13 results about "Fucosidase" patented technology

The present invention relates to a cartilage tissue single cell extraction kit and an extraction method thereof, the kit comprises: (1) a first enzyme for enzymatic hydrolysis, the first enzyme for enzymatic hydrolysis comprising one or more of trypsin, chondroitinase ABC and fucosidase; (2) a second enzyme for enzymolysis, wherein the second enzyme for enzymolysis comprises collagenase II, neutral protease II, hyaluronidase and Dnase I; and (3) a basic solution, wherein the basic solution comprises a basic culture medium and / or a buffer solution. The invention also provides a method for extracting single cells from cartilage tissues by using the kit. The cartilage tissue single-cell suspension prepared by the method has the advantages of high cell yield, high activity and the like, and has a wide application prospect.

The present invention relates to use of an enzyme that cleaves both alpha- and beta-1,4-glycosidic bonds and, more particularly, the present invention provides α-L-fucosidase which has α-1,4-glucosidase and β-1,4-galactosidase activities and thus is capable of producing glucose and / or galactose, which are monosaccharides, from non-fucosylated saccharides. The enzyme has transglycosylation activity and thus produces maltooligosaccharides having a higher degree of polymerization than maltooligosaccharides used as a substrate, thereby being applicable to the development of high-value-added new materials in the food industry.

The invention provides a preparation method of an antibody conjugate containing an sLeX tetrasaccharide structure, which comprises the following steps: S1, sequentially adding Endo S2 enzyme and alpha-L-fucosidase into a target antibody, and respectively analyzing glycosylation to avoid modification site conflict or carbohydrate chain structure interference; s2, adding the accurately represented antibody in the S1, B4GalT1 and UDP-Gal into a Tris-HCl system containing Mn < 2 + >, and incubating to obtain a reaction solution I; s3, the reaction liquid I, ST3Gal3, CMP-Neu5Ac and Tris-HCl containing Mg < 2 + > are taken and mixed, and incubation is carried out; adding CMP-N-acyl neuraminic acid phosphodiesterase for reaction, and purifying Protein A to obtain a purified antibody I; s4, the purified antibody I, alpha1, 3 FucT-2HR, GDP-Fuc and Tris-HCl containing Mg < 2 + > are taken and mixed, and incubation is performed; and purifying Protein A to obtain a target product. According to the modified antibody obtained by the invention, while the targeting property is improved, the binding affinity of the antibody and a tumor cell surface antigen can be enhanced, so that the immune blocking activity of the antibody is further enhanced, and dual improvement of the targeting property and the activity is realized.

The present invention relates to the technical field of functional enzymes, and in particular to an α-L-fucosidase mutant and an application thereof. The amino acid sequence of the mutant has at least 80% homology to the amino acid sequence of fucosidase SEQ ID NO: 1; the mutant has at least one characteristic sequence of the fucosidase, and the amino acid sequence of the fucosidase is modified, substituted, deleted or added with one or more amino acids to obtain the amino acid sequence of the mutant; the mutant has an amino acid sequence encoded by a nucleotide sequence shown in SEQ ID NO: 2 or its complementary sequence, or a sequence that is different from the nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 2 or its complementary sequence due to the degeneracy of the genetic code. The mutant provided by the present invention is beneficial to reducing the cost and reaction time in the catalytic reaction of fucosylated compound synthesis.

This invention belongs to the field of plant protection and discloses the application of α-L-fucosidase MoFco1 in screening drugs for the control of rice blast, as well as the application of compound 0989 in the control of rice blast. This invention identifies α-L-fucosidase MoFco1, which is active against XXFG in rice blast fungus. Located in the apoplast space, it regulates the pathogenicity of the fungus by degrading hemicellulose oligosaccharides in rice cell walls, inhibiting host ROS accumulation, and regulating defense responses. Based on virtual screening of the MoFco1 structure predicted by AlphaFold3, compound 0989 was obtained. This compound specifically binds to MoFco1 and effectively inhibits the pathogenicity of rice blast fungus. This invention provides new targets and new agents for the green control of rice blast and has significant application value.

The present application relates to a kind of extraction kit of cartilage tissue single cell and its extraction method, the kit includes: (1) first enzymolysis enzyme, the first enzymolysis enzyme includes one or more of trypsin, chondroitinase ABC and fucosidase;(2) second enzymolysis enzyme, the second enzymolysis enzyme includes collagenase II, neutral protease II, hyaluronidase and Dnase I;(3) base liquid, the base liquid includes base medium and / or buffer solution.The present application also provides the method for extracting single cell from cartilage tissue using the kit.The cartilage tissue single cell suspension prepared by the present application has the advantages of high cell yield and high activity, and has wide application prospect.

A method for preparing fucose and intermediates thereof and applications thereof. In the method for synthesizing GDP-fucose, GDP-fucose is generated by catalytic reaction of sucrose synthase, CDP-threose 2-epimerase, GDP-mannose dehydratase and isomerase with guanosine diphosphate and sucrose as substrates. In the method for synthesizing fucosyllactose, fucosyllactose is obtained by catalytic action of Hp13FucT2 enzyme with GDP-fucose and pNP-lactose as initial substrates. In the method for preparing L-fucose by enzyme, L-fucose is generated by catalytic reaction of fucosidase with fucosyllactose as substrate. The E. coli expression system has the characteristics of high expression efficiency, large expression amount, low cost and easy operation, and the expressed fucosidase has the advantages of high purity and high yield.

The application discloses an alpha-L-fucosidase detection kit and relates to the field of kits.The alpha-L-fucosidase detection kit comprises a box body, a box cover is arranged on the top of the box body, a placing cavity is formed in the inside of the box body, a placing seat is fixedly arranged in the inside of the placing cavity, a moving seat is arranged above the placing seat, a plurality of reagent tubes are inserted between the moving seat and the placing seat, and a lifting assembly is arranged at the bottom of the moving seat.When a plurality of reagent tubes need to be used for experiment, the end of the inserting rod is separated from the upper inserting hole by pulling the inserting rod outward, the moving seat can be pulled upward at the moment, a plurality of reagent tubes can be moved upward by the moving strip at the moment, the identification on the plurality of reagent tubes is convenient to check, and the internal reagents in the plurality of reagent tubes can be detected.

The invention discloses an alpha-L-fucosidase mutant and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. According to the alpha-L-fucosidase mutant and the application thereof disclosed by the invention, based on sequence analysis, a geobacterium alpha-L-fucosidase female parent gene is subjected to rational design, primers are designed, and ten mutants are obtained by adopting a site-directed mutagenesis PCR (Polymerase Chain Reaction) technology; the method comprises the following steps of: performing random mutation on a female parent gene by using error-prone PCR (Polymerase Chain Reaction), constructing a random mutation library, performing high-throughput screening on the mutation library, screening six mutation sites capable of improving the heat resistance of alpha-L-fucosidase through two schemes, and integrating the six mutation sites onto a coding sequence through a complete sequence synthesis method, thereby obtaining the alpha-L-fucosidase. And obtaining the combined mutant. The thermal stability of the combined mutant is obviously improved compared with that of a female parent, and the combined mutant shows potential application value in industries such as biological energy, feed, health food and medicine.

The present invention discloses a method based on solid-phase fucosylated glycoprotein enrichment and fucosylation enzymatic digestion analysis, comprising the steps of: extracting proteins, then hydrolyzing the proteins to obtain polypeptides and glycopeptides, and then treating with endoglycosidase, including: ① treating the peptide segments with endoglycosidase, then oxidizing and binding the glycopeptides to hydrazide resin, and obtaining fucosylation sites or defucosylated glycopeptides after hydrolysis by fucosidase; ② treating the glycopeptides with endoglycosidase, and then obtaining intact fucosylated glycopeptides using hydrophilic interaction chromatography. The glycoprotein or enzymatically digested glycopeptides can also be enzymatically digested to obtain branched α1,3-fucosylated glycopeptides after oxidation and binding to hydrazide. Finally, mass spectrometry is used to analyze the fucosylated peptides and the peptides containing fucosylation sites. This method is of great significance for the discovery of fucosylated disease markers in diseased cells, clinical tissues and human body fluid specimens, as well as the research on early disease diagnosis and prognosis markers, etc.

The invention discloses acid-resistant alpha-L-fucosidase IDSFuc29-2 and application thereof, and belongs to the field of protease, the amino acid sequence of the alpha-L-fucosidase IDSFuc29-2 is as shown in SEQ ID No.2, the alpha-L-fucosidase IDSFuc29-2 is derived from a metagenome of bottom mud of the East China Sea, and recombinase with definite catalytic activity can be obtained through a prokaryotic expression system. The enzyme shows good reaction characteristics under the acidic condition, the optimum reaction pH of the enzyme is about 4.0, high relative enzyme activity can still be kept within the pH range of 4.0-6.0, and meanwhile, the enzyme has acceptable stability under the condition of 20-40 DEG C. The application conditions of the GH29 family enzyme in degradation and related application of fucoidin are expanded, and the GH29 family enzyme has important research significance and application value.

The present invention relates to the field of biological preparation technology, and more particularly to a kind of α-L-fucosidase detection reagent or kit and its preparation method. Which include: substrate, buffer solution with pH 4.5~5.5, metal ion chelator, inorganic compound containing metal ion, glycerol, Gemini surfactant, Tween and PEG. The present invention provides an α-L-fucosidase single reagent that improves substrate stability and resists the influence of multiple interfering substances. The single reagent can be stably stored for at least 20 months at 2-8 ° C, effectively removes the interference of multiple serum interfering substances such as bound bilirubin, free bilirubin, vitamin C, chyle and hemoglobin, and has high accuracy. Moreover, the single reagent of the present invention is convenient, simple and easy to operate, reduces the use of instrument reagent positions, and reduces detection costs.