CN101602790A - A kind of deep-sea fish skin collagen oligopeptide and preparation method thereof with antihypertensive function - Google Patents
A kind of deep-sea fish skin collagen oligopeptide and preparation method thereof with antihypertensive function Download PDFInfo
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- CN101602790A CN101602790A CNA2009100995080A CN200910099508A CN101602790A CN 101602790 A CN101602790 A CN 101602790A CN A2009100995080 A CNA2009100995080 A CN A2009100995080A CN 200910099508 A CN200910099508 A CN 200910099508A CN 101602790 A CN101602790 A CN 101602790A
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Abstract
The invention discloses a kind of deep-sea fish skin collagen oligopeptide and preparation method thereof, belong to the Food Engineering Development field of functional food, particularly have the deep-sea fish skin collagen oligopeptide of antihypertensive function with antihypertensive function.It is raw material with the used for deep-sea fish skin, utilize the complex enzyme zymohydrolysis technology fish-skin to be degraded into polypeptide, oligopeptides and the total free aminoacids mixed solution that contains molecular weight and differ in size, adopt membrane separation technique then, hold back with the nanofiltration membrane of different size respectively and remove total free aminoacids and polypeptide, obtain the deep-sea fish skin collagen oligopeptide of oligopeptides content 50～80%; This deep-sea fish skin collagen oligopeptide is pollution-free, security good, and its molecular weight ranges is at 200～1300D, and its product has the function that reduces hyperpietic's blood pressure.
Description
Technical field
The invention belongs to the Food Engineering Development field of functional food, particularly have deep-sea fish skin collagen oligopeptide of antihypertensive function and preparation method thereof.
Technical background
In recent years along with the development of bio-science technology, it is found that protein except that human body multiple amino acids and energy are provided, some little peptides (oligopeptides) that discharge from the original sequence of protein also have different physiological roles, these little peptides (oligopeptides) can directly be absorbed by the body by the degraded of enteron aisle plurality of enzymes system, have from the isolating functional peptides of natural protein at present varied, according to the function branch: the calcium absorption of promotion peptide is arranged; Blood pressure lowering peptide; The research of immunomodulatory peptides etc., particularly blood pressure lowering peptide is the focus of bioactive peptide research especially, studies show that blood pressure lowering peptide (oligopeptides) is the inhibition drug of a kind of angiotensin-converting enzyme (angiotension I cemverting enzyme ACE).
Blood pressure lowering peptide (oligopeptides) mainly contains the blood pressure effect that reduces the hyperpietic, its hypotensive effect is to realize by the activity that suppresses blood in human body in angiotensin saccharase (ACE), the intravital blood pressure of normal people is regulated by multiple factor, wherein most important regulation system is renin-angiotensin system and kallikrein kinin system, the former be the rising regulation system its, the latter is the reduction regulation system, (ACE) enzyme that plays an important role in the renin-angiotensin system, it makes it slough C-terminal two amino acid (His-Len) to be transformed into Angiotensin II by acting on the nervous plain I of this system medium vessels, and Angiotensin II is a strongest active contraction drug in the renin-angiotensin regulation system, and it can act on arteriole and make vascular smooth muscle contraction causing rapidly pressor effect.Blood pressure lowering peptide (oligopeptides) hypotensive effect is to realize by the competitive inhibition to ACE, but under normal circumstances very little to the effect of ACE.
The peptide of preparation step-down at present raw material has: utilize the hairtail spine to make with fermentation process; Utilize rice separated protein to make through protease hydrolyzed; Utilize yolk protein to make with the Sumizyme MP enzymolysis; Utilize animal skin glue collagen to make etc. through protease hydrolyzed in addition, the product that makes is referred to as the step-down peptide, all contains polypeptide in its product, and oligopeptides and total free aminoacids come down to polypeptide, oligopeptides and free amino acid mixture, and molecular weight differs in size.The main component that really plays the step-down effect in these step-down peptide products is an oligopeptides, molecular weight ranges between 200～1000D, can be in these products the content of oligopeptides lower.
The preparation method of oligopeptides has: enzymolysis process, extraction method, autolysis method, modal is enzymolysis process, the raw material of selecting for use has vegetable-protein, animal proteinum, wherein animal proteinum is based on terrestrial animal, if produce the Pi Weiduo that collagen oligopeptide is generally also selected the terrestrial animal domestic animals for use, because of these raw materials cheap and easy to get, but there is potential safety hazard in these raw materials, as mad cow disease, foot and mouth disease, and the raising of these domestic animals animals is subject to Effect of Environmental, so people have caused worry to its security.Why we have selected used for deep-sea fish skin is the feedstock production collagen oligopeptide, and this is because these fish growths among the deep-sea, can not be subjected to Effect of Environmental, can thoroughly eliminate the worry of security.
Being seen in the market collagen protein series products comes down to protein, polypeptide, oligopeptides, amino acid whose mixture, based on protein, polypeptide, molecular weight differs in size, be mainly used in the raw material of food service industry as food, protective foods, small part is used for cosmetic industry, as the raw material of producing beauty and skin care class makeup, the content of oligopeptides that plays active function in these raw materials is not high, and the index of oligopeptide-free content, have protein content only, these indexs of molecular-weight average can not embody the quality scale of product.Proteinic enzymolysis is a complicated biochemical reaction process, be subjected to the influence of multiple condition element, we are according to the mechanism of action of its various condition elements, select suitable enzyme and enzymatic hydrolysis condition, the higher enzymolysis solution of oligopeptides content can be obtained, the content of oligopeptides can be further improved again by the nanofiltration membrane separation means.
Summary of the invention
The objective of the invention is to adopt used for deep-sea fish skin is raw material, select suitable enzymatic hydrolysis condition, obtain the higher enzymolysis solution of oligopeptides content, hold back separation means by nanofiltration membrane then and further improve oligopeptides content in the enzymolysis solution, make deep-sea fish skin collagen oligopeptide product through vacuum concentration, spraying drying again with antihypertensive function, oligopeptides content is 50～80% in the product, molecular weight ranges is at 200D～1300D, this deep-sea fish skin collagen oligopeptide product is pollution-free, security good, has hypotensive effect.
The manufactured materials of this product is a used for deep-sea fish skin, carry out enzymolysis with compound protease, the fish-skin concentration of enzymolysis is 10～20%, utilize enzyme (prozyme) technology of separating that the fish-skin enzymolysis is become polypeptide, oligopeptides that molecular weight differs in size and the mixed solution that contains total free aminoacids, adopt membrane separation technique then, hold back with the nanofiltration membrane of different size respectively and remove total free aminoacids and molecular weight polypeptide, obtain the deep-sea fish skin collagen oligopeptide product of oligopeptides content 50～80% greater than 1300D.
The preparation method of deep-sea fish skin collagen oligopeptide of the present invention, be that freezing used for deep-sea fish skin is thawed, make fish-skin be dispersed into loose sheet or diffusing strip, drop into and add the water that 5-10 doubly measures in the enzymolysis pot, add the 0.1-0.5% compound protease, adjust PH, stirring is heated, temperature is controlled at 45 ℃～65 ℃, 2～8 hours time, is warming up to 85 ℃ after enzymolysis finishes and keeps 15 minutes, enzyme goes out, going out adds 1～2% activated carbon behind the enzyme and slowly stirs and kept 30～60 minutes, takes off raw meat, decolouring adds 4%～8% diatomite then and adds while stirring, carry out coarse filtration, smart filter, the smart filtrate that obtains is held back by nanofiltration membrane, obtains the trapped fluid of oligopeptides content 50～80%, and trapped fluid is carried out vacuum concentration, reach at 15～20% o'clock in solid content, carry out spraying drying, obtain pulverous deep-sea fish skin collagen oligopeptide with antihypertensive function, the finished product that makes is packed.
The enzymolysis solution that the present invention obtains adopts decolorizing with activated carbon, diatomite to help filter, and its consumption is 1～3 (unit).
Enzymolysis solution that the present invention obtains adopts nanofiltration membrane to hold back separation, remove molecular weight greater than the polypeptide of 1300D and molecular weight less than the 200D total free aminoacids, the trapped fluid oligopeptides content 50～80% that obtains.
Oligopeptides liquid after the present invention separates carries out vacuum concentration, and vacuum tightness 0.08～0.1 MPa, 50 ℃～65 ℃ of temperature, solid content reach at 15～20% o'clock and carry out spraying drying or lyophilize.
The deep-sea fish skin collagen oligopeptide product of the present invention's preparation, oligopeptides content reaches 50～80%, has the function of bringing high blood pressure down.
Used for deep-sea fish skin raw material of the present invention, because of the solubility of deep-sea fish skin collagen good, with respect to the Mammals collagen protein, its collagenous fiber bundle of collagen of fish skin is thicker, easier to reactions such as enzyme, heat, easily become small-molecular peptides (oligopeptides), can improve the oligopeptides content in the product by the animal protease hydrolyzed.
The enzymolysis of used for deep-sea fish skin adopts composite animal albumen enzyme, enzymolysis PH5.0-9.0; Hydrolysis temperature 45-60 ℃, time 2-8 hour, the enzymolysis solution that obtains through enzymolysis was warming up to 85 ℃ and keeps the enzyme that goes out half an hour, adds activated carbon, diatomite simultaneously, stirs and takes off raw meat, decolouring, obtains clear filtrate after filtration.The clear filtrate that obtains adopts the nanofiltration membrane segmentation to hold back separation, be that the nanofiltration membrane of 1300D is held back polypeptide and the protein of removal molecular weight greater than 1300D earlier with molecular weight cut-off, leave and take filtered solution, selecting molecular weight cut-off then for use is that the above nanofiltration membrane of 200D is held back, remove total free aminoacids by nanofiltration membrane, hold back by the different nanofiltration membrane of two kinds of molecular weight cut-offs, the oligopeptides ratio in the trapped fluid is improved, content reaches 50～80%.
Adopt vacuum concentration by the filtrate that nanofiltration membrane is held back after the separation, carry out spraying drying or lyophilize when making solid content reach 15-20%, inlet temperature 190-220 ℃; Temperature out 90-105 ℃; Make deep-sea fish skin collagen oligopeptide.
Deep-sea fish skin collagen oligopeptide is the preparation method comprise the steps:
1) the freezing used for deep-sea fish skin that will go out freezer carries out ice-melt and thaws, and makes to be dispersed into strip or sheet, drops in the enzymolysis pot, adds the water of 4～8 times of amounts, slowly stirs and melts.
2) add fish-skin amount 0.1～0.3% compound protease, regulate PH6.0～8.0, temperature slowly stirs enzymolysis at 45～65 ℃, 4～8 hours time.
3) enzymolysis time is warming up to 80～90 ℃ after arriving, and keeps 10～30 minutes, and enzyme goes out
4) enzymolysis solution behind the enzyme that goes out adds 1～3% activated carbon, stirs 20～40 minutes, adds 1～3% diatomite again and slowly stirs 15 minutes, puts into whizzer and carries out coarse filtration, and the coarse filtration liquid that obtains carries out the smart filter of flame filter press again, obtains the smart filtrate of clear;
5) the smart filtrate that obtains is held back with the nanofiltration membrane of molecular weight cut-off＞1300D, leaves and takes the filtrate of passing through.
6) filtrate that will pass through is held back for＞200 nanofiltration membrane with molecular weight cut-off again, leaves and takes and is trapped liquid.
7) trapped fluid that obtains carries out vacuum concentration, vacuum tightness 0.08～0.1MPa, and 50～70 ℃ of temperature are concentrated into solid content and reach at 15～20% o'clock and stop;
8) concentrated solution that obtains is carried out spraying drying, (190～215 ℃ of inlet temperatures, 95～105 ℃ of temperature outs) obtains pulverous deep-sea fish skin collagen oligopeptide.
The deep-sea fish skin collagen oligopeptide product that the present invention makes is pollution-free, security good, and its molecular weight ranges is at 200～1300D, and its product of oligopeptides content has the effect that reduces hyperpietic's blood pressure.
Description of drawings
Fig. 1 is the deep-sea fish skin collagen oligopeptide preparation technology schema with antihypertensive function.
Embodiment
Embodiment 1
1) the freezing used for deep-sea fish skin that will go out freezer carries out ice-melt and thaws, and makes to be dispersed into strip or sheet, drops in the enzymolysis pot, adds the water of 6 times of amounts, slowly stirs, and is warming up to 56 ℃.
2) regulate PH to 6.0, add fish-skin amount 0.15% compound protease, controlled temperature was held time 6 hours at 53～56 ℃.
3) enzymolysis time is warming up to 85 ℃ after arriving, and keeps 15 minutes, and enzyme goes out;
4) enzymolysis solution behind the enzyme that goes out adds 2% activated carbon, stirs 30 minutes, adds 2% diatomite again and slowly stirs 20 minutes, and enzymolysis solution is put into whizzer and carried out coarse filtration, and the coarse filtration liquid that obtains carries out the smart filter of flame filter press again, obtains the smart filtrate of clear;
5) the smart filtrate that obtains is held back with the nanofiltration membrane of molecular weight cut-off 1300, leaves and takes the filtrate of passing through.
6) filtrate that will pass through is that 200 nanofiltration membrane is held back with molecular weight cut-off again, leaves and takes trapped fluid.
7) trapped fluid that obtains carries out vacuum concentration, vacuum tightness 0.08MPa, and 65 ℃ of temperature are concentrated into solid content and reach termination in 15～20% o'clock;
8) concentrated solution that obtains is carried out spraying drying, 200 ℃ of inlet temperatures, 95 ℃ of temperature outs obtain pulverous deep-sea fish skin collagen oligopeptide.
Embodiment 2
1) the freezing used for deep-sea fish skin that will go out freezer carries out ice-melt and thaws, and with the making beating of mincer strand, drops in the enzymolysis pot, adds the water of 6 times of amounts, mixes heat temperature raising to 58 ℃.
2) regulate PH to 6.0～7.0, add fish-skin amount 0.1% compound protease, controlled temperature adds 0.5% compound protease at 55～58 ℃ after 3 hours again, continues to keep 2 hours.
3) after enzymolysis finishes, add 2% activated carbon, warming while stirring to 85 ℃, kept 20 minutes, the enzyme that goes out adds 2% diatomite again and slowly stirred 20 minutes, enzymolysis solution is put into whizzer and is carried out coarse filtration, and the coarse filtration liquid that obtains carries out the smart filter of flame filter press again, obtains the smart filtrate of clear.
4) the smart filtrate that obtains is held back with the nanofiltration membrane of molecular weight cut-off 1300, leaves and takes the filtrate of passing through.
5) filtrate that will pass through is that 200 nanofiltration membrane is held back with molecular weight cut-off again, leaves and takes and holds back filtrate.
6) trapped fluid that obtains carries out vacuum concentration, vacuum tightness 0.1MPa, and 60 ℃ of temperature are concentrated into solid content and reach termination in 15～20% o'clock;
7) concentrated solution that obtains is carried out spraying drying, 205 ℃ of inlet temperatures, 95 ℃ of temperature outs obtain pulverous deep-sea fish skin collagen oligopeptide.
Embodiment 3
1) the freezing used for deep-sea fish skin that will go out freezer carries out ice-melt and thaws, and makes to be dispersed into strip or sheet, drops in the enzymolysis pot, adds the water of 8 times of amounts, slowly stirs, and is warming up to 56 ℃.
2) regulate PH to 6.8, add fish-skin amount 0.2% compound protease, controlled temperature was held time 5 hours at 53～56 ℃.
3) enzymolysis is warming up to 85 ℃ after stopping, and keeps 20 minutes, and enzyme goes out;
4) enzymolysis solution behind the enzyme that goes out is cooled to the fishy-smell removing agent of 28～35 ℃ of addings 0.1%, stirred 30 minutes, and added 2% diatomite again and slowly stirred 20 minutes, enzymolysis solution is put into whizzer and is carried out coarse filtration, the coarse filtration liquid that obtains carries out the smart filter of flame filter press again, obtains the smart filtrate of clear;
5) the smart filtrate that obtains is held back with the nanofiltration membrane of molecular weight cut-off 1300, leaves and takes the filtrate of passing through.
6) filtrate that will pass through is that 200 nanofiltration membrane is held back with molecular weight cut-off again, leaves and takes and holds back filtrate.
7) trapped fluid that obtains carries out vacuum concentration, vacuum tightness 0.08MPa, and 65 ℃ of temperature are concentrated into solid content and reach termination in 15～20% o'clock;
8) concentrated solution that obtains is carried out spraying drying, 200 ℃ of inlet temperatures, 100 ℃ of temperature outs obtain pulverous deep-sea fish skin collagen oligopeptide.
Deep-sea fish skin collagen oligopeptide capsule formula one:
Deep-sea fish skin collagen oligopeptide powder 75-85%
Tea-polyphenol 5-10%
Aloe 5-10%
Semen Ginkgo extrac 4-6%
Mix dry granulation, can capsule.
Deep-sea fish skin collagen oligopeptide granular recipe two:
Deep-sea fish skin collagen oligopeptide powder 70-80%
Skim-milk 15-20%
Broken pollen 10-15%
Honey 5-10%
Mix, granulate, dry, aluminium foil is packed makes.
Claims (8)
1. deep-sea fish skin collagen oligopeptide with antihypertensive function, it is characterized in that preparing raw material is used for deep-sea fish skin, carry out enzymolysis with compound protease, utilize zymolysis technique that fish-skin is degraded into polypeptide, oligopeptides that molecular weight differs in size and the mixed solution that contains total free aminoacids, adopt membrane separation technique then, hold back with the nanofiltration membrane of different size respectively and remove total free aminoacids and molecular weight polypeptide, obtain the deep-sea fish skin collagen oligopeptide product of oligopeptides content 50～80% greater than 1300D.
2. according to claims 1 described deep-sea fish skin collagen oligopeptide, the fish-skin concentration that it is characterized in that enzymolysis is 10～20%.
3. the preparation method of collagen of fish skin oligopeptides according to claim 1, it is characterized in that freezing used for deep-sea fish skin is thawed, fish-skin is dispersed in diffusing sheet or the diffusing strip input enzymolysis pot, add the water that 5-10 doubly measures, the 0.1-0.5% compound protease, adjust PH, stirring is heated, temperature is controlled at 45 ℃～65 ℃, 2～8 hours time was warming up to 85 ℃ after enzymolysis finishes and kept 15 minutes, and enzyme goes out, go out and add 1～2% activated carbon behind the enzyme and slowly stir and kept 30～60 minutes, take off raw meat, decolouring adds 4%～8% diatomite then and adds while stirring, carries out coarse filtration then, smart filter, the smart filtrate that obtains is held back by nanofiltration membrane, obtain the trapped fluid of oligopeptides content more than 70%, trapped fluid is carried out vacuum concentration, reach at 15～20% o'clock in solid content, carry out spraying drying, obtain pulverous deep-sea fish skin collagen oligopeptide.
4. deep-sea fish skin collagen oligopeptide preparation method according to claim 3 is characterized in that the enzymolysis solution that obtains adopts decolorizing with activated carbon, diatomite to help filter, and its consumption is 1～3 (unit).
5. deep-sea fish skin collagen oligopeptide preparation method according to claim 4, the PH that it is characterized in that enzymolysis solution is 5.0～9.0.
6. deep-sea fish skin collagen oligopeptide preparation method according to claim 4, it is characterized in that the enzymolysis solution that obtains adopts nanofiltration membrane to hold back separation, remove molecular weight greater than the polypeptide of 1300D and molecular weight total free aminoacids, the trapped fluid oligopeptides content 50～80% that obtains less than 200D.
7. collagen of fish skin preparation method of oligopeptide according to claim 3, oligopeptides liquid after it is characterized in that separating carries out vacuum concentration, vacuum tightness 0.08～0.1 MPa, 50 ℃～65 ℃ of temperature, solid content reach at 15～20% o'clock and carry out spraying drying or lyophilize.
8. deep-sea fish skin collagen oligopeptide according to claim 1 is characterized in that the deep-sea fish skin collagen oligopeptide for preparing has the function that reduces hyperpietic's blood pressure.
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| CNA2009100995080A CN101602790A (en) | 2009-06-11 | 2009-06-11 | A kind of deep-sea fish skin collagen oligopeptide and preparation method thereof with antihypertensive function |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974588A (en) * | 2010-09-07 | 2011-02-16 | 青岛海健堂生物科技有限公司 | Method for preparing fish collagens |
| CN102845586A (en) * | 2012-09-19 | 2013-01-02 | 百洋水产集团股份有限公司 | Method for decolorizing, debitterizing and deodorizing fishskin protein liquid |
| CN103388017A (en) * | 2013-08-06 | 2013-11-13 | 国家海洋局第三海洋研究所 | Preparation technology of fish skin collagen oligopeptide |
| CN103805665A (en) * | 2014-02-14 | 2014-05-21 | 青岛华科生物技术有限公司 | Preparation method of deep-sea fishskin collagen polypeptide |
-
2009
- 2009-06-11 CN CNA2009100995080A patent/CN101602790A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974588A (en) * | 2010-09-07 | 2011-02-16 | 青岛海健堂生物科技有限公司 | Method for preparing fish collagens |
| CN101974588B (en) * | 2010-09-07 | 2013-01-16 | 青岛海健堂生物科技有限公司 | Method for preparing fish collagens |
| CN102845586A (en) * | 2012-09-19 | 2013-01-02 | 百洋水产集团股份有限公司 | Method for decolorizing, debitterizing and deodorizing fishskin protein liquid |
| CN103388017A (en) * | 2013-08-06 | 2013-11-13 | 国家海洋局第三海洋研究所 | Preparation technology of fish skin collagen oligopeptide |
| CN103388017B (en) * | 2013-08-06 | 2014-12-17 | 国家海洋局第三海洋研究所 | Preparation technology of fish skin collagen oligopeptide |
| CN103805665A (en) * | 2014-02-14 | 2014-05-21 | 青岛华科生物技术有限公司 | Preparation method of deep-sea fishskin collagen polypeptide |
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Open date: 20091216 |