62 results about "Glycosidase activity" patented technology

The present invention provides a method for increasing the activity of a mutant or wild-type α-glucosidase enzyme in vitro and in vivo by contacting the enzyme with a specific pharmacological chaperone which is a derivative of 1-deoxynojirimycin. The invention also provides a method for the treatment of Pompe disease by administration of chaperone small molecule compound which is a derivative of 1-deoxynojirimycin. The 1-deoxynojirimycin derivative is substituted at the N or C1 position. Combination therapy with replacement α-glucosidase gene or enzyme is also provided.

A method is provided for determining the activity of an enzyme which releases methylumbelliferone (MU) from an MU-containing substrate wherein the enzyme has a pH optimum below the pKa of MU comprising: contacting a sample suspected of containing the enzyme with the MU-containing substrate at a pH suitable for activity of the enzyme to allow release of MU by the enzyme; contacting the sample with light of a wavelength in the range of about 310 nm to about 350 nm; determining fluorescence produced by the released MU, thereby determining the activity of the enzyme. This real time method provides improved diagnostic methods, for example for diseases associated with an abnormal level of activity of a glycosidase. The real time assay also can be used to screen compounds for their ability to modulate enzyme activity using MU-containing substrates.

The invention discloses a separation and purification method of main catechin components in tea polyphenol and glycosidase activity thereof. In the separation and purification method, tea polyphenol extracts are absorbed by macroporous adsorption resin of nonpolar or weak polar polystyrene; water and ethanol are used for the elution; alcohol eluates, through polyamide columns for the chromatography and are eluted respectively by water and ethanol. The water recrystallization is carried out on the alcohol eluates of the polyamide columns to obtain EGCG pure products with the purity larger than 95 percent and the yield larger than 50 percent; reversed-phase C18 filled columns are used for the chromatography of water eluates of the polyamide columns to obtain EC and EGC; the ethanol of 50-80 percent is used for the recrystallization to obtain pure products of the EC and EGC with the purity all larger than 96 percent. Tests in vitro show that EGCG, EGC and EC all have inhibitory activity on alpha-glucosidase and alpha-amylase and can be used for preparing weight reduction products reducing the absorption of carbohydrates or drugs or health products lowering the postprandial blood glucose.

The invention provides a beta-glucosidase, a preparation method and application of beta-glucosidase, belonging to the field of genetic engineering technology and biological medicine. The beta-glucosidase is selected from the protein shown as (1) or (2) as follows: (1) the protein with amino acid sequence shown as SEQ ID NO.1 and (2) the protein with glycosidase activity and with one / more amino acid residue substituted and / or deleted and / or added in the amino acid residue sequence. The beta-glucosidase provided by the invention has high heat stability and wide pH enzymolysis scope, can be used for producing rare ginsenoside and mixture thereof, has relatively high hydrolysis capacity, is capable of converting the ginsenoside into the rare ginsenoside which has relatively high biological activity, is capable of being easily efficiently absorbed by a human body and has trace volume in the plants such as ginseng, and can be applied to various fields of food, medicine health, and the like.

The invention relates to a pharmaceutical composition having alpha-glucosidase inhibition activity, wherein the pharmaceutical composition comprises a flavone compound and an alpha-glucosidase inhibitor, the flavone compound is at least one selected from a monomer such as baicalein, quercetin, luteolin, baicalein-7-O-glucoside and catechin, an organic salt of the monomer, and an inorganic salt of the monomer, and the alpha-glucosidase inhibitor is at least one selected from a monomer such as acarbose, voglibose and miglitol, an organic salt of the monomer, and an inorganic salt of the monomer. According to the present invention, the pharmaceutical composition can effectively reduce postprandial blood glucose, can inhibit the activity of alpha-glucosidase adopting starch, maltose and sucrose as substrates, and less uses the alpha-glucosidase inhibitor, such that the efficacy can be improved, the side effect of the alpha-glucosidase inhibitor can be effectively reduced, and hypoglycemia and other problems easily caused by drug combination are effectively solved.

The invention provides a method for treating an animal suffering from a disease associated with reduced activity of a lysosomal hexosaminidase by administering to the animal an effective amount of a compound which increases the activity of the hexosaminidase.

The invention discloses a beta-glycosidase mutant and a coding gene thereof, and an application thereof in producing ginsenoside CK. The protein disclosed by the invention is named as lacS-mut, and is one of defined as the following items: 1) the protein is composed of amino acid residue sequence of a sequence 4 in the sequence table; 2) the protein is derived from the protein as defined in the item 1) by substituting and / or deleting and / or adding of one or several amino acid residues in the amino acid residue sequence of the sequence 4 in the sequence table, and has beta-glycosidase activity. Experiments prove that the beta-glycosidase mutant is obtained by mutating a wild type beta-glycosidase gene; compared with the wild type, the beta-glycosidase mutant has the advantage that the activity of the beta-glycosidase mutant in synthesizing the ginsenoside CK is obviously improved, and therefore the beta-glycosidase mutant has better industrial application prospect.

A method for generating human milk oligosaccharides (HMOs) or precursors thereof, compounds obtainable by the method, and uses and compositions involving such compounds. The method comprising the steps of a) providing at least one donor selected from the group of compounds of any of formulae 5 to 10, b) providing at least one acceptor from a group of lactose, LNT, LNnT and derivatives thereof, c) providing at least one enzyme comprising a transglycosidase activity and / or a glycosynthase activity, d) preparing a mixture of the at least one donor, at least one acceptor and at least one enzyme provided in steps a), b) and c); and e) incubating the mixture prepared according to step d).

The invention provides a processing method of Huangguanyin black tea. The processing method comprises the following steps of (1) selecting materials; (2) performing sunlight withering; (3) performinglight rocking of green leaves; (4) performing indoor withering; (5) performing rolling; (6) performing fermentation; (7) performing shaping and strip tidying; (8) performing baking; and (9) performingaroma increasing. According to the processing method disclosed by the invention, a manner of combining sunlight withering with indoor withering is adopted, so that the enzymatic effects of polyphenols substances can be promoted, the activity of glycosidases is also improved, and the production of fragrance of the black tea is facilitated; besides, a manner of performing light rocking of green leaves is adopted, so that mutual friction between tea green leaves is facilitated, leaf margin cells are destructed due to friction, enzymatic oxidation is accelerated, water drainage of tea green leaves is promoted, water and effective substances in the leaves are combined, and many rich fragrance substances are converted; and besides, through evaporation and motion of moisture, transformation of fragrance and taste is facilitated, and high-quality tea leaves can be processed. The tea leaves processed by the processing method are tight and direct in strips of product appearance, tidy and uniform in length, thick, rich, sweet and fragrant in fragrance, and fresh, mellow and refreshing in taste.

The invention discloses saccharomyces cerevisiae and application thereof in the preparation of an externally-applied agent for skin. The saccharomyces cerevisiae is collected in CCTCC (China Center for Type Culture Collection) and has the collection number of CCTCC No: M 2015725. The saccharomyces cerevisiae CCTCC No: M 2015725 has the characteristics of high ultraviolet ray resisting capability and high beta-glycosidase activity and can be applied to the preparation of the externally-applied agent for the skin, so as to achieve the aims of resisting oxidation and anti-aging.

The invention provides applications of four kaurane diterpene compounds in preparation of glycosidase inhibitor medicines. The four compounds capable of highly inhibiting alpha-glycosidase activity are natural compounds which are high in safety and capable of rapid natural degradation without residue in the environment. The four compounds are obtained by separation from wedelia trilobata, and other plant materials. The plant materials are rich in source. A preparation process is easy in operation. Monomers of the four compounds are stable and liable to store. The alpha-glucosidase inhibition activity of the four compounds is obviously higher than or equivalent to that of acarbose that is a clinic medicine. The four compounds are extremely likely developed into effective and safe alpha-glucosidase inhibitor medicines used for preventing and treating type II diabetes, and have good prospects.

The invention relates to an oenococcus oeni strain and an application thereof, and belongs to the technical field of bioengineering. The strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation name is (Oenococcus oeni)JQ88, the preservation date is 28th, June, 2013, the preservation number is CGMCC NO.7800, and the address is Beijing, China. The oenococcus oeni thallus is obtained by high-density fermenting, and then the oenococcus oeni thallus is uniformly mixed with a protective agent to obtain the thallus serum, the thallus serum is frozen and dried to obtain the freeze-dried powder; and the strain and the freeze-dried powder thereof can be applied to the secondary fermentation of fruit wine or grape wine, and can be applied to producing yogurt in the fermenting method of plant juice or animal milk. The oenococcus oeni strain malic acid-lactic acid is strong in fermentation capacity, the related beta-D-glycosidase is high in activity, the strain or the freeze-dried powder is applied to the secondary fermentation of the fruit wine and grape wine, and capable of endowing rich fruity and flowery for the fruit wine and grape wine product, and improving the taste thereof.

The invention provides a preparation method of spermacoce latifolia triterpenoids and an application of the spermacoce latifolia triterpenoids in preparation of a glycosidase inhibitor medicine. Six compounds for intensively inhibiting the activity of alpha-glycosidase provided by the invention are natural compounds which are high in safety, and can be naturally degraded rapidly without a residue in the environment. The compounds can be separated from plant materials such as spermacoce latifolia; the plant materials are abundant in source; and the preparation process is easy to operate. Monomers of the six compounds are relatively stable and easy to store; the alpha-glycosidase inhibiting activity is obviously higher than that of clinical medicine acarbose; and the compounds are extremely likely to be further developed into the effective and safe alpha-glycosidase inhibitor medicines for preventing and treating type 2 diabetes mellitus, and have relatively good prospects.

The present invention relates to compositions, methods and use of a mixture of enzymes having DNase activity and an enzyme having hexosaminidase activity.

The invention belongs to the technical field of natural polymers and discloses a mallotus furetianus homopolysaccharide with antioxidant activity and alpha-glucosidase inhibition activity as well as apreparation method and application thereof. The mallotus furetianus homopolysaccharide disclosed by the invention is named as MFP-2A, and the structural formula of the mallotus furetianus homopolysaccharide is as shown in the specification. The mallotus furetianus homopolysaccharide MFP-2A in the invention is an acid polysaccharide, simultaneously contains six glucosidic bonds of -4,6)-Galp-(1-,-3,6)-Manp-(1-,-5)-Araf-(1-,T-Araf-(1-,-4)-GalpA-(1- and -4)-Glc-(1-, particularly contains alpha-(1,4) glucosidic bonds, has high-efficiency abilities of scavenging DPPH (diphenyl-1-picrylhydrazyl) free radicals, ABTS (azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) free radicals and superoxide radicals, also has properties of resisting oxidation and inhibiting alpha-glucosidaseactivity, can serve as a natural antioxidant, and is applied to preparation of hypoglycemic drugs or health foods.

The invention provides an extraction and purification method of water-soluble total alkaloid with an alpha-glycosidase activity inhibition function from mulberry branch bark and application of the water-soluble total alkaloid. The purification method comprises the following steps: based on the mulberry branch bark as a raw material, adding an extraction solvent (water and ethanol at a ratio of 100-20 percent to 0-80 percent), extracting, heating and refluxing, filtering, repeatedly extracting twice, combining two filtrates, and concentrating; adding 75-100 percent methanol or ethanol by volume, and performing alcohol precipitation at a temperature of between 4 and 30 DEG C; centrifuging or filtering to remove residues and insoluble substances, collecting the supernatant after alcohol precipitation and concentration, purifying through a cation exchange resin column, washing non-adsorbed impurities with deionized water, and eluting with 0.2 to 2mol of ammonia water; passing the eluate through an anoin exchange resin, washing with deionized water, collecting the non-adsorbed part, and regulating the pH to 8; filtering with a membrane, drying the filtrate to prepare powder, namely light-yellow water-soluble total alkaloid extract, wherein the extract can inhibit the activity of alpha-glycosidase, and the content of DNJ (1-deoxynojirimycin) is between 50 and 70 percent.

The invention discloses a seminal plasma α-glucosidase activity quantitative detection kit, which includes R1 reagent and R2 reagent; the R1 reagent includes the following components: 90-110mmol / L pH6.8-7.0 buffer solution, 1.0-3.0 U / L glucose oxidase, 1.0-2.5U / L peroxidase and 0.01-0.1% preservative; R2 reagent includes the following components: 90-110mmol / L pH6.8-7.0 buffer, 100-115mmol / L color former, 70-100mmol / L maltose, 0.2-1.0mmol / L 4-aminoantipyrine and 0.01-0.1% preservative. The invention also discloses the application of the seminal plasma alpha-glucosidase activity quantitative detection kit in measuring seminal plasma alpha-glucosidase activity. The present invention only uses two kinds of liquid reagents, and the types and dosage of reagents are greatly reduced; the rate method is used for detection, and the sample does not need to be pre-diluted, which shortens the reaction time; it is used in conjunction with a fully automatic biochemical instrument, which simplifies the steps and saves reagents. The human error is reduced, and the automatic and large-scale detection greatly improves the detection efficiency while improving the accuracy and reliability of the detection results.

The invention relates to a method for checking uracil DNA glycosidase activity taking molecule beacon as substrate. It makes use of UDG to cut the N- glycosidic bond between uracil (U) base and glycosyl in molecule beacon, then treats base- loss substrate with alkali to make product release fluorescent signal, and determines UDG activity according to fluorescent signal. The method comprises following steps: designing substrate molecule beacon, determining the first- class structure squence according to lowest free energy principle, inserting one deoxyuracil base into the molecule beacon stem of stem-loop structure, treating reacting mixing liquid with alkali of proper concentration after a certain time of thermal insulation, and separating stem-loop and emitting fluorescent. The fluorescent signal can be detected out on fluorescence detector directly without product separation. The invention can not only sensitivly detect pure UDG, but aslo can detect UDG content in coarse raffinate.

The invention discloses a method for detecting an activity of an endoglycosidase. The method includes providing the endoglycosidase with a substrate of the endoglycosidase and detecting cleavage of the substrate. The method further includes at least partly inhibiting the transglycosidase activity of the endoglycosidase. The transglycosidase activity may be inhibited by chemically modifying the substrate such that transglycosylation of the substrate by the endoglycosidase is at least partly inhibited while the endoglycosidase is still capable of cleaving the substrate. In one embodiment, the substrate comprises an oligosaccharide chain. Compounds and kits suitable for use in a method of the invention are also disclosed. Methods involving competitive inhibitors are also disclosed as are methods for the synthesis of glycosylated substrates involving the transglycosidase activity of an endoglycosidase.

The present invention discloses one kind of hypoglycemic polypeptide from silkworm and its preparation process and use as well as composition, medicine, food, tonic and health product connecting the hypoglycemic polypeptide. The hypoglycemic polypeptide of the present invention can combine with glycosidase inhibitor in intestinal tract competitively to inhibit the activity of glycosidase partially, lower the saccharide decomposing speed of glycosidase, lower the amount of glucose produced in unit time and lower the sugar absorbing amount unit time, reaching hypoglycemic effect.

Disclosed herein are polypeptides and variants thereof comprising a polypeptide sequence having substantial identity to ricin A chain (RTA) that lack detectable N-glycosidase-rRNA activity or exhibit reduced N-glycosidase-rRNA activity as compared to controls and methods of making and using thereof. The polypeptides and variants have a greater solubility in aqueous solutions of physiological pH and ionic strength than RTA and also retain the integrity of the neutralizing immunological epitope of wild type RTA. Also disclosed are immunogenic compositions that may be used to immunize a subject against ricin intoxication. Methods of immunizing against, treating, and preventing ricin intoxication are disclosed.

The invention relates to a thermophilic enzyme having β-glycosidase activity which comprises the amino acid sequence of SEQ ID NO: 2 in which one or a plurality of amino acid residues may be deleted, replaced or added.

The invention provides application of a compound 2alpha,3beta-dyhydroxy-23-formyl -olive-12-ene-28-acid in preparation of glycosidase inhibitor medicine. The compound with highalpha-glycosidase inhibiting activity can be obtained from akebia trifoliate and other plants, monomers of the compound 2alpha,3beta-dyhydroxy-23-aldehyde-olive-12-alkene-28-acid are stable, storage is easy, the alpha-glycosidase inhibiting activity of the compound is remarkably higher than that of clinical medicine acarbose (about 8.5 times of the activity of acarbose), and the compound is highly likely to be further developed into the effective and safe alpha-glycosidase inhibitor medicine for preventing and treating II diabetes, and has good prospects.

Provided is a method for diagnosing Pompe disease in a patient by measuring acid α-glucosidase activity in a sample from the patient. The invention also provides a method for monitoring the treatment of Pompe disease with specific pharmacological chaperones by measuring acid α-glucosidase activity in a sample from the patient.

Disclosed herein ricin toxin A chain polypeptides having an engineered disulfide bond ((SS)RTA) and compositions thereof. The disclosed (SS)RTA polypeptides retain the immunological epitope of wild type RTA, lack detectable N-glycosidase activity or exhibit reduced N-glycosidase activity as compared to controls, and exhibit increased solubility, thermal stability and a lower tendency to self-aggregate as compared to RTA 198 and / or RTA 1-33 / 44-198. Also disclosed are immunogenic compositions that may be used to immunize a subject against ricin intoxication. Methods of immunizing against, treating, inhibiting, reducing and / and preventing ricin intoxication are disclosed.

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 a method for detecting an activity of an endoglycosidase. The method includes providing the endoglycosidase with a substrate of the endoglycosidase and detecting cleavage of the substrate. The method further includes at least partly inhibiting the transglycosidase activity of the endoglycosidase. The transglycosidase activity may be inhibited by chemically modifying the substrate such that transglycosylation of the substrate by the endoglycosidase is at least partly inhibited while the endoglycosidase is still capable of cleaving the substrate. In one embodiment, the substrate comprises an oligosaccharide chain. Compounds and kits suitable for use in a method of the invention are also disclosed. Methods involving competitive inhibitors are also disclosed as are methods for the synthesis of glycosylated substrates involving the transglycosidase activity of an endoglycosidase.

The present invention provides a method for alpha-N-acetylglucosaminidase (EC3.2.1.50) activity detection, and further provides a substrate and reagents for alpha-N-acetylglucosaminidase activity determination. The method, the substrate and the reagents can be used for analyzing and determining alpha-N-acetylglucosaminidase activity in a sample (including human body fluid or tissue or cell samples) requiring determination, and are mainly used in the field of clinical laboratory. The determination reagent prepared by using the method has characteristics of convenience, rapidness and high sensitivity, and is easily promoted and applied.

The invention discloses a preparation method of polyphenol extracts in different states of palm fruit, and belongs to the technical field of food or natural medicine. The preparation method comprisesthe steps that firstly super high pressure is adopted to perform pre-processing on a palm fruit, and then extracting is carried out under assistance of ultrasonic wave, and finally the polyphenol extracts in the different states of the palm fruit are obtained after extraction, rotary evaporation and drying; compared with a traditional extraction method, a total phenol content measurement result shows that super high pressure treatment can extremely and significantly improve the extraction rate of phenolic substances and the available rate of the polyphenol extracts in the different states of the palm fruit, and meanwhile, the bioactivity of the polyphenol extracts in three different three states of the palm fruit is all improved significantly verified by experiments, the inhibiting rate ofpancreatic lipase activity, and the inhibiting rate of <alpha>-glycosidase activity and the protective capability of cell oxidative damage are all significantly improved by the polyphenol extracts ofthe palm fruit. The polyphenol extracts of the palm fruit referred have the functions of better lipid lowering and weight losing, blood sugar reducing and prevention and treatment of body oxidative damage, and can be used for preparing relevant health food or medicine.