78 results about "Proteolytic enzymes" patented technology

The invention discloses keratinase Phyto-Keratinase and application of the keratinase Phyto-Keratinase. The keratinase Phyto-Keratinase comprises the following components: (1) protease which is coded by a nucleotide sequence as shown in SEQ ID NO.1; and / or (2) protease which is derived from bacillus subtilis, is coded by a nucleic acid sequence with more than 98% of identity with the SEQ ID NO.1 nucleotide sequence and has the characteristic of hydrolyzing keratin. According to the plant keratinase Phyto-Keratinase disclosed by the invention, the Phyto-Keratinase (plant keratinase) produced by utilizing microbial fermentation is a serine proteolytic enzyme, is also a biocompatible cutinase, and is obtained by fermenting saccharomyces cerevisiae. Phyto-Keratinase provides a nitrogen source for the growth of plants (beans), and contains various amino acids (functional peptides) and special proteolytic enzymes derived from the plants at the same time. The Phyto-keratinase can provide the effects of resisting inflammation, removing cutin and providing nutrition for skin regeneration at the same time, can relieve various inflammations by inhibiting PAR-2 receptors, and has potential value in the fields of cosmetics, beauty medicine and environment.

The present disclosure relates to: a block polymer conjugate comprising an antibody-linker-polypropylene oxide, in which block polymer comprising polypropylene oxide is conjugated to an antibody through a linker; and pharmaceutical composition for treating an autoimmune disease, comprising the conjugate. The block polymer conjugate comprising an antibody-linker-polypropylene oxide, according to the present disclosure, exhibits the effects of maintaining specific reactions of conventional antibodies, simultaneously, increasing the stability from proteolytic enzymes and improving in vivo half-life. When the conjugate according to the present disclosure is administered to a disease site, effects can be exhibited for longer than those of a conventional antibody and biostability and anti-inflammatory effects greater than those of a conventional antibody are exhibited.

Use of a bitter melon exosome and an oral medicament prepared therefrom. The bitter melon exosome obtained from the bitter melon juice can be used for preparing a pharmaceutical formulation embedding a protein, a polypeptide, a small molecule peptide, a nucleic acid, or a small molecule compound. The compositions of the oral medicament of the present invention comprise the bitter melon exosome and an active pharmaceutical ingredient encapsulated in the bitter melon exosome. The bitter melon exosome is suitable for preparing a drug that is easily damaged by the gastrointestinal tract and is mostly administered by means of an injection route into an oral agent, and the oral agent can produce a similar therapeutic effect to that of the injection administration route by oral administration, and is convenient to use. The extraction method for the bitter melon exosome is simple and has a low cost. Moreover, the bitter melon exosome has no immunogenicity, has relatively high biocompatibility and biosafety, and has relatively strong tolerance to gastric acid and digestive tract proteolytic enzymes, and intestinal barrier permeability.

The invention relates to the field of blood products, in particular to a preparation method of human activated C1 esterase. The preparation method comprises the following steps: carrying out contact adsorption on cryoprecipitate-removed human plasma and anion exchange resin, washing and eluting to obtain an eluent containing C1 proesterase, carrying out membrane filtration, and carrying out ultrafiltration desalination treatment; adding an anionic substance into the eluent, incubating in vitro to effectively activate the C1 esterase, and filtering by a membrane to obtain a feed liquid containing the activated C1 esterase; and carrying out purification treatment on the feed liquid to obtain the human activated C1 esterase, and carrying out cryopreservation. According to the method, an anion substance is introduced into an eluent system obtained through anion exchange, mild and effective activation of the C1 proesterase is achieved under the condition of not depending on foreign proteolytic enzyme, the risk of non-specific hydrolysis and self-cutting inactivation is remarkably reduced, good synergy is formed with a follow-up purification process, and the method is suitable for industrial production. The human activated C1 esterase product with relatively high activity and good stability is obtained.

The present invention relates to a dual functional domain structure and its uses. The dual functional domain according to the present invention includes a first domain that binds to a disease-site-specific expression protein and a second domain that binds to a disease-specific expression protein. In this case, the first domain serves to mask the drug's function at a non-disease site, thereby enabling disease-site-specific drug delivery. In addition, the first domain is cleaved by a proteolytic enzyme specifically expressed at the disease site to induce drug activity and exhibit drug activity in a disease-site-specific manner. Furthermore, the multispecific fusion protein comprising the dual functional domain structure can be designed in various forms to encompass various functions.Therefore, the dual functional domain structure and the multispecific fusion protein comprising the dual functional domain structure according to the present invention can be used for various applications as an alternative to conventional antibodies.

The invention belongs to the technical field of cultural relic protection, and particularly relates to a preparation method and application of a wheat starch adhesive suitable for repairing paper cultural relics. The preparation method of the wheat starch adhesive suitable for repairing the paper cultural relics comprises the following steps: S1, dissolving plant proteolytic enzyme in water, and uniformly stirring to prepare a plant proteolytic enzyme solution; s2, adding wheat flour into the vegetable protein hydrolase solution, and uniformly stirring; s3, further performing centrifugal separation, and washing, dehydrating and drying an obtained solid product for multiple times to obtain wheat starch; and S4, gelatinizing wheat starch to obtain the wheat starch adhesive. According to the method, the wheat flour is treated by adopting the vegetable protein hydrolase, so that protein combined with wheat starch can be efficiently removed, and the wheat starch with relatively high purity and dispersed particle morphology is released; and the formed wheat starch adhesive can significantly improve the folding resistance and tensile strength of paper in the repair of paper cultural relics.

The invention relates to the technical field of daily chemicals, and provides an oat amino acid surfactant and a preparation method thereof. The preparation method comprises the following steps: dissolving oat protein polypeptide in deionized water, heating to 50-70 DEG C, adjusting the pH value of a system to 4-10, adding proteolytic enzyme, stirring for 4-6 hours, maintaining the pH value of the system to 4-10, cooling to room temperature, and drying to obtain the oat oligopeptide. The preparation method comprises the following steps: dissolving a sodium hydroxide solution into deionized water, controlling the temperature to be 50-70 DEG C, adjusting the pH value of a system to be 11-12, dropwise adding fatty acyl chloride, keeping the pH value of the system to be 11-12, continuously keeping the pH value of the system to be 11-12 after dropwise adding, and reacting for 0.5-1.5 hours; performing curing; and cooling to 40-60 DEG C, decolorizing, and adjusting the pH value of the system to 6.5-7.5 to obtain the oat amino acid surfactant. The preparation method is simple in process, and the obtained oat amino acid surfactant is light in chromaticity and low in smell.

Provided herein are cytokines or functional fragments thereof that, in some embodiments, are engineered to be masked by a masking moiety at one or more receptor binding site(s) of the cytokine or functional fragment thereof. In some embodiments, the cytokines are engineered to be activatable by a protease at a target site, such as in a tumor microenvironment, by including a proteolytically cleavable linker. In some embodiments, the proteolytically cleavable linker links the cytokine to the masking moiety, links the cytokine to a half-life extension domain, and / or links the masking moiety to a half-life extension domain. The masking moiety blocks, occludes, inhibits (e.g., decreases) or otherwise prevents (e.g., masks) the activity or binding of the cytokine to its cognate receptor or protein. Upon proteolytic cleavage of the cleavable linker at the target site, the cytokine becomes activated, which renders it capable of binding to its cognate receptor or protein with increased affinity.

The invention discloses a method for determining activity of host cell proteolytic enzyme of hydrolyzed polysorbate, and belongs to the technical field of biological detection. The method comprises the following steps: adding a solid-phase reaction substrate (containing a polysorbate analogue) and a to-be-detected sample into a buffer solution to react, removing the solid-phase reaction substrate by utilizing a magnetic adsorption method after the reaction is finished, and carrying out fluorescent quantitative PCR amplification reaction on the residual reaction solution to determine the content of free DNA in the reaction solution. Host cell proteolytic enzyme activity to hydrolyze polysorbate is determined by free DNA content. The method effectively solves the problems of insufficient sensitivity, complex operation and unstable result in the traditional detection method, obviously improves the detection precision and the experimental operation efficiency, enhances the quality monitoring in the drug research and development stage and the safety guarantee of the final product, improves the quality control level of the protein biological drugs, and has a wide application prospect. The effectiveness and the safety of the protein biological medicine in clinical medication are ensured.

The present invention relates to a composition comprising a solid carrier, a proteinase or a fragment thereof immobilized on the surface of the solid carrier, a protective layer for protecting the proteinase or the fragment thereof by embedding the proteinase or the fragment thereof, and a functional component immobilized on the surface of the protective layer, wherein the functional component fixed on the surface of the protective layer is a polymer comprising repeating units, and each repeating unit comprises at least one amino group and / or at least one mercapto group. The invention also relates to a method for producing said composition.

The present disclosure relates to formulations comprising catmint oil, a proteolytic enzyme, an antioxidant, a surfactant; and a solvent, as well as to pharmaceutical compositions comprising the formulations, methods and uses of the formulations to treat or prevent respiratory infections and methods of reducing infectivity of a virus using the formulations.

The present invention provides, among other things, a masked cytokine comprising an interleukin 2 (IL-2) polypeptide, a VHH masking moiety, an anti-PD1 targeting moiety, and an engineered Fc domain comprising a tumor-associated protease cleavage site. In such masked cytokine, the IL-2 polypeptide is engineered to be activatable by a protease at a target site, such as in a tumor microenvironment. The VHH masking moiety blocks, occludes, inhibits (e.g., decreases) or otherwise prevents (e g masks) the activity or binding of the cytokine to its cognate receptor or protein. Upon proteolytic cleavage of the cleavage site in the Fc domain, the IL-2 polypeptide becomes activated, which renders it capable of binding to its cognate receptor or protein with increased affinity.

The present invention provides a novel proteolytic enzyme composition more particularly an orally administered pro¬teolytic enzyme composition comprising of one or more acid proteases, one or more alkaline protease and one or more plant proteases. More particularly, the composition comprises of microbial (fungal, bacterial or other microbes) protease enzymes, proteases from plant and animals proteases thereof.

To enable high-sensitivity detection of banana that might cause allergies contained in food ingredients and products even if the amount is minute.SOLUTION: A method of detecting banana is provided, comprising extracting protein from a sample, obtaining an enzyme digest of the extracted protein using a protease, and analyzing the enzyme digest to detect at least one peptide of SEQ ID NO: 1 using a mass spectrometer so as to qualitatively or quantitatively determine the presence or absence of banana protein in the sample.SELECTED DRAWING: Figure 1

The invention discloses a method for simultaneously extracting DNA (deoxyribonucleic acid) and RNA (ribonucleic acid), and relates to the technical field of molecular biology, the method comprises the following steps: sample lysis incubation: co-incubating a biological sample with a lysis buffer solution, a reducing agent and proteolytic enzyme; adding a binding solution, namely adding the binding solution into the mixture subjected to lysis incubation, wherein the binding solution at least comprises a chaotropic agent, a nonionic surfactant and alcohol; according to the method, the biological sample is incubated together with the cracking buffer solution which does not contain chaotropic agent and chelating or complexing agent, the reducing agent and the proteolytic enzyme, and the anionic surfactant and the buffer compound in the cracking buffer solution have a synergistic effect and are matched with the reducing agent and the proteolytic enzyme, so that the biological sample can be efficiently cracked without physical crushing treatment; and even complex samples such as fiber tissues and high cell content can be fully cracked, so that DNA and RNA released from the samples are more complete, and nucleic acid loss and breakage caused by insufficient cracking are reduced.

The invention relates to bacillus subtilis as well as a construction method and application thereof, and the bacillus subtilis is a gram-positive bacterium from soil, also belongs to one kind of probiotics, and has multiple benefits to a human body. The Irisin is a novel muscle factor which is generated by shearing a transmembrane fibronectin type III domain protein 5 (fibronectin type III domain-conforming protein 5, FNDC5) through a proteolytic enzyme, and has the effects of alleviating obesity, improving insulin resistance, resisting inflammation and the like. According to the invention, a strain of bacillus subtilis capable of expressing human active irisin is constructed, and meanwhile, a probiotic oral embedding method is provided, so that the in-vivo activity and effect of the bacillus subtilis are improved. The expression strain not only can express and produce the tectoridin protein with activity and without endotoxin, but also can exert the activity of probiotics through oral administration after embedding.

The invention relates to the technical field of pollution treatment, and particularly discloses a composite bio-enzyme preparation for polluted water treatment and a preparation method of the composite bio-enzyme preparation. The invention relates to a composite biological enzyme preparation for polluted water treatment. The composite biological enzyme preparation comprises the following raw materials in parts by weight: 25-30 parts of an amylolytic enzyme compound, 20-25 parts of a proteolytic enzyme compound, 15-20 parts of lipase, 10-15 parts of cellulose hydrolase, 15-20 parts of flora freeze-dried powder, 10-15 parts of porous calcium alginate microspheres, 5-8 parts of chitosan modified silicon dioxide and 3-5 parts of an enzyme stabilizer. According to the application, the content of total microcystic toxins and the content of refractory organic matters in the polluted water can be obviously reduced, the sewage treatment effect is facilitated, and secondary pollution in the sewage treatment process is reduced.

Combinations of cannabinoids, vitamins, trace elements, bioactive components, bioflavonoid polyphenols, proteolytic enzymes, amino acids, and antioxidants which work independently and synergistically for the prevention, treatment, and inhibition of excess cytokines caused by the immune response to extreme viral and bacterial infections, including the inhibition of COVID-19, Rhinovirus and / or other virus spike proteins and mRNA vaccine spike proteins from attaching to body organ cells, penetrating cell membranes, and the replication of virus particles and spike proteins inside the cells. Inhibition of inflammation and prevention of long-term side effects, health issues (long-haulers), and disease caused by the SARS-COV-2 and its variants, Rhinovirus, other viruses, bacteria, and mRNA vaccines are provided by the nutraceutical compositions. Specific combinations of these compounds work synergistically to increase the efficacy of the independent nutraceuticals in treatment.

The present application provides the use of proteolytic enzymes to reduce or eliminate interfering antibodies in immunoassays for antigens and / or antibodies. The present disclosure utilizes biological methods or biological methods in combination with chemical methods to disrupt the structure of interfering antibodies, thereby attenuating or even eliminating the signal (false positive) or masked signal (false negative) from interfering antibodies, which can significantly improve the accuracy of reagent detection.

Combinations of cannabinoids, vitamins, trace elements, bioactive components, bioflavonoid polyphenols, proteolytic enzymes, amino acids, and antioxidants which work independently and synergistically for the prevention, treatment, and inhibition of excess cytokines caused by the immune response to extreme viral and bacterial infections, including the inhibition of COVID-19, Rhinovirus and / or other virus spike proteins and mRNA vaccine spike proteins from attaching to body organ cells, penetrating cell membranes, and the replication of virus particles and spike proteins inside the cells. Inhibition of inflammation and prevention of long-term side effects, health issues (long-haulers), and disease caused by the SARS-CoV-2 and its variants, Rhinovirus, other viruses, bacteria, and mRNA vaccines are provided by the nutraceutical compositions. Specific combinations of these compounds work synergistically to increase the efficacy of the independent nutraceuticals in treatment.

The invention belongs to the technical field of biological pharmacy, and particularly relates to a purification method of lumbrukinase suitable for an injection. Affinity chromatography and ion exchange chromatography are adopted, and impurities such as viruses and 45k brown lumbrukinase are removed; the lumbrukinase obtained by purification is identified as proteolytic enzyme, has dual pharmacological effects of thrombolysis and fibrinogen reduction, is free of adverse reactions such as viruses, heat sources and allergy, can be prepared into powder injection and water injection, and can be used as a medicine for treating thrombolysis and fibrinogen reduction. The pharmaceutical composition can be used for thrombolysis treatment of acute myocardial infarction and acute cerebral infarction in a large dose of 1,000,000-2,400 million units, and can also be used for defibrination treatment of acute cerebral infarction in a small dose of 25,000 units. In addition, raw materials are rich in source, low in price and suitable for industrial production.

The present invention provides a chimeric polynucleotide comprising: a first nucleic acid sequence encoding a first polypeptide selected from: alpha S1 casein, alpha S2 casein, and beta casein, a whey polypeptide, or a functional analog thereof, a second nucleic acid sequence encoding a cleavage site of a proteolytic enzyme; and, a third nucleic acid sequence encoding a kappa casein macropeptide, Further provided are a chimeric polypeptide, as well as compositions comprising same, and methods of using same, such as for producing an edible composition comprising casein and / or whey polypeptide aggregates, such as a dairy substituent curd.

Provided is a method for measuring glycated hemoglobin in a sample containing hemoglobin and plasma, the method including reacting glycated hemoglobin in a sample containing hemoglobin and plasma, a proteolytic enzyme, and a fructosyl peptide oxidase to generate hydrogen peroxide, and measuring the generated hydrogen peroxide, wherein the reaction is performed in the presence of ferrocyanide and a ferrocene derivative.

The present invention relates to a composition for producing chitin and a method for producing chitin, comprising a Bacillus subtilis EG1 strain or a culture solution thereof as an active ingredient. Since the Bacillus subtilis EG1 strain or a culture solution thereof according to the present invention has proteolytic and lipolytic enzyme activities, it can be effectively utilized for producing chitin from a mealworm larva substrate.

This invention discloses a nanopore peptide sequencing method based on host-guest interactions. The method includes the following steps: 1) sequentially hydrolyzing the peptide to be sequenced into monomeric amino acids using a proteolytic enzyme; 2) linking the monomeric amino acids to a load molecule containing the guest molecule structure; 3) performing single-channel recording in a nanopore after the host molecule and guest molecule have bound. This invention provides a de novo sequencing method that utilizes the sequential hydrolysis of peptides by exopeptidases to obtain the peptide sequence information; by linking the load molecule to a single amino acid, the current blocking ratio of the characteristic signals generated after the binding of 20 encoding amino acids to the load molecule is accurately measured. The characteristic current is contributed by a single amino acid, eliminating decoding difficulties; the target peptide dosage is small, and the detection limit is low; the experimental operation is safe and simple, with most reactions carried out in an aqueous phase, without involving toxic reagents, making it green and safe.

Use of caseinolytic protease P (ClpP) function and / or concentration as a biomarker for predicting the response of a neoplastic disease, preferably cancer or another disease where enhancing ClpP activity may provide a therapeutic benefit, to a compound of Formula I. In other aspects it relates to methods and kits, as well as methods of treatment involving the use of the biomarker.

Disclosed is a dissolvable, gel-forming film, and methods for its use, comprising a water-soluble cellulose ether, a hydrophilic rheological modifying agent, and an active proteolytic enzyme or other drug substance. The gel-forming film has a water content of less than 15% w / w and is capable of forming a hydrogel when contacted with water or other aqueous medium. The disclosed films achieve delivery of stable proteolytic enzymes to the desired site of action in a manner that provides uniform delivery of the enzymes.

The present invention is in the field of tissue and organ bioengineering and refers to an enzyme-free process to produce a tissue-specific extracellular matrix solution, based on a decellularized extracellular matrix, comprising: (a) decellularizing a tissue or organ to obtain the isolated extracellular matrix; and (b) hydrolyzing the isolated extracellular matrix in an acidic environment, in the absence of proteolytic enzymes, optionally followed by purification of the solution. Still, the present invention refers to an enzyme-free process to produce a tissue-specific hydrogel. Thus, the present invention also relates to hydrogels produced according to the current processes, and to ancillary inventions, having the tissue-specific extracellular matrix solution, the hydrogel, and / or the products derived from the tissue-specific extracellular matrix solution as the core that connects the inventive aspects disclosed in this document.

The present invention addresses the problem of providing: a method for confirming that a proteolytic enzyme has been removed from a protein-containing composition; a method for producing a protein-containing composition from which a proteolytic enzyme has been removed; and a method for confirming that an autolysate of a proteolytic enzyme is not contained in a final product. This method for confirming that a proteolytic enzyme has been removed comprises: using an ultrafiltration membrane having a molecular weight cutoff of at most 10 kDa to exclusively ultrafilter a proteolytic enzyme; and measuring the proteolytic enzyme activity and / or protein content of a permeated liquid and / or a non-permeated liquid.