222 results about "Collagenase" patented technology

The invention provides a preparation method of non-denatured type II collagen capable of increasing bone mineral density and improving bone elasticity, and belongs to the technical field of biologication.The preparation method comprises the steps that an EDTA-citric acid buffer solution containing beta-mercaptoethanol is adopted for decalcifying cartilage, a supercritical CO2 extraction technology is adopted for degreasing, and the collagen is obtained; alkaline protease and collagenase II are adopted for substrate directional enzymolysis and gradient digestion; then the non-denatured type II collagen is separated by adopting three-phase centrifugal separation, and the non-denatured type II collagen with high extraction rate, high purity and high structural integrity can be obtained by adopting bionic H column chromatography purification.

The invention discloses bacillus subtilis for secretory expression of collagenase and application of the bacillus subtilis. The invention provides bacillus subtilis for secretory expression of collagenase and application of the bacillus subtilis in fat cell extraction, and after optimization of an expression element promoter and a signal peptide and large-scale fermentation of a fermentation tank, the highest enzyme activity can reach 669U. ML <-1 >. After purification, adipose-derived cells are successfully extracted and separated from mouse adipose cells, and after passage, the adipose-derived cells present the form of adipose cells. The method shows that the method has a good application prospect in the aspect of fat cell extraction.

The invention discloses starch-collagen composite hydrogel as well as a preparation method and application thereof. Collagen is catalyzed by microbial transglutaminase to form a covalent cross-linked network, and meanwhile, hydroxypropyl starch is introduced to construct a physical interpenetrating network through molecular chain penetration and hydrogen-bond interaction; and chondroitin sulfate is further integrated to strengthen the network structure and biological activity through electrostatic interaction. The method is mild in condition and does not need a toxic chemical cross-linking agent, the obtained composite hydrogel has an interpenetrating double-network structure and has excellent mechanical properties, enzymatic degradation resistance and cell affinity, the compression modulus of the composite hydrogel is remarkably improved, the composite hydrogel can keep structural stability for a long time in a collagenase environment, cell adhesion and proliferation can be effectively promoted, and the composite hydrogel has a good application prospect. The hydrogel can be widely applied to tissue engineering scaffolds, wound dressings, drug sustained-release carriers and cartilage repair materials.

The present invention discloses a livestock and poultry manure biochar-based fertilizer, comprising the following raw materials in parts by weight: 10-42% of dilute protein solution and collagenase-modified livestock and poultry manure biochar, 20-40% of urea, 7-25% of diammonium phosphate, 7-24% of potassium chloride or potassium sulfate, 0.5-1% of modified starch, and 6-23% of bentonite. The livestock and poultry manure is selected from one of chicken manure, cow manure, and pig manure. After the biochar is modified with collagenase, the surface of the biochar has more phenolic hydroxyl groups, carboxyl groups, aldehyde groups, and hydroxyl functional groups, which are effective for NH4 + , PO4 3‑ , K + , has a stronger adsorption effect, collagenase modification makes the protein distribution more uniform, and the generated groups form a stable chemical bond with the biochar surface: it improves the dispersibility of the modified biochar in aqueous solution and reduces agglomeration; it has better chemical stability in acidic and alkaline environments.

The invention relates to the technical field of cell separation and culture, in particular to a method for separating and culturing haematochrome cells of plectropomus leopardus. The separation method of the plectropomus leopardus haematochrome cells is created for the first time, the EDTA digestive juice is used for pre-digesting epidermis cells on the outer layers of the plectropomus leopardus fins, then the collagenase digestive juice is used for digesting connective tissues (pigmented cells) of internal dermis, digestive enzymes such as trypsin and other mechanical methods which have great damage to the cells are not used, and the method is suitable for large-scale production of the plectropomus leopardus haematochrome cells. The plectropomus leopardus haematochrome cell can reduce cell damage, can be effectively separated to obtain the plectropomus leopardus haematochrome cell which is high in purity, good in activity and capable of being cultured in vitro, and lays a foundation for subsequent experiments such as single cell sequencing, cell in-vitro culture and molecular function verification.

This invention describes a wound dressing product for active continuous debridement of devitalized tissues in non-healing wounds including diabetic ulcers, pressure ulcers, burn injuries and other etiologies. The present invention pertains to the principle of continuous wound debridement which makes necrotic tissue more susceptible for removal and hence enhances progressive wound healing. The dressing contains an active ingredient, such as collagenase which serves to debride wounds in-situ. In the present invention purified Collagenase (90% pure) was deposited onto several wound dressing materials. A key feature of this invention is that the activity level of the Collagenase used was substantially preserved.

The invention relates to the technical field of medical cosmetology, in particular to a porous hollow glass bead capable of slowly releasing collagen and application of the porous hollow glass bead. The porous hollow glass bead capable of slowly releasing the collagen comprises a porous hollow glass bead body, the outer wall of the porous hollow glass bead body is provided with a pore channel structure penetrating through the exterior and an inner cavity of the hollow glass bead body, and the pore channel and the inner cavity of the porous hollow glass bead body are filled with collagen gel. The compressive strength of the hollow glass beads is utilized to improve the mechanical supporting effect, the collagen gel is subjected to catalytic decomposition under the action of collagenase in a body so as to achieve the slow-release effect, the outer walls of the hollow glass beads slow down the reaction time through physical isolation, and then the slow-release time is prolonged; meanwhile, ions such as Si and Ca released by the hollow glass beads can promote cells to secrete collagen, the biological activity is improved, and the three beneficial effects of physical support, collagen slow release and biological safety are achieved.

The invention relates to a method for extracting high-purity extracellular vesicles from liver tissues of pelteobagrus vachelli, which comprises the following steps of: firstly, mixing collagenase II and collagenase IV, performing enzymolysis on the tissues, adding a CaCl2 solution, incubating, and digesting the liver tissues into a tissue suspension; then removing cells and fragments through low-speed centrifugation, and filtering to remove impurities by using needle filters of 0.8 [mu] m and 0.22 [mu] m; and finally, further purifying by combining ultracentrifugation with a high-precision iodixanol density gradient centrifugation method, and merging density layers with high purity to obtain high-purity EVs. The method has the characteristics of high extraction purity, high efficiency and simplicity and convenience in operation, and has a wide application prospect.

The invention discloses an eggshell membrane peptide for enhancing bone mineral density as well as a preparation method and application thereof, belongs to the technical field of egg by-product processing, and particularly relates to a preparation method of the eggshell membrane peptide for enhancing bone mineral density, which comprises the following steps: carrying out vacuum freeze drying and crushing on an eggshell membrane to obtain eggshell membrane powder; mixing the eggshell membrane powder with deionized water to obtain an eggshell membrane solution; adding keratinase, performing enzymolysis, and taking supernate; adding collagenase, carrying out enzymolysis, and centrifuging to take supernate, so as to obtain primary enzymatic hydrolysate; adding alkaline phosphatase, performing enzymolysis and centrifugation, and taking supernate to obtain enzymatic hydrolysate; and performing ultrafiltration and drying to obtain the eggshell membrane peptide. According to the eggshell membrane peptide for enhancing the bone mineral density, keratin cross-linking is destroyed through enzymolysis of keratinase, active peptide capable of stimulating osteoblast proliferation is released through enzymolysis of collagenase, phosphopeptide calcium binding sites are further activated through enzymolysis of alkaline phosphatase, calcium salt deposition is promoted, and the eggshell membrane peptide for enhancing the bone mineral density is provided.

The present invention relates to a cartilage tissue single cell extraction kit and an extraction method thereof, the kit comprises: (1) a first enzyme for enzymatic hydrolysis, the first enzyme for enzymatic hydrolysis comprising one or more of trypsin, chondroitinase ABC and fucosidase; (2) a second enzyme for enzymolysis, wherein the second enzyme for enzymolysis comprises collagenase II, neutral protease II, hyaluronidase and Dnase I; and (3) a basic solution, wherein the basic solution comprises a basic culture medium and / or a buffer solution. The invention also provides a method for extracting single cells from cartilage tissues by using the kit. The cartilage tissue single-cell suspension prepared by the method has the advantages of high cell yield, high activity and the like, and has a wide application prospect.

The invention discloses a novel colorectal cancer organoid culture method, which obviously improves the activity of primary cells (more than or equal to 90%) and the forming efficiency of the organoid (the success rate is more than 95%) through alcohol gradient cleaning, collagenase-hyaluronidase combined digestion and stent-free culture technologies. The improved culture medium takes R-Spondin1 as a core, nicotinamide and a small-molecule inhibitor are combined, and long-term amplification of organoids (passage is more than or equal to 10 times) is supported. The method is suitable for individualized drug screening, tumor microenvironment simulation and drug resistance mechanism research, has the advantages of high efficiency, stability and low cost, and provides a reliable model for precise medical treatment of colorectal cancer.

The invention belongs to the technical field of biology, and particularly relates to bacillus velezensis HALZ676 as well as application and a method of the bacillus velezensis HALZ676 to cowhide fermentative degradation, the bacillus velezensis HALZ676 is separated from preserved pig trotters, and by analyzing biological characteristics, whole genome and metabonomics differences of the bacillus velezensis HALZ676, the cowhide can be efficiently degraded, and the cowhide can be efficiently degraded. It is found that peptide components smaller than 3000 Da in cow leather degradation liquid obtained through fermentation of the strain HALZ676 account for 68% or above, and the cowhide degradation liquid has remarkable antioxidant capacity and antibacterial capacity; the cowhide fermentation liquor is mainly used for promoting conversion and enrichment of antioxidant, anti-inflammatory and anti-cancer metabolites through a linoleic acid metabolic pathway, so that the antioxidant, anti-inflammatory and anti-cancer capacities of the cowhide fermentation liquor are enhanced; the cowhide fermented by the strain HALZ676 can enrich functional active components, and a theoretical basis and a promising strategy are provided for high-valued production of processing byproducts of livestock and poultry skin fermented by probiotic bacillus with high yield of collagenase.

The invention discloses a cod bone-derived high-activity calcium chelating peptide and a preparation method thereof, and belongs to the technical field of functional biological product preparation. The calcium chelating peptide provided by the invention is derived from a cod bone collagen enzymatic hydrolysate and is obtained through affinity chromatography separation of hydroxyapatite, the calcium chelating activity is 4.9-5.2 mu g / mg, and the calcium chelating peptide contains aspartic acid and glutamic acid enriched peptide fragments, through deep processing of cod bones, high-valued utilization of cod bone resources is realized, the high-activity calcium chelating peptide is prepared, and the calcium chelating peptide has a wide application prospect. And a high-quality raw material is provided for the development of a novel calcium supplement.

The invention provides a preparation method of a single-cell sequencing suspension fixedly preserved in situ. The invention provides digestive juice for preparing a single-cell suspension. The digestive juice is a solution composed of collagenase II and neutral protease. The invention also provides a kit containing the digestive juice. The invention provides application of the digestive juice to preparation of a kit and application of the kit to in-situ fixed preservation of cells and preparation of a single-cell suspension. The invention also provides a preparation method of the in-situ immobilized preserved cell and the single-cell suspension, and the single-cell suspension prepared by the preparation method.

The invention belongs to the field of biological material extraction, and particularly relates to a method for extracting collagen peptide from marine organisms, which comprises the following steps: S1, crushing marine organism raw materials, soaking the crushed marine organism raw materials in a citric acid buffer solution containing alginate oligosaccharide for swelling treatment, and then performing high-voltage pulse electric field treatment to obtain pretreated collagen slurry; and S2, carrying out first-stage enzymolysis treatment on the pretreated collagen slurry through alkaline protease, and then carrying out second-stage enzymolysis treatment through collagenase and flavourzyme. S3, performing separation treatment through a first ultrafiltration membrane to obtain first filtrate, performing deodorization treatment on the first filtrate through a first nanofiltration membrane of which the surface is coated with a beta-cyclodextrin adsorbent to obtain second filtrate, and performing separation treatment on the second filtrate through a second nanofiltration membrane to obtain third filtrate. And S4, concentrating, desalting and drying the third filtrate to obtain collagen peptide powder. Through the synergistic effect of multiple steps, the concentration of the molecular weight of the collagen peptide and the efficient deodorization are realized on the basis of retaining the active structure of the collagen.

The invention provides a bovine type I collagen terminal end peptide characteristic polypeptide, the bovine type I collagen terminal end peptide characteristic polypeptide comprises an N terminal end peptide characteristic polypeptide and a C terminal end peptide characteristic polypeptide, the amino acid sequence of the N terminal end peptide characteristic polypeptide is Q * LSYGYDE, and the amino acid sequence of the C terminal end peptide is LSFLPQPPQE. The invention also provides application of the bovine type I collagen terminal peptide marker polypeptide in detection of N-terminal peptide and C-terminal peptide of bovine type I collagen. The invention also provides a preparation method of a collagen enzymolysis product containing the bovine type I collagen terminal peptide characteristic polypeptide. The bovine type I collagen tail end terminal peptide marker polypeptide provided by the invention is a special terminal peptide at the tail end of bovine type I collagen. The terminal peptide marker polypeptide is used as a marker and can be used for quickly identifying the bovine type I collagen terminal peptide and accurately and quantitatively analyzing the bovine type I collagen terminal peptide. The qualitative and quantitative detection methods are simple to operate and high in sensitivity.

The present invention provides a composition comprising collagenase as an enzyme; calcium, histidine, and glycine as coenzymes, and a method for stabilizing enzymes, the method comprising the preparation of the composition by adding histidine and glycine to collagenase. According to the present invention, the enzyme composition and method for stabilizing enzymes can minimize or inhibit the agglutination of enzymes, maintain the concentration of freeze-dried enzymes even during long-term storage, and provide a formulation that has excellent rehydration time and hygroscopicity when restored to a liquid state, as well as superior enzyme activity.

The invention relates to a collagenase-loaded LDH (layered double hydroxide) / EDTA (ethylene diamine tetraacetic acid) layered double hydroxide nano material as well as a preparation method and application thereof. The collagenase-loaded LDH / EDTA layered double hydroxide nano material is prepared from a zinc-aluminum layered double hydroxide LDH carrier, ethylenediamine tetraacetic acid EDTA inserted between layers of the LDH carrier through anion exchange, and collagenase loaded on the surface of the LDH carrier through an electrostatic adsorption effect. The invention further discloses a preparation method of the collagenase-loaded LDH / EDTA layered double hydroxide nano material for preparing the collagenase-loaded LDH / EDTA layered double hydroxide nano material for preparing the collagenase-loaded LDH / EDTA layered double hydroxide nano material.

The present application relates to a kind of anti-aging composition and its application of brightening and shrinking pores.The composition includes the following components: halocyprid extract, purple magnolia extract, desert rose leaf cell extract, rain lily labiatae flower extract and white lily extract, and the mass ratio of the halocyprid extract, purple magnolia extract, desert rose leaf cell extract, rain lily labiatae flower extract and white lily extract is (0.2-2):(0.001-0.1):(0.001-0.1):(0.001-0.1):(0.001-0.1).The composition of the present application can effectively improve the clearance rate of collagenase, maintain the elasticity and tightness of skin, have good anti-aging and pore-shrinking effect, and can also effectively remove hydroxyl radicals, have good whitening and brightening effect.

The invention relates to a low-temperature gradient enzymolysis technology for collagen peptide in the body wall of a sea cucumber, and discloses mild decomposition and efficient extraction of collagen in the body wall of the sea cucumber by virtue of a multi-section temperature zone reaction, enzyme seed compounding control and on-line regulation system. And the collagen peptide with concentrated molecular weight, strong biological activity and high terminal product quality is prepared by the low-temperature gradient enzymolysis technology. The method is characterized by comprising the following steps: cutting the body wall of the sea cucumber to obtain a particle size suitable for enzymolysis, adding the cut body wall of the sea cucumber into a neutral buffer solution, stirring at normal temperature, and adjusting the pH value to 6.5-7.5, so that the tissue of the body wall of the sea cucumber is fully expanded and uniformly dispersed to be more suitable for the subsequent enzymolysis process; the method comprises the following steps: carrying out staged reaction on the body wall of the sea cucumber and an enzyme at different temperatures, carrying out fine separation and terminal treatment on an enzymatic hydrolysate, and immediately cooling a system to 4 DEG C after the reaction of the enzymatic hydrolysate is finished.

The application provides a preparation method of non-denatured type II collagen for increasing bone density and improving bone elasticity, and belongs to the technical field of biotechnology. The application adopts EDTA-citric acid buffer containing beta-mercaptoethanol to decalcify cartilage, adopts supercritical CO2 extraction technology to defat, adopts alkaline protease and collagenase II to perform matrix directional enzymolysis, and performs gradient digestion; then three-phase centrifugal separation is adopted to separate non-denatured type II collagen, and biomimetic H column chromatography purification is adopted, so that non-denatured type II collagen with high extraction rate, high purity and high structural integrity can be obtained.

A multi-part processed human amniotic composition configured to treat Peyronie's disease in a subject in need thereof by intracorporeal injection of the composition into the corpus cavernosum of the subject to reduce plaque size associated with Peyronie's disease. The multi-part processed human amniotic composition includes a micronized human amnion composition; and an aqueous human amniotic fluid filtrate configured to reconstitute and suspend the micronized human amnion composition therein. In certain aspects, the multi-part processed human amniotic compositions are not processed with exogenous enzymes during production thereof and do not include exogenous enzymes, such as collagenase, added thereto.

The invention discloses an in-vitro biochemical test method for evaluating the anti-wrinkle effect of cosmetics and raw materials. The in-vitro biochemical test method comprises the following steps: step 1, preparing a working solution; 2, preparing a sample solution; step 3, adding an enzyme working solution; step 4, loading samples in groups; 5, adding a substrate and reacting; step 6, developing treatment; step 7, determining absorbance; 8, calculating, analyzing and processing data; and step 9, result judgment: analyzing and evaluating the anti-wrinkle effect of the sample through a statistical means. According to the method, based on a collagenase inhibition test, when peptide bonds between GLY and GLY in Z-GLY-PRO-GLY-GLY-PRO-ALA-OH are specifically hydrolyzed by collagenase, GLY-PRO-ALA is released, and the activity of the collagenase is indirectly evaluated by measuring the change of a light absorption value at 565 nm after the GLY-PRO-ALA reacts with a color developing agent, so that whether a sample to be detected has an anti-wrinkle effect or not is judged. The method is economical and fast, the test process is controllable, the test system is standardized, and result deviation caused by individual difference can be avoided.

1. The name of this design product: packaging box (collagenase outer box). 2. Purpose of this design product: product packaging. 3. The key design points of this product are the combination of shape, pattern and color. 4. The picture or photo that best illustrates the key points of the design: Design 1 expansion diagram. 5. The design for which protection is sought includes color. 6. Designate Design 1 as the base design.

The invention relates to a preparation method of a placenta trophoblast-derived transporter and application of the placenta trophoblast-derived transporter in a medicine for treating spontaneous abortion, which comprises the following steps of: taking out a placenta of a suckling mouse, cutting the placenta into fine tissue blocks, resuspending by using digestive juice, digesting in a water bath kettle at 37 DEG C for 20 minutes, and then stopping digestion by using 1640 culture containing high-temperature heat-inactivated fetal calf serum; diluting and neutralizing collagenase digestive juice according to 5 times of volume; and centrifuging the collagenase digestive juice for multiple times to obtain a precipitate, namely the exosome. And carrying out density gradient centrifugation on the coarse migration body by using Optiprep as a density medium, washing, precipitating and centrifuging by using PBS on an ultra-high-speed centrifuge by using a horizontal rotor to obtain the placental trophoblast source migration body. The invention finds that the generation of the placental trophoblast cell source migration body is related to the physiological process of pregnancy for the first time, and the placental trophoblast cell source migration body can enter the circulatory system and reach the placenta through in-vitro supplementation.

The content disclosed herein relates to a method of treating subcutaneous cellulite on the thighs or buttocks of a human subject by administering an effective amount of collagenase and evaluating a reduction in the severity of subcutaneous cellulite through one or more scales.

The invention relates to the technical field of neutrophile granulocyte separation, in particular to a mouse neutrophile granulocyte separation method which comprises the following steps: (1) anesthetizing and disinfecting a mouse; (2) tissue sampling; (3) tissue digestion; (4) preparing a cell suspension; (5) purifying cells; (6) cell culture; according to the method disclosed by the invention, isoflurane is adopted in the anesthesia disinfection step, so that central nervous activity of mice can be rapidly inhibited, tissue damage caused by stress reaction is avoided, and chemical stimulation can be reduced while body surface microorganisms are cleared through combination of ethanol disinfection and sterile normal saline flushing, and aortic tissues with intact activity are provided for subsequent material taking. In the compound enzyme digestive juice, collagenase I can specifically degrade aortic wall collagenous fibers, trypsin dissociates adhesion connection between cells, neutral protease assists in decomposing elastic fibers, and DNA enzyme I is supplemented to inhibit cell aggregation, so that the contradiction between incomplete digestion and large cell damage of the traditional enzyme juice is effectively solved.

The content disclosed herein relates to a method of treating subcutaneous cellulite on the thighs or buttocks of a human subject by administering an effective amount of collagenase and evaluating a reduction in the severity of subcutaneous cellulite through one or more scales.

The present application relates to cell preparation process in the field of bioengineering, disclose a kind of high-efficiency preparation mouse primary hepatocyte method, comprising: perfusion needle is placed in reverse direction to liver direction through inferior vena cava, utilize the blood vessel clamp with elastic micro-tooth damping surface to lock the perfusion needle and inferior vena cava wall;The overflow damping of hepatic portal vein outflow end is dynamically regulated by blood vessel clamp, to construct controlled positive pressure permeation environment in liver vascular bed;Collagenase solution is radially permeated to liver parenchyma using the environment, the present application makes enzyme solution radial deflection and cross blood sinus barrier into di's cavity by constructing micro-pressure environment, eliminate the digestion dead zone originally due to flow path preference, ensure that enzyme molecule and extracellular matrix are uniformly contacted in global dimension, realize the collaborative promotion of tissue cutting efficiency and total amount of cell release, ensure that the output cell population has highly consistent physiological state.

The invention provides cross-linked recombinant collagen gel. The cross-linked recombinant collagen gel consists of the following components in percentage by mass: 0.4-1.6% of recombinant III-type humanized collagen; 0.2%-2% of a D-core; 0.1% to 0.5% of carbomer; 0.1% to 0.6% of triethanolamine; 1.0%-10% of glycerol; the collagen gel disclosed by the invention has the advantages of low immunogenicity, high safety, no toxicity, no stimulation, good biocompatibility, structural stability, collagen enzymolysis resistance and the like, and meanwhile, the invention also provides a preparation method and application of the cross-linked recombinant collagen gel.