32 results about "Enzyme action" patented technology

To provide a glycan preparation method for preparing labeled glycans quickly from glycoproteins.SOLUTION: A glycan preparation method comprises: a releasing step in which a glycosidase is allowed to act on a glycoprotein immobilized on a solid phase in a container to obtain a released product containing glycans; and a labeling step in which a labeling reagent is added to the released product in the container to obtain a labeled product containing labeled glycans, wherein the glycoprotein is an antibody, the solid phase has on its surface a ligand selected from the group consisting of protein A, protein G, protein L, protein H, protein D, and protein Arp, and the labeling reagent comprises a reducer, a solution and at least one selected from the group consisting of 8-aminopyrene-1,3,6-trisulfonic acid, a sodium salt thereof, and 9-aminopyrene-1,4,6-trisulfonic acid.SELECTED DRAWING: None

The invention relates to a circular DNA probe for recognizing vibrio vulnificus and a preparation method of the circular DNA probe, specifically, a linear probe carrying DNAzyme is mixed with a looping primer, the mixture is connected under the action of T4DNA ligase to form the circular DNA probe, the circular DNA probe can be specifically recognized with a target and then is subjected to rolling circle amplification to form long-chain DNA, the long-chain DNA contains a large amount of DNAzyme formed by amplification and repeated GC-rich regions, and the long-chain DNA can be used for detecting the vibrio vulnificus. Intramolecular or intermolecular hydrogen bonds are spontaneously formed and are self-assembled to form the DNA hydrogel in a porous state, so that the structure promotes region limitation, and the detection sensitivity is greatly improved; target concentration gradient verification shows that the linear relation of the kit is good, R2 is equal to 0.9915, clinical sample verification shows that the specificity of the kit is high, and the kit can be used for visual and portable detection of vibrio vulnificus.

The present invention provides a novel means for degrading PEF. Provided is a method for degrading polyethylene furanoate that includes causing an enzyme comprising a polypeptide which has an amino acid sequence having at least 90% identity with the amino acid sequence of SEQ ID NO: 1 and has one or more amino acid substitutions selected from the group consisting of H129W, F208S, A172C, and A209C to act on polyethylene furanoate.

[Problems] To provide plant-base milk with reduced powdery feel and altered taste.[Solution] The present technique provides an enzyme agent for reducing the powdery feel of a plant-base beverage or food or a plant-base beverage or food material, which contains a cell wall polysaccharide degrading enzyme, and a plant-base beverage or food or a plant-base beverage or food material using the enzyme agent for reducing powdery feel. The present technique also provides a method for producing a plant-base beverage or food or a plant-base beverage or food material, and a method for reducing the powdery feel of a plant-base beverage or food or a plant-base beverage or food material, which include a step of allowing a cell wall polysaccharide degrading enzyme to act on a plant-base raw material.

The invention discloses a preparation method of a proteoglycan protein conjugate carrying a specified connection site. The method comprises the following steps: carrying out recombinant expression on a recombinant tetanus toxin heavy chain C fragment rTTHc containing a Sortase A 7M ligase recognition sequence, introducing O, O '-(ethane-1, 2-diyl) bis (hydroxylamine) as a nucleophilic reagent into the C tail end of the rTTHc under the action of ligase Sortase A 7M, and carrying out specific condensation reaction on the C tail end of the rTTHc as a unique chemical reaction site and polysaccharide carrying an aldehyde group functional group to obtain the tetanus toxin heavy chain C fragment rTTHc. Therefore, polysaccharide protein conjugates with definite polysaccharide and protein connection sites are produced. According to the invention, the problem that the key antigen epitope of the carrier protein and the heterogeneity of the product are possibly damaged by the traditional random modification strategy is solved, the antigen epitope of the carrier protein is prevented from being damaged to a great extent, and a reliable technical scheme is provided for preparing the polysaccharide conjugate vaccine with high quality and high immunogenicity.

The present invention relates to a single base editor using adenosine deaminase acting on RNA (ADAR enzyme) and a single base editing method using same. Specifically, the present invention relates to a single base editor using an ADAR enzyme and a single base editing method using same, wherein the single base editor solves the problem of not only a target single base but also all single bases around the target single base being replaced, which is the biggest limitation of conventional base editing tools, and can selectively replace just a single base.

To provide a method for efficiently and accurately distinguishing two similar diseases that differ in the presence or absence of a mutant allele in the same gene region.SOLUTION: A method for distinguishing between a genetic disease caused by a codon mutation in a gene and another disease includes the steps of: performing an amplification reaction to amplify a genetic region containing the codon, the amplification reaction including adding a DNA primer for amplifying the genetic region containing the codon and a PNA that inhibits the amplification of a normal allele containing the codon; and treating a product obtained by the amplification reaction with a restriction enzyme that specifically recognizes and cleaves either a base sequence containing the codon with the mutation or a base sequence containing the codon without the mutation, thereby obtaining a restriction enzyme-treated product.SELECTED DRAWING: Figure 3

This invention relates to the field of biodetection technology, specifically to a non-isolated digital multiplex detection method based on a DNA walker and TdT enzyme amplification. The method first utilizes coding microspheres with capture elements coupled to their surface to specifically bind to target miRNAs or proteins in the sample; then, specific guide DNA (gDNA) corresponding to the target molecule is released through enzymatic digestion or thermal denaturation; the gDNA then... Pf Under the action of Ago endonuclease, the DNA walker on the surface of the encoded microspheres is activated, cyclically cutting the orbital strand to generate multiple DNA fragments with 3'-hydroxyl ends. Finally, TdT enzyme is used to extend fluorescently labeled nucleotides at the hydroxyl ends, achieving in-situ signal amplification, and digital quantitative analysis is performed on the fluorescence signal of the encoded microspheres. This invention achieves high-sensitivity detection of multiple targets in a single reaction without chamber isolation, with detection limits for each target molecule reaching the fM or fg / mL level, providing an effective technical means for the early screening of major diseases such as Alzheimer's.

The invention belongs to the technical field of bioactive protein peptides / protein hydrolysates, and particularly relates to an anti-aging soybean protein peptide and a preparation method thereof. Soybean protein is used as a raw material, the soybean protein is firstly subjected to thermal denaturation treatment to change the secondary structure of the soybean protein, and then the alkaline subunit amount of the thermal denaturation soybean protein is increased by utilizing high static pressure physical treatment, so that the exposure of hydrophobic groups in soybean protein molecules and the increase of enzyme action sites are facilitated, and the proteolysis rate is further increased. And finally, carrying out enzymolysis on the treated soybean protein by using alkaline protease to obtain the soybean protein peptide. The soybean protein peptide obtained by the preparation method disclosed by the invention has very strong antioxidant activity and a good anti-aging effect.

Devices and methods that can detect and control an individual polymer in a mixture is acted upon by another compound, for example, an enzyme, in a nanopore are provided. The devices and methods also determine (˜>50 Hz) the nucleotide base sequence of a polynucleotide under feedback control or using signals generated by the interactions between the polynucleotide and the nanopore. The invention is of particular use in the fields of molecular biology, structural biology, cell biology, molecular switches, molecular circuits, and molecular computational devices, and the manufacture thereof.

To provide a technique for reducing a saccharide in a plant-base food or beverage.The technique provides an enzymatic agent for reducing a saccharide in a plant-base food or beverage, which contains a cyclodextrin-producing enzyme, and a plant-base food or beverage using the enzymatic agent for reducing a saccharide. The technique also provides a method for manufacturing a plant-base food or beverage, which includes a step of allowing a cyclodextrin-producing enzyme to act in a liquefaction process of a plant-base raw material, and a method for reducing a saccharide in a plant-base food or beverage, which includes a step of allowing a cyclodextrin-producing enzyme to act on a plant-base raw material.

A simpler method for measuring enzyme activity is provided. The-method for estimating enzyme activity uses a base on which spots of immobilized substrates are formed, and includes (a) enabling an enzyme to act on the substrate on the spot to obtain an enzyme product; (b) setting one or more compartments on the spot and obtaining density of the substrate in each compartment; (c) measuring the density of the enzyme product in the each compartment; and (d) estimating the enzyme activity of the enzyme based on a relationship between the density of the substrate in the each compartment obtained by step (b) and the density of the enzyme product in the each compartment obtained by step (c).

This invention discloses a method for synthesizing chiral γ-acyloxybutenoic acid lactones, characterized by the following steps: (1) dissolving the substrate 2-furoic acid in an organic solvent and carrying out a photocatalytic oxidation reaction under the action of a photocatalyst and under light and oxygen conditions; (2) after the substrate in step (1) is completely converted, adding an acyl donor, and carrying out an enzymatic acylation reaction in the resulting reaction solution under the action of lipase to obtain γ-acyloxybutenoic acid lactones. This invention solves the problem of solvent compatibility between photocatalysis and enzyme catalysis systems, supporting both efficient photocatalytic oxidation and excellent medium for enzyme catalytic acylation. It realizes a one-pot conversion from the substrate 2-furoic acid to the product chiral γ-acyloxybutenoic acid lactones, with advantages such as mild reaction conditions, environmental friendliness, high synthesis efficiency, excellent selectivity, easy scale-up, and no need for separation and purification of intermediate products.

The invention relates to the technical field of extract refining, and particularly discloses a method for preparing stevioside RM. The method comprises the following steps: (1) crushing dry stevia rebaudiana leaves, mixing with water, extracting, stirring, extracting, centrifugally filtering, performing column chromatography purification, washing with water, eluting with ethanol to obtain an eluent, and concentrating the eluent to obtain a stevioside solution; (2) adding an auxiliary stable mixed aid, cane sugar, cane sugar synthase, beta-1, 2-UDP-glucosyltransferase from dendrobenthamia grandiflora and beta-1, 3-UDP-glucosyltransferase from walnut into the stevioside solution obtained in the step (1), carrying out decoloration and desalination treatment after catalytic reaction, eluting with an ethanol-water solution, and carrying out vacuum drying to obtain the stevioside. And finally, carrying out vacuum concentration, recrystallization and drying treatment to obtain the product. According to the method, the stability of an enzyme action system in the stevioside RM preparation process can be remarkably improved.

To provide a technique for improving the shape-retainability of a plant-base ice cream.The technique provides an enzyme preparation for improving the shape-retainability of a plant-base ice cream, which contains one or more enzymes selected from the group consisting of maltotriose-producing amylase, lipase, and β-amylase, and a plant-base ice cream using the enzyme preparation for improving the shape-retainability. The technique also provides a method for producing a plant-base ice cream and a method for improving the shape-retainability of a plant-base ice cream, which include one or more steps selected from a maltotriose-producing amylase action step of allowing maltotriose-producing amylase to act on a carbohydrate-containing plant-base material, a lipase action step of allowing lipase to act on a fat-containing plant-base material, and a β-amylase action step of allowing β-amylase to act.

The invention provides a synthesis method of a single-chain aptamer and application of the single-chain aptamer in synthesis of a C5 complement inhibitor, and relates to the technical field of aptamer synthesis, the synthesis method comprises the following steps: cutting off the single-chain aptamer into two fragments, pairing the two fragments through complementary bases carried by the two fragments to form a double-chain secondary structure with a single gap, and under the enzymatic action, synthesizing the C5 complement inhibitor by using the double-chain secondary structure to obtain the C5 complement inhibitor. A phosphodiester bond is formed at a notch of the double-chain secondary structure to obtain a single-chain aptamer, and the base length ratio of the two fragments is (14-26): (25-13). The self-complementary nucleic acid fragment of the to-be-synthesized single-stranded aptamer is used as a substrate, specific connection is realized under the action of ligase, an exogenous template chain is not needed, and the final purity and yield of the single-stranded aptamer are improved. The technical problems that in the prior art, enzymatic connection of single-stranded oligonucleotides needs to depend on an exogenous template, so that an experimental system is complex, mismatching is likely to happen, and connection position errors are likely to happen are solved.

Provided is a novel technique for improving the freezing resistance of a starch-containing food. Provided is a method for producing a starch-containing composition for freezing, the method comprising an enzyme action step for causing one or more enzymes selected from the group consisting of a microorganism-derived beta-amylase, a soybean-derived beta-amylase, and a 4-alpha-glucanotransferase to act on a starch-containing material. The invention also provides a preparation method of the starch-containing composition, which comprises a freezing process, and a freezing step for freezing an enzyme-treated starch-containing composition obtained by causing one or more enzymes selected from the group consisting of a microorganism-derived beta-amylase, a soybean-derived beta-amylase, and a 4-alpha-glucanotransferase to act on the starch-containing material.

The application discloses a synthesis method and application of a fluorescent probe for lipase detection, and the fluorescent probe has the following structural formula; the synthesis method comprises the following steps: under the action of inert gas protection, a catalyst and alkali, triphenylol compound and a fatty chain or an aromatic ring organic acid are reacted in an organic solvent at 25-100 DEG C for 8-24h; after the reaction is stopped, dichloromethane and water are added to carry out extraction; organic phases are combined, dried, filtered, and the solvent is removed under reduced pressure; and the product is separated and purified; the fluorescent probe is basically non-fluorescent by itself, releases bright fluorescence after reacting with lipase, and can realize rapid detection of bacteria by using the characteristics that most bacteria secrete lipase in the growth and reproduction process.

The invention discloses a glycosyl transferase gene for biosynthesis of polygonatum cyrtonema steroid saponin and application of the glycosyl transferase gene. The glycosyl transferase gene directly related to steroid saponin synthesis in the UGT85A subfamily is cloned and functionally verified from polygonatum cyrtonema for the first time, and gene resources of steroid saponin biosynthesis are enriched; the activity of the PcUGT85As gene for catalyzing the glycosylation of the sitenogenin is defined, and the key enzyme action of the PcUGT85As gene is proved; a key target spot is provided for realizing metabolic regulation and synthetic biological production of the polygonatum cyrtonema active saponin; the gene can be applied to microbial cell factory construction, transgenic plant breeding or biological manufacturing of high-yield saponin, and has a good application prospect and industrial value.

The application relates to the field of nano drug loading technology, and discloses application of beta-glucan in preparation of liver-targeted drugs for treating liver cancer and the liver-targeted drugs, wherein the beta-glucan is derived from black fungus, and a chemical structural formula of the beta-glucan is that two beta-(1, 6)-glucose residues are arranged on the side chain of every three beta-(1, 3)-glucose main chains. The application provides a new use of black fungus beta-glucan in preparation of liver-targeted drugs for treating liver cancer. The black fungus beta-glucan has the abilities of liver targeting, enzymatic release and activation of macrophages, so that the black fungus beta-glucan can be used as a carrier of the liver-targeted drugs, the drugs can be stably delivered to the liver, and the release of the drugs is realized under the action of liver enzymes in the liver, so that the liver cancer (hepatocellular carcinoma and intrahepatic cholangiocarcinoma, etc.) can be effectively treated, the toxic side effect is small, and the drug carrier can have a synergistic effect with the drugs loaded thereon, so that the intrahepatic cholangiocarcinoma can be better treated.

This invention relates to the field of enzymatic hydrolysis technology, specifically to a method and application for improving the degree of hydrolysis of dehulled soybean meal. The method includes: steaming the dehulled soybean meal; mixing a first mixed enzyme with the reactants after the steaming treatment and performing a first enzymatic hydrolysis reaction; mixing a second mixed enzyme with the reactants after the first enzymatic hydrolysis treatment and performing a second enzymatic hydrolysis reaction and enzyme inactivation treatment. This method improves the degree of hydrolysis of the dry matter in dehulled soybean meal by optimizing the enzymatic hydrolysis process. The physical treatment of the dehulled soybean meal raw material by steaming alters the properties of the raw material, making it more conducive to enzyme action. A two-stage enzymatic hydrolysis method is adopted, and considering the physicochemical properties of the enzyme preparation, the pH and temperature of the hydrolysate are adjusted. The first stage degrades a small amount of cellulose and protein in the dehulled soybean meal; the second stage increases the solubility of protein while reducing the fat content, converting the remaining substances into small-molecule soluble substances, which are more easily digested and absorbed by animals.

The invention provides an exponential rolling circle amplification method based on Argonaute protein. The exponential rolling circle amplification method comprises the following steps: providing a 5'end phosphorylated single-chain gDNA; the method comprises the following steps: carrying out hybridization annealing on target RNA (Ribonucleic Acid) and a Padlock probe to form an RNA-Padlock compound; under the action of ligase, cyclizing the RNA-Padlock compound to form a circular DNA (Deoxyribose Nucleic Acid) template; the gDNA and TtAgo are subjected to pre-incubation, and a TtAgo / gDNA compound is formed; the method comprises the following steps: mixing a circular DNA template, a TtAgo / gDNA compound, DNA polymerase, dNTPs and a fluorescent dye in a same reaction system, and carrying out an exponential rolling circle amplification reaction. According to the exponential rolling circle amplification method based on the Argonaute protein, accurate shearing can be achieved at any preset position of an RCA product, a specific enzyme cutting site does not need to be introduced, system construction is greatly simplified, and exponential amplification is achieved.

Devices and methods that can detect and control an individual polymer in a mixture is acted upon by another compound, for example, an enzyme, in a nanopore are provided. The devices and methods also determine (˜>50 Hz) the nucleotide base sequence of a polynucleotide under feedback control or using signals generated by the interactions between the polynucleotide and the nanopore. The invention is of particular use in the fields of molecular biology, structural biology, cell biology, molecular switches, molecular circuits, and molecular computational devices, and the manufacture thereof.

The main purpose of the present invention is to provide a novel technique for producing a beta-glucan-containing vegetable liquid composition in a simple and low-cost manner. In the present technology, provided is an enzyme preparation for producing a plant liquid composition containing a beta-glucan, said enzyme preparation containing an alpha-amylase and a beta-glucanase. Also provided in the present technology is an agent for increasing the beta-glucan content of a vegetable liquid composition, said agent containing an alpha-amylase and a beta-glucanase. The present technology further provides a method for producing a beta-glucan-containing vegetable liquid composition, the method including an enzyme action step for causing an alpha-amylase and a beta-glucanase to act on a starch-containing vegetable material.

This invention provides a method for defluorinating fluoroalkane compounds containing hydrophilic groups. [Solution] An enzyme and microorganism that defluorinate hydrophilic group-containing fluoroalkane compounds have been isolated. A method for defluorinating fluoroalkane compounds is disclosed, comprising acting an enzyme that defluorinates hydrophilic group-containing fluoroalkane compounds onto a fluoroalkane compound. The enzyme that defluorinates hydrophilic group-containing fluoroalkane compounds is an enzyme containing a specific amino acid sequence. This enzyme was isolated from an environmental microorganism (a strain of Pandoraea pnomenusa) and has the characteristic of defluorinating not only the α-position but also the β-position of hydrophilic group-containing fluoroalkane compounds.

The invention discloses a method for accurately determining the activity of swine-derived co-lipase, which comprises the following steps: emulsifying tributyrin by using gum arabic powder and glycerol together, and performing enzymolysis on the tributyrin into free fatty acid according to the coenzyme action of co-lipase and the combined action of co-lipase and lipase, thereby obtaining the activity of the swine-derived co-lipase. Meanwhile, the blank that the tributyrin is independently enzymolyzed by the same amount of lipase is taken, finally, the activity of the co-lipase is calculated according to the calculation formula by utilizing the volume difference value of the sodium hydroxide solutions consumed for generating the free fatty acid under the two conditions, and the method for designing the activity of the pig-derived co-lipase is stable and accurate in detection result.

Provided is a technique for improving the foamability of a vegetable food or beverage. The present technology provides a method for improving the foamability of a vegetable food or beverage, the method comprising: a cyclodextrin-producing enzyme action step for causing a cyclodextrin-producing enzyme to act on a composition containing a vegetable raw material; and / or a cyclodextrin addition step in which cyclodextrin is added to a composition containing a plant starting material. In the present technology, provided is a method for improving the foam quality of a foamable vegetable food or beverage, said method comprising a cyclodextrin-producing enzyme action step for causing a cyclodextrin-producing enzyme to act on a composition containing a vegetable raw material. In addition, the present technology provides a foamability-improving agent for a vegetable food or beverage, said foamability-improving agent containing a cyclodextrin-generating enzyme and / or a cyclodextrin. In addition, the present technology provides a foamability-improving agent for foamability vegetable foods and drinks, the foamability-improving agent containing a cyclodextrin-generating enzyme. In addition, the present technology provides a vegetable food or beverage using the foamability-improving agent and / or the foam quality-improving agent according to the present technology.

The application provides a high-gel-quality surimi gel and a preparation method thereof. After frozen white silver carp surimi is thawed, ice temperature vacuum dehydration and ultrahigh pressure homogenization treatment are carried out, the myofibrillar protein molecular chain is stretched by mild shearing force, and the TG enzyme action site is fully exposed; then, the surimi is chopped and mixed with salt, starch, TG enzyme porous starch-pectin oligosaccharide nanosphere dispersion and ice water at low temperature, wherein the nanosphere shields the TG enzyme at low temperature to avoid early activation, and is accurately released to catalyze cross-linking when heated; finally, the surimi is gelled by step heating, the strength and uniformity of the surimi gel are significantly improved, the flavor is improved, and the surimi gel is suitable for industrial production.

The invention belongs to the technical field of biology, and relates to a synthesis method of a fluorescent probe for alpha-amylase activity detection. The invention provides a synthesis method of a fluorescent probe for detecting activity of alpha-amylase. The synthesis method of the fluorescent probe comprises two steps of preparing a compound as shown in a formula (A1) and preparing a compound as shown in a formula (1). The invention also provides a fluorescence detection method of alpha-amylase activity, the method innovatively utilizes alpha-amylase to act on alpha-1, 4-glucosidic bond to hydrolyze gamma-cyclodextrin, so that the fluorescence probe wrapped by gamma-cyclodextrin is released to change fluorescence intensity, which is physical and chemical correlation, and the fluorescence detection method is used for detecting the activity of alpha-amylase. The change of optical signals caused by the change of fluorescence intensity is measured by adopting a method for testing the fluorescence of a sample, so that the activity value of the alpha-amylase is quantitatively given. According to the method, on the basis of ensuring that the accuracy is equivalent to that of the prior art, the test result can be given at a higher speed and a lower cost.

Provided is a technique for improving the foamability of a vegetable food or beverage. The present technology provides a method for improving the foamability of a vegetable food or beverage, the method comprising: a cyclodextrin-producing enzyme action step for causing a cyclodextrin-producing enzyme to act on a vegetable raw material; and a protein deamidase action step for causing a protein deamidase to act on the plant raw material.