CN104745555A - Preparation method of snail protease - Google Patents
Preparation method of snail protease Download PDFInfo
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- CN104745555A CN104745555A CN201510188191.3A CN201510188191A CN104745555A CN 104745555 A CN104745555 A CN 104745555A CN 201510188191 A CN201510188191 A CN 201510188191A CN 104745555 A CN104745555 A CN 104745555A
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- snail
- helicopepsin
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6402—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals
- C12N9/6405—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals not being snakes
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Abstract
The invention relates to a preparation method of snail protease and solves the technical problem that the existing snail protease extraction method is complex in process, poor in operability, low in extraction rate and high in cost. The preparation method comprises the following steps: selecting fresh and alive achatina fulica; cleaning; collecting mucus; filtering for the first time; centrifugalizing; filtering for the second time; freeze-drying in vacuum; milling glue; and storing. The preparation method provided by the invention is widely applied to preparing snail protease.
Description
Technical field
The present invention relates to a kind of preparation method of enzyme, specifically a kind of preparation method of helicopepsin.
Background technology
Helicopepsin is the biological activity protein prepared from the mucus that the pedal gland of snail is secreted, and it is rich in the requisite materials of maintenance skin such as natural wallantoin, calcium, glycolic acid, osso-albumin, protein and VITAMIN.In addition, snail mucus or a kind of natural antibiotics, can kill some bacterium on people's skin, and can eliminate skin speckle and acne, because its 3rd aspect that can penetrate to the skin.The chemical composition that the mucus of snail secretion contains has firming effect, can make skin rapid regeneration.The wallantoin extracted from snail mucus can make skin regeneration; Protein and VITAMIN can increase the nutrition of skin, make skin finer and smoother and smooth.And the extracting method of helicopepsin exists the low defect of complex process, operability difference and purification rate at present, the high helicopepsin that makes of preservation condition holds at high price, and can not realize volume production.
Summary of the invention
The present invention is exactly to solve existing helicopepsin extracting method complex process, operability is poor, purification rate is low, cost is high technical problem, provides the preparation method of the helicopepsin that a kind of technique is simple, strong operability, purification rate are high, cost is low.
The invention provides a kind of preparation method of helicopepsin, comprise the following steps:
(1) snail is chosen: select fresh and alive African Snail;
(2) snail cleaning: snail is placed in the clear water flushing that Cultivating box temperature is 18 DEG C ~ 28 DEG C, washes ight soil and foreign material off;
(3) snail mucus is collected: get the fresh and alive snail cleaned up, leave standstill on the table, treat that its health stretches out shell, with small-sized charge power supply adjustment open circuit voltage >900 volt, load current <4 milliampere, shock by electricity snail foot instantaneously, and one touches namely from repetition 5 times, draws in beaker after snail secreting mucus with dropper;
(4) first time filters: by the mucus that gathers by 450 order filtering nets, be collected in centrifuge tube;
(5) centrifugal: the mucus after being filtered first time with refrigerated centrifuge at 0 DEG C ~ 5 DEG C centrifugal 10 minutes, centrifugal rotational speed is 11000 revs/min, gets supernatant liquor;
(6) second time is filtered: filtered through bacterial filter by supernatant liquor;
(7) vacuum-freeze-dry: filtrate carries out vacuum-freeze-dry 20 hours with plastic containers, and helicopepsin water content is below 6%;
(8) glue mill: the helicopepsin after freeze-drying is clayed into power in rubber mill, is divided in 200g ampulla;
(9) preserve: 200g ampulla is bled tube sealing, vanning is placed on dry shady and cool place and preserves.
Preferably, the weight of the fresh and alive African Snail selected in step (1) is more than 15 grams, and in described step (2), the clear water temperature of rinsing is 25 DEG C; In described step (5), temperature time centrifugal is 3 DEG C; In described step (7), helicopepsin water content is 6%.
The invention has the beneficial effects as follows, present method technique is simple, strong operability, purification rate are high, and can realize the volume production of helicopepsin, helicopepsin finished product price is low, and every 100 kilograms of snail mucus can produce 1.2 kilograms of helicopepsins.The helicopepsin that the present invention extracts carries out enzymic activity experiment through SB/T 10317-1999 protease activity amylograph, and its enzymic activity is 7.5 × 10
-3u/g.
Accompanying drawing explanation
Fig. 1 is the procedure chart of preparation method of the present invention.
Embodiment
As shown in Figure 1:
(1) snail is chosen:
Select the fresh and alive African Snail (formal name used at school: Achatina Fulica) of more than 15 grams.
(2) snail cleaning:
Snail is placed in the clear water flushing that Cultivating box temperature is 18 DEG C ~ 28 DEG C, wash ight soil and foreign material off, each cleaning also carefully should choose the snail except having died.For the clear water rinsed, its temperature is preferably 25 DEG C.
(3) snail mucus is collected:
Get the fresh and alive snail that 50KG cleans up, leave standstill on the table, treat that its health stretches out shell, with small-sized charge power supply adjustment open circuit voltage >900 volt, load current <4 milliampere, shock by electricity snail foot instantaneously, and one touches namely from repetition 5 times, draw in beaker with dropper after snail secreting mucus, 50 kilograms of snails can collect 5000 grams of mucus.
(4) first time filters:
By the mucus that gathers by 450 order filtering nets, be collected in centrifuge tube.
(5) centrifugal:
Mucus refrigerated centrifuge at 0 DEG C ~ 5 DEG C centrifugal 10 minutes, centrifugal rotational speed is 11000 revs/min, gets supernatant liquor.For centrifuging temperature, be preferably 3 DEG C.
(6) second time is filtered:
Supernatant liquor is filtered through bacterial filter.
(7) vacuum-freeze-dry:
Filtrate carries out vacuum-freeze-dry 20 hours with plastic containers, and helicopepsin water content is below 6%, and the helicopepsin of freeze-drying is xln, and in rice white, 5000 grams of mucus can obtain 60 grams of helicopepsin lyophilized powders.For helicopepsin water content, be preferably 6%.
(8) glue mill:
Helicopepsin after freeze-drying is clayed into power in rubber mill, is divided in 200g ampulla.
(9) preserve:
200g ampulla is bled tube sealing, and vanning is placed on dry shady and cool place and preserves.
Above-mentioned African Snail can extract after needing to feed 5 days with clear water and concentrated feed after collecting mucus again.
Claims (2)
1. a preparation method for helicopepsin, is characterized in that comprising the following steps:
(1) snail is chosen: select fresh and alive African Snail;
(2) snail cleaning: snail is placed in Cultivating box, rinses with the clear water that temperature is 18 DEG C ~ 28 DEG C, washes ight soil and foreign material off;
(3) snail mucus is collected: get the fresh and alive snail cleaned up, leave standstill on the table, treat that its health stretches out shell, with small-sized charge power supply adjustment open circuit voltage >900 volt, load current <4 milliampere, shock by electricity snail foot instantaneously, one touch namely from, repeat 5 times, after snail secreting mucus, draw in beaker with dropper;
(4) first time filters: by the mucus that gathers by 450 order filtering nets, be collected in centrifuge tube;
(5) centrifugal: the mucus after being filtered first time with refrigerated centrifuge at 0 DEG C ~ 5 DEG C centrifugal 10 minutes, centrifugal rotational speed is 11000 revs/min, gets supernatant liquor;
(6) second time is filtered: filtered through bacterial filter by supernatant liquor;
(7) vacuum-freeze-dry: filtrate carries out vacuum-freeze-dry 20 hours with plastic containers, and helicopepsin water content is below 6%;
(8) glue mill: the helicopepsin after freeze-drying is clayed into power in rubber mill, is divided in 200g ampulla;
(9) preserve: 200g ampulla is bled tube sealing, vanning is placed on dry shady and cool place and preserves.
2. the preparation method of helicopepsin according to claim 1, is characterized in that:
The weight of the fresh and alive African Snail selected in described step (1) is more than 15 grams, and in described step (2), the clear water temperature of rinsing is 25 DEG C; In described step (5), temperature time centrifugal is 3 DEG C; In described step (7), helicopepsin water content is 6%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510188191.3A CN104745555A (en) | 2015-04-20 | 2015-04-20 | Preparation method of snail protease |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510188191.3A CN104745555A (en) | 2015-04-20 | 2015-04-20 | Preparation method of snail protease |
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| CN104745555A true CN104745555A (en) | 2015-07-01 |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2734725B1 (en) * | 1995-06-02 | 1997-08-14 | Beltchev Stoyan | TECHNICAL PROCESS FOR THE EXTRACTION OF SNAIL MUCUS |
| CN102199582A (en) * | 2010-03-23 | 2011-09-28 | 叶阿彬 | Method for extraction and preservation of helicase |
| CN102846519A (en) * | 2012-08-11 | 2013-01-02 | 叶阿彬 | Method for extracting snail mucus |
| CN103468663A (en) * | 2013-08-12 | 2013-12-25 | 叶阿彬 | Snail protease extraction method |
| CN103961797A (en) * | 2013-02-05 | 2014-08-06 | 台湾小品蜗牛生技有限公司 | Method for promoting secretion of snail mucus and extraction method thereof |
-
2015
- 2015-04-20 CN CN201510188191.3A patent/CN104745555A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2734725B1 (en) * | 1995-06-02 | 1997-08-14 | Beltchev Stoyan | TECHNICAL PROCESS FOR THE EXTRACTION OF SNAIL MUCUS |
| CN102199582A (en) * | 2010-03-23 | 2011-09-28 | 叶阿彬 | Method for extraction and preservation of helicase |
| CN102846519A (en) * | 2012-08-11 | 2013-01-02 | 叶阿彬 | Method for extracting snail mucus |
| CN103961797A (en) * | 2013-02-05 | 2014-08-06 | 台湾小品蜗牛生技有限公司 | Method for promoting secretion of snail mucus and extraction method thereof |
| CN103468663A (en) * | 2013-08-12 | 2013-12-25 | 叶阿彬 | Snail protease extraction method |
Non-Patent Citations (1)
| Title |
|---|
| 蔡方顺: "白玉蜗牛的人工养殖技术", 《当代畜牧》 * |
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Application publication date: 20150701 |