CN1414095A - Preparation method of high actirity earthworm kinase dry powder - Google Patents
Preparation method of high actirity earthworm kinase dry powder Download PDFInfo
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- CN1414095A CN1414095A CN 02129969 CN02129969A CN1414095A CN 1414095 A CN1414095 A CN 1414095A CN 02129969 CN02129969 CN 02129969 CN 02129969 A CN02129969 A CN 02129969A CN 1414095 A CN1414095 A CN 1414095A
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Abstract
A process for preparing high-activity dried earthworm kinase powder includes such steps as rinsing the living earthworm, dripdrying, mixing with saline, stirring, centrifual separation several times, collecting supernatant, adding solution of sodium polyacrylate, laying aside at 0-4 deg.C for 6-24 hr, centrifugal separation for 10-30 min, collecting supernatant, adding solution of sodium polyacrylate, laying aside at 0-4 deg.C for 6-20 hr, centrifugal separation, collecting supernatant, dissolving in water, superfilter, low-temp drying and Co60 radiating. Its advantages are high activity, high output rate and low cost.
Description
Technical field
The present invention relates to a kind of preparation method of the method for utilizing earthworm to produce Lumbrukinase, particularly a kind of high actirity earthworm kinase dry powder.
Background technology
Put down in writing from TCM Document, earthworm has multiple pharmacological effect, as step-down, relieving asthma, analgesic, diuresis, calm anticonvulsion, vein relaxing, kill essence, excited uterus, thrombolysis detoxify and promote granulation, antianaphylaxis, antibiotic and anticancer etc.Adopting fresh and alive earthworm is raw material, and the proteolytic enzyme of the biologically active of extraction is commonly referred to as Lumbrukinase.Lumbrukinase has the scleroproein in the degraded blood, improves functions such as platelet aggregation, has significant anti-freezing, fibrinolytic, antithrombotic and thrombus dissolving effect, is adapted to the prevention and the treatment of cardiovascular and cerebrovascular disease.It is a lot of to utilize earthworm to produce the method for earthworm dry powder and earthworm acid enzyme at present, and as " preparation method of active dried earthworm powder " patent application of Patent Office of the People's Republic of China's bulletin on April 12nd, 2000, Granted publication number is CN1051229C.This patent application discloses a kind of method for preparing dried earthworm powder, this method is by being frozen into ice cube behind the fresh and alive earthworm edulcoration, be placed in the salt brine solution then and dissolve, and pulping is stirred in self-dissolving, after the homogenate that obtains carries out centrifugal treating at least, after supernatant liquor carried out pre-filtering sterilization and ultra-filtration and concentrate dry powder, this method main purpose has provided a kind ofly can thoroughly remove earthworm ight soil and harmful element, fully keep the earthworm effective ingredient, but the composition of the earthworm dry powder that this method is made is the mixing composition of earthworm, be not single composition, and its product activity is lower.
Summary of the invention
The inventive method has that operational path is short, and facility investment is little, is easy to the characteristics of suitability for industrialized production, and the Lumbrukinase that this method is produced has active high, with the urea kinases be control activity up to 260uk/mg, producing yield can reach about 3.5%.
The present invention is achieved by the following technical solutions:
A kind of preparation method of high actirity earthworm kinase dry powder is characterized in that:
A. fresh and alive earthworm rinsing is clean, after draining the water, add fresh and alive earthworm 1-4 salt solution doubly and mix, stir pulping;
The earthworm liquid that b. will stir into pulpous state carried out centrifugation 10-30 minute, and is stand-by after supernatant liquor is collected;
C. after the precipitation that centrifugation gone out adds the salt water washing, carry out centrifugation 10-30 minute after, collect supernatant liquor;
D. after the supernatant liquor that step b and c are collected mixes, add sodium polyacrylate solution, 0 °-4 ℃ leave standstill 6-24 hour after, centrifugation 10-30 minute, and collect supernatant liquor;
E. in the supernatant liquor of collecting, add sodium polyacrylate solution, 0 °-4 ℃ leave standstill 6-24 hour after, carry out centrifugation 10-30 minute, and the collecting precipitation thing;
F. throw out is soluble in water, use the membrane separating method ultrafiltration;
G. with the filtrate after the ultrafiltration after cryodrying, promptly get Lumbrukinase dry powder.
Adding brinish concentration is 0.75-2%; The concentration that adds sodium polyacrylate solution is 0.5-40%.The preferred vacuum lyophilization of cryodrying.
Embodiment
Get the fresh and alive earthworm of 100kg, fresh and alive earthworm is clean with the clear water rinsing, behind the drain well, the salt solution 100kg that adds 1% concentration mixes with the fresh and alive earthworm of clean drain well, the container of putting into the band stirring simultaneously stirs into pulpous state, then the earthworm liquid of pulpous state was put into the whizzer centrifugation after 10 minutes, stand-by after supernatant liquor is collected; The precipitation that centrifugation is gone out adds an amount of salt solution of 1% concentration again, after the washing of precipitate thing is washed, carries out centrifugation 15 minutes once more, collects supernatant liquor; After the supernatant liquor of above collection mixed, add concentration and be 10% sodium polyacrylate solution, 4 ℃ leave standstill 24 hours after, carry out centrifugation, the centrifugation time is 15 minutes and collects supernatant liquor; In the supernatant liquor of collecting, add concentration again and be 10% sodium polyacrylate solution, 4 ℃ leave standstill 12 hours after, carry out centrifugation, and the collecting precipitation thing; Throw out is dissolved in the clear water, uses the membrane separating method ultrafiltration; Ultrafiltrated after the ultrafiltration after vacuum lyophilization, promptly behind cobalt 60 radiation sterilizations, is got Lumbrukinase dry powder and is finished product.
Claims (5)
1. the preparation method of a high actirity earthworm kinase dry powder is characterized in that:
A. fresh and alive earthworm rinsing is clean, after draining the water, add fresh and alive earthworm 1-4 salt solution doubly and mix, stir pulping;
The earthworm liquid that b. will stir into pulpous state carries out centrifugation, and is stand-by after supernatant liquor is collected;
C. after the precipitation that centrifugation gone out adds the salt water washing, carry out centrifugation after, collect supernatant liquor;
D. after the supernatant liquor that step b and c are collected mixes, add sodium polyacrylate solution, 0 °-4 ℃ leave standstill 6-24 hour after, centrifugation, and collect supernatant liquor;
E. in the supernatant liquor of collecting, add sodium polyacrylate solution, 0 °-4 ℃ leave standstill 6-24 hour after, carry out centrifugation, and the collecting precipitation thing;
F. throw out is soluble in water, use the membrane separating method ultrafiltration;
G. with the filtrate after the ultrafiltration after cryodrying, promptly get Lumbrukinase dry powder.
2. the preparation method of a kind of high actirity earthworm kinase dry powder according to claim 1, it is characterized in that adding brinish concentration is 0.75-2%.
3. the preparation method of a kind of high actirity earthworm kinase dry powder according to claim 1, the concentration that it is characterized in that adding sodium polyacrylate solution is 0.5-40%.
4. according to the preparation method of the described a kind of high actirity earthworm kinase dry powder of claim 1, it is characterized in that the preferred vacuum lyophilization of cryodrying.
5. according to the preparation method of the described a kind of high actirity earthworm kinase dry powder of claim 1, it is characterized in that the centrifugation time was at 10-30 minute.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02129969 CN1414095A (en) | 2002-08-30 | 2002-08-30 | Preparation method of high actirity earthworm kinase dry powder |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02129969 CN1414095A (en) | 2002-08-30 | 2002-08-30 | Preparation method of high actirity earthworm kinase dry powder |
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| Publication Number | Publication Date |
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| CN1414095A true CN1414095A (en) | 2003-04-30 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| CN 02129969 Pending CN1414095A (en) | 2002-08-30 | 2002-08-30 | Preparation method of high actirity earthworm kinase dry powder |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102433316A (en) * | 2011-11-27 | 2012-05-02 | 甘肃华羚生物技术研究中心 | Preparation method for lumbrokinase dry powder |
| CN110897982A (en) * | 2019-10-26 | 2020-03-24 | 银川凤仪堂生物工程有限公司 | Lumbrukinase freeze-dried powder and application thereof in skin care products |
| CN112889756A (en) * | 2019-12-03 | 2021-06-04 | 泰州市春光生态农业发展有限公司 | Novel high-efficiency method for extracting living earthworm mucus |
| CN115068363A (en) * | 2022-08-02 | 2022-09-20 | 银川凤仪堂生物工程有限公司 | Earthworm polypeptide freeze-dried powder and application thereof in skin care products |
-
2002
- 2002-08-30 CN CN 02129969 patent/CN1414095A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102433316A (en) * | 2011-11-27 | 2012-05-02 | 甘肃华羚生物技术研究中心 | Preparation method for lumbrokinase dry powder |
| CN110897982A (en) * | 2019-10-26 | 2020-03-24 | 银川凤仪堂生物工程有限公司 | Lumbrukinase freeze-dried powder and application thereof in skin care products |
| CN112889756A (en) * | 2019-12-03 | 2021-06-04 | 泰州市春光生态农业发展有限公司 | Novel high-efficiency method for extracting living earthworm mucus |
| CN115068363A (en) * | 2022-08-02 | 2022-09-20 | 银川凤仪堂生物工程有限公司 | Earthworm polypeptide freeze-dried powder and application thereof in skin care products |
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