CN101948898A - Method for preparing nano oligopeptide collagen - Google Patents
Method for preparing nano oligopeptide collagen Download PDFInfo
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- CN101948898A CN101948898A CN 201010282386 CN201010282386A CN101948898A CN 101948898 A CN101948898 A CN 101948898A CN 201010282386 CN201010282386 CN 201010282386 CN 201010282386 A CN201010282386 A CN 201010282386A CN 101948898 A CN101948898 A CN 101948898A
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Abstract
The invention relates to a method for preparing nano oligopeptide collagen. The method comprises the following steps of: mincing chicken toes and adding a certain amount of water and lipase into the minced chicken toes for degreasing under the conditions of temperature of between 30 and 50 DEG C and pH of between 8.0 and 10.0; placing the degreased chicken toes in an enzymolysis tank, adding a certain amount of water into the tank, raising the temperature to 70 to 90 DEG C and keeping the temperature for 10 to 30 minutes; adding a certain amount of papain under the conditions of temperature of between 40 and 50 DEG C and pH of between 6.0 and 7.0 to hydrolyze for 1 to 2 hours; adding a certain amount of complex protease under the conditions of temperature of between 50 and 60 DEG C and pH of between 6.0 and 8.0 to hydrolyze for 1 to 2 hours; continuously adding a certain amount of flavourzyme to hydrolyze for 2 to 4 hours, raising the temperature to 75 to 100 DEG C after the hydrolysis and keeping the temperature for 10 to 30 minutes; and performing post-processing to obtain the nano oligopeptide collagen. The nano oligopeptide collagen prepared by the method of the invention has advantages on molecular weight distribution and hydroxyproline content.
Description
Technical field
The present invention relates to a kind of extracting method of collagen protein, be specifically related to a kind of preparation method of nanometre collagen oligopeptide.
Background technology
Collagen protein is the spirality fiber shape protein that is twisted into by three peptide chains, is a kind of glycoprotein, and molecule contains sugar, reaches a large amount of glycine, proline(Pro), oxyproline etc., exist in all multicellular animals bodies, and be the maximum class protein of in-vivo content.In human body, account for 1/4th of total protein content, be present in nearly all tissue, it is a kind of extracellular protein, exist with the insoluble fiber form, formation to animal and human's body skin, blood vessel, bone, muscles and bones, cartilage is very important, being the important substance of reticular tissue, is decision reticular tissue flexible principal element, plays the part of the role of conjunctive tissue in zooblast.With advancing age, human body is after 25 years old, and intravital collagen protein just begins to run off gradually, especially the women.Therefore keep bone health and skin elasticity and just need replenishing collagen in time.Containing multiple biologically active peptides in the collagen protein, as suppressing hypertensin conversion enzyme activity, suppressing agglutinate activity of blood platelet, anti-oxidant activity, anti-tumor activity etc.
Collagen oligopeptide is to be raw material with various collagens (skin of animal, bone, joint etc.), and through the directed gradient hydrolyzed product of multiple protein enzyme, its molecular weight is below 3000Da.Collagen oligopeptide can be dissolved in water (cold water is solubilized also) fully, and aqueous solution low viscosity also has flowability under 60% high density, and acid resistance is good, does not all have precipitation under the situation of acid, alkali; Resistance to elevated temperatures is good, and 200 ℃ of heating also do not have precipitation, because its molecular weight is less, so easier being absorbed by the body.
China is foster fowl big country in the world, and chicken is cultured had become the rapidest most typical industry of agriculture industrialization already, and with annual 5%～10% speed sustainable growth.Continuous development along with market, in the chicken course of processing, many chicken byproducts such as chicken organ, shank are used as food raw material more and are processed into cooked product, the utilization ratio and the added value of product are lower, nearly 1,000,000 tons of the output of shank in 2008, except can be made into prepared food, most shank all is not fully utilized.
China shank deep processing enterprise is less at present, and most of enterprise carries out elementary processing with regard to list marketing, and the enterprise that has does not even process with regard to list marketing, existing deep processing lags behind, scientific and technological content is few, and up-to-dateness is hanged down inferior outstanding problem, seriously restricts the lifting of shank added value of product.
At present; external a lot of bibliographical informations about some researchs of physiological function of shank collagen; Preliminary study arrived its improve skeleton intensity, protection stomach mucous membrane, suppress increased blood pressure, anti-ageing and promote the effect of aspect such as skin collagen metabolism; reported that shank collagen not only has wide practical use on food, makeup; but also has preventing hypertension, application medically such as anti-treating rheumatic arthritis.Therefore, utilize shank to extract collagen protein and be applied on food, makeup, the medicine, have important scientific meaning and huge applicating and exploitation value as raw material.
In the method for preparing collagen peptide of prior art, degreasing need be used a large amount of organic solvents or alkali and be deviate from fat in the raw material, and organic solvent and alkali degreasing all exist serious potential safety hazard and environmental pollution; It is the combinative enzyme hydrolysis method that enzymolysis process is used maximum, but the main drawback of existing enzymolysis process method is: enzymolysis process is difficult to control, and the wide and product of the molecular weight distribution of the collagen peptide of preparation has bitter taste; The producer that has also adopts ultrafiltration afterwards with the filtrate evaporation concentration, and evaporation concentration need consume a large amount of heats, and does not reach the purpose of desalination and removal total free aminoacids.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of nanometre collagen oligopeptide overcomes enzymolysis process of the prior art and is difficult to control, and the molecular weight distribution of collagen peptide has problems such as bitter taste than wide and product.
In order to solve the problems of the technologies described above, the present invention adopts following scheme:
A kind of preparation method of nanometre collagen oligopeptide comprises:
(1) rubs: shank is cleaned up the back rubbing obtain the shank material;
(2) degreasing: described shank material being placed degreasing tank, add a certain amount of water and lipase, is 30-50 ℃ in temperature, and pH carries out degreasing under the condition of 8.0-10.0, finishes the after-filtration washing in degreasing and obtains the degreasing shank;
(3) enzymolysis: described degreasing shank is placed enzymatic vessel, add a certain amount of water,, added a certain amount of papain hydrolysis 1-2 hour under the condition of pH 6.0-7.0 at temperature 40-50 ℃; At temperature 50-60 ℃, add a certain amount of conjugated protein enzymic hydrolysis 1-2 hour under the condition of pH 6.0-8.0; Continue to add a certain amount of flavor protease hydrolysis 2-4 hour, after hydrolysis finishes, be warming up to 75-100 ℃ and kept 10-30 minute, filter and obtain shank filtrate just;
(4) aftertreatment: the first filtrate of described shank is carried out aftertreatment obtain described nanometre collagen oligopeptide.
By technique scheme of the present invention, adopt lipase to carry out degreasing operation, avoided organic solvent and alkali degreasing exist in the prior art serious potential safety hazard and environmental pollution, omitted the residual operations such as alkali of follow-up removal, thereby have very high biological safety.By at first adopting specific two kinds of restriction endonucleases (papoid and compound protease) enzymolysis, utilize suitable specific excision enzyme (flavor protease) that hydrophobic amino acid is modified again, the nanometre collagen oligopeptide for preparing does not have any bitter taste, has improved the quality of nanometre collagen oligopeptide.Add three kinds of proteolytic ferments (papoid, compound protease and flavor protease) successively by employing and carry out enzymolysis, make enzymolysis process be easy to control, the nanometre collagen oligopeptide for preparing has more advantage on molecular weight distribution and oxyproline HYP content.In accordance with the present production process, technology is simple, and uniqueness is with short production cycle.
According to a preferred embodiment of the present invention, the quality of the water that adds in the described degreasing operation is 2-10 a times of shank quality, is preferably 2-5 doubly, is preferably 4-5 doubly, more preferably 4 times.
According to a preferred embodiment of the present invention, the quality of the lipase that adds in the described degreasing operation accounts for the 0.05%-0.5% of shank quality, is preferably 0.05%-0.15%, is preferably 0.15%.
According to a preferred embodiment of the present invention, the temperature in the described degreasing operation is preferably 35-40 ℃, is preferably 37 ℃.
According to a preferred embodiment of the present invention, the pH in the described degreasing operation is preferably 9.0-10.0, is preferably 9.0, is preferably 9.5.
According to a preferred embodiment of the present invention, described degreasing was carried out 1-2 hour, was preferably 1.5 hours, was preferably 1 hour.
According to a preferred embodiment of the present invention, described degreasing operation finishes after-filtration, and the shank material that leaches was washed 30 minutes, is preferably and is washed till neutrality at normal temperatures.
According to a preferred embodiment of the present invention, the quality of the water that adds in the described enzymolysis operation is 1-5 a times of shank quality, is preferably 2-4 doubly, is preferably 4 times.
According to a preferred embodiment of the present invention, before adding papain hydrolysis, be warming up to 70 ℃-90 ℃ earlier and kept 10-30 minute in the described enzymolysis operation.By technique scheme of the present invention, before adding protease hydrolysis, be warming up to 70 ℃-90 ℃ earlier in the described enzymolysis operation and kept 10-30 minute, make pretreated albumen can expose more enzyme point of application, the proteolysis reaction after quickening.
According to a preferred embodiment of the present invention, the quality of described papoid accounts for the 0.1%-1% of shank quality, is preferably 0.1%-0.5%, is preferably 0.2%-0.3%, is preferably 0.2%; The temperature of reaction of described papoid is preferably 40-50 ℃, is preferably 45 ℃; The reaction pH of described papoid is preferably 6.0-7.0, is preferably 6.5; The reaction times of described papoid is preferably 1-2 hour, is preferably 1 hour.
According to a preferred embodiment of the present invention, the quality of described compound protease accounts for the 0.1%-1% of shank quality, is preferably 0.1%-0.5%, is preferably 0.2%-0.3%, is preferably 0.2%; The temperature of reaction of described compound protease is preferably 50-60 ℃, is preferably 50 ℃; The reaction pH of described papoid is preferably 6.0-8.0, is preferably 7; The reaction times of described papoid is preferably 1-2 hour, is preferably 1 hour.
According to a preferred embodiment of the present invention, the quality of described flavor protease accounts for the 0.2%-2% of shank quality, is preferably 0.2%-1%, is preferably 0.4%-0.6%, is preferably 0.4%; The temperature of reaction of described compound protease is preferably 50-60 ℃, is preferably 50 ℃; The reaction pH of described papoid is preferably 6.0-8.0, is preferably 7; The reaction times of described papoid is preferably 2-4 hour, is preferably 2 hours.
According to a preferred embodiment of the present invention, be warming up to 75-100 ℃ behind the described enzymolysis EO and kept 10-30 minute, be preferably 85 ℃ and kept 15 minutes, centrifuging goes to obtain crude extract behind upper strata grease, the sub-cloud bone slag.
According to a preferred embodiment of the present invention, described post-processing operation comprises: add the stirring of 0.5-2% gac and decoloured in 30-60 minute, and then carry out Plate Filtration, obtain the smart filtrate of clear; Ultra-filtration membrane by molecular weight 3000-10000Da filters collagen oligopeptide solution, working pressure 0.1～1MPa, service temperature 20-50 ℃; Adopt the nanofiltration membrane separation technology that filtrate is carried out desalination and concentrated working pressure 0.8～5MPa, service temperature 20-50 ℃; Spraying drying.By technique scheme of the present invention, adopt the ultra-filtration membrane isolation technique that shank collagen enzymolysis solution has been carried out molecular-weight gradation, solved the wide problem of collagen peptide molecular weight distribution, obtained the nanometre collagen oligopeptide that a kind of molecular weight distribution is concentrated.By technique scheme of the present invention, adopt the nanofiltration membrane separation technology that ultrafiltrated is concentrated and desalination, nanofiltration membrane hold back relative molecular mass between ultrafiltration and reverse osmosis, be particularly suitable for low-molecular-weight oligopeptides is carried out desalination and concentrated, and do not influence the biological activity of separate substance, so nanofiltration is applied to concentrating of oligopeptides material and desalination has good effect, can realize industrialization.
According to a preferred embodiment of the present invention, the prepared according to the methods of the invention nanometre collagen oligopeptide is at medicine, healthcare products, the application on the makeup.
Description of drawings
Fig. 1 shows according to nanometre collagen oligopeptide graph of molecular weight distribution of the present invention.
Embodiment
Specific embodiments of the invention below are described in detail in detail.
Be that Licpun (lipase degreasing Lipase degrease-protease hydrolyzed Complex protease hydrolysis-ultra-filtration membrane the separates Ultrafiltration separation-nanofiltration membrane separation Nanofiltration separation) prepared that the independent development of employing company goes out obtains nanometre collagen oligopeptide according to a preferred embodiment of the invention.
The shank that adopts in the present embodiment is purchased white domestic common fryer factory, and lipase, proteolytic enzyme are all purchased the letter Bioisystech Co., Ltd in Denmark Novi.
Embodiment 1
1) the freezing shank that will go out freezer carries out ice-melt and thaws, and with clear water shank is cleaned up, and drains.With warm water that the shank rinsing is clean, the shank after the cleaning is smashed to pieces with tissue mashing machine.Shank material after smashing to pieces moves into degreasing tank.
2) add the water of 4 times of amounts to degreasing tank, regulate pH to 9.0, the lipase that adds shank amount 0.1% carries out degreasing, controlled temperature keeps 1.5h to carry out the degreasing enzymolysis at 35-40 ℃, and degreasing finishes after-filtration, the shank material that leaches was washed 30 minutes, washed twice, normal-temperature water is washed till neutrality, squeezes into enzymatic vessel again.
3) add 4 times deionized water, controlled temperature keeps 30 minutes postcooling to 40-50 ℃ at 90 ℃, regulate pH to 6.5, add the papain papoid of shank amount 0.2% and the protamex compound protease of shank amount 0.2%, hydrolysis was cooled to 50-60 ℃ after 2 hours; Press shank amount 0.4% again and add the Flavourzyme flavor protease, continue hydrolysis and finished in 2 hours, slowly stir during hydrolysis, be warming up to 75-100 ℃ after hydrolysis finishes and kept 10-30 minute, centrifuging goes to obtain crude extract behind upper strata grease, the sub-cloud bone slag.
4) enzymolysis solution after the enzyme cooling of going out is cooled to 35-40 ℃ and adds 1% gac and decolour, and stirs 30-120 minute, and then carries out Plate Filtration, obtains clear essence filtrate.
5) ultra-filtration membrane of smart filtrate by molecular weight 5000Da filtered collagen oligopeptide solution, working pressure 0.1～1MPa, 25 ℃ of service temperatures obtain ultrafiltrated.
6) ultrafiltrated is adopted the nanofiltration membrane separation technology filtrate is carried out desalination and concentrate, working pressure 0.8～5MPa, 25 ℃ of service temperatures obtain the shank collagen oligopeptide concentrated solution behind the purifying.
7) concentrated solution that nanofiltration is obtained carries out spraying drying, promptly gets shank nanometre collagen oligopeptide powder.
The molecular weight distribution that the nanometre collagen oligopeptide powder that obtains is measured the shank collagen oligopeptide by gel permeation chromatography is as shown in table 1:
Table 1
(instrument: Waters 600 high performance liquid chromatographs (are joined 2487 UV-detector and Empower workstation GPC software.Chromatographic condition: chromatographic column: TSKgel 2000SW
XL300mm * 7.8mm; Moving phase: acetonitrile/water/trifluoroacetic acid, 45/55/0.1 (V/V); Detect: UV220nm; Flow velocity: 0.5ml/min; Column temperature: 30 ℃; Specimen preparation: draw sample 2ml in the 10ml volumetric flask, be diluted to scale, with supplying sample introduction behind the millipore filtration membrane filtration with moving phase.)
As seen from the above table, add papoid and compound protease at first simultaneously, add flavor protease subsequently separately, obtain the molecular weight of shank nanometre collagen oligopeptide mainly be distributed in 180-2000Da, the content of the oligopeptides of molecular weight below 1000 accounts for 72% of total content, and its molecular-weight average is about 900Da.
Embodiment 2:
1) the freezing shank that will go out freezer carries out ice-melt and thaws, and with clear water shank is cleaned up, and drains.With warm water that the shank rinsing is clean, the shank after the cleaning is smashed to pieces with tissue mashing machine.Shank material after smashing to pieces moves into degreasing tank.
2) add the water of 4 times of amounts to degreasing tank, regulate pH to 9.0, the lipase that adds shank amount 0.1% carries out degreasing, controlled temperature keeps 1.5h to carry out the degreasing enzymolysis at 35-40 ℃, and degreasing finishes after-filtration, the shank material that leaches was washed 30 minutes, washed twice, normal-temperature water is washed till neutrality, squeezes into enzymatic vessel again.
3) add 4 times deionized water, controlled temperature keeps 30 minutes postcooling to 40-50 ℃ at 90 ℃, regulates pH to 6.5, the papain papoid of adding shank amount 0.2%; After the hydrolysis 1 hour, be warming up to 47-53 ℃, regulate pH to 7.0, press shank amount 0.2% and add the protamex compound protease, continue hydrolysis after 1 hour, be cooled to 50-60 ℃; Press shank amount 0.4% again and add the Flavourzyme flavor protease, continue hydrolysis and finished in 2 hours, slowly stir during hydrolysis, be warming up to 75-100 ℃ after hydrolysis finishes and kept 10-30 minute, centrifuging goes to obtain crude extract behind upper strata grease, the sub-cloud bone slag.
4) enzymolysis solution after the enzyme cooling of going out is cooled to 35-40 ℃ and adds 1% gac and decolour, and stirs 30-120 minute, and then carries out Plate Filtration, obtains clear essence filtrate.
5) ultra-filtration membrane of smart filtrate by molecular weight 5000Da filtered collagen oligopeptide solution, working pressure 0.1～1MPa, 25 ℃ of service temperatures obtain ultrafiltrated.
6) ultrafiltrated is adopted the nanofiltration membrane separation technology filtrate is carried out desalination and concentrate, working pressure 0.8～5MPa, 25 ℃ of service temperatures obtain the shank collagen oligopeptide concentrated solution behind the purifying.
7) concentrated solution that nanofiltration is obtained carries out spraying drying, promptly gets shank nanometre collagen oligopeptide powder.
Embodiment 3:
1) the freezing shank that will go out freezer carries out ice-melt and thaws, and with clear water shank is cleaned up, and drains.With warm water that the shank rinsing is clean, the shank after the cleaning is smashed to pieces with tissue mashing machine.Shank material after smashing to pieces moves into degreasing tank.
2) water that adds 5 times of amounts is regulated pH to 9.5 to degreasing tank, and the lipase that adds shank amount 0.15% carries out degreasing, and controlled temperature keeps 1h to carry out the degreasing enzymolysis at 37 ℃, and degreasing finishes after-filtration, will squeeze into enzymatic vessel behind the shank material thorough washing that leach.
3) add 3 times deionized water, controlled temperature is at 85 ℃, keeps 40 minutes postcooling to 45 ℃, regulates pH to 6.5, the papain papoid of adding shank amount 0.3%; After the hydrolysis 1 hour, be warming up to 50 ℃, regulate pH to 7.0, press shank amount 0.3% and add the protamex compound protease, continue hydrolysis after 1 hour, be cooled to 50 ℃, press shank amount 0.6% again and add the Flavourzyme flavor protease, continue hydrolysis and finished in 2 hours, slowly stir during hydrolysis, be warming up to 85 ℃ after hydrolysis finishes and kept 15 minutes, centrifuging goes to obtain crude extract behind upper strata grease, the sub-cloud bone slag.
4) enzymolysis solution after the enzyme cooling of going out is cooled to 37 ℃ and adds 2% gacs and decolour, and stirs 30 minutes, and then carries out Plate Filtration, obtains clear essence filtrate.
5) ultra-filtration membrane of smart filtrate by molecular weight 5000Da filtered collagen oligopeptide solution, working pressure 0.2MPa, 25 ℃ of service temperatures obtain ultrafiltrated.
6) ultrafiltrated is adopted the nanofiltration membrane separation technology filtrate is carried out desalination and concentrate, working pressure 0.8～5MPa, 25 ℃ of service temperatures obtain the shank collagen oligopeptide concentrated solution behind the purifying.
7) concentrated solution that nanofiltration is obtained carries out spraying drying, promptly gets shank nanometre collagen oligopeptide powder.
By detecting, embodiment 2 is roughly the same with the result of embodiment 3.With embodiment 3 is example, and the nanometre collagen oligopeptide powder that obtains is measured molecular weight distribution such as Fig. 1 of shank collagen oligopeptide by gel permeation chromatography, and concrete molecular weight distribution is as shown in table 2:
Table 2
As seen from the above table, successively add papoid, compound protease and flavor protease successively, the molecular weight of the shank nanometre collagen oligopeptide that obtains mainly is distributed in 180-1000Da, the content of the oligopeptides of molecular weight below 1000 accounts for 80% of total content, its molecular-weight average is about 600Da, belong to collagen oligopeptide, on molecular weight distribution, have more advantage, its partial amino-acid series such as table 3.
The nanometre collagen oligopeptide powder that obtains according to a preferred embodiment of the invention is by HPLC-MS-MS qualitative analysis (Institute of Analysis of Southern Yangtze University that possesses CMA and CNAS authentication credential), and the protein peptide that result's demonstration obtains mainly is made up of nanometre collagen oligopeptide.
Table 3
The nanometre collagen oligopeptide powder that obtains according to a preferred embodiment of the invention adopts automatic analyzer for amino acids, carries out determined amino acid by OPA column front derivation high performance liquid chromatography.The amino acid that above-mentioned two embodiment obtain is formed roughly the same, and its amino acid is formed (Institute of Analysis of Southern Yangtze University provides report) as shown in table 4:
Table 4
| The amino acid title | Content (mg/100mg) | The amino acid title | Content (mg/100mg) |
| Aspartic acid | 5.50 | Gelucystine | 0.22 |
| L-glutamic acid | 10.89 | Xie Ansuan | 3.25 |
| Serine | 2.63 | Methionine(Met) | 1.35 |
| Histidine | 0.92 | Phenylalanine | 2.68 |
| Glycine | 20.02 | Isoleucine | 2.32 |
| Threonine | 2.10 | Leucine | 4.28 |
| Arginine | 7.03 | Methionin | 3.86 |
| L-Ala | 8.06 | Proline(Pro) | 10.84 |
| Tyrosine | 1.20 | Oxyproline | 12.01 |
| Total amino acid | 100 |
(instrument: the Agilent1100 high performance liquid chromatograph, be furnished with UV-detector, automatic sampler and chromatographic working station.Chromatographic condition: chromatographic column: other C18 posts of the same type that Hypersil ODS C18 125mm * 4.0mm or performance are close therewith.Moving phase: A phase: take by weighing 4.0 gram crystallization sodium acetates in 1000 ml beakers, add 1000 ml waters and be stirred to all crystal water dissolvings, add 225 microlitre triethylamines again, stir the also acetic acid of Dropwise 5 %, PH is transferred to 7.20 ± 0.05; Add 5 milliliters of tetrahydrofuran (THF)s, standby after mixing.B phase: take by weighing 4.0 gram crystallization sodium acetates in 500 ml beakers; Add 200 milliliters and be stirred to all crystallization dissolvings; Drip 2% acetic acid PH is transferred to 7.20 ± 0.05; This solution is added 400 milliliters of acetonitriles and 400 ml methanol, standby after mixing.Detect wavelength: UV338nm; Flow velocity: 1.0mL/min; Column temperature: 30 ℃; Sampling volume: 10 μ L.)
As seen from the above table, consist of according to shank nanometre collagen oligopeptide product feature amino acid: glycine (glycine) 20.02%, L-Ala (alanine) 8.06%, proline(Pro) (proline) 10.84%, oxyproline (hydroxyproline) 12.01%.
Above-mentioned two embodiment (embodiment 2 and embodiment 3) are the preferred embodiments of the present invention, by a large amount of experimental results show that: when the shank raw material, zymin and enzymatic hydrolysis condition, when the membrane sepn condition is constant, the molecular weight distribution of resulting nanometre collagen oligopeptide is metastable, shows that its feature graph of molecular weight distribution and amino acid composition is relatively stable with molecular weight distribution and amino acid composition among the embodiment 3.
Above-described, be preferred embodiment of the present invention only, be not in order to limiting scope of the present invention, the above embodiment of the present invention can also be made various variations.Be that every simple, equivalence of doing according to the claims and the description of the present patent application changes and modification, all fall into the claim protection domain of patent of the present invention.The present invention not detailed description be the routine techniques content.
Claims (8)
1. the preparation method of a nanometre collagen oligopeptide comprises:
(1) rubs: shank is cleaned up the back rubbing obtain the shank material;
(2) degreasing: described shank material being placed degreasing tank, add a certain amount of water and lipase, is 30-50 ℃ in temperature, and pH carries out degreasing under the condition of 8.0-10.0, finishes the after-filtration washing in degreasing and obtains the degreasing shank;
(3) enzymolysis: described degreasing shank is placed enzymatic vessel, add a certain amount of water,, added a certain amount of papain hydrolysis 1-2 hour under the condition of pH 6.0-7.0 at temperature 40-50 ℃; At temperature 50-60 ℃, add a certain amount of conjugated protein enzymic hydrolysis 1-2 hour under the condition of pH 6.0-8.0; Continue to add a certain amount of flavor protease hydrolysis 2-4 hour, after hydrolysis finishes, be warming up to 75-100 ℃ and kept 10-30 minute, filter and obtain shank filtrate just;
(4) aftertreatment: the first filtrate of described shank is carried out aftertreatment obtain described nanometre collagen oligopeptide.
2. the preparation method of nanometre collagen oligopeptide as claimed in claim 1 is characterized in that, the quality of the water that adds in the described degreasing operation is 4-5 a times of shank quality.
3. the preparation method of nanometre collagen oligopeptide as claimed in claim 1 is characterized in that, the quality of the lipase that adds in the described degreasing operation is the 0.05%-0.15% of shank quality.
4. the preparation method of nanometre collagen oligopeptide as claimed in claim 1 is characterized in that, the quality of the water that adds in the described enzymolysis operation is 2-4 a times of shank quality.
5. the preparation method of nanometre collagen oligopeptide as claimed in claim 1 is characterized in that, before adding papain hydrolysis, is warming up to 70 ℃-90 ℃ earlier and keeps 10-30 minute.
6. the preparation method of nanometre collagen oligopeptide as claimed in claim 1 is characterized in that, the quality of described papoid accounts for the 0.2%-0.3% of shank quality; The quality of described compound protease accounts for the 0.2%-0.3% of shank quality; The quality of described flavor protease accounts for the 0.4%-0.6% of shank quality.
7. the preparation method of nanometre collagen oligopeptide according to claim 1 is characterized in that described post-processing operation comprises: add the 0.5-2% gac and stir and decoloured in 30-60 minute, and then carry out Plate Filtration, obtain the smart filtrate of clear; Ultra-filtration membrane by molecular weight 3000-10000Da filters collagen oligopeptide solution, working pressure 0.1～1MPa, service temperature 20-50 ℃; Adopt the nanofiltration membrane separation technology that filtrate is carried out desalination and concentrated working pressure 0.8～5MPa, service temperature 20-50 ℃; Spraying drying.
8. the nanometre collagen oligopeptide of the method for claim 1 preparation is at medicine, healthcare products, the application on the makeup.
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| CN102367464A (en) * | 2011-10-18 | 2012-03-07 | 得利斯集团有限公司 | Method for extracting pigskin collagen peptide |
| CN102559820A (en) * | 2011-12-22 | 2012-07-11 | 上海东锦食品集团有限公司 | Preparation method and applications of nano collagen with oxidation resistance capacity |
| CN103393191A (en) * | 2013-07-09 | 2013-11-20 | 广州中医药大学 | Crocodile meat oral liquid and preparation method thereof |
| CN108371255A (en) * | 2018-02-08 | 2018-08-07 | 青岛彬利宠物用品有限公司 | A kind of dog food cat food and preparation method thereof preparing OCO collagen small peptides using enzymolysis process |
| US10765714B2 (en) * | 2014-08-20 | 2020-09-08 | Goo Whan KIM | Collagen component |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1552889A (en) * | 2003-12-18 | 2004-12-08 | 四川铭让生物科技有限公司 | Use and preparation of collagen protein and collagen polypeptide |
-
2010
- 2010-09-15 CN CN 201010282386 patent/CN101948898B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1552889A (en) * | 2003-12-18 | 2004-12-08 | 四川铭让生物科技有限公司 | Use and preparation of collagen protein and collagen polypeptide |
Non-Patent Citations (1)
| Title |
|---|
| 《国立中兴大学农业暨自然资源学院动物科学系动科硕博士论文》 20091128 林亮全 以废弃鸡爪开发具抗氧化与降高血压效果酵素水解物之研究 摘要 1-8 , * |
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| CN102367464A (en) * | 2011-10-18 | 2012-03-07 | 得利斯集团有限公司 | Method for extracting pigskin collagen peptide |
| CN102559820A (en) * | 2011-12-22 | 2012-07-11 | 上海东锦食品集团有限公司 | Preparation method and applications of nano collagen with oxidation resistance capacity |
| CN103393191A (en) * | 2013-07-09 | 2013-11-20 | 广州中医药大学 | Crocodile meat oral liquid and preparation method thereof |
| CN103393191B (en) * | 2013-07-09 | 2015-08-12 | 广州中医药大学 | Crocodile meat oral liquid and preparation method thereof |
| US10765714B2 (en) * | 2014-08-20 | 2020-09-08 | Goo Whan KIM | Collagen component |
| CN108371255A (en) * | 2018-02-08 | 2018-08-07 | 青岛彬利宠物用品有限公司 | A kind of dog food cat food and preparation method thereof preparing OCO collagen small peptides using enzymolysis process |
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|---|---|
| CN101948898B (en) | 2013-06-26 |
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