CN106636267A - Extracting method of small-molecular sea cucumber-oyster polypeptide - Google Patents
Extracting method of small-molecular sea cucumber-oyster polypeptide Download PDFInfo
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- CN106636267A CN106636267A CN201610896521.9A CN201610896521A CN106636267A CN 106636267 A CN106636267 A CN 106636267A CN 201610896521 A CN201610896521 A CN 201610896521A CN 106636267 A CN106636267 A CN 106636267A
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- 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
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- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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Abstract
The invention discloses an extracting method of a small-molecular sea cucumber-oyster polypeptide. A biological enzymolysis technology and a membrane technology are scientifically and reasonably integrated, and raw materials are subjected to biological enzymolysis through specific compound enzyme; active carbon can have the effects of removing color and fishiness and removing heavy metal and harmful substances; a concentrated solution containing the small-molecular sea cucumber-oyster polypeptide is obtained by sequentially separating, concentrating and purifying through an ultra-filtration membrane and a nano-filtration membrane, which have specific molecular weight cutoff; and finally, the concentrated solution is sprayed and dried to obtain a small-molecular sea cucumber and oyster polypeptide finished product. The method disclosed by the invention has high operability and the extracting rate on the small-molecular sea cucumber-oyster polypeptide is 1.0%-3.8%; the prepared small-molecular sea cucumber-oyster polypeptide has high purity and the bioactivity of the small-molecular sea cucumber-oyster polypeptide can be effectively guaranteed; and the small-molecular sea cucumber-oyster polypeptide cab be used as the raw material of various high-additional-value products in future, and has a wide market space and economic value.
Description
Technical field
The present invention relates to a kind of extracting method of marine product small molecular polypeptide, specifically a kind of small molecule ginseng oyster polypeptide
Extracting method.
Background technology
With the popularization developed rapidly with nutrient knowledge of science and technology, and increase and the level of consumption of per capita income level
Improve, the mankind have been transitioned into trophic level for the requirement of food from having enough to eat and wear, natural, healthy, nutrition, safety have become
The main themes of 21 century development of food industry.Collagen as a kind of important bio-tissue structural proteins polypeptide, more
To get over the concern and interest that cause people, market potential is huge, and economic worth is higher.Compared with macro-molecular protein, collagen egg
White micromolecule active polypeptide not only has high synthesis low cost, utilization rate, toxicity and the low feature of immunogenicity, also with efficient
Anti-oxidant, antibacterial and the pharmacological action such as anticancer, be widely used in medicine, beauty and health care and food service industry.
Biological species in ocean are various, and resource is extremely enriched, and biological chain perfects substantially, and power of regeneration is strong.Ocean
Protein, unrighted acid, taurine of the biology containing the abundant health that is beneficial to man, its micro- pedigree is also significantly
It is the fabulous source of human health food better than terrestrial life.Marine environment is very different with terrestrial environment, such as high pressure,
Low temperature, high temperature (submarine volcano mouth and near) and high salt etc..In order to be suitable for marine environment, marine protein matter no matter amino acid
Composition or the sequence of amino acid all make a big difference with terrestrial life protein.Meanwhile, marine protein resource without
By being all far longer than land protein resource in species or quantity.Miscellaneous protein amino acid sequence, with special
Protease hydrolytic, can release the Functional Polypeptides with physiological function.Therefore, these resources are opened using modern separation technology
Send out and utilize, will be one of current scientific research field most with prospects.
From different material extract polypeptide traditional handicraft can be divided mainly into five classes, i.e. acid system, alkaline process, enzyme process, salt method with
And Hot water extraction, its general principle is all the characteristic according to polypeptide, changes the external environment of protein, polypeptide from other eggs
Separate in white matter.Acid system and alkaline process are the methods for respectively extracting polypeptide under acid and alkaline environment, and acid system exists
Apply than wide in the extraction process of polypeptide, and alkaline process report is little；Salt method be by raw material in certain density salting liquid
Extract, the salt for mainly using has sodium chloride, potassium chloride and sodium acetate.Although three of the above method process is simple, cost
It is relatively low, but due to needing to use substantial amounts of fresh water and acid-alkali salt in extraction process, belong to the extracting method of high energy oyster, discharge
Acid-alkali salt can have a negative impact to ecological environment around, not meet green economy and sustainable development theory.And hot water soaks
Formulation needs to use substantial amounts of fresh water and the energy, does not meet the main flow of country and development of world economy.Biological enzyme is to utilize enzyme
The gentle high efficiency method that lifts out from different raw materials by polypeptide under certain environmental conditions of reaction, can overcome on
The deficiency of other techniques is stated, is a kind of technique of upgrading.
Rich in protein, biological function peptide, glycogen, taurine, EPA, DHA and vitamin in oyster, zinc, calcium, selenium,
The bioactive ingredients such as iron, for its application in food and medicine field huge support is provided.China is oyster culture
Big country, oyster yield accounts for 40% of cultivated shellfish or so, and resource is extremely enriched.Oyster culture in Ninghai County of Ningbo City Xi Dian towns
There is 700 years history.Ancients think it is " most your person of aquatic products ", and roman is described as " marine delicious-holy fish ", west it
People is referred to as " god grants magic stone ", " marine milk ", and what Japanese then praised is " source of root ".Oyster is that ministry of Health of China is announced
One of food of a collection of 87 kinds of integration of drinking and medicinal herbs.Contain rich in protein, functional polypeptide, various ammonia needed by human in oyster meat
The nutrient contents such as base is sour, various unrighted acids and abundant glycogen, sufficient vitamin and mineral matter, with huge
Edibility and medical value.However, with the fast development of oyster culture technology, oyster industry has occurred that drug on the market and valency
Lattice downward tendency, trace it to its cause mainly due to：1) industrialization level of development of resources is very low, and scope of the enterprise is little, industry input
Wretched insufficiency, resource utilization is very low；2) simple production process, product is mainly based on preliminary working, and scientific and technological content is not high, production
Cycle is long, high cost；Added value is few, and product competitiveness in the market is low；3) existing product quality controllability, stability, homogeneity compared with
Difference；4) scientific research and development ability not enough, are especially a lack of following the trail of market comsupton trend and trend, are designed according to consumer demand
With the ability of research and development product, technology and market can not find corresponding point.These factors cause that the oyster technological element of a product is low, and market accounts for
There is rate very little, resources advantage is not converted to economic advantages.Realize that oyster higher value application is that oyster culture industry health is persistently sent out
One of prerequisite of exhibition.At present, oyster mainly with directly eat and preliminary working product based on, such as dry, salt marsh, tank processed and carry
Refining oyster sauce etc., and the high value added product with oyster as raw material is less.Sea cucumber is a kind of traditional marine products treasure, containing abundant egg
White matter, amino acid and various trace elements, are all considered as excellent tonic product by people since ancient times, are of high nutritive value, physiologically active
Content of material enriches.Compared with the protein of macromolecule, polypeptide have little molecular weight, easy absorption, no antigen, with various
The features such as BA, increasingly approved by people.In recent years, in sea cucumber deep processing, the preparation skill of sea cucumber polypeptide
Art becomes focus.High value added product at present with sea cucumber and oyster as raw material is less, not yet develops for small molecule in the industry
The effective side that ginseng oyster polypeptide (i.e. by small molecule sea cucumber polypeptide and the polypeptide of small molecule oyster polypeptides in combination) is extracted
Small molecule ginseng oyster polypeptide is extracted in method, industrialization, in technology and application or blank.
The content of the invention
The technical problem to be solved is：The extracting method that a kind of small molecule joins oyster polypeptide, the extraction side are provided
Method is workable, and its recovery rate for joining oyster polypeptide to small molecule is 1.0～3.8%, the small molecule ginseng oyster polypeptide for preparing
Purity it is high, can effectively ensure that small molecule joins the biologically active of oyster polypeptide.
The present invention solve the technical scheme that adopted of above-mentioned technical problem for：A kind of small molecule joins the extraction side of oyster polypeptide
Method, comprises the following steps：
(1) clean：With fresh sea cucumber and oyster as raw material, with clear water most sea cucumber and oyster surface silt are washed；
(2) oyster shell is removed：In the environment that temperature is less than 5 DEG C, oyster meat is taken out into stand-by with special oyster knife, deposited
Time is less than 24 hours；
(3) crush：Oyster meat moisture is drained to not dripping, oyster meat and sea cucumber is crushed with industrial beating crusher, powder
Broken is less than 2 centimetres of particle；
(4) digest：By the oyster meat after crushing and sea cucumber with running water or pure water with 1:2～8 weight ratio adds to life
In thing enzymatic vessel, and alkali protease, Collagenase and papain and activated carbon, alkali are added in biological enzymolysis tank
Property protease addition be the 0.1～1% of oyster meat after crushing and sea cucumber weight, the addition of Collagenase is to crush
Oyster meat and the 0.4～4% of sea cucumber weight afterwards, the addition of papain is oyster meat and sea cucumber weight after crushing
0.2～2%, the addition of activated carbon is the 1～8% of the oyster meat after crushing and sea cucumber weight, with 40～60 DEG C of temperature enzyme
Solution 8～20 hours, obtains enzymolysis product；
(5) it is centrifuged：Enzymolysis product is centrifuged repeatedly for several times by centrifuge with the rotating speed of 5000 turns/hour, centrifugation
After remove supernatant, obtain be centrifuged product；
(6) separate：By the centrifugation product Ultra filtration membrane of the dalton of aperture 6000～100000, clear liquid is removed；
(7) concentrate：Lower clear liquid to obtaining in step (6) is concentrated with aperture less than the NF membrane of 60 dalton, takes supernatant
Liquid；
(8) it is dried：Supernatant to obtaining in step (7) is dried using spray dryer or freeze dryer, that is, obtain
Small molecule joins oyster polypeptide finished product.
Preferably, described activated carbon is the activated carbon of 200 mesh.
Preferably, the EAT that described spray dryer is dried is 120～145 DEG C, described freeze dryer is dried
Temperature be subzero 60 DEG C.
Compared with prior art, it is an advantage of the current invention that：A kind of small molecule that the present invention is provided joins the extraction of oyster polypeptide
Method, by biological enzymolysis technology and membrane technology scientific and reasonable integration is carried out, and biology is carried out to raw material by specific complex enzyme
Digest, and activated carbon can play a part of except color deodorization and go removing heavy metals and harmful substance, then successively through specific section
The milipore filter and nanofiltration membrane separation concentration purification of molecular weight is stayed, obtains joining the concentrate of oyster polypeptide containing small molecule, finally to this
Concentrate is spray-dried, that is, obtain small molecule ginseng oyster polypeptide finished product.Extracting method of the present invention is workable, and it is to little point
The recovery rate of son ginseng oyster polypeptide is 1.0～3.8%, and the purity of the small molecule ginseng oyster polypeptide for preparing is high, can effectively ensure that little
Molecule joins the biologically active of oyster polypeptide, and the small molecule ginseng oyster polypeptide subsequently can have as the raw material of various high value added products
The wide market space and economic worth.
Description of the drawings
Fig. 1 joins the chromatogram of oyster polypeptide for the small molecule of embodiment 1.
Specific embodiment
The present invention is described in further detail below in conjunction with accompanying drawing embodiment.
The small molecule of embodiment 1 joins the extracting method of oyster polypeptide, comprises the following steps：
(1) clean：With fresh sea cucumber and oyster as raw material, with clear water most sea cucumber and oyster surface silt are washed；
(2) oyster shell is removed：In the environment that temperature is less than 5 DEG C, oyster meat is taken out into stand-by with special oyster knife, deposited
Time is less than 24 hours；
(3) crush：Oyster meat moisture is drained to not dripping, oyster meat and sea cucumber is crushed with industrial beating crusher, powder
Broken is less than 2 centimetres of particle；
(4) digest：By the oyster meat after crushing and sea cucumber with running water or pure water with 1:2 weight ratio adds to biology enzyme
In solution tank, and the activity of alkali protease, Collagenase and papain and 200 mesh is added in biological enzymolysis tank
Charcoal, the addition of alkali protease is the 0.2% of the oyster meat after crushing and sea cucumber weight, and the addition of Collagenase is powder
0.4% of oyster meat and sea cucumber weight after broken, the addition of papain is oyster meat and sea cucumber weight after crushing
0.2%, the addition of activated carbon is the 3% of the oyster meat after crushing and sea cucumber weight, is digested 15 hours with 45 DEG C of temperature, is obtained
To enzymolysis product；
(5) it is centrifuged：Enzymolysis product is centrifuged repeatedly for several times by centrifuge with the rotating speed of 5000 turns/hour, centrifugation
After remove supernatant, obtain be centrifuged product；
(6) separate：By the centrifugation product Ultra filtration membrane of the dalton of aperture 6000～100000, clear liquid is removed；
(7) concentrate：Lower clear liquid to obtaining in step (6) is concentrated with aperture less than the NF membrane of 60 dalton, takes supernatant
Liquid；
(8) it is dried：Supernatant to obtaining in step (7) is dried using spray dryer, and spray dryer is dried
EAT be 140 DEG C, that is, obtain embodiment 1 small molecule ginseng oyster polypeptide finished product.After testing, the small molecule ginseng of embodiment 1
The recovery rate of oyster polypeptide is 3.8%.
The small molecule of embodiment 2 joins the extracting method of oyster polypeptide, comprises the following steps：
(1) clean：With fresh sea cucumber and oyster as raw material, with clear water most sea cucumber and oyster surface silt are washed；
(2) oyster shell is removed：In the environment that temperature is less than 5 DEG C, oyster meat is taken out into stand-by with special oyster knife, deposited
Time is less than 24 hours；
(3) crush：Oyster meat moisture is drained to not dripping, oyster meat and sea cucumber is crushed with industrial beating crusher, powder
Broken is less than 2 centimetres of particle；
(4) digest：By the oyster meat after crushing and sea cucumber with running water or pure water with 1:5 weight ratio adds to biology enzyme
In solution tank, and the activity of alkali protease, Collagenase and papain and 200 mesh is added in biological enzymolysis tank
Charcoal, the addition of alkali protease is the 0.3% of the oyster meat after crushing and sea cucumber weight, and the addition of Collagenase is powder
0.5% of oyster meat and sea cucumber weight after broken, the addition of papain is oyster meat and sea cucumber weight after crushing
0.2%, the addition of activated carbon is the 2% of the oyster meat after crushing and sea cucumber weight, is digested 10 hours with 55 DEG C of temperature, is obtained
To enzymolysis product；
(5) it is centrifuged：Enzymolysis product is centrifuged repeatedly for several times by centrifuge with the rotating speed of 5000 turns/hour, centrifugation
After remove supernatant, obtain be centrifuged product；
(6) separate：By the centrifugation product Ultra filtration membrane of the dalton of aperture 6000～100000, clear liquid is removed；
(7) concentrate：Lower clear liquid to obtaining in step (6) is concentrated with aperture less than the NF membrane of 60 dalton, takes supernatant
Liquid；
(8) it is dried：Supernatant to obtaining in step (7) is dried using freeze dryer, and the temperature that freeze dryer is dried is
Subzero 60 DEG C, that is, obtain the small molecule ginseng oyster polypeptide finished product of embodiment 2.After testing, the small molecule of embodiment 2 joins carrying for oyster polypeptide
Rate is taken for 3.5%.
The small molecule of embodiment 3 joins the extracting method of oyster polypeptide, comprises the following steps：
(1) clean：With fresh sea cucumber and oyster as raw material, with clear water most sea cucumber and oyster surface silt are washed；
(2) oyster shell is removed：In the environment that temperature is less than 5 DEG C, oyster meat is taken out into stand-by with special oyster knife, deposited
Time is less than 24 hours；
(3) crush：Oyster meat moisture is drained to not dripping, oyster meat and sea cucumber is crushed with industrial beating crusher, powder
Broken is less than 2 centimetres of particle；
(4) digest：By the oyster meat after crushing and sea cucumber with running water or pure water with 1:7 weight ratio adds to biology enzyme
In solution tank, and the activity of alkali protease, Collagenase and papain and 200 mesh is added in biological enzymolysis tank
Charcoal, the addition of alkali protease is the 0.1% of the oyster meat after crushing and sea cucumber weight, and the addition of Collagenase is powder
0.6% of oyster meat and sea cucumber weight after broken, the addition of papain is oyster meat and sea cucumber weight after crushing
1.1%, the addition of activated carbon is the 5% of the oyster meat after crushing and sea cucumber weight, is digested 12 hours with 50 DEG C of temperature, is obtained
To enzymolysis product；
(5) it is centrifuged：Enzymolysis product is centrifuged repeatedly for several times by centrifuge with the rotating speed of 5000 turns/hour, centrifugation
After remove supernatant, obtain be centrifuged product；
(6) separate：By the centrifugation product Ultra filtration membrane of the dalton of aperture 6000～100000, clear liquid is removed；
(7) concentrate：Lower clear liquid to obtaining in step (6) is concentrated with aperture less than the NF membrane of 60 dalton, takes supernatant
Liquid；
(8) it is dried：Supernatant to obtaining in step (7) is dried using spray dryer, and spray dryer is dried
EAT be 133 DEG C, that is, obtain embodiment 3 small molecule ginseng oyster polypeptide finished product.After testing, the small molecule ginseng of embodiment 3
The recovery rate of oyster polypeptide is 3.0%.
For the small molecule of embodiment 1 joins oyster polypeptide finished product, by high performance liquid chromatograph and according to GB/T 22729-
2008《Ocean fish oligopeptide powder》Detection peptide molecular weight distribution, the chromatogram for obtaining is shown in Fig. 1, and molecular weight detection the results are shown in Table 1.
The small molecule of the embodiment 1 of table 1 joins the molecular weight detection result of oyster polypeptide finished product
As seen from Table 1, in the small molecule ginseng oyster polypeptide finished product of embodiment 1, molecular weight is less than the polypeptide of 2000 dalton
Weight ratio is 99.26%.
Claims (3)
1. a kind of small molecule joins the extracting method of oyster polypeptide, it is characterised in that comprise the following steps：
（1）Cleaning：With fresh sea cucumber and oyster as raw material, with clear water most sea cucumber and oyster surface silt are washed；
（2）Remove oyster shell：In the environment that temperature is less than 5 DEG C, oyster meat is taken out into stand-by with special oyster knife, during storage
Between be less than 24 hours；
（3）Crush：Oyster meat moisture is drained to not dripping, oyster meat and sea cucumber are crushed with industrial beating crusher, crushed and be
Particle less than 2 centimetres；
（4）Enzymolysis：By the oyster meat after crushing and sea cucumber with running water or pure water with 1:2～8 weight ratio adds to biology enzyme
In solution tank, and alkali protease, Collagenase and papain and activated carbon, alkaline egg are added in biological enzymolysis tank
The addition of white enzyme is 0.1～1 % of the oyster meat after crushing and sea cucumber weight, and the addition of Collagenase is after crushing
0.4～4 % of oyster meat and sea cucumber weight, the addition of papain is the 0.2 of the oyster meat after crushing and sea cucumber weight
～2 %, the addition of activated carbon is 1～8 % of the oyster meat after crushing and sea cucumber weight, and with 40～60 DEG C of temperature 8 are digested
～20 hours, obtain enzymolysis product；
（5）Centrifugation：Enzymolysis product is centrifuged repeatedly for several times by centrifuge with the rotating speed of 5000 turns/hour, is gone after centrifugation
Except supernatant, obtain that product is centrifuged；
（6）Separate：By the centrifugation product Ultra filtration membrane of the dalton of aperture 6000～100000, clear liquid is removed；
（7）Concentration：To step（6）In the lower clear liquid that obtains concentrated less than the NF membrane of 60 dalton with aperture, take supernatant；
（8）It is dried：To step（7）In the supernatant that obtains be dried using spray dryer or freeze dryer, that is, obtain little point
Son ginseng oyster polypeptide finished product.
2. a kind of small molecule according to claim 1 joins the extracting method of oyster polypeptide, it is characterised in that described activated carbon
For the activated carbon of 200 mesh.
3. a kind of small molecule according to claim 1 and 2 joins the extracting method of oyster polypeptide, it is characterised in that described spraying
The EAT that drying machine is dried is 120～145 DEG C, and the temperature that described freeze dryer is dried is subzero 60 DEG C.
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| CN201610896521.9A CN106636267A (en) | 2016-10-14 | 2016-10-14 | Extracting method of small-molecular sea cucumber-oyster polypeptide |
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| CN201610896521.9A CN106636267A (en) | 2016-10-14 | 2016-10-14 | Extracting method of small-molecular sea cucumber-oyster polypeptide |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113456561A (en) * | 2021-07-09 | 2021-10-01 | 广西和桂集团有限公司 | Preparation method and application of essence containing polypeptide and plant extract |
| CN114350733A (en) * | 2021-12-29 | 2022-04-15 | 广东冠龙生物科技有限公司 | Oyster active protein peptide composite precise enzymolysis process |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104263788A (en) * | 2014-09-25 | 2015-01-07 | 冯群力 | Preparation method of sea cucumber polypeptide |
| CN105296584A (en) * | 2015-10-09 | 2016-02-03 | 烟台源力德海洋生物有限公司 | Preparation method of marine biological health care peptide preparation |
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2016
- 2016-10-14 CN CN201610896521.9A patent/CN106636267A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104263788A (en) * | 2014-09-25 | 2015-01-07 | 冯群力 | Preparation method of sea cucumber polypeptide |
| CN105296584A (en) * | 2015-10-09 | 2016-02-03 | 烟台源力德海洋生物有限公司 | Preparation method of marine biological health care peptide preparation |
Non-Patent Citations (1)
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| 许庆陵 等: "牡蛎水解蛋白制备及脱腥技术研究", 《食品研究与开发》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113456561A (en) * | 2021-07-09 | 2021-10-01 | 广西和桂集团有限公司 | Preparation method and application of essence containing polypeptide and plant extract |
| CN114350733A (en) * | 2021-12-29 | 2022-04-15 | 广东冠龙生物科技有限公司 | Oyster active protein peptide composite precise enzymolysis process |
| CN114350733B (en) * | 2021-12-29 | 2024-01-26 | 广东冠龙生物科技有限公司 | Oyster active protein peptide composite accurate enzymolysis technology |
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Application publication date: 20170510 |