34 results about "Galactosyltransferase" patented technology

The invention relates to the technical field of biology, in particular to a beta-1, 4-galactosyl transferase mutant and a method for biologically synthesizing lactose-N-neotetraose by using the beta-1, 4-galactosyl transferase mutant. The invention discloses a protein, which is a protein HpgalT-186 or a protein HpgalT-146-186, a protein HpgalT-146 and a protein HpgalT-146. The protein HpgalT-186 is a protein obtained by mutating an amino acid residue at the 186th site of a wild type HpgalT; the protein HpgalT-146-186 is a protein obtained by carrying out mutation on an amino acid residue at the 186 site and an amino acid residue at the 146 site of wild type HpgalT. On the basis, a series of escherichia coli is constructed and optimized, the yield of the LNnT synthesized through shake flask fermentation of the finally obtained strain reaches 1.73-4.1 g / L, the highest yield in a 5L fermentation tank reaches 20-45.2 g / L, and efficient synthesis of the LNnT is achieved.

The present invention belongs to the field of biotechnology, and specifically relates to a GmSGT2 gene related to plant height and branch development, and a mutant thereof and the use thereof. A Glycine max mutant having fewer branches and decreased plant height is obtained by means of EMS mutagenesis, and a target gene thereof that is located by means of a map-based cloning technique is a GmSGT2 gene of Hedou 12. It is found through searching that the gene encodes soyasapogenol B glucuronide galactosyltransferase, and can galactosylate soyasapogenol B monoglucuronide, thereby affecting the plant height of Glycine max and reducing the branches of Glycine max. Therefore, the GmSGT2 mutant can be used for cultivating dwarf high-yield Glycine max varieties, and is of great significance in the breeding of ideal plant types of Glycine max and research on important agronomic traits of plants. The mutant has broad application prospects and great research value with regard to understanding the plant height and branch regulation mechanism of Glycine max and improving the process of Glycine max breeding; and can provide an excellent germplasm resource stock for regulating the close planting and high-yield breeding of Glycine max.

The application discloses a galactosyltransferase mutant and application thereof in preparation of flavonoid compounds, and belongs to the technical field of enzymology. The application provides a UDP-galactosyltransferase mutant, which is obtained by site-directed mutation of one amino acid in the amino acid sequence of wild-type galactosyltransferase VcUFGT into alanine (A). The effects of a series of mutants H82A, V139A, G141A, P186A, N245A, V282A and S307A are verified in the examples, the mutants can perform enzyme catalysis reaction with quercetin and UDP-galactoside as substrates, and generate flavonoid compound hyperoside, and the specific enzyme activity of the mutants is more than 1.5 times that of the wild type, thereby providing an effective application basis for biosynthesis of flavonoid compounds.

The invention discloses a beta 1, 3-galactosyl transferase Pm beta 3GalT-S207H and an application thereof, and belongs to the technical field of functional enzymes. And the amino acid sequence of the beta 1, 3-galactosyl transferase Pm beta 3GalT-S207H is as shown in SEQ ID NO. 3. The invention further discloses the application of the beta 1, 3-galactosyl transferase Pm beta 3GalT-S207H in the preparation of lactose-N-tetrasaccharide. According to the invention, 1, 3-galactosyl transferase Pm beta 3GalT is subjected to mutation modification to obtain a mutant enzyme, and compared with a wild enzyme, the enzyme activity of the mutant enzyme is improved by 157.21% and is obviously improved, and the thermal stability and alkali resistance are also obviously improved. The beta 1, 3-galactosyl transferase Pm beta 3GalT-S207H disclosed by the invention has important theoretical value and application value on the industrial production of LNT (Lipoxystrobin).

This invention discloses a high-yield lactyl-3-4 ...5-4-4-4-4-4- N This invention relates to plasmid-free genetically engineered strains of tetrasaccharides and their applications, belonging to the field of microbial genetic engineering. The invention screened strains with higher activity. β -1,3-galactosyltransferase gene PmgalT Computer-aided enzyme function modification was used to achieve a shake-flask yield of 10.05 g / L for the constructed mutant. Subsequently, genome integration was performed... breast Multi-copy integration PmgalT * V27I / V39I / Y147F ‑galE‑galU A plasmid-free strain was constructed, enabling it to grow in shake flasks with lactoyl- N The yield of tetrasaccharides reached 14.4 g / L. Using glucose as the sole carbon source, the highest yield of 68.19 g / L was achieved at 64.5 h; using glycerol as the sole carbon source, the highest yield of 62.88 g / L was achieved at 60.5 h, providing a basis for its industrial production.

The invention provides a preparation method of an alpha-2, 3 sialic acid glycosyl transferase mediated sugar chain fixed-point coupling antibody drug conjugate, which comprises the following steps: S1, dissolving an antibody, adding UDP-Gal, Mn < 2 + > and beta-1, 4 galactosyl transferase for reaction, and purifying after reaction to obtain an antibody I with galactose at the tail end of a sugar chain; s2, dissolving the antibody I, adding an azide-modified CMP sialic acid derivative, alpha-2, 3 sialic acid glycosyltransferase and alkaline phosphatase for reaction, and purifying after reaction to obtain an antibody II of which the tail end of a carbohydrate chain is connected with the sialic acid derivative; s3, dissolving toxin molecules in DMSO, adding the dissolved toxin molecules into the antibody II for reaction, and performing ultrafiltration after the reaction to obtain ADC molecules; or the ADC molecule is directly prepared by adopting a one-pot enzyme method. The screened ST3Gal3 is used for preparing the carbohydrate chain fixed-point coupling ADC, the DAR value is controllable, batch-to-batch stability is achieved, uniformity is high, repeatability is high, quality control and CMC are better, and then the drug effect of the ADC can be better improved.

The invention discloses beta 1, 4-galactosyl transferase Ha beta 4GalT-I211V and application of the beta 1, 4-galactosyl transferase Ha beta 4GalT-I211V in synthesis of pLNnH, and belongs to the technical field of functional enzymes. And the amino acid sequence of the beta 1, 4-galactosyl transferase Ha beta 4GalT-I211V is as shown in SEQ ID NO. 3. The invention further discloses the application of the beta 1, 4-galactosyl transferase Ha beta < 4 > GalT-I211V in the synthesis of pLNnH. The beta 1, 4-galactosyl transferase Ha beta 4GalT-I211V provided by the invention is obtained by mutation modification, and the beta 1, 4-galactosyl transferase Ha beta 4GalT-I211V can be used for preparing straight-chain lactoyl-N-neohexasaccharide by taking UDP-galactose as a donor and straight-chain lactoyl-N-pentasaccharide as a receptor. The beta 1, 4-galactosyl transferase Ha beta 4GalT-I211V disclosed by the invention has a wide application prospect, and is of great significance to industrial preparation of linear-chain lactyl-N-neohexaose.

The application discloses a beta 1,4-galactosyltransferase Ha beta 4GalT-S233P and application thereof in synthesis of LNnT, and belongs to the technical field of functional enzymes. The amino acid sequence of the beta 1,4-galactosyltransferase Ha beta 4GalT-S233P is shown as SEQ ID NO. 3. The application discloses the application of the beta 1,4-galactosyltransferase Ha beta 4GalT-S233P in synthesis of LNnT. The beta 1,4-galactosyltransferase Ha beta 4GalT-S233P is obtained by modification of the beta 1,4-galactosyltransferase Ha beta 4GalT, has higher glycosyltransfer activity, and the conversion rate of synthesis of LNnT can reach 99.80%. The application lays a foundation for metabolic engineering of Escherichia coli for production of LNnT.

The application provides a genetically engineered bacterium for producing lactose-N-neotetraose, a construction method and application thereof. The genetically engineered bacterium satisfies the following conditions: (1) exogenous expression of MFS transporter protein, beta-1, 3-N-acetylglucosamine transferase and beta-1, 4-galactosyltransferase; (2) overexpression of galactoside permease and UDP-galactose-4-epimerase; and (3) no expression or weakened expression of setA transporter protein. The applicant of the application finds through research that exogenous introduction of MFS transporter protein, beta-1, 3-N-acetylglucosamine transferase and beta-1, 4-galactosyltransferase in the genetically engineered bacterium, enhancement of the expression amount of galactoside permease and UDP-galactose-4-epimerase in the genome, and reduction of the expression amount of setA transporter protein can effectively reduce the residual amount of LNT II in the LNnT synthesis pathway and improve the fermentation yield of LNnT.

The application belongs to the technical field of genetic engineering, and discloses a kind of synthetic lactosyl-N-neotetraose and safe recombinant bacillus subtilis and its construction method and application, the recombinant bacillus subtilis is obtained by transforming plasmid containing beta-1,3-N-acetylglucosamine transferase, beta-1,4-galactosyltransferase gene and lactose permease gene in bacillus subtilis 168.The application aims at the huge demand of lactosyl-N-neotetraose in the market in prior art, and the lactosyl-N-neotetraose produced by most existing engineering bacteria is not safe, provides an engineering bacteria for producing lactosyl-N-neotetraose and application, to meet the demand of lactosyl-N-tetraose in the market.

Arrhythmogenic cardiomyopathy (ACM) is a familial heart disease characterized by progressive myocardial fibrofatty infiltration. Compositions in the treatment of fibrofatty accumulation and its pathogenesis in ACM include activators of glycosphingolipid (GSL) glycosyltransferase, β-1,4-galactosyltransferase- V (β-1,4-GalT-V).

The invention discloses plasmid-free recombinant escherichia coli for synthesizing lactoyl-N-neotetraose as well as a construction method and application of the plasmid-free recombinant escherichia coli, and belongs to the field of gene engineering. An engineering strain MG-LNT-07 is used as a starting strain, beta-1, 4-galactosyl transferase Hplex2B is introduced, the copy number of the engineering strain is optimized, a multidrug efflux transporter gene mdfA is integrated, the expression of a glycoside donor pathway gene is fine-tuned, and a rapZ gene is knocked out, so that a plasmid-free strain for efficiently synthesizing the lactoyl-N-neotetraose is constructed, finally, the extracellular titer of LNnT is 7.76 g / L, and the expression of the lactoyl-N-neotetraose can be efficiently synthesized. And only 0.58 g / L of precursor LNTriII is remained, so that the method has a good industrial prospect.

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.

Provided herein are genetically modified rodents (e.g., mice or rats) with a human or humanized Fc epsilon receptor 1 alpha (FcεR1α) that do not express a functional α-1,3, -galactosyltransferase (GGTA1). Also provided herein are methods of making and / or using such rodents, cells from such rodents, and ES cells comprising the same genetic modifications.

This invention discloses the application of β-1,4-galactosyltransferase 1 and its inhibitors in liver diseases, particularly acute liver injury and liver failure. The application of β-1,4-galactosyltransferase 1 and its inhibitors in acute liver injury and liver failure provides the correlation between β-1,4-galactosyltransferase 1 and acute liver injury and liver failure, confirming that inhibiting the activity of β-1,4-galactosyltransferase 1 can alleviate acute liver injury and liver failure. β-1,4-galactosyltransferase 1 can serve as a drug target for screening acute and chronic hepatitis, liver injury, fatty liver, liver fibrosis, and acute and chronic liver failure. This invention also confirms the alleviating effect of β-1,4-galactosyltransferase 1 inhibitors on acute liver failure. β-1,4-galactosyltransferase 1 inhibitors improve acute liver failure by reducing the enzyme activity or protein expression of β-1,4-galactosyltransferase 1.

The application belongs to the technical field of biological medicine, and discloses a preparation method and application of an antibody drug conjugate with site-specific glycosyl coupling. The application discloses a recombinant cell containing a nucleic acid molecule encoding trastuzumab, and the FUT8 gene and the B4GALT1 gene are deleted. The application provides a recombinant cell which can efficiently and stably prepare trastuzumab with a uniform A2 glycosylation type. Based on the trastuzumab, 2, 4 or 6 azido groups can be introduced at specific sites of the antibody by using alpha 1, 6-fucosyltransferase and / or beta 1, 4-galactosyltransferase, and a toxin molecule is further connected, so that an antibody conjugate with uniformity and DAR value = 2, DAR value = 4 or DAR value = 6 is prepared.

The application discloses application of beta 1,4-galactosyltransferase 1 (B4GALT1) in preparation of a medicine for resisting mycobacterium tuberculosis infection and belongs to the technical field of biotechnology. The application first determines that a host functional protein B4GALT1 can improve the serum IgG galactosylation level, promote macrophage phagocytosis and killing of mycobacterium tuberculosis, and reduce the bacterial load. The B4GALT1 can be used as a new potential medicine for resisting tuberculosis infection. The application achievement can provide a new tool and thought for clinical treatment of tuberculosis, especially has a broad prospect in development and clinical application of an anti-tuberculosis medicine and can be directly applied to the scientific research field. The application has important application value for obtaining a new drug target for resisting tuberculosis and screening a new medicine.

The invention discloses a construction method and application of genetically engineered bacteria for producing lactoyl-N-neotetraose, and belongs to metabolic engineering and food fermentation technologies. A gene lacZ, a gene wecB, a gene ushA and a gene setA in escherichia coli BL21 (DE3) are knocked out, a gene SmSET is integrated in situ, expression of beta-1, 3-acetyl glucosamine transferase and beta-1, 4-galactosyl transferase is regulated and controlled, efficient synthesis of lactoyl-N-neotetraose is achieved, and the capacity of producing LNnT in a shake flask is improved to 1.85 g / L from initial 0.79 g / L; in a 3L fermentation tank, the yield of the LNnT reaches 39.85 g / L. The recombinant escherichia coli is used for preparing the LNnT, so that the method has the advantages of simple culture medium, cheap substrate, fast strain growth, genetic stability, high expression level and the like, and has remarkable industrial production potential.

This invention belongs to the field of biotechnology, specifically relating to the GmSGT2 gene and its mutants related to plant height and branching development, and their applications. Specifically, this invention obtains a soybean mutant with fewer branches and reduced plant height through EMS mutagenesis, and locates the target gene as the GmSGT2 gene of soybean variety 'He Dou 12' using map-based cloning technology. Searching reveals that this gene encodes soybean saponin B glucuronide galactosyltransferase, which can galactosylate soybean saponin B monoglucuronide, thus affecting soybean plant height and reducing branching. Therefore, the gmsgt2 mutant can be used to breed dwarf, high-yielding soybean varieties, which is of profound significance in breeding ideal soybean plant architecture and research on important agronomic traits. It has broad application prospects and high research value for understanding the regulation mechanism of soybean plant height and branching, improving the soybean breeding process, and can provide excellent germplasm resources for regulating soybean dense planting and high-yield breeding.

The present invention provides certain donor animals, tissues and cells that are particularly useful for xenotransplantation therapies. In particular, the invention includes porcine animals, as well as tissue and cells derived from these, which lack any expression of functional alpha 1,3 galactosyltransferase (aGT) and express one or more additional transgenes which make these animals suitable donors for xenotransplantation of vascularized xenografts and derivatives thereof. Methods of treatment and using organs, tissues and cells derived from such animals are also provided.

The invention discloses beta 1, 4-galactosyl transferase Ha beta 4GalT-S233P and application of the beta 1, 4-galactosyl transferase Ha beta 4GalT-S233P in synthesis of LNnT, and belongs to the technical field of functional enzymes. And the amino acid sequence of the beta 1, 4-galactosyl transferase Ha beta 4GalT-S233P is as shown in SEQ ID NO. 3. The invention further discloses the application of the beta 1, 4-galactosyl transferase Ha beta < 4 > GalT-S233P in the synthesis of LNnT. The beta 1, 4-galactosyl transferase Ha beta 4GalT-S233P provided by the invention is obtained by modifying the beta 1, 4-galactosyl transferase Ha beta 4GalT, and has higher transglycosylation activity, and the conversion rate of the synthesized LNnT can reach 99.80%. The invention lays a foundation for producing LNnT by transforming escherichia coli through metabolic engineering.

The invention relates to a method for increasing the yield of lactose-N-tetraose based on a dynamic regulation and control system, and belongs to the technical field of biological engineering. UDP-N-acetylglucosamine-2-epimerase, beta-galactosidase and 6-phosphofructokinase coding genes are knocked out from escherichia coli, glucosamine 6-phosphate synthase, N-acetyl glucosamine 1-phosphate uridinetransferase / glucosamine 1-phosphate acetyltransferase and glucosamine phosphomutase are expressed, and the glucosamine 6-phosphate synthase, the N-acetyl glucosamine 1-phosphate uridinetransferase and the glucosamine phosphomutase are respectively expressed in beta-1, 2, 3, 4, 5, 6-tetramethyl-1, 3, 4-tetramethyl-1, 3, 4-tetramethyl-1, 3, c ends of the beta-1, 3-N-acetylglucosamine transferase and the beta-1, 3-galactosyl transferase are connected with polypeptide tags, and a stable-phase promoter is used for expressing a micro-interval protein and a tag ligand, so that the polypeptide tags and the tag ligand are specifically combined. In order to further increase the yield of a target product, a growth-coupled promoter is used for expressing 6-phosphofructokinase and a degradation tag, uncoupling of cell growth and a product synthesis process is realized, the final yield of lactose-N-tetrasaccharide reaches 2.89 g / L, the production intensity is 0.04 g / L / h, and the method has relatively good application prospects and industrial potential.

The present invention provides transgenic animals (e.g., transgenic porcine animals), organs, tissues, and cells derived from the transgenic animals that are particularly useful for xenotransplantation therapies. In particular, the present invention provides transgenic porcine animals, as well as organs, tissues and cells derived from the transgenic porcine animals, which lack any expression of a functional alpha 1,3 galactosyltransferase (GTKO) gene and comprise at least six transgenes under the control of at least three promoters within a single multi-gene expression vector, and further comprise at least four additional genetic modifications. Also provided are methods of making the transgenic animals (e.g., transgenic porcine animals), and methods of using the transgenic animals, organs, tissues, and cells derived from the transgenic animals for xenotransplantation therapies and treating a disease or condition.

The application provides a construction method and application of recombinant escherichia coli for efficiently preparing high-purity sialyllactose-N-tetraose a. The application precisely integrates genes encoding beta-1,3-N-acetylglucosamine glycosyltransferase, beta-1,3-galactosyltransferase and alpha-2,3-sialic acid glycosyltransferase in the genome of escherichia coli BL21 (DE3), knocks out endogenous competitive metabolic genes, and optimizes the metabolic pathways of precursor substances UDP-acetylglucosamine, UDP-galactose and CMP-N-acetylneuraminic acid, effectively reduces the generation of non-target byproducts including lactose-N-triose, lactose-N-tetraose and 3'-sialyllactose, and successfully constructs a recombinant chassis strain capable of efficiently producing high-purity sialyllactose-N-tetraose a. Fermentation culture of the constructed engineering bacteria can efficiently prepare high-purity sialyllactose-N-tetraose a under the premise of guaranteeing biosafety, and is suitable for the production demand in the food field.

The present invention relates to an expression system for the extracellular production of recombinant tri-fucosylated N-glycoproteins in insect cells. The invention also relates to an expression cassette comprising a glyco-module comprising polynucleotide sequences of a nematode galactosyltransferase, fucosyltransferase, and mannosidase; and a polynucleotide sequence encoding one or more tri-fucosylated N-glycoproteins derived from a nematode; a recombinant baculovirus comprising said expression cassette, a method for producing said tri-fucosylated N-glycoproteins in transfected insect cells, and a transfected insect cell. The invention also relates to an immunological composition comprising said tri-fucosylated N-glycoproteins, which can be administered to ruminants to induce protective anti-nematode immunity.

The present invention relates to a vector, such as a viral vector, comprising a first nucleic acid sequence encoding a glycosyltransferase (such as a galactosyltransferase), an enzymatically active variant or truncated form thereof; and a second nucleic acid sequence encoding a Helicobacter pylori neutrophil-activating protein (NAP), an immunologically equivalent variant or fragment thereof. The invention also relates to a vector system, an isolated cell population, a polynucleotide and relating gene therapy systems comprising said first and said second nucleic acid sequence. Related methods, kits, pharmaceutical compositions and medical uses, in particular, in the treatment of cancer, such as pancreatic cancer, are also disclosed.

The present application relates to the technical field of molecular biology, in particular to a kind of recombinant alpha-1,3-galactosyltransferase and its preparation method and application.The recombinant protein is recombinant alpha-1,3-galactosyltransferase, and the amino acid sequence of the recombinant protein is shown as SEQ ID NO.1.The sequence of alpha-GT gene of the present application realizes the aggregation of high concentration of protein in cell after expression in engineering bacteria, but at the same time, it does not occur with the combination of cell membrane, and this characteristic helps to improve the yield of target protein in the fermentation process of engineering bacteria, reduces the operation difficulty in the process of purification, improves protein purity and activity.

The invention discloses a nicotiana benthamiana heterologous expression recombinant draytumeukumab as well as a preparation method and application of the recombinant draytumeumab. The method comprises the following steps: by taking nicotiana benthamiana as a production chassis, co-expressing, separating and purifying the dalratuloumab and human beta-1, 4-galactosyl transferase for the first time, so as to obtain the recombinant dalratuloumab for heterologous expression of nicotiana benthamiana. According to the present invention, the biological activity determination is performed on the recombinant drayukumab subjected to the heterologous expression of the nicotiana benthamiana, and the results show that the antigen binding capacity and the anti-tumor cell proliferation capacity of the recombinant drayukumab are equivalent to the animal source drayukumab; according to the invention, a scientific basis is provided for production of the recombinant draytumeumab from the plant chassis, and a reference basis is provided for research on difference and feasibility of production of the recombinant antibody from a plant platform instead of animal cells.

The invention discloses a cotton drought-resistant related protein galactosyl transferase GhNGalT1 positioned in chloroplast and application of the cotton drought-resistant related protein galactosyl transferase GhNGalT1. According to the invention, a galactosyl transferase gene GhNGalT1 which is induced by ABA and is positioned in chloroplast in cotton is cloned and identified for the first time, a new potential drought-resistant regulation pathway of ABA signal-GhNGalT1-chloroplast function is disclosed, and the understanding of the plant glycosyl transferase function, especially the abiotic stress response function of the plant glycosyl transferase gene in chloroplast, is broadened. Functional verification shows that GhNGalT1 is a positive regulation factor for drought resistance of cotton. Overexpression of the gene can significantly enhance the drought resistance of cotton, and silence or knockout of the gene causes the cotton to be more sensitive to drought. The GhNGalT1 gene has an important application value in main varieties of cotton, can be used for molecular design breeding and cultivation of new varieties of drought-resistant, water-saving, stable-yield and high-quality cotton, and has practical significance for guaranteeing the safety of cotton production in China.

The present invention provides transgenic animals (e.g., transgenic porcine animals), organs, tissues, and cells derived from the transgenic animals that are particularly useful for xenotransplantation therapies. In particular, the present invention provides transgenic porcine animals, as well as organs, tissues and cells derived from the transgenic porcine animals, which lack any expression of a functional alpha 1,3 galactosyltransferase (GTKO) gene and comprise at least six transgenes under the control of at least three promoters within a single multi-gene expression vector, and further comprise at least four additional genetic modifications. Also provided are methods of making the transgenic animals (e.g., transgenic porcine animals), and methods of using the transgenic animals, organs, tissues, and cells derived from the transgenic animals for xenotransplantation therapies and treating a disease or condition.