58 results about "Chymase" patented technology

The invention belongs to the field of bioactive peptides. The invention provides an angiotensin converting enzyme inhibitory peptide as well as a screening method and application thereof. The amino acid sequence of the inhibitory peptide is GKGLW and / or GDGLKW. According to the screening method of the ACE inhibitory peptide, the soybean protein is taken as a raw material, and the ACE inhibitory peptide is prepared and screened by adopting a targeted alkaline protease and chymotrypsin double-enzymolysis technology, so that rapid screening of the ACE inhibitory peptide is realized, and an experimental basis is provided for the structure-function relationship of the ACE inhibitory peptide. The inhibitory peptide provided by the invention has strong antihypertensive activity, high safety and no toxic or side effect. Digestive tract stability determination shows that the inhibitory peptide has good gastrointestinal tract stability in gastrointestinal tract digestion, and can still keep good activity after entering blood.

The invention discloses application of a low-toxicity broad-spectrum antibacterial peptide taking WW as a symmetric center, WK is repeated twice and symmetrically distributed on two sides, WW is taken as the symmetric center to obtain an antibacterial peptide WKWKWWKWKW-NH2 (N2W2), and tryptophan at the C terminal and the N terminal of the N2W2 is replaced with leucine and / or isoleucine to obtain the antibacterial peptide. The antibacterial peptide can basically maintain the excellent antibacterial activity of the parent peptide, and the hemolytic activity of the antibacterial peptide is greatly reduced, so that the safety is improved. Wherein the antibacterial activity of IL-N2W2 is the highest, but the stability of IL-N2W2 needs to be improved, so that D-IL-N2W2 is obtained by carrying out D-type amino acid substitution on IL-N2W2. The D-IL-N2W2 keeps the high antibacterial activity and low toxicity of IL-N2W2, can still completely inhibit the growth of bacteria after being incubated with high-concentration trypsin or chymotrypsin, has great potential in the aspect of relieving the drug resistance of antibiotics, and has good application prospects in preparation of clinical antibacterial drugs.

The invention relates to a medicine for skin regeneration and repair and a preparation method thereof, and belongs to the technical field of biological medicine, the medicine comprises cyperus esculentus extract peptide, brooklet anemone root tremella extract, rhodomyrtus tomentosa root-suberect spatholobus stem-radix cyathulae composite extract, pomegranate peel polyphenol and the like. The cyperus esculentus extracted peptide is a product which is more than 1kDa and is obtained by performing enzymolysis on cyperus esculentus through cellulase, alpha-chymotrypsin and ficin protease in sequence and then performing purification through a G-50 sephadex column; the tremellodon gelatinosum tremella extract is a product which is less than 3kDa and is obtained by performing common enzymolysis on tremellodon gelatinosum tremella sporocarp through cellulase and bromelain; the rhodomyrtus tomentosa root-suberect spatholobus stem-radix cyathulae composite extract is a product obtained by performing gradient extraction on rhodomyrtus tomentosa root, suberect spatholobus stem and radix cyathulae through an ethanol water solution to obtain an extract, and performing polyamide resin column adsorption and LH-60 sephadex column purification in sequence. A good repairing system is constructed by multiple components, and wound repairing and healing are accelerated.

The invention discloses a tilapia skin DPP-IV inhibitory peptide as well as a preparation method and application thereof, and belongs to the technical field of food-borne active peptide development. The preparation method comprises the following steps: taking tilapia skin as a raw material, extracting gelatin from the tilapia skin through an acid-alkali treatment method, and then performing simulated gastrointestinal digestion on the gelatin by utilizing simulated digestive juice prepared from pepsin, trypsin and chymotrypsin to obtain a digestion product with DPP-IV inhibitory activity; the tilapia skin active peptide with relatively high DPP-IV inhibitory activity is obtained through further ultrafiltration and identification, amino acid sequences of the tilapia skin active peptide are AKVELLK, VPGPMGPMGPR and DGIR respectively, and the three inhibitory peptides have good DPP-IV inhibitory activity and can be used for developing food, health care products or medicines for assisting in regulating blood sugar.

The invention relates to the technical field of food, in particular to a preparation method of low-sensitization hydrolyzed whey protein nanoscale microcapsule powder, which comprises the following steps: S1, preparing a whey protein solution, performing microfiltration, and treating with cysteine hydrochloride; s2, trypsin hydrolysis; s3, inactivating and cooling; s4, adding chymotrypsin, and synchronously applying ultrasonic waves; s5, bromelain is added, ultrafiltration is performed, and peptide fragments are collected; s6, preparing a capsule wall solution; s7, adopting a micro-fluidic chip to form a nano emulsion; s8, the mixture is added into a CaCl2 solution to be stirred, multi-layer self-assembly is conducted, and micro-capsule particles are obtained; and S9, carrying out ultrahigh-pressure micro-jet homogenization, vacuum freeze drying and low-temperature plasma assisted grinding on the micro-capsule particles to obtain the nano-scale micro-capsule powder. The stability of active ingredients is maintained through multi-enzyme cooperation and ultrasonic-assisted efficient elimination of allergens, multi-layer nano-embedding under microfluidic control and low-temperature plasma grinding, and the limitations of low single enzymolysis efficiency, insufficient embedding rate and thermal denaturation risk in the prior art are overcome.

The present invention encompasses the use of general chymotrypsin inhibitors, and more specifically, substituted bicyclic substituted uracil of general formula (I) as described and defined herein, and 3-methylbenzo-[b] thiophene)-2-sulfonamido derivatives of general formula (II) for the manufacture of pharmaceutical compositions for the treatment or prevention of stroke, pulmonary embolism, pulmonary embolism, and the like. The traditional Chinese medicine composition can be used for treating thrombosis, deep vein or superficial vein thrombosis, thrombotic microangiopathy, infection, inflammation, thrombotic microangiopathy in a hypercoagulable state after transplantation, disseminated intravascular coagulation, vaccine-induced immune thrombotic thrombocytopenia, and thrombosis or occlusion at a vascular access part.

The invention discloses a urate oxidase mutant (rAfUOX) with trypsin and chymotrypsin tolerance and application of the urate oxidase mutant (rAfUOX). The invention provides a urate oxidase mutant with trypsin and chymotrypsin tolerance by modifying a key amino acid residue in a wild type aspergillus flavus urate oxidase molecule through a protein engineering technology. The activity half-life period of the recombinant aspergillus flavus urate oxidase mutant rAfUOXK49Q after trypsin digestion is increased by 244% compared with the activity half-life period of the wild type rAfUOX, and the activity half-life period of the recombinant aspergillus flavus urate oxidase mutant rAfUOXK49Q after chymotrypsin digestion is increased by 191% compared with the activity half-life period of the wild type rAfUOX. Other enzymatic properties are basically consistent with those of the wild type rAfUOX.

System (100) for the production of biomimetic magnetic nanocomposites for the immobilization of proteolytic enzymes and the targeted treatment of bovine uterine infections, comprising: a) a unit for producing a carrier matrix (102) configured for the production of magnetic nanoparticles; b) an enzyme preparation unit (104) configured to provide proteolytic enzymes selected from the group consisting of protease, trypsin, chymotrypsin, and papain; c) an immobilization unit (106) configured to immobilize the proteolytic enzymes on the modified renewable carrier matrix; and d) a characterization unit (108) configured to perform toxicological tests, comprising: - Equipment for testing antimicrobial activity, and - DPPH assay analysis system.

The present invention relates to a pharmaceutical composition for treating peptic gastric ulcer and a preparation method thereof, belonging to the field of biotechnology. The pharmaceutical composition contains chicken gizzard-membrane peptide, pumpkin seed extract, complex bacteria, white hyacinth bean flower extract and bletilla striata extract; the chicken gizzard-membrane peptide contains the products obtained by successively subjecting chicken gizzard-membrane to fractional enzymolysis with pepsin, chymotrypsin and keratinase; the pumpkin seed extract contains the products obtained by subjecting pumpkin seeds to co-enzymolysis with cellulase and pectinase and then successively to fractional enzymolysis with pepsin, nattokinase and aminopeptidase; the complex bacteria contain the co-fermentation bacteria of Lactobacillus acidophilus, Bifidobacterium and Lactobacillus casei; the white hyacinth bean flower extract is an ethanol percolation extract; the bletilla striata extract contains the products obtained by subjecting bletilla striata tubers to co-enzymolysis with cellulase and pectinase and then successively to fractional enzymolysis with pepsin, bromelain and serrapeptase. The various components are used in combination, having a good therapeutic effect on gastric ulcer, quickly and efficiently repairing gastric mucosa, and being safe and free of side effects.

The disclosure relates to pharmaceutical formulations and method of using chymase inhibitors for the prevention and treatment of mast cell-related disease states in which chymase is involved. More particularly the present disclosure is in the field of treating or preventing or reducing the symptoms of mast cell-related diseases, including atopic dermatitis (AD), chronic urticaria (CU), Eosinophilic Gastrointestinal Disorders (EGIDs), and prurigo nodularis (PN), using a chymase inhibitor, including a chymase inhibitor that prevents cleavage of stem cell factor (SCF) and its interaction with receptor tyrosine kinase (c-kit), thereby reducing mast cell activation and proliferation.

The invention relates to a self-cutting-resistant and high-specific-activity chymotrypsin mutant, in particular to a self-cutting-resistant and high-specific-activity chymotrypsin mutant obtained through molecular modification and screening and a coding gene and application of the self-cutting-resistant and high-specific-activity chymotrypsin mutant, and belongs to the technical field of protein and gene engineering. The chymotrypsin mutant provided by the invention is obtained by mutation of at least one of the following sites on the basis of an amino acid sequence as shown in SEQ ID NO.1: L97A, L162I, F71I, L83I, F114I and W207H. Compared with original chymotrypsin, the chymotrypsin has better self-cutting resistance and higher specific activity, compared with wild type chymotrypsin, the self-cutting resistance is improved by 82% at most, the specific enzyme activity is improved by 37% at most, and the problem that the activity is easily reduced due to self-cutting in the storage and use process of the chymotrypsin is solved.

The application discloses a kind of low-toxicity broad-spectrum antibacterial peptide with WW as symmetry center, the antibacterial peptide is that WK is repeated twice and symmetrically distributed in two sides, and WW is used as symmetry center, obtain antibacterial peptide WKWKWWKWKW-NH2 (N2W2), then the tryptophan of C terminal and N terminal of N2W2 is replaced by leucine and / or isoleucine;The antibacterial peptide can basically maintain the excellent antibacterial activity of parent peptide, and their hemolytic activity is greatly reduced, and the safety is improved. The antibacterial activity of IL-N2W2 is highest, but the stability needs to be improved, so D-type amino acid replacement is carried out on IL-N2W2 to obtain D-IL-N2W2. D-IL-N2W2 maintains the high antibacterial activity and low toxicity of IL-N2W2, and can still completely inhibit bacterial growth after incubation with high-concentration trypsin or chymotrypsin, has very great potential in relieving antibiotic resistance, and has good application prospect in preparing clinical antibacterial drugs.

The present invention covers the use of chymase inhibitors in general and more in particular substituted bicyclically substituted uracils of general formula (I) as described and defined herein, and 3-methylbenzo-[b]thiophene)-2-sulfonamido derivatives of general formula (II) for manufacturing pharmaceutical compositions for the treatment or prophylaxis of stroke, pulmonary embolism, deep or superficial vein thrombosis, thrombotic microangiopathy, thrombotic microangiopathy in hypercoagulable states after infection, inflammation, transplantation, disseminated intravascular coagulation, vaccine-induced immune thrombotic thrombocytopenia, vascular access site thrombosis or occlusion.

The application provides a polypeptide with angiotensin converting enzyme inhibitory activity and a preparation method and application thereof, and relates to the technical field of active peptide preparation. The abalone is pulverized, a complex enzyme is added for primary enzymolysis, and then a specific protease mixture containing prolyl endopeptidase, chymotrypsin and protease K is added for two-step enzymolysis, so that the obtained polypeptide solution has high ACE inhibitory activity. The obtained polypeptide is analyzed and screened to obtain polypeptides with amino acid sequences as shown in SEQ ID NO. 1-5. The polypeptide has high ACE inhibitory activity, and the IC 50 0.05mg / mL-0.40mg / mL, and therefore can play a blood pressure lowering effect as an angiotensin converting enzyme inhibitor.

The invention belongs to the technical field of comprehensive utilization of oyster mushrooms, and provides an oyster mushroom active peptide for inhibiting foaming of macrophages and a preparation method of the oyster mushroom active peptide. The method comprises the following steps: taking oyster mushroom as a raw material, preparing oyster mushroom protein through alkali extraction and acid precipitation, preparing a crude product of the oyster mushroom active peptide for inhibiting foaming of macrophages through compound enzymatic hydrolysis of the oyster mushroom protein by pepsin, trypsin and chymotrypsin, and separating and purifying the crude product by adopting ultrafiltration membrane ultrafiltration and Sephadex G-15 gel column chromatography to obtain the peptide with good activity and high purity. The bioactive peptide can reduce lipid accumulation, ROS level, TNF-alpha content and IL-6 content in RAW264.7 foamed macrophages, and effectively improve lipid accumulation, oxidative stress and inflammatory state of the foamed macrophages, thereby inhibiting foaming of the macrophages.

The present invention belongs to the field of polypeptide drugs, and specifically relates to a glycosylated bicyclic stapled peptide that can improve the anti-drug-resistant bacteria activity, improve the serum stability, and has good enzymatic hydrolysis stability on the basis of a template polypeptide, and its application. The present invention modifies the amino acid sequence of the template polypeptide SLP-0: Ac-LKRVWKRVFKLLKRYWRQLKKPVR-NH2 to obtain a glycosylated bicyclic stapled peptide. Compared with the template polypeptide SLP-0, its antibacterial effect is significantly improved, especially the antibacterial performance against Klebsiella pneumoniae (KP) is increased by 31 times compared with the template peptide SLP-0. Among them, SLP-51 has significant stability against trypsin, chymotrypsin and serum.

The invention belongs to the field of polypeptide drugs, and particularly relates to a glycosylated double-ring stapled peptide which can improve drug-resistant bacterium resistance activity and serum stability on the basis of template polypeptide and is good in enzymolysis stability and application. The amino acid sequence of the template polypeptide SLP-0: Ac-LKRVWKRVFKLLKRYWRQLKKPVR-NH2 is modified and transformed to obtain the glycosylated double-ring stapled peptide, compared with the template polypeptide SLP-0, the antibacterial effect of the glycosylated double-ring stapled peptide is remarkably improved, and particularly, the antibacterial performance of the glycosylated double-ring stapled peptide on klebsiella pneumoniae (KP) is improved by 31 times compared with that of the template peptide SLP-0. Wherein the SLP-51 has remarkable stability on trypsin, chymotrypsin and serum.

The invention provides a polypeptide with angiotensin converting enzyme inhibition as well as a preparation method and application thereof, and relates to the technical field of active peptide preparation. According to the method, the abalone is crushed, the compound enzyme is added for primary enzymolysis, then the specific protease mixture containing prolyl endopeptidase, chymotrypsin and protease K is added for secondary enzymolysis, and the obtained polypeptide liquid has high ACE (angiotensin converting enzyme) inhibition. The obtained polypeptide is analyzed and screened to obtain the polypeptide with the amino acid sequence as shown in SEQ ID NO.1-5. The polypeptide disclosed by the invention has high ACE (angiotensin converting enzyme) inhibition, and IC50 (50% of IC50) of the polypeptide to ACE is 0.05 mg / mL-0. 40mg / mL, so that the polypeptide can be used as an angiotensin converting enzyme inhibitor to play a role in reducing blood pressure.

The invention discloses a micro-ecological protective oral cleaning composition for whole-course control of dental calculus and application of the micro-ecological protective oral cleaning composition, and belongs to the technical field of oral care products. The composition contains citrate, a multienzyme mixture (at least two of lysozyme, xylanase and chymotrypsin) and a matrix additive. Citrate is chelated with calcium ions to soften and dissolve calcified dental calculus; enzymolysis of a multi-enzyme mixture inhibits early bacterial attachment and sediment accumulation and cooperates with citrate to decompose calcified dental calculus; the matrix additive ensures stable release of active ingredients. The components are synergistic, the whole process from the initial stage of dental calculus formation to calcification maturation intervenes, inhibits deposition and dissolves a calcification structure, and meanwhile, the composition can protect the micro-ecology of the oral cavity and has no obvious destructive effect on oral flora. The composition is applied to preparation of oral care products, can solve the problems of one-sided cleaning effect, poor hidden area effect and dysbacteriosis caused by sterilization of existing products, achieves the dual purposes of whole-course dental calculus control and micro-ecological maintenance, and is wide in application prospect.

The invention relates to the technical field of antibacterial peptide application, in particular to an antibacterial peptide CAMP502NC3 with high stability and application of the antibacterial peptide CAMP502NC3. The compound not only shows good antibacterial activity on staphylococcus aureus, but also can be applied to wide pH environments with pH of 2-8 and various protease environments, such as pepsin, trypsin and chymotrypsin; the compound can maintain a certain bactericidal ability under the conditions of high temperature and high salt concentration as high as 250 mM, shows high stability, and has the potential of preparing environment-stable antibacterial drugs.

Provided herein are compositions, methods, and kits related to chymotrypsin production for characterizing proteins, assessing, and monitoring disease stages and phases, predicting the likelihood of disease progression, predicting and monitoring responses to disease therapies, and treating disease conditions.

The present invention discloses an anti - enzymolysis antibacterial peptide FRL, its preparation method and application, belonging to the field of bioengineering. The sequence of the antibacterial peptide FRL is shown as SEQ ID No.1. This antibacterial peptide FRL exhibits narrow - spectrum antibacterial activity, can effectively inhibit Gram - negative bacteria, and has the potential to be applied in drugs for treating Gram - negative bacterial infectious diseases. This antibacterial peptide has very low hemolytic activity and eukaryotic cell toxicity. This antibacterial peptide fails to cause 10% erythrocyte hemolysis at a concentration of 128 μmol / L, and the survival rate of mouse macrophage RAW264.7 reaches more than 80%. This antibacterial peptide shows good stability against trypsin, pepsin, chymotrypsin, and proteinase K (0.4 mg / mL; 0.5 h) at pH = 5 and 37°C. In summary, the antibacterial peptide FRL has the development potential to become an alternative to antibiotics.

The present invention covers the use of chymase inhibitors in general and more in particular substituted bicyclically substituted uracils of general formula (I) as described and defined herein, and 3-methylbenzo-[b]thiophene)-2-sulfonamido derivatives of general formula (II) for manufacturing pharmaceutical compositions for the treatment or prophylaxis of stroke, pulmonary embolism, deep or superficial vein thrombosis, thrombotic microangiopathy, thrombotic microangiopathy in hypercoagulable states after infection, inflammation, transplantation, disseminated intravascular coagulation, vaccine-induced immune thrombotic thrombocytopenia, vascular access site thrombosis or occlusion.

The invention belongs to the technical field of biology, and particularly relates to separation and application of proteasome-rich human placenta-derived vascular stem cell outer vesicles. The extracellular vesicles provided by the invention are derived from human placenta source blood vessel promoting stem cells, and the human placenta source blood vessel promoting stem cells show that markers PAI-1 are positive, MECOM is positive, CD201 is positive, CD44 is positive, CD73 is positive, HLA-ABC is positive, CD34 is negative, CD31 is negative and CD326 is negative. Compared with human umbilical cord mesenchymal stem cell outer vesicles, the human placenta-derived vascular stem cell outer vesicles provided by the invention are rich in proteasomes and angiogenic protein factors, wherein the proteasomes have high chymotrypsin-like activity. Moreover, in-vivo and in-vitro experimental data show that the human placenta-derived vascular stem cell outer vesicles provided by the invention are rich in proteasomes, and can degrade accumulation of HIF and inhibit cell apoptosis; the medicine has a treatment effect on myocardial infarction.

The present application belongs to the technical field of preparation of sheep placenta freeze-dried powder, and provides a method for preparing sheep placenta freeze-dried powder by biological enzymolysis, comprising: sheep placenta pretreatment, enzymolysis and cooking, enzyme inactivation, filtration, concentration, plating, quick freezing, reduced pressure drying, etc., wherein the enzyme added to the enzymolysis and cooking tank is: one or more of alkaline protease, papain, lipase, neutral protease, and chymotrypsin; the enzymolysis temperature is 37°C to 62°C, the amount of the enzyme is 0.5% of the mass of sheep placenta minced meat, and the enzymolysis pH is 6 to 8.5; the acidic pH regulator is a vitamin C aqueous solution, and the alkaline pH regulator is a sodium bicarbonate aqueous solution. The present application uses vitamin C to adjust the pH value of the enzymolysis reaction, which is safer and non-toxic compared to using inorganic compounds such as acetic acid as pH regulators.

The invention provides an HSA fluorescent probe as well as synthesis and application thereof. The fluorescent probe is an enzymatic HSA fluorescent probe and has a structure as shown in a formula (I). The fluorescent probe has high selectivity and high sensitivity, can distinguish HSA from other enzymes (such as chymotrypsin, lipase, protease, beta-galactosidase, pepsin, acetylcholin esterase, butyrylcholine esterase, carboxylesterase, trypsin, DNA enzyme, alkaline phosphatase, lysozyme and the like), and has the detection limit as low as 0.27 mu g / mL, so that the fluorescent probe has the advantages of high selectivity, high sensitivity, high sensitivity and the like. The fluorescent probe provided by the invention can realize qualitative and quantitative detection of HSA in serum, urine or cells.

The present application relates to the technical field of antibacterial peptide application, and particularly relates to an antibacterial peptide CAMP502NC3 with high stability and application thereof.The antibacterial peptide CAMP502NC3 not only exhibits good antibacterial activity on staphylococcus aureus, but also can maintain certain bactericidal capacity in a wide pH environment of pH 2-8, a variety of protease environments, for example, pepsin, trypsin, chymotrypsin, and a salt concentration condition of up to 250 mM, and shows high stability and has the potential to prepare environment-stable antibacterial drugs.

The present application belongs to the field of bioactive peptides. The present application provides an angiotensin converting enzyme inhibitory peptide and a screening method and application thereof, and the amino acid sequence of the inhibitory peptide is GKGLW and / or GDGLKW. The screening method of the inhibitory peptide of the present application uses soybean protein as raw material, adopts targeted alkaline protease and chymotrypsin double enzymolysis technology for preparation and screening, realizes rapid screening of ACE inhibitory peptides, and provides experimental basis for structure-activity relationship. The inhibitory peptide has strong blood pressure lowering activity, high safety and no toxic side effects. The determination of the stability of the digestive tract shows that the inhibitory peptide has good gastrointestinal stability in the gastrointestinal digestion, and can still maintain good activity after entering the blood.