1588results 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 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.

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.

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.

The invention discloses an N-acetylglucosamine transferase AcetRS-R346D and an application thereof, and belongs to the technical field of functional enzymes. And the amino acid sequence of the N-acetylglucosamine transferase AcetRS-R346D is as shown in SEQ ID NO. 3. The invention further discloses the application of the N-acetylglucosamine transferase AcetRS-R346D in the preparation of the chitosan oligosaccharide. The N-acetylglucosamine transferase AcetRS-R346D disclosed by the invention is obtained by carrying out mutation modification on the N-acetylglucosamine transferase AcetRS, and can be used for producing the chitosan oligosaccharide with the polymerization degree of 4 to 7, especially (GlcNAc) 7. The product spectrum of the N-acetylglucosamine transferase is widened, and the N-acetylglucosamine transferase can be used for preparing high-polymerization-degree chitosan oligosaccharide and has a wide application prospect.

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.

The invention discloses a light-driven synthetic flora and application thereof. The light-driven synthetic flora comprises phototrophic bacteria and heterotrophic bacteria, the phototrophic bacteria are genetically modified synechococcus slender, and the gene modification comprises construction and expression of heterologous sucrose permease, glyceraldehyde dehydrogenase and sucrose synthase in the synechococcus slender; and the heterotrophic bacteria are the P.adami. According to the light-driven synthetic flora disclosed by the invention, carbon dioxide is converted into cane sugar through photosynthesis of phototrophic bacteria, and the cane sugar is secreted out of cells to provide a carbon source for heterotrophic bacteria; carrying out cell growth and polyhydroxyalkanoate synthesis on the heterotrophic bacteria by using cane sugar; therefore, the light-driven synthetic flora provided by the invention can realize direct conversion of carbon dioxide into polyhydroxyalkanoate without additional carbon sources, has the advantage of environmental friendliness, and provides a new economical and feasible scheme and approach for production of polyhydroxyalkanoate.

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 is a glycosyltransferase UGTSL2 mutant, which is produced through any one or more selected from the group consisting of mutations based on an amino acid sequence shown in SEQ ID NO: 1. Also provided is a glycosyltransferase mutant, which is the following A1) or A2): A1) a protein produced through an amino acid residue substitution in an amino acid sequence shown in SEQ ID NO: 66, where the protein has an identity of 90% or more with and the same function as the amino acid sequence shown in SEQ ID NO: 66; and A2) a fusion protein produced by linking a tag to an N-terminus and / or a C-terminus of the protein in the A1).

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 invention provides an ATP phosphoribosyltransferase mutant and its application in histidine production. The mutant is obtained by performing point mutations at positions 84 and / or 205 on the amino acid sequence of the wild-type ATP phosphoribosyltransferase derived from Acinetobacter baumannii. The mutant, the nucleic acid molecule encoding the mutant, the expression cassette containing the nucleic acid molecule, the recombinant vector or the recombinant microorganism are used for the production of L-histidine, and the yield of L-histidine is significantly increased, laying a good foundation for the large-scale industrial production of L-histidine.

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 a method for constructing an enzyme polymer to improve enzyme catalytic activity. The method comprises the following steps: modifying a phage sponge protein Acb2 at a C terminal or an N terminal of glucosidase, glucuronidase, sucrose phosphorylase and the like through a genetic engineering means; the enzyme polymer with better catalytic activity is obtained by screening and optimizing the types of connecting peptides between the sponge protein and the natural enzyme and the number of amino acids. The specific enzyme activity of the sponge protein driven clustered enzyme aggregate enzyme designed and constructed in the invention is improved by 3.43 times compared with that of a wild enzyme, and kcat is improved by 32.3 times. The sponge protease polymer is high in catalytic activity, simple in preparation process and wide in application range, and has the potential of being applied to large-scale industrial production.

The invention provides recombinant escherichia coli for producing chondroitin sulfate A and application of the recombinant escherichia coli, and relates to the technical field of genetic engineering. According to the recombinant escherichia coli disclosed by the invention, chondroitin synthase, chondroitin 4-O-sulfotransferase, N-acetyl glucosamine epimerase, UDP-glucose dehydrogenase and chondroitin sulfate degrading enzyme are overexpressed; wherein the amino acid sequence of the chondroitin sulfate degrading enzyme is as shown in SEQ ID NO. 5. The recombinant escherichia coli is inoculated into a culture medium for fermentation culture, the average molecular weight of the obtained chondroitin sulfate A is 800Da, and the yield is about 12.5 g / L. Therefore, low-molecular-weight chondroitin sulfate A can be obtained and the yield of chondroitin sulfate A can be increased by fermenting the recombinant escherichia coli disclosed by the invention.

The invention provides construction and application of a hyaluronic acid recombinant bacterium, and belongs to the fields of biochemical engineering, synthetic biology, genetic engineering and metabolic engineering, the recombinant corynebacterium glutamicum contains a hyaluronic acid synthase gene, a UDP-glucose dehydrogenase gene and a glutamide-6-phosphate fructose transaminase gene; the hyaluronic acid synthase gene, the UDP-glucose dehydrogenase gene and the glutamine-6-phosphate fructose transaminase gene are respectively connected with a sequence RBS1, a sequence RBS2 and a sequence RBS3; the recombinant bacterium provided by the invention also contains a dCas9 protein gene dCas9 and any one of genes sgRNA1, sgRNA2, sgRNA3, sgRNA4, sgRNA5 and sgRNA6, and the recombinant bacterium provided by the invention is high in hyaluronic acid yield and low in production cost.

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 relates to a method for producing glycosylglycerol. The method comprises the following steps: S1, culturing trichocephalus microcoleus to a logarithmic phase; and S2, adding glycerol and glucose into the culture environment of the coleus microcoleus to induce the coleus microcoleus to accumulate the glycosylglycerol. According to the invention, by optimizing the culture of the coleus microcoleus BL0902, the production of the glycosylglycerol by using the strain is realized, and on the basis, the strain is subjected to metabolic engineering transformation, so that the yield of the glycosylglycerol is greatly improved by the obtained engineering strain.

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 invention belongs to the technical field of bioengineering, and particularly relates to a sucrose phosphorylase mutant and application thereof in synthesis of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid. The sucrose phosphorylase mutant is obtained by simultaneously mutating multiple sites of amino acids at the 91st site, the 154th site, the 185th site, the 323rd site and the 424th site of wild sucrose phosphorylase derived from Bifidobacterium pseudolongum, so that the efficiency of producing 2-O-alpha-D-glucopyranosyl-L-ascorbic acid by taking sucrose and vitamin C as raw materials is remarkably improved; only 15 hours are needed for completely catalyzing 1.2 M of vitamin C to produce 2-O-alpha-D-glucopyranosyl-L-ascorbic acid; and the yield of the 2-O-alpha-D-glucopyranosyl-L-ascorbic acid reaches 358 g / L. Therefore, the 2-O-alpha-D-glucopyranosyl-L-ascorbic acid has an important application value in the field of production of the 2-O-alpha-D-

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 invention relates to a nicotinamide ribose phosphate transferase mutant, a genetically engineered bacterium and application of the nicotinamide ribose phosphate transferase mutant and the genetically engineered bacterium. Compared with a wild enzyme strain NMN, the production capacity of a mutant enzyme strain is greatly improved. The saccharomyces cerevisiae genetically engineered bacterium capable of stably inheriting and efficiently synthesizing the NMN is obtained, and safe, efficient and green synthesis of the NMN is achieved.

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.