68 results about "Subtilisin" patented technology

The invention relates to an shRNA (short hairpin Ribonucleic Acid) interference sequence of a targeted silence PCSK9 gene and a construction method and lipid-lowering application of a recombinant adeno-associated virus vector of the shRNA interference sequence. Hyperlipidaemia is a metabolic disease characterized by abnormal rising of cholesterol and triglyceride levels in blood, and the design of lipid-lowering drugs is the focus of attention to improvement of hyperlipidaemia. Proprotein convertase subtilisin / kexin type 9 (PCSK9) can be combined with a low-density lipoprotein receptor (LDL-R) and degrade the LDL-R, so that accumulation of LDL-C in blood is further promoted, and hyperlipidemia is caused. Aiming at the key target PCSK9, a specific shRNA interference sequence is designed, and a recombinant adeno-associated virus vector (rAAV) carrying the sequence is constructed by an enzyme digestion-connection method. In-vitro experiments prove that the vector can remarkably reduce the expression level of PCSK9 protein, so that the cyclic utilization of a low-density lipoprotein receptor (LDL-R) is promoted, and the concentration of low-density lipoprotein cholesterol (LDL-C) in plasma is reduced. The rAAV vector provided by the invention has the characteristics of low production cost, high transfection efficiency, lasting action time and the like, and provides a new thought for gene therapy of hyperlipidemia.

Disclosed herein is one or more subtilisin variant, nucleic acid encoding same, and compositions and methods related to the production and use thereof, including one or more subtilisin variant that has improved stability compared to one or more reference subtilisin.

The invention provides an anti-aging and skin-tightening composition and a preparation method thereof, and belongs to the technical field of anti-aging and skincare, and the composition comprises a hovenia acerba infructescence extract, an okra seed extract, tetrapeptide-4, acetyl octapeptide-1, a hill-pit snail extract, paeonol and a carnosine-zinc gluconate chelate. And carrying out aerobic fermentation on the raisin tree infructescence powder through aspergillus oryzae, and purifying the centrifuged supernatant through a composite filler resin column to obtain the raisin tree infructescence extract. The preparation method comprises the following steps: sequentially carrying out enzymolysis on okra seeds by virtue of compound enzyme, ficin and subtilisin, adding absolute ethyl alcohol for extraction, centrifuging supernate, and purifying by virtue of an LH-20 sephadex column, so as to obtain the okra seed extract. The preparation method comprises the following steps: sequentially carrying out enzymolysis on the mountain snail meat through trypsin and lumbrukinase, centrifuging the enzymatic hydrolysate, and carrying out ultrafiltration on the supernate to obtain the mountain snail extract with the molecular weight of less than 3kDa. The components are matched for use, so that the balance of high-efficiency anti-aging and low-irritation can be realized through synergism of three major pathways of inhibiting collagen degradation, promoting collagen synthesis and resisting sugar and relieving.

The utility model relates to a device for removing impure protein in crude polysaccharide of traditional Chinese medicine by utilizing immobilized protease, which comprises a protein removal reactor loaded with the immobilized protease, a dissolved oxygen tester and a constant-temperature magnetic stirrer, and a dissolved oxygen electrode of the dissolved oxygen tester is connected to the protein removal reactor. The papain immobilized on the microcrystalline cellulose is used for carrying out enzymolysis reaction on impure protein in the traditional Chinese medicine crude polysaccharide, and the device has the characteristics of simplicity in operation, mild conditions and environment friendliness. The immobilized papain is limited between the two layers of screens, so that the problem of separation of the immobilized papain in the traditional Chinese medicine crude polysaccharide solution can be avoided, and the method has the characteristics of rapidness and convenience. The immobilized papain in the device can be expanded to immobilize other proteases, such as bromelain, subtilisin, protease K and the like, on cellulose according to protein types in different traditional Chinese medicine polysaccharide solutions.

The invention provides a pharmaceutical composition for improving bone mineral density and a preparation method thereof, and belongs to the technical field of biological medicine, the pharmaceutical composition comprises lactarius vomica extract peptide, winged yam rhizome peel extract, rainbow trout extract, calcium citrate-glycine chelate, vitamin K2 and vitamin D3. The lactarius volemus extract peptide is obtained by sequentially carrying out enzymolysis on lactarius volemus by bromelain and subtilisin and purifying and extracting enzymatic hydrolysate through an LH-60 sephadex column. The winged yam rhizome peel extract is obtained by extracting the peels of winged yam rhizome tubers with an ethanol water solution and purifying and extracting the extracting solution with an AB-8 macroporous adsorption resin column. The rainbow trout extract is a product below 5kDa obtained by sequentially performing enzymolysis on fish heads, fish skins and fish scales of rainbow trout through trypsin and lumbrukinase. All the components are compounded according to a specific proportion, and the core effect of improving the bone mineral density is achieved through two-way regulation and control of bone metabolism, optimization of calcium utilization and improvement of safety.

Disclosed herein are one or more subtilisin variants, nucleic acids encoding the same, and compositions and methods relating to the production and use thereof, including one or more subtilisin variants having improved stability and / or soil removal compared to one or more reference subtilisin.

This invention provides an antigen-binding protein that binds to proprotein converterase subtilisin kexin type 9 (PCSK9), as well as a method for using and producing the antigen-binding protein. [Solution] The present invention provides an isolated neutralizing antigen-binding protein that interacts with proprotein converterase subtilisin kexin type 9 (PCSK9), and which contains a specific amino acid sequence that binds to the PCSK9 protein and reduces the LDLR-reducing effect of PCSK9 on LDLR.

Disclosed herein is one or more subtilisin variant, nucleic acid encoding same, and compositions and methods related to the production and use thereof, including one or more subtilisin variant that has improved stability and / or soil removal compared to one or more reference subtilisin.

Disclosed herein is a detergent composition comprising a subtilisin variant that has improved stability and / or soil removal compared to one or more reference subtilisin.

The application provides a preparation method, composition and application of tree orchid flower oil, and relates to the technical field of cigarette flavors. The preparation method of the tree orchid flower oil comprises the following steps: step one, drying and crushing tree orchid flowers to obtain tree orchid flower powder; step two, uniformly mixing the tree orchid flower powder and water, adding enzyme preparation, and performing enzymolysis reaction under the condition that the temperature is 40-50 DEG C and the pH value is 4-7 for 1-5 h to obtain a treatment liquid; the enzyme preparation is composed of hemicellulase, pectinase, xylanase, beta-glucanase and subtilisin with an enzyme activity ratio of (4-6):(3-5):(3-5):(1-3):(1-2); and step three, performing supercritical carbon dioxide fluid extraction on the treatment liquid to obtain tree orchid flower oil. The application provides tree orchid flower oil suitable for cigarettes, the tree orchid flower oil has high extraction rate, obvious flower fragrance and tea fragrance, no bitter and astringent traditional Chinese medicine smell, and no miscellaneous smell.

The invention belongs to the technical field of protein peptide powder. The invention provides a preparation method of ovalbumin peptide powder, which comprises the following steps: S1, transferring protein liquid into a fermentation tank, adjusting the pH value to 6.0-6.9, adding dry yeast, stirring and fermenting to obtain fermentation liquid; s2, transferring the fermentation liquor obtained in the S1 into an enzymolysis tank, heating to 50-55 DEG C, adding a compound enzyme preparation, carrying out enzymolysis for 7-10 hours, and stirring for 10 minutes per hour to obtain enzymatic hydrolysate; the compound enzyme preparation comprises papain, subtilisin, trypsin and aminopeptidase; and S3, transferring the enzymatic hydrolysate obtained in S2 back to a fermentation tank, cooling, adding dry yeast, stirring, fermenting, spray-drying, and placing in a hot chamber for 3-7 days to obtain the product. According to the ovalbumin peptide powder, the oxidation resistance and the heat resistance are improved at the same time, so that the ovalbumin peptide powder with more excellent functionality and wider application range is obtained.

The invention relates to the field of bioengineering, in particular to a perch collagen peptide as well as a preparation method and application thereof. The invention provides a preparation method of perch collagen, which comprises the following steps: carrying out acid extraction on pretreated perch scales, hydrolyzing with a first enzyme, filtering, salting out filtrate, centrifuging, collecting precipitate, redissolving, purifying, hydrolyzing with a second enzyme, and heating to obtain the perch collagen, enzymes adopted by the first enzyme hydrolysis and the second enzyme hydrolysis can be the same or different; the enzyme comprises one or more of alkaline protease, papain, subtilisin and pepsin. The collagen peptide extracted from sea bass scales and processed into the sea bass scales is systematically researched and developed, and the collagen peptide has an excellent anti-photoaging effect and high replicability. The collagen peptide is low in molecular weight, high in oxidation resistance and wide in application range, and can be applied to the fields of cosmetics, oral collagen supplements and the like.

The present invention relates to methods of treating or preventing cholesterol related disorders, such as hypercholesterolemia, hyperlipidemia or dyslipidemia, using antibodies against proprotein convertase subtilisin / kexin type 9 (PCSK9). Formulations and methods of producing said formulations are also described.

The present invention relates to a dual-targeting siRNA agent comprising two distinct siRNAs targeting two different genes or their pharmaceutically acceptable salts, wherein the two distinct siRNAs or their salts are linked by a pharmaceutically acceptable ligand. The siRNA is a dsRNA composed of a sense strand and an antisense strand, and the two different genes are selected from a group consisting of angiotensinogen (AGT), proprotein convertase subtilisin / kexin type 9 (PCSK9), and human angiopoietin-like protein 3 (ANGPTL3). The present invention provides the application of the dual-targeting siRNA agent in the preparation of drugs for preventing or treating diseases associated with hypertension and / or dyslipidemia. The dual-targeting siRNA agent described in the present invention can effectively inhibit the expression of two target genes simultaneously in vivo, offering the advantages of strong non-antagonistic activity and high safety. The present invention also provides the siRNAs targeting corresponding genes for the aforementioned dual-targeting siRNA agent and their use for preventing or treating diseases associated with hypertension and / or dyslipidemia.

The present application relates to a kind of multifunctional pharmaceutical composition of dental comprehensive repair and its preparation method, belong to biopharmaceutical technical field, and pharmaceutical composition includes keratin peptide, coral fungus extract, eggshell derived nano calcium phosphate, catechin zinc complex, hydroxyapatite, modified chitosan-PEG gel and other components.Keratin peptide is the product between 3kDa-10kDa obtained by keratinase enzymolysis to white goose velvet;Coral fungus extract is the product obtained by cellulase, subtilisin and bromelain enzymolysis to coral fungus, and purified by G-25 dextran gel column;Modified chitosan-PEG gel is the product obtained by sodium cyanoborohydride stabilization and glycerol plasticization to the reactant of chitosan and dialdehyde polyethylene glycol.Integrated multiple functions such as antibacterial, mineralization and tissue repair, through optimizing component and dosage form, realize efficient, minimally invasive dental damage repair, and good biocompatibility and long-acting.

The invention belongs to the technical field of biology, and particularly relates to a preparation method of collagen peptide, a nutrition powder composition for promoting wound healing and a preparation method, a product and application of the nutrition powder composition. The preparation method of the collagen peptide comprises the following steps: firstly, adding water into bovine bone particles, heating, soaking and filtering to obtain bovine bone protein liquid; laccase, subtilisin and flavourzyme are sequentially added into the bovine bone protein liquid for enzymolysis, centrifugation is conducted, and supernate is collected; and finally, adding chitosan into the supernate, and carrying out ultrafiltration and drying, so as to obtain a finished product. The prepared collagen peptide is high in yield, good in taste and beneficial to human body absorption, the invention further provides a nutrition powder composition which can accelerate wound healing by promoting wound angiogenesis, collagenous fiber deposition and epidermis regeneration, and therefore the collagen peptide has good application prospects in skin tissue wound repairing.

The invention provides a kidney-tonifying and anti-fatigue polypeptide composition and a preparation method thereof, and belongs to the technical field of medicinal biological peptide complexes, the composition comprises oyster polypeptide, stichopus japonicus polypeptide, a blue ear grass-dilleniad fruit composite extract, a cistanche-dendrobium officinale composite extract, a ginseng extract and a penis et testis cervi extract; the oyster polypeptide is a common enzymolysis product of oyster meat through trypsin and bovine chymotrypsin; the stichopus japonicus polypeptide is a common enzyme product of enzymolysis of stichopus japonicus body walls by alkaline protease and collagenase type II and subtilisin; the blue ear grass-dilleniad fruit composite extract is a product obtained by carrying out enzymolysis on blue ear grass and dilleniad fruit, extracting with an ethanol water solution and purifying with an SA sulfonic group silica gel strong cation exchange column; the cistanche-dendrobium officinale composite extract is a product obtained by carrying out enzymolysis on cistanche and dendrobium officinale, extracting with an ethanol water solution and purifying with a phenylboronic acid affinity column-polyamide column tandem column; and the synergistic effect of tonifying kidney and resisting fatigue is realized.

The application provides a blood-activating and stasis-removing medicinal composition containing fermented douchi and a preparation method thereof, belongs to the technical field of biological medicine, and the medicinal composition comprises fermented douchi, drynaria extract, extract of pereskia aculeata, tanshinone II A, notoginsenoside R1, ligustrazine, rhodioside, total flavones of hawthorn leaves and total flavones of mulberry leaves. The fermented douchi is prepared from soybeans and black beans through fermentation by bacillus subtilis and bacillus coagulans. The drynaria extract is prepared from drynaria powder through common enzymolysis by cellulase, chitinase and beta-glucanase, extraction by a two-phase solvent, and purification by a dextran gel column. The extract of pereskia aculeata is prepared from pereskia aculeata powder through common enzymolysis by pectinase, cellulase and xylanase, common enzymolysis by bromelain and subtilisin, and purification by a macroporous adsorption resin column. The medicinal components are prepared in a specific proportion to relieve blood stasis from two dimensions of "improving blood flow dynamics + preventing thrombus formation".

Disclosed are, inter alia, combinations comprising a first antisense (e.g., first double stranded RNAi (dsRNAi)) agent for inhibiting expression of proprotein convertase subtilisin kexin 9 (PCSK9), for example, human PCSK9, and a second antisense agent (e.g., second dsRNAi agent) for inhibiting expression of lipoprotein (a) (Lp(a)); pharmaceutical compositions including the same, and methods of treatment using the same.

The invention relates to a composition for treating thyroid nodules and a preparation method thereof, and belongs to the technical field of biological medicine, the composition comprises trepang body wall fibrinolytic small peptides, kelp small molecule active components, a selfheal-saussurea lappa composite extract and a safflower-ligusticum wallichii composite extract; the sea cucumber body wall fibrinolytic small peptide is a product obtained by performing common enzymolysis on sea cucumber body wall powder through alkaline pectinase, bromelain and trypsin and then performing common enzymolysis through subtilisin and lumbrukinase. The kelp micromolecular active component is a product obtained by performing enzymolysis extraction on kelp powder through alginate lyase, performing purification through an X-5 macroporous adsorption resin column and performing ultrafiltration. The selfheal-saussurea lappa composite extract is a product obtained by extracting selfheal powder and saussurea lappa powder with an ethanol water solution, purifying with a polyamide resin column and performing ultrafiltration; the safflower-Szechuan lovage rhizome composite extract is a product obtained by extracting safflower powder and Szechuan lovage rhizome powder with an ethanol aqueous solution, purifying with an NKA-9 macroporous resin column and performing ultrafiltration.

This provides a method to reduce cardiovascular risk in patients with a genetic profile associated with a response to proprotein converter subtilisin / kexin type 9 (PCSK9) inhibitor therapy. [Solution] A method for treating a patient at risk of major cardiovascular adverse events (MACE) includes: determining the patient's polygenetic risk score (CAD-PRS), which includes a weighted sum of multiple gene variants associated with coronary artery disease; identifying the patient as being at high risk of MACE if the patient has a CAD-PRS greater than a threshold CAD-PRS determined from a reference population; and administering a PCSK9 inhibitor to the patient if the patient has been identified as being at high risk of MACE.

A mutant subtilase cytotoxin B subunit protein is provided which can bind glycans having α2-3-linked N-glycolylneuraminic acid and glycans having α2-6-linked N-glycolylneuraminic acid. The mutant SubB protein has deletions of one or more of the amino acid sequence TTSTE and has a previously undescribed ability to bind glycans having α2-6-linked N-glycolylneuraminic acid, while not losing the ability to bind glycans having α2-3-linked N-glycolylneuraminic acid.

The invention relates to a dual-targeting siRNA agent, which comprises two different siRNAs targeting two different genes or pharmaceutically acceptable salts thereof, the two different siRNAs or salts thereof are connected through pharmaceutically acceptable ligands, the s-RNA is dsRNA composed of a sense strand and an antisense strand, and the s-RNA is dsRNA composed of a positive-sense strand and a negative-sense strand. The two different genes are selected from two of angiotensinogen (AGT), proprotein convertase subtilisin 9 (PCSK9) and human angiopoietin-like protein 3 (ANGPTL3). The invention provides application of the dual-targeting s iRNA agent in preparation of drugs for preventing or treating diseases related to hypertension or / and lipid disorder. The dual-targeting siRNA agent can effectively inhibit expression of two target genes in vivo at the same time, and has the advantages of being high in mutual non-antagonistic activity and high in safety. The invention also provides the s iRNA of the corresponding gene of the dual-targeting s iRNA agent and application of the s iRNA to prevention or treatment of diseases related to hypertension or / and lipid disorder.

Disclosed herein is a cleaning composition comprising at least one subtilisin variant and at least one surfactant, and a method of cleaning a surface with the cleaning composition.

The invention relates to a multifunctional pharmaceutical composition for comprehensive tooth repair and a preparation method thereof, and belongs to the technical field of biological pharmacy, the pharmaceutical composition comprises keratin peptide, a coral fungus extract, eggshell derived nano calcium phosphate, a catechin zinc complex, hydroxyapatite, modified chitosan-PEG gel and other components. The keratin peptide is a product between 3kDa and 10kDa obtained by performing enzymolysis on white goose down through keratinase; the coral fungus extract is a product obtained by carrying out enzymolysis on coral fungus through cellulase, subtilisin and bromelain and purifying through a G-25 sephadex column; the modified chitosan-PEG gel is a product obtained by stabilizing a reactant of chitosan and dialdehyde polyethylene glycol through sodium cyanoborohydride and plasticizing through glycerol. Multiple functions of antibiosis, mineralization, tissue repair and the like are integrated, efficient and minimally invasive tooth injury repair is achieved by optimizing components and dosage forms, and biocompatibility and long-term effectiveness are good.

This invention relates to compounds of Formula I and their isotopic forms, stereoisomers, tautomers, pharmaceutically acceptable salts, pharmaceutically acceptable solvates, hydrates, prodrugs, and polymorphs. The compounds of Formula I can be used as inhibitors of proprotein convertase subtilisin 9 (PCSK9), effectively inhibiting PCSK9 activity and reducing the concentration of low-density lipoprotein cholesterol in plasma, and have good application prospects in the preparation of drugs for the treatment or prevention of cardiovascular diseases.

The application provides a kidney-tonifying and anti-fatigue polypeptide composition and a preparation method thereof, belongs to the technical field of medicinal biological peptide complexes, and the composition comprises oyster polypeptide, holothurian polypeptide, spider silk and blue ears grass-pentamerous fruit composite extract, cistanche-dendrobium officinale composite extract, ginseng extract and deer penis extract; the oyster polypeptide is a common enzymolysis product of oyster meat by trypsin and bovine chymotrypsin; the holothurian polypeptide is a common enzymolysis product of holothurian body wall by alkaline protease, collagenase type II and subtilisin; the spider silk and blue ears grass-pentamerous fruit composite extract is a product obtained by the following steps: subjecting spider silk and blue ears grass and pentamerous fruit to enzymolysis, extracting with an ethanol aqueous solution, and purifying with a SA sulfonic acid silica gel strong cation exchange column; the cistanche-dendrobium officinale composite extract is a product obtained by the following steps: subjecting cistanche and dendrobium officinale to enzymolysis, extracting with an ethanol aqueous solution, and purifying with a phenylboronic acid affinity column-polyamide column series connection column; and the composition realizes the synergistic effect of kidney tonification and anti-fatigue.

The invention belongs to the technical field of cosmetics, and particularly discloses a sebum-like film composition and application thereof, and the sebum-like film composition comprises the following components in parts by weight: 25-30 parts of a natural fermented oil composition, 1-2 parts of tocopheryl acetate, 0.05-5 parts of ceramide NP, 5-10 parts of glycerol, 0.5-2 parts of 1, 2-hexanediol, 2.5-3 parts of inulin lauryl carbamate, 0.05-0.5 part of sodium surfactin and the balance of deionized water, totaling 100 parts. The natural fermented oil composition is prepared by fermenting phyllanthus emblica kernel oil, sea buckthorn fruit pulp oil and pongamia pinnata seed oil. The sebum-like film composition disclosed by the invention shows excellent compatibility to dry, oily and sensitive skin, and can be used for rapidly repairing barrier injury and reducing percutaneous moisture loss.

This application provides a penetration-enhancing composition for anti-wrinkle and firming active ingredients and an anti-wrinkle and firming cosmetic. The penetration-enhancing composition for anti-wrinkle and firming active ingredients comprises: an anti-wrinkle and firming active ingredient, including one or more of arginine / lysine peptides, tripeptide-1 copper, and sturgeon caviar extract; and a penetration-enhancing composition, including 1,2-pentanediol, inositol, and sodium subtilisin. This application can promote the penetration of anti-wrinkle and firming active ingredients, reduce the amount of anti-wrinkle and firming active ingredients added to cosmetics, and help reduce production costs. Furthermore, the resulting anti-wrinkle and firming cosmetic can better inhibit the expression of MMP-9, promote the expression of type I collagen, and reduce the area of ​​wrinkles.

This specification provides a gene repressor system comprising a fusion protein, such as a fusion protein containing a non-catalyzed class 2, type V CRISPR protein useful for repressing the precursor protein-converting enzyme subtilisin / kexin type 9 (PCSK9) gene, and a guide nucleic acid (gRNA). Methods for constructing and using such a system for repressing PCSK9 transcription are also provided.