82 results about "Fibrinolytic enzyme" patented technology

An isolated Bacillus amyloliquefaciens Ba-BPD1 having an Accession No. of DSM 21836 is provided. This novel strain has unique 16S ribosomal RNA sequenced as SEQ ID NO:1 and produces amylase, protease, cellulase and lipase, fibrinolytic enzyme to show their biodegradation capacities. Further, B. amyloliquefaciens Ba-BPD1 produces the antibiotic substances, such as iturin, fengycin and surfactin, and has antimicrobial capacity for inhibiting the fungal or bacterial growth. In conclusion, the novel strain of Bacillus amyloliquefaciens Ba-BPD1 and its products can be applied in agriculture, wastewater treatment, food industry and chemical industry.

An isolated Bacillus amyloliquefaciens Ba-BPD1 having an Accession No. of DSM 21836 is provided. This novel strain has unique 16S ribosomal RNA sequenced as SEQ ID NO:1 and produces amylase, protease, cellulase and lipase, fibrinolytic enzyme to show their biodegradation capacities. Further, B. amyloliquefaciens Ba-BPD1 produces the antibiotic substances, such as iturin, fengycin and surfactin, and has antimicrobial capacity for inhibiting the fungal or bacterial growth. In conclusion, the novel strain of Bacillus amyloliquefaciens Ba-BPD1 and its products can be applied in agriculture, wastewater treatment, food industry and chemical industry.

The present invention relates to a novel microorganism Streptomyces megasporus SD5. The present invention also relates to a process for the isolation of said Streptomyces megasporus SD5. The invention also relates to a novel fibrinolytic enzyme actinokinase extracted from said microorganism and to a process for the extraction of said enzyme. In another aspect, the invention also pertains to a method for the treatment of thrombolytic disorders using said enzyme.

The invention provides bacillus velezensis as well as a separation screening method and application thereof, and belongs to the field of microbe technology and food processing. The invention providesthe bacillus velezensis with high fermentation capability; the bacillus velezensis is obtained through screening; during the fermentation, the capability of high yielding protease, amylase and fermented bean plasmin is realized; the bacillus velezensis can be applied to food fermentation in a large scale when being prepared into a food fermentation strain; fermentation food obtained through fermentation by using the bacillus velezensis, the contents of free amino acid and douchi fibrinolytic enzyme are high; the nutrition is rich; the characteristics of easy digestion, thrombus prevention andthe like are realized.

The invention discloses a fungal fibrinolytic enzyme and a preparation method thereof. The fungal fibrinolytic enzyme is prepared by taking a Cordyceps militaris strain as a strain and performing liquid fermentation culture, separation and purification. The relative molecular weight of the fibrinolytic enzyme is about 21,000. The Cordyceps militaris fibrinolytic enzyme has good thrombolytic performance, is free from obvious acute toxicity, has prospects for clinical trial, development and application, and adds a new member to a rare fibrinolytic enzyme family. Particularly, a preferable fibrinolytic-enzyme Cordyceps militaris strain C.LSG-1 obtained through severe screening is rough in growth conditions, short in enzyme production cycle and capable of harvesting a large amount of Cordyceps militaris mycelium during enzyme production, and is the strain excellent in production performance. The Cordyceps militaris fibrinolytic enzyme is an extracellular enzyme which is particularly beneficial to subsequent separation and purification during preparation. The fungal fibrinolytic enzyme takes corn protein with extensive sources as a main culture medium, thereby having low cost for raw materials and providing a novel way for developing and utilizing the prior resources.

The invention discloses an armillaria luteo-virens fibrinolytic enzyme and a production method thereof. Fermentation and cultivation are made to armillaria luteo-virens ZJUQH CGMCC No.1884 to get fermented liquid and the armillaria luteo-virens fibrinolytic enzyme is separated and purified from the fermented liquid. The invention first realizes that artificial cultivated armillaria luteo-virens ZJUQH CGMCC No.1884 can be used for producing the fibrinolytic enzyme. The produced armillaria luteo-virens fibrinolytic enzyme takes agricultural and sideline products such as bean pulp, soybean flour and wheat bran with low cost as the main raw materials, thereby being safe and nonpoisonous. Being produced with a liquid fermentation method, the armillaria luteo-virens fibrinolytic enzyme is a hyper strain with relatively higher yield among the reports of epiphytes. Extraction and purification can be carried out easily and the armillaria luteo-virens is harmless, frozen-dry health products that contain thalli and enzyme liquid can be directly developed, thus providing a new thought and an evidence for developing and using novel edible fungus used in foods and medicines.

A medical device includes a base material having an immobilized fibrinolytic enzyme and dextran sulfate. The dextran sulfate has a molecular weight that is less than 40 kilo dalton (kDa). The medical device is formed from at least a base material. A fibrinolytic enzyme is immobilized on the base material. The fibrinolytic enzyme is stabilized with a dextran sulfate having a molecular weight of less than 40 (kDa).

The invention relates to fibrinolytic enzyme separated and prepared from a marine invertebrate, wherein 7 amino acid sequences on the N-terminal are NH2-IVGGRDV-OH, the molecule weight is about 35 kDa, and experimental study results show that the fibrinolytic enzyme and drug compositions thereof provide significant effects for anti-thrombus and stroke prevention and treatment, and further provide good effects in the field of tumor treatment.

The invention discloses a cDNA sequence and an amino acid sequence of coding lugworm fibrinolytic enzyme which are respectively shown in SEQIDNo.1 and SEQIDNo.2 in a sequence table, and belongs to the field of medical biological engineering. The cDNA sequence has the beneficial effects that according to the cDNA sequence of the lugworm fibrinolytic enzyme, the coding gene of the fibrinolytic enzyme can be cloned from digestive tract tissue of lugworm, and can be expressed in a prokaryotic expression system or a eukaryotic expression system through a gene recombination technology. The recombinant lugworm fibrinolytic enzyme has plasminogen activation activity, and can be used as a new thrombolytic medicament in genetic engineering.

The invention discloses a fibrinolytic enzyme-producing Bacillus subtilis and a fermentation method and application thereof. The Bacillus subtilis is isolated and sieved from a traditional Chongqing fermented food, i.e., fermented soya beans. The Bacillus subtilis DC27 is collected in China Center for Type Culture Collection under CCTCC NO: M 2015573. The invention further provides a liquid fermentation method of the Bacillus subtilis, and the fibrinolytic enzyme in obtained fermentation broth has an activity of up to 4506 IU / ml. The Bacillus subtilis is from foods and can secrete the high-activity fibrinolytic enzyme; the Bacillus subtilis is safe and nontoxic, and is applicable to the development of functional foods or drugs for preventing or relieving thrombotic diseases; and the Bacillus subtilis has a promising application prospect.

A new Bacillus subtilis strain and its use in preparing medicine for treating thrombosis are provided. Particularly, the use of fibrinolytic enzyme produced by Bacillus subtilis in preparing medicine for treating thrombosis is provided. The present invention also discloses a method for producing said fibrinolytic enzyme (nattokinase). It is efficient and cost-effective to prepare nattokinase by using the new Bacillus subtilis strain of the invention, suitable for mass industrial production.

The present invention discloses an armillaria luteo-virens fibrinolytic enzyme and a production method thereof. Fermentation and cultivation are made to armillaria luteo-virens ZJUQH CGMCC No.1884 to get fermented liquid and the armillaria luteo-virens fibrinolytic enzyme is separated and purified from the fermented liquid. The armillaria luteo-virens fibrinolytic enzyme and the production method thereof of the present invention first realizes that artificial cultivated armillaria luteo-virens ZJUQH CGMCC No.1884 can be used for producing the fibrinolytic enzyme. The produced armillaria luteo-virens fibrinolytic enzyme takes agricultural and sideline products such as bean pulp, soybean flourand wheat bran with low cost as the main raw materials, thereby being safe and nonpoisonous. Being produced with a liquid fermentation method, the armillaria luteo-virens fibrinolytic enzyme is a hyper strain with relatively higher yield among the reports of epiphytes. Extraction and purification can be carried out easily and the armillaria luteo-virens is harmless, frozen-dry health products that contain thalli and enzyme liquid can be directly developed, thus providing a new thought and an evidence for developing and using novel edible fungus used in foods and medicines.

The present invention relates to a novel microorganism Streptomyces megasporus SD5. The present invention also relates to a process for the isolation of said Streptomyces megasporus SD5. The invention also relates to a novel fibrinolytic enzyme actinokinase extracted from said microorganism and to a process for the extraction of said enzyme. In another aspect, the invention also pertains to a method for the treatment of thrombolytic disorders using said enzyme.

The invention discloses a rapid production method of lactic acid bacteria fermented soya beans capable of dissolving fibrous protein. According to the rapid production method, two-stage solid-state fermentation is adopted to produce functional fermented soya beans. The rapid production method comprises following steps: soybean is selected, and impurities are removed; soybean is immersed in warm water at a ratio of 1:2-3 for 6 to 9h, wherein the temperature of the warm water ranges from 25 to 30 DEG C; steaming is carried out for 8 to 15min at 110 to 121 DEG C, after cooling, an obtained product is inoculated with 3 to 5% (V / W) of SYL-1 Bacillus subtilis; fermentation is carried out for 42 to 54h at 26 to 32 DEG C so as to obtain a fermented soya beans semi-finished product, and the fermented soya beans semi-finished product is inoculated with 3 to 5%(V / W) of SYL-2 Lactobacillus brevis, is stirred to be uniform, and is subjected to fermentation for 42 to 54h at 32 to 37 DEG C; after fermentation, an obtained product is subjected to 72h of post-fermentation in a refrigerator at 4 DEG C so as to obtain a finished product. According to the rapid production method, bioengineering technology is adopted; two-stage fermentation, including bacillus subtilis primary fermentation and lactic acid bacteria post-fermentation, is carried out via combination with traditional fermentation method; flavor components such as fermented soya beans ester substances are increased; nutritional ingredients such as peptone, polypeptides, and amino acids are increased; functional ingredients such as douchi fibrinolytic enzyme, oligosaccharides, organic acids, and lactic acid bacteria are increased; the lactic acid bacteria fermented soya beans is capable of adjusting intestinal flora balance, dissolving fibrous protein, treating constipation, preventing oxidation, and preventing cardiovascular diseases.

The invention discloses a bacillus licheniformis fibrinolytic enzyme produced by the method: bacillus licheniformis ZJUEL31410 CGMCC 1397 is fermented and cultured to get fermented liquid, then separation and purification are carried out to the fermented liquid to get the bacillus licheniformis fibrinolytic enzyme. The invention firstly discovers that the bacillus licheniformis ZJUEL31410 CGMCC 1397 can be used for producing fibrinolytic enzyme. The bacillus licheniformis fibrinolytic enzyme products produced by the invention take cheap agricultural by-products of soybean pulp, expanded soybeans and bran as raw materials, thereby being nontoxic. Besides, a liquid fermentation method is adopted in the production, resulting in short technique process, low cost, high industrial production efficiency and high activity of enzyme, thereby being with development value.

This invention discloses a method for manufacturing a fibrinolytic enzyme from whole-body or coelomic fluid of Urechis unicinctus, which comprises the steps of: (1) grinding and / or separating the tissues, (2) centrifuging to obtain the clear supernatant, (3) ultrafiltrating, (4) freeze-drying to obtain freeze-dried powders, or precipitating with acetone at a low temperature and vacuum filtrating to obtain the precipitate, (5) dissolving, purifying through a gel permeation column, and freeze-drying to obtain the fibrinolytic enzyme. The enzyme has good anticoagulation effects, and can be used to manufacture drugs for treating cardiovascular and cerebrovascular diseases.

The invention provides a thrombolytic active substance extracted from a Guizhou fermented blank bean and a product thereof. The fermented blank bean is a folk favorite conventional food in Guizhou. The substance is prepared by taking soybeans as a raw material and carrying out microorganism fermentation, and the flavor tastes sweet but smells smelly. People think that the food stimulates the appetite and is favorable for digestion, and the substance is generally edible and a favorite food of people in Guizhou for a long time. The natural thrombolytic active substance extracted from the Guizhou conventional fermented blank bean and the product thereof have obvious thrombolytic activity, can reduce blood viscosity, promote blood circulation, and are used for treating and preventing heart cerebrovascular diseases such as thrombus diseases and the like. The thrombolytic active substance has the biochemistrical characteristics that: molecular is 28KD; the optimal pH is 8.0; the substance is stable below 50 DEG C, and is not stable in the stomach environment; and the substance has the hrombolytic effects of activating plasminogen to be converted into fibrinoclase, and directly dissolving fibrin in the thrombus.

The invention discloses a preparation method of a sea cucumber egg fermentation product with functions of lowering blood pressure and resisting blood coagulation. The preparation method of the sea cucumber egg fermentation product with the functions of lowering blood pressure and resisting blood coagulation comprises the following steps: washing sea cucumber eggs, and freeze-drying the washed sea cucumber eggs, so that freeze-dried sea cucumber egg powder is prepared for later use; mixing the freeze-dried sea cucumber egg powder with water, adding glucose, and adjusting the initial pH, so that a sea cucumber egg basic culture medium is prepared; sterilizing the basic culture medium; adding a bacillus natto bacterial suspension, and carrying out shaking culturing, so that a sea cucumber egg fermentation broth is prepared; centrifuging the sea cucumber egg fermentation broth, and collecting the obtained supernatant; and then, concentrating the supernatant, and freeze-drying the concentrate, so that the sea cucumber egg fermentation product freeze-dried powder with the functions of lowering blood pressure and resisting blood coagulation is prepared. The sea cucumber eggs are fermented with the bacillus natto according to the preparation method, so that fibrinolytic enzymes with the effect of dissolving thrombus are produced; moreover, polypeptides which are easy to absorb and have the functions of resisting blood coagulation and lowering blood pressure are also produced. The sea cucumber egg fermentation product can be directly eaten as a food, or prepared into health-care functional products after further developments, including concentrating, separating and so on.

The invention discloses a method for preparing fibrinolytic enzyme. The method comprises the steps of utilizing fibrinolytic enzyme strains, namely bacillus to produce the fibrinolytic enzyme and performing liquid fermentation through bacteria. The method is low in production cost and strong in feasibility, and can be used for mass production. The fibrinolytic enzyme is directly made into capsules, medicinal granules, oral liquid or oral buccal tablets, or is used as functional food ingredient in health-care food or common food. The method improves the prior method of screening fibrinolytic-enzyme producing strain, adopts a liquid medium for enrichment culture and a screening method with panel primary screening which takes fibrin as main matrix and shake-flask secondary screening, and simultaneously determines fibrinolytic activity. The enzyme activity of obtained fermentation broth is higher than 588.4 u / ml.

The present invention discloses an application of a bacillus subtilis fibrinolytic enzyme in treating bronchitis. The bacillus subtilis fibrinolytic enzyme is prepared by fermenting by bacillus subtilis QK02 strain, the strain is preserved in China Center for Type Culture Collection, and the strain number is CCTCCNO:M203078. The bacillus subtilis fibrinolytic enzyme has a very good therapeutic effect on the bronchitis and has a good cough-stopping effect.

This invention relates generally to a method of preparation of a phyto-percolate having enzymatic, specifically fibrinolytic, activity in the body of mammals, including humans. The phyto-percolate is derived from fresh water mixture including algae that produces an enzyme having proteolytic activity. The invention further relates to the use off the phyto-percolate in a variety of disease states.

The invention relates to a synthesis method of a mutant venin fibrinolytic enzyme gene, in particular to a full-length target gene segment prepared by the segmented synthesis of target genes and the artificial synthesis of the overlapping polymerase chain reaction (PCR) technology. The amino acid sequence (GenBank No.P28891) which is published by the national center for biotechnology information (NCBI), of the venin Fibrolase protein of the Agkistrodon contortrix is utilized to design 14 segments of amino acid sequences of which average length is 60bp, wherein the sequences are shown in SEQ ID No.3-16; and the extension and amplification of the synthetized segments are performed to obtain the complete target gene. By adopting the preparation method in the invention, the coding genes of Alfimeprase is easier to obtain, the complicated step that mRNA is converted to cDNA through reverse transcription is eliminated and the corresponding mutant sites can be designed according to needs.

The invention relates to a preparation method of an actinomycetes fibrinolytic enzyme. Specifically, the method comprises the following steps: (1) adding ammonium sulfate powder to a fermentation supernatant containing actinomycetes fibrinolytic enzyme until a saturation degree is 25-35%W / V, standing by for a whole night at 4 DEG C and centrifuging so as to obtain a supernatant, adding ammonium sulfate powder to the supernatant once again until a saturation degree is 55-65%W / V, standing by for a whole night at 4 DEG C, centrifuging so as to obtain a precipitate, and dissolving the precipitate by virtue of a PB buffer solution so as to obtain crude enzyme liquid; and (2) centrifuging the crude enzyme liquid obtained from the step (1) at low temperature so as to remove impurities; purifying an obtained supernatant subsequently through Octyl-Sepharose FF hydrophobic interaction chromatography and Phenyl-Sepharose HP hydrophobic interaction chromatography so as to obtain a chromatographically pure enzyme; dialyzing and desalting the enzyme, and freeze-drying the enzyme so as to obtain fibrinolytic enzyme freeze-dried powder which can be preserved for a long time at minus 20 DEG C. The separation and purification method of the actinomycetes fibrinolytic enzyme disclosed by the invention has the advantages of simple operation steps and high recovery rate.

The invention discloses active bio-enzyme conditioning extract. The active bio-enzyme conditioning extract comprises, by mass, 14%-16% of lactobacillus acidophilus, 1.35%-15.5% of bifidobacterium longum, 12.5%-14.5% of lactobacillus casei, 11.8%-13.8% of oviform bifidobacteria, 11%-13% of lactobacillus jensenii, 8%-10% of Raman lactobacillus, 6.5%-8.5% of bifidobacterium thermophilum, 1.5%-2.5% of pullulan, 1.5%-2.5% of bacillus subtilis fibrinolytic enzyme, 1.5%-2.5% of lysozyme, 1.5%-2.5% of lysozyme beta-glucan, 1.5%-2.5% of plankton extract, 1.3%-1.7% of bletilla striata root extract, 0.8%-1.2% of scutellaria baicalensis extract, 1%-1.4% of centella asiatica extract, 0.4%-0.6% of licorice extract, 0.7%-0.9% of polygonum cuspidatum extract and 0.3%-0.5% of matricaria extract. The active bio-enzyme conditioning extract is an active bio-enzyme and plant ferment formula developed specially for people with acne-prone and sensitive skin, and is capable of fundamentally improving sensitive, dark and rough skin, removing acne, and recovering white and bright skin.

The invention provides a method for separating and purifying recombinant bacillus subtilis fibrinolytic enzyme. The method comprises the following steps: carrying out hydrophobic chromatography: filling a chromatographic column with phenyl-agarose-series hydrophobic filler, washing and balancing the chromatographic column with liquid A, detecting when an ultraviolet absorption wavelength is 280 nanometers, removing thallus of fermentation liquor of the recombinant bacillus subtilis fibrinolytic enzyme and then directly loading a sample into the hydrophobic chromatographic column, continuously adding the liquid A in a flowing way until a flow-through peak flows through completely, and replacing the liquid A with liquid B for eluting until the recombinant bacillus subtilis fibrinolytic enzyme combined with the filler is completely eluted so as to obtain the high-purity recombinant bacillus subtilis fibrinolytic enzyme. A bacillus subtilis fibrinolytic enzyme separation and purification method with a pilot scale test level is established in a fermentation culture solution on the basis of recombinant yeast fermentation of the bacillus subtilis fibrinolytic enzyme; after the method is used, a great deal of high-purity fibrinolytic enzyme can be obtained, and the purity of the obtained fibrinolytic enzyme reaches up to 95%.

The invention relates to the technical field of health food, particularly to a method for enriching effective components such as tetramethylpyrazine and fibrinolytic enzyme by fermenting coix seed. According to the present invention, coix seed is fermented with Bacillus subtilis, such that fibrinolytic enzyme with high fibrinolysis activity can be obtained, and tetramethylpyrazine as a calcium ionantagonist and acetoin with wide uses can be obtained; and the tetramethylpyrazine content in the fermented coix seed is more than 150 times the tetramethylpyrazine content in the soybean fermented with the same strain, the acetoin content in the fermented coix seed is more than 4 times the acetoin content in the soybean fermented with the same strain, and the fibrinolytic enzyme content in the fermented coix seed is equal to the fibrinolytic enzyme content in the soybean fermented with the same strain.

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 discloses actinomycete fibrinolytic enzyme and a preparation method thereof. According to the actinomycete fibrinolytic enzyme, a actinomycete strain YY21 is adopted as a strain, and culture fermentation, 30-60% W / V saturation degree ammonium sulfate salting-out, Octyl-SepharoseFF hydrophobic interaction chromatography, and Phenyl-Sepharose HP hydrophobic interaction chromatography are performed to prepare the actinomycete fibrinolytic enzyme, wherein a molecular weight of the actinomycete fibrinolytic enzyme is about 32000 daltons, and the actinomycete fibrinolytic enzyme has a fibrin dissolving function. According to the present invention, the used strain has characteristics of short enzyme production period and excellent performances, and can produce other substances having bacterial inhibition activity while producing the enzyme; the actinomycete fibrinolytic enzyme is an extracellular enzyme, such that subsequent separation and purification during preparation are easily performed; and only two step hydrophobic interaction chromatography with advantages of rapid flow rate, high capacity and good sample activity storage are required to prepare the high purity product.

The present invention provides Bacillus subtilis HA KCCM-10775P having a high productivity of protease, fibrinolytic enzyme and mucilage, a method preparing fermented soybeans (e.g., Cheonggukjang) using the same strain, in which bean fragments, soybeans and black soybeans are fermented, and fermented soybeans (e.g., Cheonggukjang) produced according to the said method. The Bacillus subtilis HA KCCM-10775P isolated from traditional fermented soybeans (e.g., Cheonggukjang) and identified is the superior strain having a high productivity of protease, fibrinolytic enzyme and mucilage. When preparing fermented soybeans (e.g., Cheonggukjang) using the strain of the present invention, it has good sensory properties by removing a peculiar smell thereof, as well as a high productivity of fibrinolytic enzyme and mucilage.

The invention belongs to the technical field of traditional Chinese medicine preparations, and specifically discloses a preparation with the effect of reducing blood glucose and blood fat and a preparation method of the preparation. The preparation mainly comprises the following raw materials in parts by weight: 3 to 10 parts of hawthorn, 5 to 15 parts of Chinese yam, 3 to 5 parts of fruit of Chinese wolfberry, 5 to 15 parts of sweet Chinese catalpa grass, 5 to 20 parts of root of kudzu vine, 10 to 20 parts of gynostemma pentaphylla, 0.1 to 0.5 part of corn stigma, 1 to 3 parts of vernoniaamygdalina del leaves, 2 to 6 parts of butterflybush flower, 0.001 to 0.005 part of eleutheroside, 0.03 to 0.07 part of soy isoflavone, 1 to 5 parts of oxidized konjac glucomannan and 0.05 to 0.5 part offermented soya bean fibrinolytic enzyme. According to the traditional Chinese medicine preparation provided by the invention, the blood viscosity can be effectively improved, the blood glucose and blood fat are reduced, and the metabolic activity of fat is strengthened; partial raw materials come from food, the preparation is safe without side effect, and the preparation is convenient to take andgood in taste; and the preparation can be mixed with flour and buckwheat powder in a certain proportion, so that a health food capable of reducing blood glucose and blood fat is prepared.