1874 results about "Glycosylation" patented technology

According to the glycosyl transferase mutant provided by the invention, the region selectivity of tyrosol is remarkably improved from 51% of a wild type to 99% or above, and when the glycosyl transferase mutant is applied to catalytic synthesis of salidroside, a single glycosylation product salidroside can be generated; besides, the catalytic activity of the microbial glycosyl transferase mutant to tyrosol is remarkably improved, the specific enzyme activity is improved to 97 mU / mg from 14 mU / mg of a wild type, and the substrate conversion rate is higher than 99%; according to the invention, the key problems of difficult glycosyl transferase expression, low regioselectivity and low activity in the salidroside preparation process are successfully solved, and the salidroside has a very great industrial application prospect.

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 invention provides a method for preparing recombinant human albumin with low O-glycosylation level, comprising taking at least one measure to improve the growth rate of the strain and reduce the O-glycosylation level of the protein during the fermentation process, and specifically comprising S1, seed liquid preparation; S2, BSM culture medium is put into a fermentation tank, the pH is adjusted to 5.0-5.8, sterilization is carried out, dissolved oxygen is corrected after cooling, temperature and ventilation volume are set, sterilized PTM1 solution is added to the culture medium, and the pH of the culture medium is adjusted to 5.0-5.8 again; S3, shaking flask seeds are connected to the fermentation tank, dissolved oxygen is corrected, and rotation speed is set; S4, when the dissolved oxygen increases suddenly, a glycerol feeding stage is entered, and glycerol feeding is added at a uniform speed; S5, glycerol feeding and methanol feeding are added simultaneously, pH is adjusted to 6.0-6.5, and the temperature is lowered; S6, methanol feeding is added; S7, after the fermentation is completed, the tank is released, the fermentation product is centrifuged, and the supernatant is collected; the method of the invention can reduce protein O-glycosylation, and the expressed recombinant protein contains a very low level of glycosylation modification.

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 provides a fusion protein with a tribasic group editing function and application thereof, and relates to the technical field of basic group editing. The fusion protein comprises adenine deaminase, N-methylpurine DNA (deoxyribonucleic acid) glycosylase and nuclease. Aiming at the problem that an existing base editing tool cannot edit by taking three bases as substrates at the same time, the invention provides a novel three-function base editor which is named as A, Camp; and G-BE. A, Camp; the G-BE can be edited by independently using A, C and G as substrates to realize the editing activity of A-G / T / C, C-T / G / A and G-T / A / C, and can also be edited by simultaneously using A, C and G as substrates on alleles.

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.

Fusion proteins are provided according to aspects of the present disclosure which include: a glycan binding component linked to a mutant E. coli biotin ligase BirA, the glycan binding component capable of specific binding to a glycosylation post-translational modification of a target protein and the mutant E. coli biotin ligase BirA having enzymatic activity to ligate biotin to proteins proximal to the target protein. Methods of detecting proteins proximal to a target protein are provided according to aspects of the present disclosure which include contacting a living cell with the fusion protein according to aspects of the present disclosure under compatible biological conditions; and detecting the biotinylated proteins, thereby detecting proteins proximal to the target protein.

The invention relates to the technical field of protein and peptide, in particular to cubilose peptide with high antioxidant activity and a preparation method of tablets of the cubilose peptide. Comprising the following steps: preparing modified cubilose zymolyte; performing multiple enzymolysis on the modified cubilose zymolyte; the antioxidant stability of the cubilose peptide is enhanced. The resveratrol is grafted on the surface of cubilose protein through free radical reaction of the resveratrol, so that not only can the antioxidant capacity of the cubilose be enhanced, but also the water solubility of the cubilose can be improved, the subsequent enzyme can be better contacted with the cubilose, the enzymolysis efficiency is improved, and the enzymolysis effect is improved. According to the preparation method, cubilose peptide is added into cubilose, reduction of antioxidant activity of the cubilose peptide due to external oxidation factors and too long enzymolysis time is avoided, then the cubilose is subjected to glycosylation modification through chitosan, the dispersion and emulsification stability of the modified cubilose zymolyte can be improved, and the contact area of subsequent enzymes and the modified cubilose zymolyte is increased; the oxidation resistance and the subsequent enzymolysis efficiency are further enhanced.

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 relates to the technical field of mulberry processing, in particular to fermented mulberry powder with sleep-aiding and sugar-resisting effects as well as a preparation method and application of the fermented mulberry powder. The preparation method comprises the following steps: adding mulberries into water, and pulping to prepare pulp; adding a compound enzyme into the slurry for enzymolysis to obtain an enzymatic hydrolysate; equally dividing the enzyme solution into two parts, and respectively adding saccharomyces cerevisiae and plant lactobacillus for first-stage fermentation to obtain a first fermentation solution; sterilizing the first fermentation liquor, and adding casei paracasei for second-stage fermentation to obtain second fermentation liquor; and filtering the second fermentation liquor, adding a solid filler into the filtrate, and then sterilizing and drying in sequence to obtain the fermented mulberry powder. Compared with common mulberry powder, the fermented mulberry powder prepared by the preparation method disclosed by the invention has the effects of aiding sleep and resisting sugar, is obvious in aiding sleep, can inhibit generation of AGEs, and has the effect of inhibiting glycosylation.

The present invention relates to a method of increasing nicotinamide mononucleotide (NMN) and / or nicotinamide adenine dinucleotide (NAD +) in the intestinal tract of a subject, the present invention relates to a method for treating a subject by administering to the subject a combination of at least one Bifidobacterium sp. Capable of growing on a sialylated HMO and having a functional de novo nicotinamide adenine dinucleotide (nad operon (nadC, nadD, and nadE) and at least one fucosylated oligosaccharide.

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 present invention relates to anti-LYPD3 antibodies that bind to LYPD3 in an O-glycosylation dependent manner. Therefore, the antibody has specificity to tumor-associated LYPD3 (LYPD3). The invention also provides a pharmaceutical composition containing the anti-LYPD3 antibody and an application of the pharmaceutical composition in cancer treatment.

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.

Conjugation of dendrimer molecules with one or more carbohydrate moieties significantly enhances targeting of tumor-associated macrophages (TAMs) and microglia by increasing brain penetration and cellular internalization, as compared with dendrimers without modification with carbohydrate molecules. Compositions of dendrimers conjugated with carbohydrate moieties, particularly glucose and / or glucosamine, and one or more active agents to prevent, treat, or diagnose a disease or disorder in a subject in need thereof, and methods of use thereof, have been developed. The compositions are particularly suited for treating and / or ameliorating brain and / or CNS tumors. Methods of treating a human subject having or at risk of brain and / or CNS tumors are provided.

The invention discloses a detection method and application of whole genome 5-hydroxymethylcytosine, and relates to the technical field of biochemical detection. Comprising the following steps: 1, closing a 3 '-OH end of DNA to be detected; 2, preparation of glycosylated 5hmC DNA (Deoxyribose Nucleic Acid); 3, carrying out TdT mediated DNA (Deoxyribonucleic Acid) amplification; and 4, fluorescence measurement. According to the invention, a homogeneous and non-template dependent type signal amplification method for detecting the 5-hydroxymethylcytosine in the whole genome is constructed. The method is based on a specific enzyme catalytic reaction and a multiple signal amplification strategy, and has the characteristics of high sensitivity, good specificity, no need of a separation step, no sequence dependence in amplification and capability of detecting 5hmC at any position in a whole genome. In addition, accurate and sensitive detection of actual clinical samples can be realized.

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 glycosylated ganoderic acid with a chemical structural formula shown in the specification, the compound can effectively improve H2O2-induced cellular morphological change, remarkably reduce generation of reactive oxygen species (ROS) and reduce accumulation of beta-galactosidase in senescent cells, and in addition, cell immunoblotting analysis shows that the compound can be used for preparing glycosylated ganoderic acid. The glycosylated ganoderic acid can significantly down-regulate the expression of H2O2-induced senescence marker proteins p16 and p21, and experimental results prove that the glycosylated ganoderic acid has potential in the aspect of reversing cell senescence, provides a new candidate compound for the development of anti-senescence drugs, and shows a wide application prospect; # imgabs0 #