1316 results about "Glycosylation" patented technology

The invention provides a high-stability recombinant XVII type collagen as well as a construction method and application thereof, and belongs to the technical field of bioengineering. According to the invention, an amino acid sequence of natural human (XVII) type collagen is selected as a basic unit and repeated for multiple times to obtain recombinant (XVII) type collagen; in the design process of the recombinant XVII type collagen, an unstable GXY triad is avoided, potential matrix metalloproteinase restriction enzyme cutting sites and glycosylation sites are abandoned, a KGD active sequence is reserved, and the probability of containing sites which are easy to degrade can be reduced; a sequence fragment of the recombinant XVII type collagen is derived from a full-length sequence of a natural collagen gene, has excellent solubility, biological activity and high stability, and can be kept stable in a high-temperature storage environment and under the condition of avoiding alkalinity; the product can be used as a raw material in the production process of medicines, medical equipment, biological materials or cosmetics, and has good practicability.

The invention discloses a method for preparing low-GI rice noodles from a white kidney bean-mulberry leaf composite extract, which comprises the following steps of: firstly, extracting functional components in white kidney beans and mulberry leaves by adopting a water extraction method, adding Arabic gum and resistant dextrin into a white kidney bean-mulberry leaf composite extracting solution, regulating acid, and performing glycosylation and embedding treatment; uniformly mixing rice flour, high amylose, fiber powder, konjac glucomannan, mono-glycerol fatty acid ester, diglycerol fatty acid ester and the white kidney bean-mulberry leaf compound extracting solution; and finally, humidifying, cooking, extruding and forming to obtain the low-GI rice noodles. According to the invention, the glucogenic index of the rice noodles is reduced by adding the white kidney bean-mulberry leaf composite extract, the heat resistance of functional components in the composite extract is improved through glycosylation and embedding treatment, and the loss of the activity of the white kidney bean-mulberry leaf composite extract in the high-temperature cooking process of the rice noodles is reduced; meanwhile, in order to improve the flavor and taste of the low-GI rice noodles, the konjac glucomannan and other auxiliary materials are added, so that the low-GI rice noodles are rich in elasticity and better in taste and boiling fastness.

The present disclosure provides methods for reducing sequencing errors comprising one or any combination of: (i) removing deaminated bases in any nucleic acid molecule throughout a library preparation workflow which includes immobilised splints which bind to the library, the use of a compaction oligonucleotide, optionally with an intervening sequence, formation of closed circular nucleic acids, creating gaps using glycosylase and lyase activities at positions with deaminated bases. The library may be sequenced using pairwise sequencing, e.g. with dark sequencing and / or sequencing using a multivalent labelled probe for the formation of an avidity molecule and soluble primer and polymerase. Method for sequencing concatemers in which the concatermers are contacted with polymerases, soluble primers and a multivalent labelled molecule which forms a complex with the polymerase. Detecting polymerase position and nucleobase bound to the polymerase in the complex. These methods generate higher quality base calls during downstream sequencing workflows.

The present disclosure relates to a low glycosylated spike protein and a vaccine designed to express the spike protein in vivo. The present disclosure also teaches a method for generating an immune response by utilizing the low glycosylated spike protein, which provides a broader protection across different variants. A method for identifying a glycan-shielded conserved peptide of a glycoprotein is also disclosed.

The invention discloses a method for preparing a high internal phase emulsion and delivering curcumin by using transglutaminase mediated casein-carboxymethyl chitosan covalent substances, aims to improve the stability and bioaccessibility of curcumin, and belongs to the field of food processing. The method comprises the following steps: firstly, carrying out ultrasonic treatment on sodium caseinate for a certain time, and carrying out transglutaminase cross-linking reaction on sodium caseinate and carboxymethyl chitosan to obtain a casein-carboxymethyl chitosan covalent substance; and then adding the mixture into the curcumin-loaded corn oil according to a certain proportion, and homogenizing to obtain the curcumin-loaded high internal phase emulsion. The emulsion has good interface characteristics and can effectively encapsulate curcumin. The preparation process is simple, efficient, green and environmentally friendly, the prepared high internal phase emulsion has high stability and good biological accessibility, a new technical means is provided for delivery of curcumin, and the high internal phase emulsion has wide application prospects in the field of food.

The invention provides an application of citrus flavonoid glycosyl transferase CitUGT73A78 in biosynthesis of flavonoid glycoside, and belongs to the technical field of gene engineering and enzyme engineering. According to the present invention, the function of the CitUGT73A78 gene is cloned and verified, and the in vitro enzyme activity test proves that the CitUGT73A78 recombinant protein has broad-spectrum catalytic activity, and can efficiently catalyze the glucosylation of a variety of flavonoids so as to generate the corresponding flavonoids 7-O-glucoside and / or flavonoids 4 '-O-glucoside. An in-vivo transient overexpression test also proves that the CitUGT73A78 gene has a promoting effect on the accumulation of the content of the citrus flavonoid 7-O-glucoside. The invention not only provides a preparation method for the production of flavonoid 4 '-O-glucoside, has the advantages of simplicity, convenience, high catalytic efficiency, low cost, greenness and safety, but also can be applied to nutritional quality improvement and germplasm innovation of citrus fruits.

The invention relates to the technical field of food processing, and discloses casein-tea polysaccharide-polyphenol composite nanoparticles and a preparation method of the casein-tea polysaccharide-polyphenol composite nanoparticles, and the preparation method comprises the following steps: mixing large yellow tea polysaccharide with a casein solution to obtain a CS / TPS mixed solution; performing glycosylation modification on casein in the CS / TPS mixed solution by adopting a Maillard reaction to prepare a casein-polysaccharide copolymer solution; the preparation method comprises the following steps: mixing a plurality of catechins, dissolving the catechins in ultrapure water to obtain a mixture stock solution, mixing the mixture stock solution and a casein-polysaccharide copolymer solution according to a volume ratio of 1: 16, stirring for 1 hour for self-assembly, pre-freezing the stirred solution at-80 DEG C, and freeze-drying for 48 hours to obtain CS-TPS-MIX nano-composite powder. According to the invention, a polyphenol delivery system with a dual antioxidant synergistic mechanism is constructed through a Maillard reaction mediated nano-composite technology, and the system is stable in structure, so that the release of polyphenol is more controllable.

The invention provides an anti-human thrombopoietin antibody AF687, a kit and application thereof. The anti-human thrombopoietin antibody disclosed by the invention can specifically recognize epitopes without glycosylation modification of natural TPO protein, and has relatively high affinity for recombinant TPO protein and specific epitopes of the recombinant TPO protein. Meanwhile, the anti-human thrombopoietin antibody disclosed by the invention can accurately detect the content of human thrombopoietin in serum, is not influenced by heterogeneity of natural protein glycosylation sites, can be used for accurately identifying and diagnosing early aplastic anemia and primary immune thrombocytopenia, and has the advantages that compared with an existing detection reagent, the anti-human thrombopoietin antibody is high in sensitivity and high in sensitivity. The false positive rate can be reduced from 13.9% to 2.7%, and a more accurate detection method is provided for differential diagnosis of the primary immune thrombocytopenia.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of a centella asiatica glycosyl transferase gene in catalyzing glycosylation of asiatic acid and madecassic acid. The nucleotide sequence of the gene is shown as SEQ ID No.1, and the protein coded by the gene can catalyze the C-28 site of asiatic acid and madecassic acid to perform glucose group modification so as to generate asiatic acid monoglucoside and madecassic acid monoglucoside, and preferentially catalyzes madecassic acid. By constructing a metabolic regulation network, the genes are widely and accurately screened, and the glycosylation process of catalyzing asiatic acid and madecassic acid is analyzed; an important reference basis is provided for improving the glycosylation efficiency of asiatic acid and madecassic acid and the yield of asiaticoside and madecassoside in the in-vitro synthesis and industrial production process of the centella asiatica triterpenoid saponin, and an important theoretical support is provided for large-scale production of the centella asiatica active saponin.

The present disclosure relates to methods for high-throughput analysis of proteins and peptides employing a set of engineered binders that recognize terminal amino acid residues of peptide analytes. In particular, disclosed herein is the analysis of occupancy of N-linked glycosylation sites in proteins and peptides. The disclosure finds utility at large-scale profiling of N-linked glycosylation sites, as well as monitoring changes in glycosylation patterns associated with numerous disease conditions.

The invention belongs to the field of enzyme engineering, and relates to a glycosyl transferase mutant and application thereof in synthesis of salidroside. On the basis of a glycosyl transferase amino acid sequence as shown in SEQ ID NO: 1, iterative saturated mutation is carried out through a molecular modification technology, and the enzymatic activity and catalytic efficiency of the modified glycosyl transferase mutant are greatly improved. The mutation at a specific site can significantly promote the glycosylation activity of a tyrosol substrate. The mutant enzyme is obtained by carrying out induced expression and protein purification on the obtained mutant strain. The substrate conversion rate of the transformed glycosyl transferase mutant for catalyzing the tyrosol glycosylation reaction can reach 95% or above, the yield reaches 55.0 g / L, and an efficient and green new way is provided for salidroside production.

The invention provides a non-disease diagnosis purpose 5-hydroxymethylcytosine 5hmC detection method and a non-disease diagnosis purpose 5-hydroxymethylcytosine 5hmC detection kit. The detection method comprises the following steps: S1, glycosylation treatment; s2, carrying out enzyme digestion treatment on the GSREs; s3, carrying out APOBEC deamination treatment; s4, carrying out hydroxymethylation detection; the kit comprises a glycosylation reaction kit, a GSREs enzyme digestion reaction kit, an APOBEC deamination reaction kit and a PCR (Polymerase Chain Reaction) kit. The glycosylation reaction kit comprises a CutSmart buffer solution, UDP-Glucose, a T4-BGT enzyme and nuclease-free water, wherein the CutSmart buffer solution, the UDP-Glucose and the T4-BGT enzyme are used as raw materials; the GSREs enzyme digestion reaction kit comprises a GSREs enzyme, an enzyme digestion reaction buffer solution and nuclease-free water, the APOBEC deamination reaction kit comprises APOBEC, BSA (Bovine Serum Albumin), an APOBEC Reactive buffer solution and nuclease-free water; the PCR kit comprises a PCR reaction solution, a primer and a probe. According to the 5-hydroxymethylcytosine 5hmC detection method and kit for non-disease diagnosis purpose provided by the invention, precise quantification of extremely high specificity of 5hmC is realized, and the detection sensitivity is also improved while the specificity of a detection signal is improved.

The present application relates to an antibody capable of specifically binding to c-Met or antigen-binding fragment thereof, and an immunoconjugate, a pharmaceutical composition and a multispecific molecule comprising the same. The present application further relates to the use of the antibody specifically binding to c-Met or an antigen-binding fragment thereof and the multispecific molecule. Compared with the control antibody, the bispecific antibody or defucosylated bispecific antibody of the present application can block HGF-dependent TKI resistance; block the proliferation and migration of tumor cells induced by HGF, induce ADCC effect, and inhibit tumor growth in vivo.

The invention discloses a haemonchus contortus subunit vaccine, and belongs to the field of veterinary vaccines. The subunit vaccine comprises an antigen protein MEP3, an antigen protein AP1, an antigen protein AP8, an antigen protein H11-2, an antigen protein H11-4 and an antigen protein AP5 of galactosylated and modified haemonchus contortus, and the antigen protein MEP3, the antigen protein AP1, the antigen protein AP8, the antigen protein H11-2, the antigen protein H11-4 and the antigen protein AP5 of galactosylated and modified haemonchus contortus. The galactosylation modification of the antigen protein is realized by jointly expressing glycosyl transferase HcGALT of haemonchus contortus and one or more of the proteins in eukaryotic cells. The antigen protein modified by galactosylation in the subunit vaccine is closer to the glycosylation modification of natural protein, and can provide effective immune protection after the goat is infected with haemonchus contortus, and the egg laying rate is reduced by 81.43%. The invention lays a foundation for development of haemonchus contortus subunit vaccines, provides an effective technical means for prevention and treatment of the haemonchus contortus subunit vaccines, and has important application value and popularization prospect.

The invention belongs to the technical field of gene editing, and discloses a base editor system and application. According to the invention, a heterologous peptide sequence is inserted into IscB protein, the obtained peptide embedded IscB is fused with deaminase or glycosylase to construct a base editor, and DNA base editing is carried out. According to the base editor system, the size limitation is overcome, and single AAV delivery is achieved; the editing efficiency is obviously improved; the broad-spectrum applicability is realized; the system optimization is more flexible; the method has efficient editing potential in vivo, and the off-target effect is reduced. The invention provides an important new strategy for hyperlipemia treatment and provides a basis for gene therapy of other hereditary diseases.

The present disclosure relates to a low glycosylated spike protein and a vaccine designed to express the spike protein in vivo. The present disclosure also teaches a method for generating an immune response by utilizing the low glycosylated spike protein, which provides a broader protection across different variants. A method for identifying a glycan-shielded conserved peptide of a glycoprotein is also disclosed.

The invention discloses a preparation method of glucosyl stevioside, and relates to the technical field of glucosyl stevioside production, rebaudioside A or stevioside is taken as a raw material, cyclodextrin is taken as an auxiliary material, cyclodextrin glucosyltransferase is added for glycosylation reaction, a glucosyl stevioside solution I is cooled, and a glucosyl stevioside solution II is obtained; the method comprises the following steps: adding sucrose, UGT glycosyl transferase, sucrose synthase and uridine diphosphate into rebaudioside A or stevioside, continuously converting unreacted rebaudioside A or stevioside into rebaudioside D or rebaudioside E, inactivating after the reaction is finished, filtering by a plate frame, removing impurities through resin, analyzing the resin by using ethanol to obtain an analysis solution, and treating the analysis solution at 78-82 DEG C for 2-4 hours to obtain the rebaudioside D or rebaudioside E; and dealcoholizing at the vacuum degree of-0.1 MPa, concentrating and drying to obtain the glucosyl stevioside. The quality of the product is improved, the cost is reduced, and the taste of the product is integrally improved.

The invention belongs to the field of food, and particularly relates to a nutritional composition containing breast milk oligosaccharide and probiotics for improving depression and application of the nutritional composition. The invention provides application of a nutritional composition in preparation of food beneficial to improving constipation and anxiety state and / or depression state caused by constipation. Wherein the nutritional composition comprises the following components (i) and (ii): (i) a neutral fucosylated breast milk oligosaccharide, and (ii) a bifidobacterium probiotic; the neutral fucosylated breast milk oligosaccharide at least comprises 2 '-fucosylated lactose, and the bifidobacterium probiotics at least comprise bifidobacterium longum subsp. Longum. According to the present invention, the substance combination of the neutral fucosylated breast milk oligosaccharide and the bifidobacterium probiotic can improve the abnormal brain-intestine interaction, such that the constipation can be easily improved, and the anxiety and depression state caused by the constipation can be easily improved;

Provided herein are isolated monoclonal antibodies to CCR8 antibodies, antigen binding fragments thereof, mutants thereof, variants thereof, and uses thereof for the treatment of diseases such as cancer. The mutant antibodies have complementarity determining region 3 modified to eliminate post-translational modifications and improve the stability and potency of the antibodies. The variant antibodies have different levels of defucosylation. Also provided herein are methods of using the anti-CCR8 antibodies, including methods of treating cancer. Also provided are polynucleotides encoding the heavy or light chain or antigen-binding portion thereof described herein, as well as vectors, particularly expression vectors, comprising the polynucleotides described herein.

The invention discloses an application of burkholderia in root system microorganisms in regulation and control of glycosylation of ginsenoside. The invention provides application of burkholderia or a composition containing the burkholderia in any one of the following applications: 1) regulating and controlling proportions of different glycosylated saponins in ginseng; 2) increasing the proportion of monoglycosylated saponin and / or the proportion of disglycosylated saponin in ginseng; 3) preparing ginseng with a high monoglycosylated saponin ratio and / or a high disglycosylated saponin ratio; and 4) improving ginseng cultivation. In a greenhouse environment, the roots of ginseng seedlings are inoculated with burkholderia, metabolome data shows that the burkholderia regulates glycosylation formation of ginsenoside, and the proportion of monoglycosylated saponin and / or the proportion of disglycosylated saponin can be increased. Further, transcriptome sequencing data analysis shows that the burkholderia regulates and controls the expression of different glycosyl transferases, so that the diversity of glycosylation of ginsenoside is influenced. The burkholderia provides a new thought for refined improvement of a ginseng cultivation technology.

The invention relates to the technical field of collagen preparation, in particular to a collagen product as well as a preparation method and application thereof. The collagen product prepared by the invention overcomes the problems of poor dissolution effect in cold water, poor fat solubility and poor emulsifying property. The preparation method comprises the following steps: crushing bovine bones, and performing collagen extraction, enzymolysis treatment, glycosylation treatment, fatty acid amidation treatment, polyphenol grafting modification and colloid compounding to obtain the collagen product. During glycosylation treatment, the cold water dissolving effect is improved by adjusting the variety of reducing sugar, the reaction temperature and time and grafting bone peptide molecules to sugar chains, and a flavor substance is generated by the Maillard reaction in the glycosylation treatment, so that the effect of covering the fishy smell of cattle bones is achieved; relevant parameters are adjusted through fatty acid amidation treatment, the bone peptide is endowed with liposolubility and emulsibility, and the solubility and emulsibility of the collagen product are synergistically improved through colloid compounding, optimization of the use amount of various colloids and control of reaction conditions.

Herein are antibodies or antigen-binding fragments specifically binding to GPRC5D. Additionally, monovalent antibodies or antigen-binding fragments specifically binding to GPRC5D are also included. The Fc region of these antibodies contains K248E and T437R mutations (designated as "RE mutations") per the EU numbering system. The described antibodies, expressed in host cells that lack fucosylation capabilities, exhibit enhanced antibody¬ dependent cellular cytotoxicity (ADCC) and enhanced complement-dependent cytotoxicity (CDC).

Provided herein are compositions, methods, and uses involving antibodies that immunospecifically bind glycosylated forms of MUC16, a tethered mucin protein. Also provided herein are uses and methods for managing, treating, or preventing disorders, such as cancer.

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 invention provides an anti-non-enzymatic glycosylation composition and its application, and a non-enzymatic glycosylation inhibitor. The anti-non-enzymatic glycosylation composition is composed of the following raw materials in parts by weight: 30 to 90 parts of sialic acid, 10 to 40 parts of ginkgo leaf extract, 10 to 40 parts of acerola extract, and 0.1 to 10 parts of rosemary extract. The composition of the present invention can achieve an anti-non-enzymatic glycosylation level comparable to that of the prior art by compounding sialic acid with ginkgo leaf extract, acerola extract, and rosemary extract in a specific ratio, while the stability is also improved. Applying it to health foods or cosmetics with anti-aging effects saves costs and is conducive to production and promotion.

The invention relates to a green preparation method and application of an anti-saccharification tea extract, and belongs to the technical field of biology. The preparation method of the tea extract comprises the following steps: extracting tea by using a deep eutectic solvent to obtain the tea extract, and carrying out deproteinization and decoloration to obtain the tea extract. The hydrogen bond acceptor of the deep eutectic solvent comprises proline or betaine, and the hydrogen bond donor of the deep eutectic solvent comprises xylitol or glycerol. By utilizing the NaDES extraction technology, effective components can flow out by efficiently breaking cell walls, so that the extraction efficiency of tea active components represented by tea polyphenol is effectively improved; the selected NaDES solvent is natural, green and easy to obtain; the tea extract disclosed by the invention has an anti-saccharification effect, blocks glycosylation reaction, reduces and inhibits generation of AGEs, improves the problems of dark skin, skin laxity and aging caused by saccharification, reduces collagen loss, keeps skin elasticity and delays skin aging.

The invention provides a fish collagen peptide with a whitening effect and a preparation method thereof, and the method comprises the following steps: performing glycosylation modification treatment on fish collagen by using glucose, and then sequentially treating the fish collagen with gastric juice and intestinal juice to obtain the fish collagen peptide with a polypeptide sequence with glycosylation sites on a peptide chain. According to the invention, the fish collagen peptide with a remarkable inhibition effect on tyrosinase is obtained.

The invention discloses a method for constructing an N-glycosylation spectrum type biological age evaluation model based on plasma immunoglobulin G and application of the biological age evaluation model. Based on a glycosyl quantitative result, a clinical biochemical detection index, a lifestyle measurement index, a multi-dimensional health score and the like, a glycosyl metabolic index, a glycosyl inflammation index, a glycosyl cardiovascular index, a glycosyl stroke index, a glycosyl liver index and a glycosyl kidney index are fitted. And inputting the obtained different glycosyl data into the model, and speculating the aging process and health risk of each organ system of the detected person according to the risk threshold in the early-stage crowd. The method can predict a'glycosyl biological age system 'which can reflect health level and disease risk better than'calendar age', has individual specificity, is very sensitive to change of inflammatory response in vivo, is relatively stable in a short time, can be used as an index for early detection and screening of senescence, and can be used as an index for early detection and screening of senescence. And thus, effective reference is provided for individualized health suggestions of the detected personnel.

The invention relates to a recombinant expression cat interferon-omega gene and a preparation method and application thereof, the nucleotide sequence of the cat interferon-omega gene is as shown in SEQ ID NO.1, and the cat interferon-omega gene is obtained by introducing an Fc fusion fragment to the C terminal and / or N terminal of the natural gene sequence of the cat interferon-omega. XTEN or PAS is introduced to prolong a peptide fragment, and original glycosylation modification sites on a natural sequence are reserved. The method comprises the following steps: cloning a cat interferon-omega gene into an expression vector containing double promoters, further transfecting into a cell, carrying out stable cloning and screening, establishing a high-expression cell strain, and carrying out fermentation culture in a bioreactor. According to the invention, the cat interferon-omega gene is subjected to multiple modification and is efficiently expressed in a CHO-K1GS system, so that the protein yield, the stability and the half-life period are remarkably improved; the obtained fusion protein is high in purity and strong in activity, can obviously enhance immune response and protection effect when being matched with cat vaccines, and is good in safety.

The invention belongs to the field of chemical synthesis, and particularly relates to four acinetobacter baumannii cell wall oligosaccharide fragments as well as a synthesis method and application thereof. Four acinetobacter baumannii core oligosaccharide fragments are synthesized by adopting a divergent synthesis strategy, compounds 1-3 take orthogonally protected alpha-(2-> 5)-Kdo-Kdo disaccharide as key precursors, protecting groups at different sites are selectively removed, glycosyl is introduced through a stereoselective glycosylation reaction, and thus a target molecular skeleton is constructed. The method disclosed by the invention and the four finally prepared acinetobacter baumannii core oligosaccharide fragments have huge industrial application potential.