CN101623321A - Thrombolytic active substance extracted from Guizhou fermented blank bean and product thereof - Google Patents
Thrombolytic active substance extracted from Guizhou fermented blank bean and product thereof Download PDFInfo
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- CN101623321A CN101623321A CN200810068814A CN200810068814A CN101623321A CN 101623321 A CN101623321 A CN 101623321A CN 200810068814 A CN200810068814 A CN 200810068814A CN 200810068814 A CN200810068814 A CN 200810068814A CN 101623321 A CN101623321 A CN 101623321A
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- semen sojae
- sojae preparatum
- active substance
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- thrombolytic
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Abstract
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.
Description
Technical field:
The present invention is that a kind of Guizhou Semen Sojae Preparatum extract thrombolytic active substance and the product that obtains.
Technical background:
Semen Sojae Preparatum are Guizhou a kind of traditional foods of liking among the people, it be a kind of be that raw material is made through microbial fermentation with the Semen sojae atricolor, it distinguishes the flavor of fragrant, smell smelly, among the peoplely think the appetizing of this food, aid digestion, be Guizhou popular for a long time food generally edible and that very like among the people.The microorganism difference that participates in during by yeast production, Semen Sojae Preparatum can be divided into Mucor type, aspergillosis type, rhizopus type and bacteria type etc.In conventional processes, if do not note technology, can cause quality to descend, even go mouldy, cause damage.The Guizhou Semen Sojae Preparatum belong to bacteria type, and are nutritious, but at present any report also do not seen in the active substance research of this food as yet.Given this plant situation, the research of Guizhou Semen Sojae Preparatum active substance is had good development prospect and huge market potential.
Summary of the invention:
The objective of the invention is to: provide a kind of Guizhou Semen Sojae Preparatum to extract active substance and the product that obtains; From the Semen Sojae Preparatum of Guizhou, extract beneficial bacteria, grope to have set up expandable Semen Sojae Preparatum preparation technology through experiment, and its active component has been carried out separation, evaluation.
The present invention constitutes like this: it mainly is to separate the active component that obtains in the Semen Sojae Preparatum that made by Semen sojae atricolor.Described Semen Sojae Preparatum are preparations like this: get Semen sojae atricolor, soak, and steaming and decocting, strain is inserted in the cooling back, places biochemical incubator to cultivate, promptly.Say accurately: described Semen Sojae Preparatum are like this preparations: get Semen sojae atricolor, soaked 6 hours, and 110 ℃ of steaming and decoctings 30 minutes, access strain in cooling back places 37 ℃ of biochemical incubators, cultivates 48 hours, promptly.
Described Semen Sojae Preparatum active substance is isolating like this: described Semen Sojae Preparatum active substance is isolating like this: get Semen Sojae Preparatum, add normal saline lixiviate in 1: 1, get supernatant, centrifugal 5000 rev/mins, 10 minutes, get bacterium mud; Get supernatant, add ethanol and make the alcohol amount of containing be 10%-60%, centrifugal 5000 rev/mins, 10 minutes, must precipitate; Get supernatant, add ethanol and make the alcohol amount of containing be 30%-90%, centrifugal 5000 rev/mins, 5 minutes, deposit B.Collecting precipitation A and B, lyophilization is preserved, promptly.Described Semen Sojae Preparatum active substance can also be isolating like this: get Semen Sojae Preparatum, add normal saline lixiviate in 1: 1, get and slowly add ammonium sulfate in the supernatant to make salinity be 10%-60%, behind 4 ℃ of placement 6h, 4 ℃, 12000 rev/mins are centrifugal 15 minutes, precipitate A, slowly adding ammonium sulfate again, to make salinity be 30%-90%, 4 ℃ of standing over night, 4 ℃ then, 12000 rev/mins are centrifugal 15 minutes, deposit B, collecting precipitation is 10000 the bag filter desalination of dialysing with molecular cut off, with gained precipitate A and B, lyophilization is preserved, promptly.
The main component of bacterium mud is a Semen Sojae Preparatum bacillus spp.; The main component of precipitate A is superoxide dismutase, isoflavone, saponins, tocopherol, dipicolinic acid, Menaquinone K6, polysaccharide etc.; The main component of deposit B is a fibrinoligase.The bacterium mud of gained can be used for preparing the medicine of treatment intestinal flora imbalance and hepatic disease.Precipitate A can be low in preparation treatment immunity, hyperglycemia, tumor, aging, osteoporosis, hyperlipidemia, hypertensive medicine.Deposit B can be used for preparing the medicine for the treatment of thrombus disease.Precipitate A and deposit B drying and crushing are mixed the back and added a small amount of Semen Sojae Preparatum powder and appropriate amount of auxiliary materials, and be encapsulated.Has stronger effect: treatment prevention blood vessel embolism, angina pectoris and myocardial infarction; Treatment prophylaxis of hypertension and osteoporosis take place; Promote that the intestines and stomach digestion and defecation are smooth and easy; But decomposing alcohol relieves the effect of alcohol; Antibioticly kill the interior bad bacterium of gastrointestinal, comprise the pylori that causes gastric cancer; Improve blood flow and ischemia apoplexy, reduce the concentration of cholesterol in serum and lipid.
The biochemical characteristic of this thrombolytic active substance is: molecular weight 28KD, optimum pH is 8.0, comparatively stable below 50 ℃, unstable in gastric environment, have activation fiber protein dissolution proenzyme and change into fibrinoclase and the direct fibrinous thrombolytic effect in the thrombus.
Prior art is compared, and the nutritive value of Guizhou tradition Semen Sojae Preparatum that the present invention is further clear and definite utilizes present biotechnology production to obtain Semen Sojae Preparatum active substance and products thereof.And Semen Sojae Preparatum have been carried out the research of extraction of active ingredients separating technology, also gained Semen Sojae Preparatum active substance has been carried out the component analysis evaluation, this Semen Sojae Preparatum active substance and products thereof has tangible thrombolytic effect, can be used for the prevention and the treatment of cardiovascular and cerebrovascular vessel, have good development prospect and huge market potential.
Concrete embodiment:
Experimental example 1
Guizhou Semen Sojae Preparatum Study on Preparation: use the plate streaking partition method from the commercially available Semen Sojae Preparatum in Guiyang City, Guizhou Province, separation screening is produced single bacterium colony of Semen Sojae Preparatum active substance, to its cultivation of going down to posterity, as supplying the examination strain.Get soy peptone 1%, Carnis Bovis seu Bubali cream 0.5%, NaCl0.5%, NaH
2PO
40.1%; Adding distil water is transferred pH to 9.0 to 6ml (2% inoculum concentration).121 ℃ of autoclavings, 30min.The strain that on superclean bench above-mentioned separation is obtained inserts culture fluid, puts into constant incubator and cultivates 24h, makes seed liquor.Get Semen sojae atricolor 300g and soaked 1-10 hour, 110 ℃ steaming and decocting 10-50 minute, the cooling back evenly is sprinkling upon seed liquor on the Semen sojae atricolor of boiling, and places biochemical incubator, cultivated 48 hours for 37 ℃, the finished product Semen Sojae Preparatum.
Experimental example 2:
Guizhou Semen Sojae Preparatum active component extracts, separation method research: with the lixiviating solution that normal saline lixiviate Semen Sojae Preparatum obtain, frozen centrifugation 5000 changeed 5-30 minute.Get supernatant and carry out fractional precipitation, add ammonium sulfate to 30% successively, 45%, 50%, 55%, 70%, 80%, 85%; And measure the activity of the thrombolysis activity component in the supernatant under each salinity with the fibrin plate method.With the solusphere diameter is vertical coordinate, and salinity 0%～85% is the abscissa mapping.Obtain best salting point 55%, separate obtaining the Semen Sojae Preparatum active substance.
Experimental example 3:
Guizhou Semen Sojae Preparatum active component thrombolysis activity is measured: get sterile petri dish, place in the levels operation platform, the preparation agarose solution, after cooling to wherein respectively adding fibrinogen solution and thrombin liquid, careful vibration is poured in two sterile petri dish, leave standstill ambient temperature overnight then, make fibrin plate, and get some apertures thereon.With the urokinase is standard substance, gets respectively in a certain amount of adding aperture; The active component of each stage gained is got a certain amount of the dropping in aperture.After putting into the cultivation of constant incubator incubation, with the solusphere diameter of slide gauge bioassay standard product and sample, the thrombolytic that utilizes standard curve to calculate the active component of each stage gained is tired.
Experimental example 4:
Guizhou Semen Sojae Preparatum active component preparation of product and usage: will from the Semen Sojae Preparatum of Guizhou, extract the active ingredient that obtains and mix, and load capsule with a small amount of Semen Sojae Preparatum dry powder and adjuvant.Oral, every day 2-3 time, each 1-2 grain can be used for treatment and prevention cardiovascular and cerebrovascular vessel thromboembolism; Treatment and prophylaxis of hypertension and osteoporosis take place; Promote that the intestines and stomach digestion and defecation are smooth and easy; But decomposing alcohol relieves the effect of alcohol; Antibioticly kill the interior bad bacterium of gastrointestinal, comprise the pylori that causes gastric cancer; Improve blood flow and ischemia apoplexy, reduce the concentration of cholesterol in serum and lipid.(production technology is seen Figure of description)
Experimental example 5:
Guizhou Semen Sojae Preparatum active component and Radix Salviae Miltiorrhizae Chinese medicine extract composition are by 1: the 1-10 compatibility makes and can supply oral preparation, and enteric coated capsule is best.Oral, every day 2-3 time, each 1-2 grain can reduce blood viscosity, and blood circulation promoting can be used for cardiovascular and cerebrovascular diseases such as treatment and prevention thrombus disease.
Experimental example 5:
Guizhou Semen Sojae Preparatum active component and Radix Astragali and Flos Carthami Chinese medicine extract composition are by 1: the 1-10 compatibility makes and can supply oral preparation, and enteric coated capsule is best.Oral, every day 2-3 time, each 1-2 grain can reduce blood viscosity, and blood circulation promoting can be used for cardiovascular and cerebrovascular diseases such as treatment and prevention thrombus disease.
Experimental example 6:
The Guizhou Semen Sojae Preparatum active component and the Radix Astragali, Radix Salviae Miltiorrhizae, Semen Persicae, Flos Carthami four flavor Chinese prescription extract compositions are by 1: the 1-10 compatibility makes and can supply oral preparation, and enteric coated capsule is best.Oral, every day 2-3 time, each 1-2 grain can reduce blood viscosity, and blood circulation promoting is used for the treatment of and prevents cardiovascular and cerebrovascular disease such as thrombus disease.
Description of drawings
Fig. 1 is that the Guizhou Semen Sojae Preparatum extract the thrombolysis activity product processes flow chart that obtains.
Claims (10)
1, a kind of thrombolytic active substance of Guizhou Semen Sojae Preparatum extraction is characterized in that: it mainly is by separating the active component that obtains in the Semen Sojae Preparatum of bacteria type Guizhou.
2, according to the preparation method of the described Guizhou of claim 1 Semen Sojae Preparatum thrombolytic active substance, it is characterized in that: described Semen Sojae Preparatum active substance is isolating like this: get Semen Sojae Preparatum, add normal saline lixiviate in 1: 1, get supernatant, centrifugal 5000 rev/mins, 10 minutes, get bacterium mud; Get supernatant, add ethanol and make the alcohol amount of containing be 10%-60%, centrifugal 5000 rev/mins, 10 minutes, must precipitate; Get supernatant, add ethanol and make the alcohol amount of containing be 30%-90%, centrifugal 5000 rev/mins, 5 minutes, deposit B.Collecting precipitation A and B, lyophilization is preserved, promptly.
3, preparation method according to the described Guizhou of claim 1 Semen Sojae Preparatum thrombolytic active substance, it is characterized in that: get Semen Sojae Preparatum, add normal saline lixiviate in 1: 1, get and slowly add ammonium sulfate in the supernatant to make salinity be 10%-60%, behind 4 ℃ of placement 6h, 4 ℃, 12000 rev/mins centrifugal 15 minutes, precipitate A, slowly adding ammonium sulfate again, to make salinity be 30%-90%, 4 ℃ of standing over night, 4 ℃ then, 12000 rev/mins centrifugal 15 minutes, deposit B, collecting precipitation, with molecular cut off is 10000 bag filter dialysis desalination, with gained precipitate A and B, lyophilization, preserve, promptly.
4, according to claim 2 or 3 described Semen Sojae Preparatum thrombolytic active substances, it is characterized in that: the main component of bacterium mud is a Semen Sojae Preparatum bacillus spp.; The main component of precipitate A is superoxide dismutase, isoflavone, saponins, tocopherol, dipicolinic acid, Menaquinone K6, polysaccharide etc.; The main component of deposit B is a fibrinoligase.
5, according to any described Semen Sojae Preparatum thrombolytic active substance among the claim 2-3, it is characterized in that: the bacterium mud of gained can be used for preparing the medicine of treatment intestinal flora imbalance and hepatic disease.
6, according to any described Semen Sojae Preparatum thrombolytic active substance among the claim 2-3, it is characterized in that: precipitate A can be low in preparation treatment immunity, hyperglycemia, tumor, aging, osteoporosis, hyperlipidemia, hypertensive medicine.
7, according to any described Semen Sojae Preparatum active substance among the claim 2-3, it is characterized in that: deposit B can be used for preparing the medicine for the treatment of thrombus disease.
8, according to claim 2-3 and the described Semen Sojae Preparatum thrombolytic active substance of 6-7, it is characterized in that: precipitate A and deposit B drying and crushing, the mixing back adds a small amount of Semen Sojae Preparatum powder and appropriate amount of auxiliary materials, and is encapsulated, gets the product of the active substance of Guizhou Semen Sojae Preparatum extraction.
9, according to claim 2-3 and the described Semen Sojae Preparatum thrombolytic active substance of 6-7, it is characterized in that: precipitate A and deposit B can further obtain the higher active substance of purity after the separation and purification again, then make the oral preparation of various confessions again, enteric coated capsule is best.
10, according to claim 2-3 and the described Semen Sojae Preparatum thrombolytic active substance of 6-7, it is characterized in that: Guizhou Semen Sojae Preparatum thrombolysis activity composition and Chinese medicine with activating blood circulation to dissipate blood stasis effect, as single medicinal material extract such as the Radix Astragali, Radix Salviae Miltiorrhizae, Semen Persicae, Flos Carthami or the compound extract formed of 2 flavor Chinese medicines, 3 flavor Chinese medicines and 4 flavor Chinese medicines wherein, Guizhou Semen Sojae Preparatum active component and Chinese medicine extract composition are by 1: the 1-10 compatibility makes and can supply oral preparation, and enteric coated capsule is best.Can reduce blood viscosity, blood circulation promoting can be used for cardiovascular and cerebrovascular diseases such as treatment and prevention thrombus disease.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104055968A (en) * | 2014-07-01 | 2014-09-24 | 王磊 | Aerosol with effects of promoting blood circulation to remove meridian obstruction for postoperative patients |
| CN105707876A (en) * | 2016-04-12 | 2016-06-29 | 通泰心脑健康管理有限公司 | Diet composition with functions of stabilizing and dissolving plaques as well as preparation method and application of diet composition |
| CN108865609A (en) * | 2018-07-03 | 2018-11-23 | 广东永兴酒业有限公司 | A kind of health liquor and preparation method thereof reducing alcoholic hepatic injury |
| CN110959676A (en) * | 2018-09-30 | 2020-04-07 | 内蒙古伊利实业集团股份有限公司 | Fermented milk product containing bifidobacterium lactis and application thereof |
-
2008
- 2008-07-07 CN CN200810068814A patent/CN101623321A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104055968A (en) * | 2014-07-01 | 2014-09-24 | 王磊 | Aerosol with effects of promoting blood circulation to remove meridian obstruction for postoperative patients |
| CN105707876A (en) * | 2016-04-12 | 2016-06-29 | 通泰心脑健康管理有限公司 | Diet composition with functions of stabilizing and dissolving plaques as well as preparation method and application of diet composition |
| CN108865609A (en) * | 2018-07-03 | 2018-11-23 | 广东永兴酒业有限公司 | A kind of health liquor and preparation method thereof reducing alcoholic hepatic injury |
| CN110959676A (en) * | 2018-09-30 | 2020-04-07 | 内蒙古伊利实业集团股份有限公司 | Fermented milk product containing bifidobacterium lactis and application thereof |
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Application publication date: 20100113 |