46 results about "Fucosidase" patented technology

The invention relates to a serum AFU detection kit. The kit comprises a reagent 1 and a reagent 2, wherein the reagent 1 comprises 50-500 mmol / L of a buffer solution (pH 4.0-4.5), 1-50 g / L of a surfactant, 1-20 g / L of an anti-interference agent and 0.1-100 g / L of a preservative; the reagent 2 comprises 50-500 mmol / L of a buffer solution (pH 7.0-7.5), 2-60 g / L of glucose, 1-20 KU / L of hexokinase, 1-20 KU / L of glucose-6-phosphate dehydrogenase, 0.3-5 g / L of magnesium sulfate, 0.2-5 g / L of a substrate, 1-100 mmol / L of a stabilizer, 0.1-40 g / L of a protective agent and 0.1-100 g / L of a preservative. With the adoption of the kit, problems of the poor substrate stability, the poor anti-interference capacity and the lower sensitivity which exist universally are solved.

The present invention relates to serum alpha-L-fucosidase activity measuring process and reagent, and the measuring process and reagent has short measuring period, high noise immunity and simple operation, and is suitable for various kinds of automatic biochemical analyzers. Of the technological scheme, the alpha-L-fucosidase activity measuring process includes adopting 6-methyl-2-thiopyridine-alpha-L-fucoside as substrate capable of producing 6-methyl-2-sulfhydyl pyridine under the action of alpha-L-fucosidase in the sample, and measuring the absorbance increasing rate of 6-methyl-2-sulfhydyl pyridine at 340 nm to find out the activity of alpha-L-fucosidase activity in the sample; and the alpha-L-fucosidase diagnosing reagent includes 6-methyl-2-sulfhydyl pyridine.

The invention discloses a liquid single reagent for detecting alpha-L-fucosidase and a preparation method thereof. The liquid single reagent contains a buffer solution, ethylenediamine tetraacetic acid, NaCl, a surface active agent, a stabilizer and a P-nitrobenzene chloride alpha-L-fucus pyranoside. The stabilizer contained in the liquid single reagent disclosed by the invention can not only effectively enhance the stability of the liquid single reagent, but also effectively eliminate the interference of sample heparin by being cooperated with the surface active agent, therefore detection accuracy and sensitivity are enhanced. A stability test result indicates that after the liquid single reagent for detecting the alpha-L-fucosidase is respectively preserved for 3 days at 42 DEG C, preserved for 14 days at 37 DEG C and preserved for 18 months at 2-8 DEG C, various properties are kept stable so as to indicate that the liquid single reagent disclosed by the invention has good stability and can resist the interference of at least 100 KU / L heparin contained in a sample.

The invention belongs to the technical field of fertilizer, and especially relates to a fertilizer containing a natural plant growth substance and a preparation technique thereof; the preparation technique of the fertilizer containing natural plant growth substance comprises S1, performing low-temperature grading leaching on smashed kelps, combining and concentrating leach liquor; S2, adding water to the leached kelps, and performing graded enzymolysis orderly by cellulase, pectinase and fucosidase to obtain enzymatic hydrolysate; S3, adjusting the pH value of enzymatic hydrolysate, combining and concentrating with leached concentrated solution to obtain the fertilizer finally. The fertilizer containing the natural plant growth substance contains rich biological active substance, and can effectively promote the development and growth of crops; meanwhile, the preparation technique of the fertilizer containing the natural plant growth substance is simple and feasible, and can effectively extract and remain the ecological active substances in kelp, the production cost is low, the production technique is easy to control, and good for large-scaled industrialized production.

Methods of diagnosing Helicobacter pylori infection or associated conditions are based in part on the correlation of the presence of a α-L-fucosidase 2 marker with the infection. Methods and compositions for treating or preventing Helicobacter pylori infection or associated conditions are based in part on administering an α-L-fucosidase 2 inhibitor to an infected subject or a subject at risk of developing the infection.

The invention discloses an alpha-L-fucosidase detection kit consists of an independent reagent 1 and an independent reagent 2, wherein the reagent 1 consists of components of a buffer agent, a preservative, a metal ion chelating agent, a stabilizing agent and water; and the reagent 2 consists of components of a buffer agent, a preservative, an enzyme action substrate, a surfactant, a stabilizing agent and water. The preparation method comprises the following steps: (1) preparing the reagent 1: dissolving the various components in distilled water or double distilled water, uniformly mixing, regulating pH to 6.5-8.5, and setting a constant volume; (2) preparing the reagent 2: dissolving various components in distilled water or double distilled water, uniformly mixing, regulating pH to 6.5-8.5, and setting a constant volume; and (3) independently sub-packaging the reagent 1 and the reagent 2, and sealing. The detection kit disclosed by the invention has the advantages of good stability, high precision, broad linear test range, good repeatability, strong anti-interference performance and the like.

The invention relates to an integrated detection reaction orifice plate and protein chip reagent box for detecting six indexes of diagnosis, forecast and prognosis of hepatitis, hepatocirrhosis and liver cancer, including hepatitis B virus surface antigen (HBsAg), hepatitis C virus antibody (HCVAb), alpha-fetoprotein (AFP), alpha-L-Fucosidase (AFU), mono amine oxidase (MAO), hyaluronic acid (HA). And the orifice plate comprises a substrate and reaction orifices on the substrate, where the reaction orifices comprise 2-384 sample orifices and 2-8 standard product orifices, and at the bottom of each reaction orifice is solid carrier coated with micro lattice of HBsAg, HCVAg, AFP, AFU,MAO,and HA antigens / antibodies or more. And the reaction plate and reagent can simply and conveniently, quickly and accurately implement simultaneous detection of the six indexes of diagnosis, forecast and prognosis of hepatitis, hepatocirrhosis and liver cancer for many persons.

The invention relates to a single chain antibody of a human liver cancer marker, a method for preparing the antibody, a detection reagent and a kit comprising the antibody, and an application thereof. More specifically, the invention relates to the single chain antibody which is specifically combined to the human liver cancer marker, and the human liver cancer marker is selected from the following components: alpha fetoprotein (AFP), glutamyltransferase isoenzyme II (GGTII or GGT2), alpha-L-fucosidase (AFU), hepatocyte growth factor (HGF) and heparin sulfate proteoglycan 3 (GPC3).

The invention discloses alpha-L-fucosidase and a related biological material and application thereof. The invention discloses a protein which is A1) a protein of which the amino acid sequence is shownin the formula SEQ ID No.4, A2) a protein of which the amino acid sequence is shown in the formula of SEQ ID No.3, A3) a fusion protein obtained by connecting the N end or / and C end of the protein ofA1) or A2) with a protein label, and A4) a protein which is obtained by substituting and / or deleting and / or adding one or more amino acid residues of the protein shown in the formula of SEQ ID No.3 or SEQ ID No.4, has 90% or more of identity with the protein shown in A1) or A2) and has the same function as the protein shown in A1) or A2). The invention further discloses a related biological material of the protein and an application of the related biological material. The protein provided by the invention can be used for efficiently synthesizing 3'-fucosyllactose and has a good application prospect in oligosaccharide synthesis.

The invention belongs to the technical field of the environmental protection, and particularly relates to an environment-friendly fertilizer containing a chlorella functional component and a preparation technology thereof. The preparation technology for the environment-friendly fertilizer containing the chlorella functional component comprises the following steps: S1 taking a chlorella raw material, adding saccharose and yeast powder, fermenting, to obtain the chlorella raw material with high content; S2 enabling the chlorella raw material with the high content to be mixed with a solvent, to obtain chlorella diluent, adding an activating agent, performing rehydration, to obtain rehydration chlorella diluent; S3 adding a protective agent to the rehydration chlorella diluent, homogenizing, to obtain wall-breaking chlorella solution; S4 successively adding cellulase, pectinase, alkaline protease and fucosidase to the wall-breaking chlorella solution, performing the classification enzymolysis, to obtain original chlorella solution; and S5 vacuum-concentrating the original chlorella solution, to obtain the fertilizer. The environment-friendly fertilizer containing the chlorella functional component is abundant in nutrition, and capable of effectively extracting and keeping bioactive substances in the chlorella, low in production cost, easy to control in production process, and beneficial to large-scale industrialized production.

The present invention provides for novel fucosidase mutants that server as fuco-ligases for core fucosylation of a range of biological glycopeptides and glycoproteins including intact therapeutic antibodies. Several mutants with mutation at the general acid / base residue E274 of the Lactobacillus casei α1,6-fucosidase, including E274A, E274S, and E274G, were able to efficiently fucosylate a wide variety of complex N-glycopeptides and intact glycoproteins. The site specific mutants enable the transfer of fucose to a core GlcNAc-Asn residue and useful for drug delivery and vaccine development.

The invention belongs to the field of sugar biology technology and engineering, and particularly relates to a novel fucosidase of core fucosidase (Cfus) as well as enzymatic activity and application thereof. An experiment provided by the invention proves that the Cfus can excise core alpha1,3 fucose on an N-glycoprotein sugar chain, which is obviously different from the fucosidase which has no enzyme digestion activity on the glycoprotein core alpha1,3 fucose in the prior art; and at the same time, the Cfus can hydrolyze fucose of other fucose conjugates. The invention provides a new tool enzyme for the research of sugar biology and biomedicine; the tool enzyme may become a potential tool for allergy treatment; and the fucosidase can be used for structural analysis of N-glycoprotein sugarchains and functional research of core fucosylated sugar chains.

The invention discloses a kit for detecting alpha-L-fucosidase. The kit comprises a reagent R1 and a reagent R2 which are independent, wherein the reagent R1 comprises the following components: a bological buffer 1, a metal ion complex, an alpha-L-fucosidase reaction substrate, a surfactant 1 and a preservative 1; and the reagent R2 comprises the following components: a bological buffer 2, 2-chloro-p-nitrophenol-alpha-L-fucosidase, an activating agent, a surfactant 2 and a preservative 2. The kit adopts a dual-reagent mode and has the advantages of high detection sensitivity, low detection limit, wide linear measurement range, high accuracy, good reproducibility, good stability and strong interference resistance. The kit can be used for semi-automatic, full-automatic biochemical analyzer and required detection instrument (biochemical analyzer) commonly used in major hospitals and test centers and is suitable for clinical popularized application, and the rapid diagnosis for emergency is especially achieved.

A method, and associated kit, for assaying the activity of lysosomal enzymes present in dried bodily fluids and cell tissue samples, such as α-L-iduronidase, β-D-galactosidase, β-D-glucosidase, chitotriosidase, total α-D-galactosidase and α-D-galactosidase A, hexosaminidase A and B, α-D-mannosidase, β-D-mannosidase, α-L-fucosidase, N-acetyl-α-galactosaminidase, arylsulfatases, sphingomyelinase, β-galactocerebrosidase, iduronate-2-sulfatase and β-D-glucuronidase. The method includes: (a) combining with a dried bodily fluid or cell tissue sample containing at least one type of lysosomal enzyme: (1) an eluent, (2) an incubation buffer and (3) a substrate or substrates capable of reacting with the assayed lysosomal enzymes and producing their corresponding enzyme product or products, (b) allowing the dried bodily fluid or cell tissue sample to react with the eluent, incubation buffer and substrate or substrates for an adequate time and temperature, and (c) applying measuring means to the enzyme product to determine the activities of the lysosomal enzymes present.

The invention discloses an alpha-L-fucosidase assay kit, and relates to the technical field of biological detection. The alpha-L-fucosidase assay kit comprises a reagent R1 and a reagent R2 which areindependent of each other; and the alpha-L-fucosidase assay kit specifically comprises the following ingredients in the following contents: the reagent R1 consisting of 7.5-8.5 g / L of disodium hydrogen phosphate, 5.0-6.0 g / L of citric acid and 0.2-0.8 g / L of sodium azide, and the reagent R2 consisting of 7.5-8.5 g / L of disodium hydrogen phosphate, 5.0-6.0 g / L of citric acid, 0.2-0.8 g / L of sodiumazide, and 4-8 g / L of 2-chloro-4 nitrobenzene-alpha-L-fucopyranoside. In the process of implementation of the alpha-L-fucosidase assay kit, arginine, asparagine and sucrose are added into the reagentR1 and the reagent R2 as stabilizers, so that accuracy, precision and stability of the kit are significantly improved; and thus, an alpha-L-fucosidase assay kit with high accuracy, excellent precision, strong stability and good linear relationship is provided.

The invention provides modified AFU (a-L-fucosidase) and a preparation method thereof, and an application of the modified AFU in AFU detection. In the modified AFU, 12 amino acids in the N-terminal nonfunctional amino acid chain of the AFU are cut off, the 101th-128th peptide fragments of AFU a-spiral are subjected to methylated modification, and peptide chains are crosslinked by a disulfide bond.A stable AFU detection kit prepared by the modified AFU comprises a reagent R and a calibration product, the reagent R includes a buffer solution, the modified AFU, a surfactant, a stabilizer and a corrosion remover; the calibration product includes a buffer solution, a surfactant, the AFU, a stabilizer and a corrosion remover. The reagent kit has the advantages of high stability, high flexibility and wide detection range.

Genetically engineered host animal cells capable of producing glycoproteins having modified glycosylation patterns, e.g., defucosylation and / or monoglycosylation. Such host animal cells can be engineered to express fucosidase, endoglycosidase or both.

The invention belongs to the technical fields of preparation and application of metal optical function nano materials and in particular relates to a fluorescent silver nanocluster as well as a simple preparation method and application thereof. The fluorescent silver nanocluster is synthesized by reacting soluble silver salt with a stabilizer, wherein the fluorescent silver nanocluster has the mean particle size of more than 1.5nm and less than 2nm; the fluorescence emitted by the fluorescent silver nanocluster has the wavelength of 610nm under excitation of light with the wavelength of 510nm; the emitted fluorescence has high illumination stability. The method is readily available in raw materials, simple in process, convenient to operate, low in biotoxicity and easy to popularize and apply. The synthesized fluorescent silver nanocluster can be applied to the fields of optical imaging, biological markers, biological or chemical sensors and the like, and especially can be used for qualitative and quantitative detection on the activity of alpha-L-fucosidase.

The present invention relates to compositions and methods for treating glycogen storage disease, such as GSD II. A human activator enzyme, termed ASA, of acid a a-glycosidase, (GAA) or acid maltase, a lysosomal enzyme, is specifically defined and characterized. The AGA has been found to increase the activity of human acid .alpha. glycosidase activity to at least 10-fold, relative to the activity of GAA in the absence of the activator protein. The invention thus also provides for a method of increasing the activity of GAA, particularly through the action of AGA. The AGA has an approximate molecular weight of 25-30 kD, and is found to be heat stable. In addition, the AGA is found to have an extended shelf life without significant loss of ability to activate GAA. The invention further reports other enzymes such as .beta.-Fucosidase, .beta.-Lactase, and .beta.-Galactosidase, that provide enhancement of enzymatic activity nine-fold, six-fold, and five-fold, for breakdown of their respective substrate protein. These enzymes are non-lysosomal enzymes. These are anticipated to be useful in treatment of disease related to reduced enzymatic activity levels in an animal.

The invention discloses alpha-L-fucosidase OUCJdch-16 with an amino acid sequence as shown in SEQ ID NO. 1, a gene for coding the alpha-L-fucosidase OUCJdch-16 and with a nucleotide sequence as shown in SEQ ID NO. 2, and application of the alpha-L-fucosidase OUCJdch-16 in the hydrolysis of pNp-Fuc / the preparation of the difucosyllactose. The invention further discloses a recombinant expression vector and a recombinant host containing the gene for coding the alpha-L-fucosidase OUCJdch-16. The alpha-L-fucosidase OUCJdch-16 has certain psychrophilicity, and has relatively high enzyme activity at the temperature of 25-45 DEG C, so that the alpha-L-fucosidase OUCJdch-16 can be industrially applied in a relatively wide temperature range. In addition, a liquid phase result shows that the OUCJdch-16 has a higher conversion rate under the optimal reaction condition, and has good biological catalytic synthesis efficiency.

The invention belongs to the field of glycobiology technology and engineering, and particularly relates to novel fucosidase and enzymatic activity and application thereof. The fucosidase can effectively act on polysaccharide structures located on the cell surfaces, and the fucosidase is named as cell surface fucosidase I (csFase I). It is found and proved that csFase I can effectively remove alpha-1,2 fucose on a H-antigen on the cell surfaces of active red blood cells, can effectively convert O-type red blood cells into rare Bombay phenotype red blood cells, and can provide a simple and fastnew way for solving the current situation of extremely difficult blood supply to patients with a Bombay phenotype blood type; and a new method is provided for the research of glycobiology and biomedicine.

The present invention discloses an alpha-L-fucosidase determination reagent. The alpha-L-fucosidase determination reagent comprises 80-200 mmol / L of a buffer, 5-10 g / L of sodium chloride, 0.1-1 g / L ofa color source-fucoside substrate, 0.5-2 g / L of a stabilizer, 0.1-2 g / L of an activator, 0.5-5 g / L of a non-ionic surfactant, and 2-5 g / L of a metal complexing agent. The stabilizer is added to solvethe problem of significant increase of a reagent blank caused by a hydrolysis reaction of the color source-fucoside substrate, and the GOOD'S biological buffer and the activator are used to improve stability and sensitivity.

The invention discloses a preparation method of alpha-L-fucosidase. The fucosidase has the characteristics of (a) or (b) or (c) or (d), wherein the amino acid sequence of (a) is shown as SEQ ID NO: 1,and (a) has the activity (EC 3.2.1.51) of alpha-L-fucosidase; the amino acid sequence of (b) has 70 % or more homology with (a); the nucleic acid sequence of (c) is SEQ ID NO: 2 or the encoded aminoacid sequence of (c) is the nucleic acid sequence of (a); and the amino acid sequence, coded by the nucleic acid sequence, of (d) has 70 % or more homology with (a). According to the method, an escherichia coli expression system is adopted; through optimization of expression conditions and purification conditions, the low-cost, high-purity, high-yield, high-stability and high-repeatability activealpha-L-fucosidase is obtained with the shortest production cycle; and the market application prospects are extremely wide.

The invention relates to the technical field of detection reagents, in particular to an anti-heparin stable alpha-L-fucosidase detection kit and application thereof. The kit comprises 0.03-0.1 g / L of streptomycin sulfate, 50-500 mmol / L of a buffer solution, 0.1-3 g / L of a substrate, 1-50 g / L of a surfactant, 0.001-0.1 g / L of a stabilizer and 0.3-3 g / L of a preservative; and the substrate is prepared from 2-chloro-p-nitrophenol-alpha-L-fucoside or M-G-2-chloro-p-nitrophenol-alpha-L-fucoside. Streptomycin sulfate with proper concentration is added into the alpha-L-fucosidase detection reagent, so that interference of heparin in a sample is effectively removed, and the problem that an AFU single reagent is not resistant to heparin is solved.

The invention relates to the technical fields of medicine and biochemistry, in particular to a kit for measuring α-L-fucosidase. The existing methods for detecting α-L-fucosidase have defects such as high detection cost, complicated and time-consuming operation, and only qualitative or semi-quantitative detection. The present invention provides a kit for measuring α-L-fucosidase, which can solve the above problems. The technical scheme of the present invention adopts the following kit reagent component formula: 2-hydroxypropane-1,2,3-tricarboxylic acid 30~90 mmol / L, sodium chloride 60~100 mmol / L, sodium azide 0.1~0.6 g / L, EDTA 12~26 mmol / L, 2-chloro-4-nitrobenzene-α-L-fucoside 18-72 mmol / L, the solvent is purified water. The kit provided by the invention produces specific hydrolyzed corresponding products, the reagents and products are non-toxic and harmless, the reaction is fast and sensitive, the detection is more accurate, the speed is fast, and there is no pollution; it can be directly detected and operated on various biochemical analyzers The method is simple and convenient, does not need to invest in expensive equipment, and saves detection costs.

The present invention provides for novel fucosidase mutants that server as fuco-ligases for core fucosylation of a range of biological glycopeptides and glycoproteins including intact therapeutic antibodies. Several mutants with mutation at the general acid / base residue E274 of the Lactobacillus casei α1,6-fucosidase, including E274A, E274S, and E274G, were able to efficiently fucosylate a wide variety of complex N-glycopeptides and intact glycoproteins. The site specific mutants enable the transfer of fucose to a core GlcNAc-Asn residue and useful for drug delivery and vaccine development.

The present disclosure provides a fusion protein comprising a fucosidase or a truncated fragment or a mutant thereof fuses with either N-terminal end or C-terminal end of the endoglycosidase or a truncated fragment of mutant thereof. The present disclosure also provides a nucleic acid molecule expressing the fusion protein and a method for remodeling a glycan of an antibody Fc region.

Genetically engineered host animal cells are capable of producing glycoproteins with modified glycosylation patterns, such as defucosylation and / or monoglycosylation. Such host animal cells can be engineered to express fucosidases, endoglycosidases, or both.