68 results about "Lumbrokinase" patented technology

The invention discloses a preparation method of Lumbrokinase, comprising the steps of grinding, deslagging, salting-out, centrifuging, ultra-filtrating, column chromatography isolating, ultra-filtrating, degerming, spray-drying and the like. The preparation method of Lumbrokinase provided by the invention is that ethanol is added as adjuvant to remove hybrid protein by physical settlement and ultrafiltration during the homogenate of earthworms, and then a column chromatography purification is performed so that the capacity of the column is reduced, thereby the yield of the Lumbrokinase is increased, the preparation cost is reduced, the waste fluid to be treated is decreased and the production capacity is increased greatly. The preparation method of Lumbrokinase has the characteristics of good quality controllability and stability of the production process.

The invention discloses a high-purity earthworm activating enzyme preparation method and take it as raw material preparation. The invention through uses the monoclonal antibody technology realization earthworm activating enzyme, this method characteristic is attains the enzyme the shift strongly, the activeness is high, achieves above each milligram protein 200,000 units; After the highly effective liquid chromatography test is the sole component; Swims in the Sds- polyacrylamide gelatin electricity presents a belt, the molecular weight is 32000+ / -2000 Dalton; The amino acid sequence analysis, the N- terminal 10 amino acids sequences are: He-Val-Gly-Gly-He-Glu-Ala-Arg-Pro-Tyr, take states the high-purity earthworm activating enzyme to be allowed to make as the raw material medicine may supply to take orally, intermuscular injection, as well as the intravenous injection and the venous transfusion many kinds of ways for the medicine preparation, uses in the thrombus disease treatment.

The invention provides a polysulfone dialysis membrane as well as a preparation method and application thereof. The polysulfone dialysis membrane is formed by connecting a bioactive molecule onto a polydopamine surface modified polysulfone membrane, wherein the bioactive molecule is any one or a combination of at least two of human serum albumin, bovine serum albumin, natto kinase, lumbrokinase, thrombomodulin and hirudin. The polysulfone dialysis membrane provided by the invention can guarantee the activity of the bioactive molecule and has a good anticoagulation function; the pure water flux is 65L / m<2>.h.bar to 140L / m<2>.h.bar, the retention rate on BSA (Bull Serum Albumin) is 95 percent or above and the retention rate on lysozyme is 25 percent to 35 percent; compared with an unmodified polysulfone membrane, the adsorption amount on the BSA, fibrinogen and blood platelets is remarkably reduced. The method provided by the invention has moderate conditions and a simple preparation process; the deterioration of membrane structure and performance and the inactivation or denaturation of bioactive macromolecules are not caused; the polysulfone dialysis membrane has a wide application prospect.

The invention relates to a method for extracting earthworm protein and lumbrokinase from earthworms. The method is characterized by utilizing an approach to combining chromatographic columns with ultrafiltration for extracting the lumbrokinase and earthworm protein from the earthworms. According to the method, a radio-frequency technique is utilized to achieve a protein extraction sterility mechanism, the appropriate temperature for occurrence of enzyme activity is controlled, and the method is safe, sanitary and free of pollution; by means of the differential control over the earthworm protein, extraction and separation of the earthworm protein are achieved, and batched production of the earthworm protein and lumbrokinase is facilitated.

The invention provides a lumbrukinase enteric-coated pellet capsule and a preparation method of the lumbrukinase enteric-coated pellet capsule. According to the preparation method, the centrifugal granulation technology is adopted, the lumbrukinase is prepared into pellet and is coated by low-temperature-filmed enteric coating and then is filled into a gastric soluble capsule; and a dried-powder wrapping method is adopted, which has the advantages of being short in preparation time, high in drug loading rate, high in yield of target pellet (20 to 30 meshes), and smooth and round in appearance of the target pellet. By adopting the lumbrukinase enteric-coated pellet capsule, the pellet can be dissolved and absorbed by the small intestine, so that the lumbrukinase can be effectively prevented from being damaged by gastric acid, and the treatment effect can be ensured; compared with common capsule, the capsule provided by the invention has the advantage that the pellet is filled in the gastric soluble capsule, so that the shortcomings that the powdered medicine flies and is adhered on the capsule can be avoided, the acting efficiency can be remarkably improved, the quality and output of the capsule can be obviously increased, and the producing cost can be reduced; compared with tablets, the capsule has the advantages that the pellet is small in granularity, hardly influenced by the food delivery frequency of the alimentary canal, and is high in bioavailability; and by adopting the lumbrukinase enteric-coated pellet capsule, the controllability of the quality is improved.

A method for extracting high-activity lumbrukinase from earthworms comprises the following steps: (1) choosing earthworms; (2) cleaning; (3) refining; (4) centrifuging with high speed centrifugal machine, removing impurities in the slurry after salting, collecting supernatant, washing the precipitate twice with salt solution, centrifuging again, collecting supernatant again, and mergingthe collected supernatant; (5) conducting first ultrafiltration on the merged supernatant to filter out high molecules insoluble in water; conducting ultrafiltration on the supernatant subjected to ultrafiltration, concentrating the effective components, and when the concentrate reaches a concentration of 5%-10%, sending the concentrate into a vacuum freeze dryer; (6) conducting vacuum freeze drying; and (7) packaging. The invention can obtain all lumbrukinase from earthworms by one step, has the characteristics of simpleness, feasibility and high effective components in the extract, and is applicable to mass production.

The invention relates to a preparation method of lumbrokinase enteric-coated pellets, comprising the following steps of: washing fresh live eisenia foetida with purified water, preparing homogenate by a colloid mill, repeatedly freezing and thawing, heating, centrifuging at high speed to obtain supernatant, separating the supernatant by an anion exchange chromatographic column, eluting, concentrating, filtering to obtain a lumbrokinase concentrated solution, mixing the concentrated solution with an adhesive, spraying onto the blank pellet cores to prepare pellets, coating the pellets with enteric coatings, and screening. The raw materials of the enteric-coated pellets prepared by the method dispense with a freeze drying process, thus simplifying the production process and lowering the production cost.

The invention discloses a rice planting soil conditioner. The rice planting soil conditioner is prepared by mixing 50 parts of a lumbrokinase stock solution, 4 parts of an earthworm cast fermentation solution, 10 parts of fulvic acid, 8 parts of wood vinegar, 30 parts of cattle manure, 2 parts of boric acid, 1 part of ammonium hydrogen carbonate, 60 parts of rice straw, 3 parts of sodium sulphite, 1 part of phosphorus lime powder and various trace elements. The rice planting soil conditioner can effectively improve soil hardening and other problems, can improve the soil structure and can effectively increase the soil nutrient content; especially during rice planting, soil nutrients are lost due to often irrigation and water discharge, and the rice planting soil conditioner can effectively improve the fertility of rice planting soil and can promote nutrient absorption of rice.

The invention relates to the technical field of fertilizer and in particular relates to earthworm biological organic fertilizer. The earthworm biological organic fertilizer is prepared from the following raw materials in parts by weight: 10 to 20 parts of earthworm protein obtained in a lumbrokinase extracting process, 50 to 60 parts of earthworm cast, 10 to 20 parts of traditional Chinese medicine dreg, 5 to 10 parts of cattle bone powder, 2 to 10 parts of anti-disease bacterium agent, 5 to 10 parts of fulvic acid, 10 to 25 parts of organic matters, 25 to 40 parts of humic acid, 0.5 to 2 parts of trace elements, 10 to 20 parts of various amino acids, cellulase and xylanase. The earthworm biological organic fertilizer is used for crops, has abundant nutrients and can be used for activatingsoil, improving soil and keeping fertility and preventing and controlling diseases.

A composite medicine used also as the additive of cosmetics and health-care food for lowering blood viscosity, dissolving thrombus, and regulating blood pressure, blood sugar and blood fat is prepared through extracting to obtain L-polybasic metallothionein (L.pMT), lumbrikinase and antibacterial peptide, proportionally mixing it with the extracts of astragalus root and red sage root, thiooctanoic acid and 1,6-pectose-zinc biphosphate, and dispensing.

The invention provides a technological method for extracting lumbrokinase by utilizing living earthworms. The technological method comprises the following technological steps: collecting the earthworms, extracting the lumbrokinase, precipitating, centrifuging, sterilizing and drying and pulverizing. The extracted lumbrokinase can be used for promoting the increasing of the quantity of the earthworms and microorganisms in soil; meanwhile, the soil can be effectively activated and loosened and the content of organic matters in the soil is increased; a soil structure and physicochemical properties are improved, and absorption of root systems on nutrients is facilitated.

The invention provides a lumbrokinase enteric-coated tablet and a preparation method thereof. The lumbrokinase enteric-coated tablet comprises following components in parts by weight: 5 to 15 parts of lumbrokinase, 7.5 to 22.5 parts of hydroxy propyl cellulose, 3.8 to 11.4 parts of powdered steatile, 1.5 to 4.5 parts of magnesium stearate, 2.3 to 6.9 parts of carboxy methyl starch sodium, and 63.9 to 185.7 part of cane sugar, wherein the titer of the lumbrokinase is between 0.9 multiply 104 and 2.7 multiple 104 unit / milligram. Compared to the conventional lumbrokinase preparation and the preparation technology thereof, the lumbrokinase enteric-coated tablet and the preparation method thereof are improved, the quality and curative effect of the lumbrokinase enteric-coated tablet are more stable and reliable, each parameter of the product reaches or exceeds the national drug standards, furthermore, the lumbrokinase stability of the active components is prominently improved, the technical defect that the titer of the thermal sensitive components reduces during the preparation and storage processes is overcome, so that the curative effect of the lumbrokinase on corresponding diseases in clinic is largely improved.

The invention relates to a method for preparing vegetable cultivation medium by utilizing earthworm cast. The vegetable cultivation medium is prepared by mixing common soil, earthworm cast, chicken manure, rice husks, rice straws, wheat straws, turfy soil, lumbrukinase original fluid, calcium superphosphate and fermentation inoculum, preparation of the vegetable cultivation medium is completed through fermentation and decomposition mainly, the problem that seedling medium is severely lacking is solved, and effective utilization ratio of organic fertilizer and crop straws is also improved. The produced vegetable cultivation medium has strong fertilizer-retaining and water-retaining capabilities, can effectively promote absorption of vegetable seedlings on nutrients and improve disease and insect resistance of vegetables and is easy to prepare, low in cost and full of nutrients.

The invention provides a pharmaceutical composition for dissolving thrombus and anti-coagulation, belonging to the technical field of anti-thrombus and anti-coagulation. 50-500 unit of hirudin, 600-6000 units of lumbrokinase, 2000-20000 unit of nattokinase, 0. 5-5g of edible gelatin, 0. 1-2g of sodium bicarbonate and 30-300mg of magnesium. A Chinese medicine in that form of leech, hirudin, an enzyme extracted from earthworms, based on lumbrokinase and nattokinase, as that biological enzyme protect agent (edible gelatin). The combination of gastric acid neutralizer (sodium bicarbonate) and enzyme accelerator (magnesium element) greatly improves the stability of the effective substance in vivo, improves the speed and efficiency of thrombolysis, has excellent thrombolytic effect, and is verysafe to the body, hardly produces side effects, is safe, convenient and effective for oral administration, and is safe, non-toxic and harmless.

The process of removing hybrid protein from crude lumbrokinase product includes the following steps: dissolving crude lumbrokinase product in alcohol-water solution to compound solution with coarse lumbrokinase concentration of 2-7 wt% and alcohol concentration of 0-40 vol% via stirring and filtering out insoluble matter; and adding 20 ml of the said mixed solution into high pressure reactor preheated to 20-40 deg.c, introducing CO2 of 20-40 deg.c temperature and 3.5-8.0 MPa pressure, regulating pH to 4.4, maintaining for 20-60 min to deposit hybrid protein while maintaining the active component in the solution, and separating the precipitate. The present invention has the features of mild operation condition, capacity of obtaining other active components, less loss of enzyme activity, etc. and the process has precisely controlled operation parameters and no fluctuation in the operation process.

The invention discloses medicament for treating cardiac neurosis and a preparation method of the medicament. The medicament solves the problem that existing medicament is poor in effect on treatment of cardiac neurosis and has side effects. The medicament for treating cardiac neurosis comprises the following main materials of schisandra chinensis, manyprickle acanthopanax roots, albizzia julibrissin durazz, American ginseng, radix polygoni multiflori preparata, magnetit, fungus ganoderma lucidum, angelica sinensis, prepared dried rehmannia, fushen, agarwood and semen ziziphi spinosae. The medicine further comprises the following auxiliary materials of lumbrokinase, polypeptide, multivitamins, water-soluble dietary fibers, acipimox, phosphatidylcholine and honey. The medicament has the good curative effect on common cardiac neurosis in cardiovascular diseases, and has wide application prospect in the fields of treatment of palpitation, precardium area pains, dyspnea, panasthenia, anxiety, sinus arrhythmia and the like.

The invention discloses health-care cigarettes, which comprise, in the proportion by weight, 1-106 parts of tobacco shreds or filters for making cigarettes, and 1-100 parts of lumbrukinase. Soluble health-care products, preferably the lumbrukinase, are sprayed to the tobacco shreds or filters of the cigarettes to be made, and accordingly a smoker smoking the cigarette can inhale the smoke with the lumbrukinase. According to the latest researches, the lumbrukinase not only can activate fibrinoclase to dissolve thrombus, but also can directly dissolve fibrous proteins. The effective rate of the lumbrukinase in clinical treatment of thrombus reaches more than 80%, and the lumbrukinase is further capable of reducing blood viscosity, inhibiting platelet aggregation, resisting blood coagulation, promoting blood circulation and the like, and has therapeutic effects on apoplexy sequela, arteriosclerosis, hypertension, hyperviscosity and the like.

The invention relates to a methoxy polyethylene glycol (mPEG)-lumbrukinase conjugate having a physiological activity, wherein the structure of the mPEG-lumbrukinase conjugate is mPEG-CH2-W-NH-Lumbrokinase, and the molecular weight of the mPEG is 5000-40000 Daltons. The in-vivo retention time of the mPEG-lumbrukinase conjugate is long, so the immunogenicity of the mPEG-lumbrukinase conjugate is reduced, and the thrombolytic efficacy of the mPEG-lumbrukinase conjugate is improved.

The invention provides paddy rice seedling culture soil containing chitosan. The paddy rice seedling culture soil is prepared from the following components by mixing: 10 parts of the chitosan, 30 parts of common soil, 30 parts of humic acid, 15 parts of attapulgite, 5 parts of a lumbrokinase stock solution, 2 parts of citric acid, 3 parts of calcium superphosphate, 2 parts of potassium sulfate and 3 parts of zinc sulfate. The paddy rice seedling culture soil containing the chitosan, provided by the invention, contains a large quantity of organic matters, macroelements and microelements, can meet nutrients needed by growth and development of paddy rice seedlings and has the effects of improving the quality of the seedlings and improving disease and insect resisting capabilities.

The invention discloses a lumbrukinase concentrated slurry production process capable of realizing normal-temperature environment storage and long-distance transportation. The method comprises the steps of pulping, deslagging, filtering, degreasing, desalting and concentrating, sterilizing and the like, impurities such as water-soluble impurities, denatured protein, earthworm grease and the like can be removed, the abundance of lumbrukinase in the lumbrukinase concentrated slurry is improved, the load of a separation medium of a production line is reduced, and the yield of lumbrukinase is increased; moreover, the produced lumbrukinase concentrated slurry is stable in physicochemical property, and the enzyme activity of the lumbrukinase concentrated slurry is maintained for a long time. Theproduction process is suitable for different lumbrukinase process routes, and the prepared lumbrukinase concentrated slurry can be directly used as an earthworm body raw material substitute of lumbrukinase refining engineering, can be stored and transported at normal temperature, has the characteristic of perennial quality guarantee, and is suitable for initial raw materials produced by lumbrukinase factories.

The invention discloses the preparation method of a medicine anti-plug enteric soluble capsule for curing anemic apoplexy, deficiency of vital energy and blood stasis. The Chinese medicines astragalus root, angelica and rhizoma ligustici wallichii are steeped into ethanol, the ethanol return flows twice, then is filtered; the filtrate is condensed at certain temperature; the dregs of a decoction are decocted in water with Chinese medicine rot of peony, semen persicae and safflower, then the liquid is filtered; the filtrate is condensed, and the concentrated solution is added by ethanol, stirred and standing; a supernatant fluid is given out and is carried on decompression condensation. Two condensates above are mixed and dried, then grinded so that an extractive is obtained; a fresh lumbricus is dried in shade and grinded, then steeped into water, an extract solution is obtained, decentered, condensed, a condensate is obtained, dried in vacuum at a certain temperature then grinded so that a lumbricus extract is obtained; the extract above is mixed with the lumbricus extract, then packed into the enteric soluble capsule. The effective ingredient of the Chinese medicine of the invention is extracted entirely, and the biological activity of the lumbrokinase of the lumbricus is kept so that the curing effect of the medicine is improved.

The invention discloses a lumbrukinase-based mineral soil remediation fertilizer and a preparation method thereof. The fertilizer comprises mineral soil, plant straws and animal manure, wherein the raw materials are proportioned by weight; the mineral soil accounts for 35 to 50 percent; the plant straws accounts for 15to 20 percent; the animal manure accounts for 30 to 50 percent; the mineral soilcomprises following components in parts by weight: 5-12 parts of diatomite, 7-14 parts of zeolite, 5-8 parts of sepiolite, 15-21 parts of calcium carbonate, and 5 to 9 parts of sodium alginate; the plant straws comprise the following components in parts by weight: 12-20 parts of rice straws, 15-18 parts of wheat straws, 5-8 parts of soybean straws and 10-15 parts of corn straws, and the animal manure comprises the following components in parts by weight: 15-20 parts of livestock manure, 10-15 parts of poultry manure and 20-30 parts of earthworms. The mineral soil, the plant straws and the animal manure are mixed to prepare the soil remediation fertilizer, the soil activity is effectively improved, and the remediation capacity is efficient.

The invention belongs to the field of soil heavy metal remediation and particularly relates to a composite bio-enzyme soil heavy metal remediation agent and a preparation method thereof. The compositebio-enzyme soil heavy metal remediation agent is composed of, by weight part, 10-15 parts of composite bio-enzymes, 5-10 parts of complex microbial agent, 50-60 parts of plant adsorbent and 20-30 parts of adsorption additives, wherein the complex microbial agent can enrich heavy metal ions through ion exchange, complexation and the like to improve the form of existence of the heavy metal ions insoil; lumbrokinase and chitinase can improve the granular structure as well as the physiochemical properties of soil and accordingly change the form of existence of the heavy metal ions; the plant adsorbent can effectively absorb the heavy metal ions in soil to achieve the aim of reducing the content of heavy metal; the adsorption additives can promote reaction of the composite microbial agent, the complex microbial agent and the heavy metal ions and accordingly can accelerate absorption and conversion and heavy metal.

The invention discloses low-immunogenicity lumbrukinase, a preparation method and application thereof, and belongs to the field of biological pharmacy. The method provided by the invention comprises the following steps of: first, extracting to obtain a lumbrukinase crude product from an earthworm, and then, passing the crude product through an ion exchange column to elute, purifying, colleting a component with fibrinolytic activity, freezing, drying, concentrating, separating a concentrated lumbrukinase solution by using a gel filtration chromatographic column, detecting the fibrinolytic activity by using an agarose-hemaleucin flat band method, collecting a second eluting peak with the fibrinolytic activity, wherein detected by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electropheresis), the molecular weight of the second eluting peak is 23500-27500 Da; and after the eluting peak is dialyzed, desalted and subjected to low-temperature freeze drying, carrying out molecular modification on the eluting peak by utilizing activated monomethoxy polyethyleneglycol and dextran to obtain the low-immunogenicity lumbrukinase. The lumbrukinase prepared by the invention has the characteristics of low immunogenicity, high enzyme activity and the like, and can be used for the research and the preparation of an antithrombotic drug and a drug for treating or preventing cardiovascular and cerebrovascular diseases.

The invention discloses a united preparation technology of earthworm elastic (collagen) protein and lumbrukinase. Earthworms are used as raw materials, by a freeze-etching method, a crust layer and anelastin layer are thoroughly separated, and collected earthworm exudate is used as pulp and can be used for producing the lumbrukinase; and the surplus parts are subjected to processing by a countercurrent flow suspension method for refining of elastic collagen protein, and then through the steps of glue cooking, decolorization, membrane separation, concentration, freeze drying and the like, theearthworm elastic collagen protein is obtained. According to the united preparation technology disclosed by the invention, the utilization degree of the earthworms is increased, no additive componentsare used in the process of producing the earthworm elastic collagen protein and the lumbrukinase, the quality of the earthworm elastic (collagen) protein and the lumbrukinase is increased, the production cost of the earthworm elastic (collagen) protein and the lumbrukinase is reduced, besides, the preparation technology is simple, the vitality of the lumbrukinase cannot be influenced, and industrialized production can be realized.

The invention discloses a method for preparing a leaf fertilizer from a lumbrokinase. The method comprises the step of mixing lumbrokinase, an earthworm cast fermentation solution, a plant ash solution, urea and dipotassium phosphate according to the ratio of 50:45:1:0.3:3 to prepare the leaf fertilizer. The method is simple in preparation and relatively low in cost; and the earthworm cast fermentation solution contains abundant nutrient substances and a great number of active substances such as beneficial microorganisms and organic matters as well as macroelements and microelements, can be used for improving the adverse-resistant characteristics such has cold resistance and drought resistance and the absorption capacity of crops to nutrients and can also be used for accelerating the absorption of plant leaves to nutrients, increasing the yield of crops and improving the quality of the crops.

The present invention provides human body sexual hormone improving health preserving food. The health preserving food is prepared from nereis succinea as a main material and prepared by compounding with sea cucumbers and fish oil, wherein the nereis succinea, sea cucumbers and fish oil have effects of mutual supports and synergistic interactions in functions. The health preserving food can safely and effectively promote human body secretion of sexual hormones, improves levels of the sexual hormones in the human body, enables eaters to restore sexual vitality and maintains reproductive health, youth and vitality. Perinereis aibuhitensis, especially obtained perinereis aibuhitensis in waters of Dongshan Island, Zhangzhou, Fujian, is preferred, and the health preserving food is prepared. The nereis succinea is rich in biosterins, amino acids, trace elements, active peptides, taurine, lumbrokinase, plasmin and other active substances, rich in nutrition, high in contents of active ingredients and free heavy metal pollution, so that the nereis succinea prepared health preserving food has advantages of being good in quality and more prominent in health preserving effects.

The invention belongs to the technical field of soil heavy metal pollution treatment, and particularly relates to a biological remediation agent for cadmium pollution and a preparation method thereof.The invention provides a biological remediation method aiming at cadmium pollution, which comprises the following steps: a compound bacterial solution is selected in the method, a cell wall containing chitin, chitosan and shell polysaccharide as main components in aspergillus versicolor can adsorb cadmium ions so as to reduce cadmium toxicity; bacillus thuringiensis can be used as microbial low-toxicity insecticide, and thermophilus thermophilus can improve soil and increase soil fertility; in addition, the lumbrokinase and earthworm dung used in the method can absorb the cadmium in the soil,thereby reducing the proportion of the cadmium in the soil and achieving the purpose of remedying the polluted soil; the method also adds waste roots, stems and leaves of diatomite, shell powder, cabbage and cauliflower which can effectively absorb high-concentration cadmium as an adsorbent, and are low in cost and easy to obtain.

The invention discloses a snailase rhinitis ointment and a preparation method thereof. The snailase rhinitis ointment comprises raw materials including snailase, lumbrokinase and traditional Chinese medicine composition ointment. The traditional Chinese medicine composition ointment comprises raw materials as follows: magnolia flowers, angelica roots, centipede minima, propolis, wild chrysanthemumflowers, mint, lanolin, white muscardine silkworm and broadleaf holly leaves. The formula is simple to combine, contains no chemical additives and has no toxic or side effects.