14 results about "Trypsin activity" patented technology

A composition for suppressing trypsin activity is disclosed. The composition contains a bacterium belonging to a genus Paraprevotella as an active ingredient.

The present disclosure provides methods of large-scale preparation of alpha-1 antitrypsin (AAT) activity modulators useful for treating alpha-1 antitrypsin deficiency (AATD), such as 4-(5-(4-fluorophenyl)-6-(tetrahydro-2H-pyran-4-yl)-1,5-dihydropyrrolo[2,3-f]indazol-7-yl)benzoic acid (Compound 1), 3-[5-(4-fluorophenyl)-6-isopropyl-1H-pyrrolo[2,3-f]indazol-7-yl]propanoic acid (Compound 2), or any pharmaceutically acceptable salt of any of the foregoing.

The application discloses an amino-functionalized magnetic mesoporous nanomaterial and a preparation method and application thereof, and the nanomaterial is Fe3O4@nSiO2@TiO2@mSiO2-NH2; from inside to outside, the nanomaterial comprises a magnetite magnetic core, a dense silica layer, a titanium dioxide layer and an amino-functionalized mesoporous silica layer. The magnetic core is synthesized by a solvothermal method, the dense silica layer and the titanium dioxide layer are coated by adopting a sol-gel method in sequence, and finally the amino-functionalized mesoporous silica layer is modified by a co-condensation method. The material can protonate the surface amino group to form a local alkaline microenvironment under an acidic condition, can maintain trypsin activity, realizes synchronous performance of rapid enzymolysis of proteins and in-situ enrichment of phosphorylated peptides, and significantly shortens the enzymolysis time. The application has excellent enrichment selectivity for the phosphorylated peptides, the sensitivity reaches 10 fmol / μL, the enrichment capacity is about 100 mg / g, and is suitable for high-throughput quantitative analysis of phosphorylated proteins in complex biological samples.

The present invention belongs to the field of biotechnology, and specifically discloses a self-cleavage site mutant of Streptomyces trypsin and its application. The trypsin amino acid sequence shown in SEQ ID NO.1 is used as a parent, and the lysine at the 81st position of the parent amino acid is mutated into serine, and the arginine at the 119th position is mutated into glutamic acid or threonine, thereby obtaining two strains of Streptomyces trypsin mutants K81S / R119E and K81S / R119T. The present invention performs mutation evolution on trypsin derived from Streptomyces in vitro, firstly scans the arginine and lysine sites in the amino acid sequence and mutates them to alanine at a specific site, determines the importance of arginine and lysine residues for maintaining trypsin activity, and then performs amino acid saturation mutation and combined mutation on the potential degradation sites where the trypsin activity is improved after mutation, thereby further improving the trypsin activity of the mutant.

The present application belongs to the field of biomedical detection technology, and particularly relates to a reaction solution for detecting trypsin-like transpeptidase activity and application thereof. The present application uses BAPNA as a specific substrate, which can produce a detectable signal change in the transpeptidation reaction, and a high-concentration amino group-glycine amide is added as a receptor molecule to enhance the transpeptidation reaction efficiency. Therefore, the present application solves the technical problem that the enzyme activity cannot be detected by the immunization method, has high reaction efficiency, stable and reliable detection process, shortened operation time (within 15 minutes), simple operation, strong practicability, can be used for evaluating the severity of acute allergic reaction, providing scientific guidance for the diagnosis of mast cell-related airway inflammation and mast cell activation syndrome, monitoring the effect of allergic treatment, and has clinical value in airway inflammation typing, differential diagnosis and efficacy monitoring through induced sputum or BAL fluid trypsin activity determination.

This disclosure provides large-scale processes for preparing a modulator of alpha-1 antitrypsin (AAT) activity that may be useful for treating alpha-1 antitrypsin deficiency (AATD), such as 4-(5-(4-fluorophenyl)-6-(tetrahydro-2H-pyran-4-yl)-1,5-dihydropyrrolo[2,3-f]indazol-7-yl)benzoic acid (Compound 1), 3-[5-(4-fluorophenyl)-6-isopropyl-1H-pyrrolo[2,3-f]indazol-7-yl]propanoic acid (Compound 2), or a pharmaceutically acceptable salt of any of the foregoing.

The invention provides application of glutathione peroxidase in feed preparation, and belongs to the technical field of feed. The method comprises the following steps: constructing recombinant pichia pastoris expressing a glutathione peroxidase mutant, adding the expressed glutathione peroxidase into crushed feed raw materials, stirring and uniformly mixing, acting at 37 DEG C for 12 hours to decompose trypsin inhibiting factors in the feed, and then preparing the feed. After the feed without the glutathione peroxidase acts with the trypsin, the activity of the trypsin is only 60.40%; after the feed prepared by adding glutathione peroxidase acts with trypsin, the trypsin activity is 95.09%. The glutathione peroxidase is applied to feed raw material treatment, trypsin inhibiting factors existing in grains such as soybean meal in feed raw materials can be effectively reduced, and therefore the digestion and absorption rate of animals on protein in feed is increased, the feed conversion ratio is reduced, and the breeding benefits are improved.

According to the method for fattening and spawning induction before spawning induction, terrestrial cabbages are adopted as one of main bait components of the purple urchins, laver and mussels are combined, and according to parent characteristics of the purple urchins before spawning induction, the purple urchins are subjected to fattening and spawning induction, and the purple urchins are subjected to spawning induction. According to the method, a multi-recipe combined feeding method adopting baits of the brassica juncea in the fattening period is creatively designed, the design of the GSI index, the egg yolk protein content and the trypsin activity of the sea urchins is combined, the fattening condition and the breeding density in the fattening period are researched and set, and through fattening in the fattening period before spawning induction, the breeding density of the brassica juncea is increased; the method has the advantages that the gonad growth and development of the mature purple urchin in the mature period are obviously improved, the intestinal flora of the purple urchin is effectively improved, the immunity of the purple urchin before spawning induction is enhanced, the spawning rate is also greatly increased, the number and survival rate of fertilized eggs after spawning induction are obviously increased, the artificial reproduction cycle is shortened, and the culture cost is reduced.

This invention provides a method for screening, separating, and identifying trypsin inhibitors, belonging to the field of biomedical technology. The invention first uses colorimetric sensor technology to rapidly identify samples with trypsin-inhibiting activity. Then, using affinity adsorption technology combined with liquid chromatography-mass spectrometry (LC-MS), specific components in the samples with trypsin-inhibiting activity can be identified. Specifically, trypsin is immobilized on Tp-Pa to form a TRY@Tp-Pa affinity chromatography carrier material. The extract to be screened is incubated with the affinity chromatography carrier material, and after removing the unadsorbed liquid, methanol is used to elute the eluent, which contains the bioactive compounds retained on TRY@Tp-Pa. These compounds are the potential active ingredients, and LC-MS is then used to identify the components. The method provided by this invention can efficiently and accurately detect trypsin activity and screen inhibitors, with high screening efficiency and a low false positive rate.

The invention provides application of phospholipid hydroglutathione peroxidase in a feed additive, and belongs to the technical field of feed. The method comprises the following steps: constructing recombinant pichia pastoris for expressing deacetylated phospholipid hydroglutathione peroxidase, adding the expressed deacetylated phospholipid hydroglutathione peroxidase into crushed feed raw materials, stirring and uniformly mixing, decomposing trypsin inhibiting factors in the feed, and then preparing the feed. The deacetylated phospholipid hydroglutathione peroxidase is not added into a control group, and the trypsin activity is only 59.13% after the feed reacts with the trypsin; after the deacetylated phospholipid hydroglutathione peroxidase group feed is added to act with the trypsin, the trypsin activity is 95.39%. The feed conversion ratio of the Chang-bai piglets fed with the feed prepared by adding the deacetylated phospholipid hydroglutathione peroxidase is reduced compared with the feed conversion ratio of the Chang-bai piglets fed with the feed without adding the deacetylated phospholipid hydroglutathione peroxidase, the weight gain of the piglets is increased by 33.06%, and the breeding benefits are effectively increased.

The invention belongs to the technical field of biomedical detection, and particularly relates to reaction liquid for detecting trypsin transpeptidase activity and application of the reaction liquid. BAPNA is adopted as a specific substrate, detectable signal change can be generated in the transpeptidation reaction, then high-concentration amino group-glycinamide is added as an acceptor molecule, and the transpeptidation reaction efficiency is enhanced. Therefore, the technical problem that the enzyme activity cannot be detected by an immunoassay is solved, the reaction efficiency is high, the detection process is stable and reliable, the operation time is shortened to be within 15 minutes, the operation is simple and convenient, and the practicability is high; the kit can be used for evaluating the severity of acute anaphylaxis, provides scientific guidance for diagnosis of mast cell related airway inflammation and mast cell activation syndrome, monitors the anaphylaxis treatment effect, and has clinical value in the aspects of airway inflammation typing, differential diagnosis and curative effect monitoring by inducing phlegm or BAL liquid trypsin activity determination.

The invention provides a trypsin inhibitor and application thereof, and relates to the technical field of molecular biology. According to the invention, it is found for the first time that zinc pyrithione, ketoconazole and / or selenium disulfide have an inhibition effect on trypsin activity, and new functional components which are convenient to obtain are provided for the trypsin inhibitor. The invention provides three compositions containing zinc hyaluronate, the zinc hyaluronate is respectively compounded with zinc pyrithione, ketoconazole or selenium disulfide according to a specific mass ratio, and it is found for the first time that the zinc hyaluronate and the zinc pyrithione, ketoconazole or selenium disulfide show the activity of synergistically inhibiting trypsin. The composition provided by the invention is simple to obtain, can be continuously compounded with other functional active components or auxiliary material components to obtain commercial products, and has a wide application prospect.

This invention discloses a composite hydrogel based on sodium alginate and citric acid, its preparation method, and its applications, belonging to the field of biomedical materials and polymer hydrogels. The composite hydrogel is formed by mixing two components, A and B, to form a gel. Component A contains an aqueous solution of sodium alginate and citric acid, while component B is a crosslinking agent solution containing soluble polyvalent cations. The polyvalent cations are selected from one or more combinations of calcium ions, barium ions, and strontium ions. This invention utilizes citric acid to competitively chelate metal cations with sodium alginate, effectively slowing down the gelation rate and optimizing the three-dimensional density of the gel, significantly improving the in-situ adhesion and sealing stability of the material. Simultaneously, the local slow-release of citric acid lowers the pH of the microenvironment to a slightly acidic range of 6.0-6.5, achieving non-competitive inhibition of trypsin activity.