134 results about "Proteolytic enzymes" patented technology

The invention relates to a recombinant pichia pastoris genetically engineered bacterium for expressing recombinant full-chain-length humanized collagen as well as a preparation method and application of the recombinant pichia pastoris genetically engineered bacterium, and belongs to the technical field of genetic engineering. The recombinant pichia pastoris genetically engineered bacterium disclosed by the invention expresses recombinant full-chain-length humanized collagen, and a ku70 gene of the recombinant pichia pastoris genetically engineered bacterium is knocked out and knocked into a recombinant 4-proline hydroxylase encoding gene. Recombinant full-chain-length humanized collagen is optimized, a 4-proline hydroxylase gene and a collagen gene are co-expressed in pichia pastoris to obtain hydroxylation modified recombinant collagen, and a corresponding recombinant pichia pastoris genetically engineered bacterium is provided, has proteolytic enzyme A defect and hydroxylation modification capability, and can be used for preparing a recombinant collagen product. The stability of the collagen in the fermentation process is improved. The recombinant full-chain-length humanized collagen provided by the invention is 100% homologous with III-type collagen derived from a human body, so that the immune risk is extremely low, and the biological activity is more perfect.

[Problem] To detect, at high sensitivity even when contained in a minute amount, almond having the risk of causing an allergic reaction when being contained in a food raw material, a product, or the like. [Solution] This almond detection method is for qualitatively or quantitatively determining whether or not an almond protein is present in a sample, by performing a step for extracting proteins from the sample and a step for obtaining enzymatic digestion products from the extracted proteins by using a protease, and analyzing the enzymatic digestion products by detecting at least one peptide selected from those represented by the group consisting of SEQ ID NO: 1-3 by using a mass spectrometer.

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.

The invention relates to the field of biological medicine, in particular to a polypeptide composition with bacteriostatic activity and a preparation method thereof.The polypeptide composition is prepared by wrapping polypeptide I with modified polyethylene glycol, then wrapping with nano-silver and finally modifying polypeptide II with the nano-silver. The outer layer is sequentially wrapped with spherical particles of modified polyethylene glycol, nano-silver and polypeptide II, and the polypeptide II and the nano-silver are promoted to be combined with a bacterial membrane and destroy the bacterial membrane through the non-specific electrostatic interaction of the polypeptide II and the nano-silver, so that the polypeptide II has relatively high antibacterial performance; the outer-layer polypeptide is degraded by proteolytic enzyme or plays a role with a bacterial membrane, so that the inner-layer polypeptide I wrapped by the modified polyethylene glycol is exposed, the antibacterial effect is further played, and the proteolytic stability of the polypeptide is improved; due to the spherical structure, the prepared polypeptide composition with the antibacterial activity has excellent antibacterial performance, meanwhile, the stability can be improved, and the polypeptide composition has wide application prospects.

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.

A nucleic acid sequence measurement method for measuring a target RNA having a specific nucleic acid sequence included in a cell suspension by hybridization, including a step of heating the cell suspension at 100° C. to 200° C. in a pressurized state to obtain an RNA extract, a step of adding a proteolytic enzyme to the RNA extract to cause the RNA extract to be reacted, thereby preparing a sample solution, a step of supplying the sample solution to a device for nucleic acid sequence measurement, the device being equipped with a fluorescent probe that hybridizes with the target RNA, a step of subjecting the target RNA and the fluorescent probe to a hybridization reaction, and a step of measuring fluorescence from the device for nucleic acid sequence measurement.

[Problem] To detect soybean, which has the potential to cause an allergic reaction, in a food ingredient, a product, or the like, at high sensitivity even when the content thereof is extremely small. [Solution] Provided is a method for detecting soybean whereby the presence or absence of soy proteins in a sample can be qualitatively or quantitatively determined by carrying out a step for extracting a protein from the sample and a step for obtaining an enzymatic digest from the extracted protein through use of a proteolytic enzyme, analyzing the enzymatic digest, and using a mass spectrometer to detect at least one peptide selected from the group consisting of SEQ ID NOs: 1-3.

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.

To detect a mackerel potentially causing an allergic reaction even with a trace quantity thereof included in a raw material of a food product or manufactured articles at high sensitivity.SOLUTION: A method for detecting a mackerel performs: a step of extracting a protein from a specimen; a step of obtaining an enzymatic digestion product from the extracted protein by a protease; and analyzing the enzymatic digestion product and detecting one or more peptides selected from a group consisting of sequence numbers 1 to 4, using a mass spectroscope. The method thus determines whether or not a mackerel protein is present in the specimen qualitatively or quantitatively.SELECTED DRAWING: Figure 1

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.

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 invention discloses a composite deodorizing agent and a preparation method thereof, the composite deodorizing agent is composed of the following components: a plant extraction component, the plant extraction component comprises a tea extract, a ginger extract, an oak leaf extract, a lavender extract, a licorice extract, a green persimmon extract, a pine bamboo extract and a schizonepeta extract; the microbial component comprises a proteolytic enzyme, an aldehyde hunting factor and nano diatomite; the auxiliaries comprise a stabilizer, titanium dioxide with a topological structure, syringol, a fermented rice product, an odor modifier, a chelating agent, an antioxidant, a plant preservative and a solvent. The deodorization effect of plants and microorganisms can be effectively integrated, the deodorization capacity after compounding is improved, and the safe and environment-friendly deodorization process is achieved.

The invention discloses a preparation method and application of a hexavalent multi-epitope fusion protein of avian pathogenic Escherichia coli. The invention preferably selects antigenic epitopes of the curli fimbriae subunit CsgA, lipoprotein SlyB, maltose porin LamB, outer membrane protein PhoE, proteolytic enzyme activator NlpD and lipoprotein Pal main structural subunit of avian pathogenic Escherichia coli, and preferably connects with the heat-labile enterotoxin LTb fusion protein in series to construct an antigenic multi-epitope fusion protein of six proteins of avian pathogenic Escherichia coli. The antigenic multi-epitope fusion protein has a wide serotype coverage, is non-allergenic, has strong immunogenicity, and produces strong specific humoral immunity and cellular immune response after immunizing animals.

To provide a passaging culture technique for cells that enables more efficient production of higher-quality cells.SOLUTION: A cell treatment device 100 includes a culture controller 53 and a vibration controller 54. The culture controller 53 applies proteolytic enzymes to cultured cells to reduce adhesion. The vibration controller 54 adds a culture medium to a culture vessel containing cultured cells with reduced adhesion and applies physical stimulation, thereby detaching the cultured cells. The culture controller 53 cultures the cultured cells detached by the vibration controller 54 in a set culture condition, thereby performing cell passaging culture.SELECTED DRAWING: Figure 8

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 present invention relates to a composition comprising an injectable mesenchymal stem cell-hydrogel and a preparation method thereof. Specifically, the injectable mesenchymal stem cell-hydrogel composition prepared by the method of the present invention has the following advantages: stem cells are attached to the scaffold of the hydrogel beads, so the stem cells are not easy to disappear or apoptosis after injection, and the paracrine effect of the stem cells is continuously exerted. The stem cells are slowly released while the hydrogel is decomposed, thereby increasing the survival rate. In addition, it can be prepared into an injectable dosage form without using proteolytic enzymes, so that healthy cells can be used without damaging the cell membrane. At the same time, the cryopreservation solution can be easily removed from the mesenchymal stem cell-hydrogel beads after freezing and thawing.

[Problem] To detect sesame (Sesamum indicum), which have the risk of causing an allergic reaction, with high sensitivity when the sesame is contained in a food raw material, a product or the like, even if the content of the sesame is very small. [Solution] Provided is a method for detecting sesame, the method comprising a step for extracting a protein from a sample, a step for allowing the extracted protein to react with a protease to produce an enzymatically digested product, and analyzing the enzymatically digested product, wherein at least one peptide selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:3 is detected using a mass spectrometer to qualitatively or quantitatively determine whether or not a sesame protein is present in the sample.

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.