CN107043800A - A kind of preparation method of collagen - Google Patents
A kind of preparation method of collagen Download PDFInfo
- Publication number
- CN107043800A CN107043800A CN201710336661.5A CN201710336661A CN107043800A CN 107043800 A CN107043800 A CN 107043800A CN 201710336661 A CN201710336661 A CN 201710336661A CN 107043800 A CN107043800 A CN 107043800A
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- CN
- China
- Prior art keywords
- collagen
- preparation
- raw material
- papain
- ficin
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
Abstract
The invention discloses a kind of preparation method of collagen, comprise the following steps：It will clean up, crushed after drying for the pigskin raw material for preparing collagen；Weigh the raw material after a certain amount of crush and be put into enzymatic vessel, according to material ratio 1:3（g/ml）‑1:5（g/ml）Purified water is added, Collagenase, papain and ficin is added according to 0.5% the 1.5% of raw material weight, Collagenase, papain, the mass ratio of ficin are 1:1:1‑5:1:1；55 DEG C of 60 DEG C of 5 8h of hydrolysis, are removed the gred using vibratory sieve, seperator, obtain filtered fluid；Degreasing, concentration are carried out to filtered fluid, is dried, collagen is obtained.By the above-mentioned means, the present invention can it is pollution-free, expeditiously prepare collagen.
Description
Technical field
The present invention relates to bioactive substance preparation field, more particularly to a kind of preparation method of collagen.
Background technology
Collagen be animal in-vivo content at most, distribution most wide protein, be distributed widely in connective tissue, bone,
The positions such as ligament, sclera, account for the 25%-33% of animal body total protein content.Because collagen has special Gly-Xaa-Yaa tri-
Peptide unit and triple helix structure, make it have the physiology such as low immunogenicity, biocompatibility, degradability, anastalsis special
Property.At present, the application of collagen is more and more extensive, has expanded to the numerous areas such as biomedical material, food, cosmetics.
The structure of pigskin is made up of epidermis, skin corium, subcutaneous connective tissue, and its main component is skin corium.Corium
Layer contains substantial amounts of collagen fabric, and its content accounts for the 99% of dry.Root it is documented that, the protein content of pigskin can
Up to 33%, wherein collagen content is 87.8%.From this, prepared using pigskin collagen have it is wide before
Scape.
The mode of production of protein hydrolysate generally has two kinds：Chemical degradation method and enzyme edman degradation Edman.Chemical degradation method master
To include acid system and alkaline process, acid system is to extract collagen under certain condition using certain density acid solution, and alkaline process is profit
Extract collagen under certain condition with certain density alkali, but acid system and alkaline process can bring environmental pollution, and it is less efficient.
The enzyme class on preparing collagen being currently known is more, how pollution-free, expeditiously prepare collagen, is current
One of collagen preparation technology urgent problem to be solved.
The content of the invention
, can pollution-free, high efficiency the present invention solves the technical problem of a kind of preparation method of collagen is provided
Ground prepares collagen.
In order to solve the above technical problems, one aspect of the present invention is：A kind of preparation of collagen is provided
Method, comprises the following steps：It will clean up, crushed after drying for the pigskin raw material for preparing collagen；Weigh a certain amount of
It is broken after raw material be put into enzymatic vessel, according to material ratio 1:3（g/ml）-1:5（g/ml）Purified water is added, according to raw material weight
0.5%-1.5% add Collagenase, papain and ficin, Collagenase, papain, without flower
The mass ratio of fruit protease is 1:1:1-5:1:1；55 DEG C of -60 DEG C of hydrolysis 5-8h, are removed the gred using vibratory sieve, seperator, obtained
Filtrate；Degreasing, concentration are carried out to filtered fluid, is dried, collagen is obtained.
Wherein, hydrolysis time is 6h.
Wherein, material ratio is 1:3（g/ml）.
Wherein, Collagenase, papain and ficin, collagen are added according to the 1% of raw material weight
Enzyme, papain, the mass ratio of ficin are 4:1:1, hydrolysis temperature is 58 DEG C.
Wherein, drying mode is spray drying.
Wherein, after hydrolysis, 86-90 DEG C of inactivation 0.5h.
Wherein, inactivation temperature is 89 DEG C.
Wherein, concentration is carried out using milipore filter.
Wherein, the collagen of acquisition is packed.
Wherein, pH value during hydrolysis is controlled in 5.0-8.0.
The beneficial effects of the invention are as follows：It is different from the situation of prior art, the preparation method bag of collagen of the invention
Include hydrolysis, slagging-off, degreasing, concentration and dry, wherein, hydrolase is Collagenase, papain and ficin
Composition, hydrolysis temperature be 58 DEG C, hydrolysis time be 6 hours.By respectively joining in the synergy of various enzymes and technological process
Several setting so that preparing for collagen is simple, efficient, pollution-free, production environment free from extraneous odour, and can also reduce and be produced into
This.
Embodiment
The present invention is described in detail below.
Embodiment 1
The preparation method of collagen, comprises the following steps：
S11, will be cleaned up for the pigskin raw material for preparing collagen, be crushed after drying.
In the present embodiment, in addition to pigskin can be as the raw material of collagen be prepared, fish-skin, fish scale or the skin of other domestic animals
Also the raw material for preparing collagen can be used as.
Wherein, raw material should be chilled pigskin, to ensure the quality of the collagen prepared.When broken, using meat cutting
Machine and meat grinder are crushed.
S12, weighs the raw material after a certain amount of crush and is put into enzymatic vessel, according to material ratio 1:3（g/ml）-1:5（g/ml）
Purified water is added, Collagenase, papain and ficin, glue are added according to the 0.5%-1.5% of raw material weight
Former protease, papain, the mass ratio of ficin are 1:1:1-5:1:1.
In hydrolytic process, Collagenase, papain and ficin are added according to the 1% of raw material weight,
Collagenase, papain, the mass ratio of ficin are 4:1:1, pass through the synergy of various enzymes so that former
Material can be hydrolyzed sufficiently, efficiently prepare collagen.In actual applications, the extraction of pigskin collagen, pancreatin
Yield is 77%, and pepsin is 64%, and Collagenase is 80%, and the yield of three kinds of enzymes of the present embodiment is 85%.
S13,55 DEG C of -60 DEG C of hydrolysis 5-8h, is removed the gred using vibratory sieve, seperator, obtains filtered fluid.
In the present embodiment, preferred feed ratio is 1:3（g/ml）, hydrolysis time be that 6h, hydrolysis temperature are 58 DEG C.In hydrolysis
During, also need pH value control in 5.0-8.0, so that the enzyme activity of Collagenase, papain and ficin
Power is maintained at maximum.Wherein, 58 DEG C of hydrolysis temperature is that cold-adapted enzyme method prepares collagen.
When enzyme digestion reaction is carried out to a certain extent, after reaching required degree of hydrolysis, it is necessary to enzymolysis reaction, otherwise
Enzyme in reaction system still keeps activity, will further hydrolyze, and influences the function and physiologically active of end product.In the present embodiment
In, inactivated using heating, specially 86-90 DEG C inactivation 0.5h, preferably inactivation temperature are 89 DEG C.
After inactivation terminates, removed the gred using vibratory sieve, seperator, obtain filtered fluid.
S14, degreasing, concentration are carried out to filtered fluid, is dried, collagen is obtained.
After degreasing, concentration is carried out using milipore filter, after the completion of concentration, done by way of spraying or freezing
It is dry.Packed after drying, complete the preparation of collagen, obtain collagen protein powder.
Embodiment 2
By the use of fish-skin as the raw material for preparing collagen, specific preparation process comprises the following steps.
A. it will clean up, crushed after drying for the fish skin raw material for preparing collagen.
Wherein, raw material should be chilled fish-skin, to ensure the quality of the collagen prepared.When broken, using meat cutting
Machine and meat grinder are crushed.
B. weigh the raw material after a certain amount of crush and be put into enzymatic vessel, according to material ratio 1:3（g/ml）-1:5（g/ml）Plus
Enter purified water, Collagenase, papain and ficin, collagen are added according to the 0.5%-1.5% of raw material weight
Protease, papain, the mass ratio of ficin are 1:1:1-5:1:1.
In hydrolytic process, Collagenase, papain and ficin are added according to the 1.2% of raw material weight
Enzyme, Collagenase, papain, the mass ratio of ficin are 2:1:1.
C. 55 DEG C of -60 DEG C of hydrolysis 5-8h, are removed the gred using vibratory sieve, seperator, obtain filtered fluid.
In the present embodiment, preferred feed ratio is 1:4（g/ml）, hydrolysis time be that 7h, hydrolysis temperature are 60 DEG C.Hydrolyzed
Cheng Zhong, is also needed pH value control in 5.0-8.0, so that the enzyme activity of Collagenase, papain and ficin
It is maintained at maximum.
When enzyme digestion reaction is carried out to a certain extent, after reaching required degree of hydrolysis, it is necessary to enzymolysis reaction, otherwise
Enzyme in reaction system still keeps activity, will further hydrolyze, and influences the function and physiologically active of end product.In the present embodiment
In, inactivated using heating, specially 86-90 DEG C inactivation 0.5h, preferably inactivation temperature are 89 DEG C.
After inactivation terminates, removed the gred using vibratory sieve, seperator, obtain filtered fluid.
D. degreasing, concentration are carried out to filtered fluid, is dried, collagen is obtained.
After degreasing, concentration is carried out using milipore filter, after the completion of concentration, done by way of spraying or freezing
It is dry.Packed after drying, complete the preparation of collagen, obtain collagen protein powder.
In summary, the preparation method of collagen of the invention includes hydrolysis, slagging-off, degreasing, concentration and dried, its
In, hydrolase is the composition of Collagenase, papain and ficin, and hydrolysis temperature is 58 DEG C, during hydrolysis
Between be 6 hours.Pass through the setting of each parameter in the synergy of various enzymes and technological process so that the preparation letter of collagen
Single, efficient, pollution-free, production environment free from extraneous odour, and can also reduce production cost.
Embodiments of the invention are the foregoing is only, are not intended to limit the scope of the invention, it is every to utilize this hair
Equivalent structure or equivalent flow conversion that bright specification is made, or other related technical fields are directly or indirectly used in,
Similarly it is included within the scope of the present invention.
Claims (10)
1. a kind of preparation method of collagen, it is characterised in that comprise the following steps：
It will clean up, crushed after drying for the pigskin raw material for preparing collagen；
Weigh the raw material after a certain amount of crush and be put into enzymatic vessel, according to material ratio 1:3（g/ml）-1:5（g/ml）Add purifying
Water, Collagenase, papain and ficin, the collagen egg are added according to the 0.5%-1.5% of raw material weight
White enzyme, papain, the mass ratio of ficin are 1:1:1-5:1:1；
55 DEG C of -60 DEG C of hydrolysis 5-8h, are removed the gred using vibratory sieve, seperator, obtain filtered fluid；
Degreasing, concentration are carried out to filtered fluid, is dried, collagen is obtained.
2. the preparation method of collagen according to claim 1, it is characterised in that hydrolysis time is 6h.
3. the preparation method of collagen according to claim 2, it is characterised in that the material ratio is 1:3（g/ml）.
4. the preparation method of collagen according to claim 3, it is characterised in that 1% according to raw material weight adds glue
Former protease, papain and ficin, the Collagenase, papain, the matter of ficin
Amount is than being 4:1:1, hydrolysis temperature is 58 DEG C.
5. the preparation method of collagen according to claim 4, it is characterised in that the drying mode is dry for spraying
It is dry.
6. the preparation method of collagen according to claim 5, it is characterised in that after hydrolysis, 86-90 DEG C of inactivation
0.5h。
7. the preparation method of collagen according to claim 6, it is characterised in that inactivation temperature is 89 DEG C.
8. the preparation method of collagen according to claim 7, it is characterised in that carried out using milipore filter at concentration
Reason.
9. the preparation method of collagen according to claim 8, it is characterised in that wrapped the collagen of acquisition
Dress.
10. the preparation method of collagen according to claim 9, it is characterised in that pH value control during hydrolysis exists
5.0-8.0。
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| CN201710336661.5A CN107043800A (en) | 2017-05-13 | 2017-05-13 | A kind of preparation method of collagen |
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| CN201710336661.5A CN107043800A (en) | 2017-05-13 | 2017-05-13 | A kind of preparation method of collagen |
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| CN201710336661.5A Pending CN107043800A (en) | 2017-05-13 | 2017-05-13 | A kind of preparation method of collagen |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109371092A (en) * | 2018-12-26 | 2019-02-22 | 河北肽都生物科技有限公司 | A kind of preparation method of collagen peptide |
| CN109567026A (en) * | 2019-01-21 | 2019-04-05 | 武汉轻工大学 | A kind of pigskin collagen and low oil squeeze cake |
| CN111249531A (en) * | 2020-01-19 | 2020-06-09 | 巴斯特医药科技（常州）有限公司 | Method for preparing collagen scaffold by using high-activity plant enzyme |
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| WO2007084769A2 (en) * | 2006-01-19 | 2007-07-26 | Solazyme, Inc. | Microalgae-derived compositions for improving the health and appearance of skin |
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2017
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| WO2007084769A2 (en) * | 2006-01-19 | 2007-07-26 | Solazyme, Inc. | Microalgae-derived compositions for improving the health and appearance of skin |
| CN102131403A (en) * | 2009-07-27 | 2011-07-20 | 成功大学 | Preparation of high purity collagen |
| CN103305575A (en) * | 2012-03-12 | 2013-09-18 | 江苏奥普莱医疗用品有限公司 | Method for preparing active collagen from compound protease |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109371092A (en) * | 2018-12-26 | 2019-02-22 | 河北肽都生物科技有限公司 | A kind of preparation method of collagen peptide |
| CN109567026A (en) * | 2019-01-21 | 2019-04-05 | 武汉轻工大学 | A kind of pigskin collagen and low oil squeeze cake |
| CN111249531A (en) * | 2020-01-19 | 2020-06-09 | 巴斯特医药科技（常州）有限公司 | Method for preparing collagen scaffold by using high-activity plant enzyme |
| CN111249531B (en) * | 2020-01-19 | 2022-03-22 | 巴斯特医药科技（常州）有限公司 | Method for preparing collagen scaffold by using high-activity plant enzyme |
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Effective date of registration: 20201202 Address after: Room 505, No.5, Keyun North Road, Tianhe District, Guangzhou City, Guangdong Province 510000 Applicant after: Guangzhou benweida Biotechnology Co.,Ltd. Address before: 510000, No. 36, middle section of Bei Tai Road, Baiyun District, Guangdong, Guangzhou Applicant before: GUANGZHOU GULUN BIOTECHNOLOGY Co.,Ltd. |
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Application publication date: 20170815 |