improve the amylopectin proportion, improve the peak viscosity and the relative crystallinity of the starch, and facilitate the improvement of the cooking quality related to the wheat starch.

[Summary] [Problems] To provide a novel technique for improving the taste of a glycosylated steviol glycoside composition. [Solution] The present technique provides a method for producing a glycosylated steviol glycoside composition and a method for improving the taste of a glycosylated steviol glycoside composition, which include a step of treating a stevia extract and dextrin with cyclodextrin glucanotransferase derived from a microorganism belonging to the genus Anoxybacillus. The production method and taste improvement method according to the present technique may further include a step of treating with a carbohydrate-processing enzyme other than the cyclodextrin glucanotransferase derived from a microorganism belonging to the genus Anoxybacillus.

The present invention belongs to the technical field of biochemistry, and in particular relates to an efficient enzymatic synthesis method for a holothurian glycosaminoglycan. The present invention establishes an efficient enzymatic synthesis route for the holothurian glycosaminoglycan, in which the holothurian glycosaminoglycan with anticoagulant activity is efficiently synthesized through a one-pot multienzymatic strategy using sulfated fucose and chondroitin sulfate as substrates and catalyzed by α-1,3-fucosyltransferase mutant and L-fucokinase / GDP-fucose pyrophosphorylase, which provides important technical support for the development of new anticoagulant drugs.

A method of synthesising a 1N-alkyl derivative of pseudouridine comprises contacting a substrate with a methyltransferase in the presence of a cofactor to produce the 1N-alkyl derivative of pseudouridine. The substrate has a structure of: Formula (I). The 1N-methyl derivative of pseudouridine has a structure of: Formula (II). R1 is a hydroxyl group, a monophosphate group, a diphosphate group, or a triphosphate group. R2 is H or a hydroxyl group. R3 is a methyl group or an ethyl group. The cofactor is a source of a methyl group or an ethyl group. The methyltransferase is configured to transfer the methyl group or ethyl group from the cofactor to the substrate. Enzymatic synthesis of 1N-methyl derivatives of pseudouridine may be more efficient and produce less waste than chemical syntheses. Also provided are methyltransferases and buffers useful in the method; and polynucleotides, expression vectors, and microorganisms useful for producing the methyltransferases.