CN104862366A - Technology for extracting collagen peptide, dermatan sulfate, hydroxyapatite and melanin from black sharkskin - Google Patents
Technology for extracting collagen peptide, dermatan sulfate, hydroxyapatite and melanin from black sharkskin Download PDFInfo
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- CN104862366A CN104862366A CN201510340198.2A CN201510340198A CN104862366A CN 104862366 A CN104862366 A CN 104862366A CN 201510340198 A CN201510340198 A CN 201510340198A CN 104862366 A CN104862366 A CN 104862366A
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- hydroxyapatite
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- 229920000045 Dermatan sulfate Polymers 0.000 title claims abstract description 36
- 102000008186 Collagen Human genes 0.000 title claims abstract description 35
- 108010035532 Collagen Proteins 0.000 title claims abstract description 35
- AVJBPWGFOQAPRH-FWMKGIEWSA-L dermatan sulfate Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@H](OS([O-])(=O)=O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](C([O-])=O)O1 AVJBPWGFOQAPRH-FWMKGIEWSA-L 0.000 title claims abstract description 34
- 229940051593 dermatan sulfate Drugs 0.000 title claims abstract description 34
- 229920001436 collagen Polymers 0.000 title claims abstract description 33
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 33
- 229910052588 hydroxylapatite Inorganic materials 0.000 title claims abstract description 32
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 title claims abstract description 32
- 238000005516 engineering process Methods 0.000 title abstract description 9
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 title abstract 8
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 102000004190 Enzymes Human genes 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 10
- 238000005119 centrifugation Methods 0.000 claims description 10
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- 238000010612 desalination reaction Methods 0.000 claims description 5
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- 206010013786 Dry skin Diseases 0.000 claims description 2
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- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
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Landscapes
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a technology for extracting collagen peptide, dermatan sulfate, hydroxyapatite and melanin from black sharkskin. The technology comprises the following steps: conducting pre-treatment on the black sharkskin; carrying out enzymolysis; carrying out centrifugal separation to separate out a clear liquid and solid sediment; conducting ultrafiltration or resin adsorption on the clear liquid to obtain collagen peptide or dermatan sulfate respectively; treating the solid sediment with NaOH; conducting centrifugal separation on the treated solid sediment to obtain hydroxyapatite and melanin. The technology has the characteristics that collagen peptide, dermatan sulfate, hydroxyapatite and melanin are obtained continuously; the comprehensive utilization of the sharkskin is realized; the discharge of waste materials in the extraction and preparation processes is reduced; the production cost is reduced; the economic efficiency is improved; the technology is simple; industrialization can be realized.
Description
Technical field
A kind of from the extraction of black shark skin collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique, belong to field of natural product extraction.
Background technology
Collagen peptide is easy to digest and assimilate, and has protection stomach mucous membrane and prevents stomach ulcer, promote that calcium absorption prevents and treats osteoporosis; promote skin collagen metabolism; suppress blood pressure to rise, and reduce many-sided physiological functions such as serum cholesterol levels, cause the demand of market to collagen peptide constantly to increase.
Dermatan sulfate is a kind of glycosaminoglycan, belongs to glycosaminoglycan, and its disaccharides is formed by connecting with β-Isosorbide-5-Nitrae or β-1,3 key primarily of N-acetylamino galactosamine and L-iduronic acid.It is the sugary segment in proteoglycan overall molecule structure, and it is connected with the nucleoprotein of proteoglycan by covalent linkage.Dermatan sulfate all has broad application prospects at food, medicine and cosmetic field.
Hydroxyapatite (HAP) is the main inorganic composition of vertebrate skeletal and tooth, and in the enamel of people, the content of hydroxyapatite is more than 96%.There is excellent biocompatibility, be acknowledged as the superior artificial bone of a kind of biological property and be widely used in clinical.Hydroxyapatite also has outstanding adsorptive power, is used as the sorbing material of stratographic analysis purifying.Also can calsium supplement etc.
Melanochrome is not only a class bright protective agent, antiradiation agent, couplant, biological anti-oxidant and immunopotentiating agent, but also is a class biological semiconductor materials.Recent study finds, melanochrome has antisnake venom, anticancer, inhibiting HIV copies, treat the effects such as parkinsonism.Some melanochrome are applied in every field gradually.As the melanochrome in Semen Sesami Nigrum, black rice, black soya bean is in the food industry applied to beverage, candy, cake, chocolate, jam etc.Research shows, natural black pigment has anti-oxidant and prevents the functions such as old and feeble, this application that will greatly promote natural black pigment in protective foods.Natural black pigment receives an acclaim at chemical industry previous night day by day, can apply in the product such as hair blackening agent, sunscreen.
Now, utilize fish-skin to produce single product and be more and more not suitable with social development, how to improve the comprehensive utilization of fish-skin, become the research topic of growing interest.In view of the collagen protein contained in black shark skin, dermatan sulfate, hydroxyapatite and melanochrome etc., make full use of collagen protein wherein, dermatan sulfate, hydroxyapatite and melanochrome, make the product that purity is higher respectively, improve the utility value of fish-skin.
At present, still not relevant coproduction patent report.
The present invention obtains fish skin collagen peptide, dermatan sulfate, hydroxyapatite and melanochrome product by adopting the technology such as enzymolysis, centrifugal, ultrafiltration or absorption from black shark skin.
Summary of the invention
The object of the present invention is to provide a kind of from the extraction of black shark skin collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique, it be with black shark skin for raw material, be prepared into small molecule active collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique.
To achieve these goals, technical scheme of the present invention is:
The present invention is a kind of from the extraction of black shark skin collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique, and concrete technology step is as follows:
(1) the black shark skin after cleaning being dried is pulverized, and adds purified water, is warmed up to 65-90 DEG C, pre-treatment 0.5-2.5 hour;
(2) by the pretreatment fluid cooling in step (1), regulate pH to 7.5-9.0 with alkali, add compound protease and carry out enzymolysis, hydrolysis temperature is 50-58 DEG C, and enzymolysis time is 4-12 hour;
(3) the enzymolysis solution pH value in step (2) is adjusted to 3.0-6.0, is warming up to 75-90 DEG C, be incubated the 10-30 minute enzyme that goes out;
(4) material in step (3) is carried out solid-liquid separation, obtain clear liquid and solid sediment respectively;
(5) by the clear liquid in step (4), utilize Ultra filtration membrane or resin absorption to be separated, obtain concentrated solution and permeate, concentrated solution is the aqueous solution of sulfur acid dermatan, and permeate is the aqueous solution containing collagen peptide;
(6) concentrated solution in step (5) is added the alcohol of its weight 3-6 times more than 90%, leave standstill 2-4 hour, obtain throw out, throw out is dissolved in water, and then regulates pH to 10.5-11.5, adds the 0.5-3% hydrogen peroxide of solution weight, leave standstill 6-8 hour, filter, by the pH regulator of filtrate to 6-7, add its volume 4-7 doubly more than 90% alcohol, leave standstill 2-6 hour, collecting precipitation, through dehydrated alcohol dehydration, obtains dermatan sulfate through drying under reduced pressure;
(7) by the permeate in step (5), through nanofiltration desalination, large aperture adsorption resin decolouring, porn star, spraying dry obtains collagen peptide;
(8) the solid sediment NaOH solution process 1-4 hour in step (4), carries out solid-liquid separation, isolated solid, through washing, high-temperature calcination, obtains hydroxyapatite; Isolated liquid, add hydrochloric acid and regulate ph value for 1-3, leave standstill 2-6 hour, throw out, through washing, is dried and is obtained melanochrome.
In step (1), the amount adding purified water is 2-6 times of shark dry hide weight.
In step (2), described compound protease is any one or combination of pancreatin, Sumizyme MP, Collagenase and papoid, and the addition of often kind of enzyme is the 0.5%-2.0% of fish-skin dry weight.
In step (3), enzymolysis solution pH value, is adjusted to 3.0-6.0 before the enzyme that goes out.
In step (4), described solid-liquid separation is centrifugation.
In step (5), it is 6000-15000 dalton that described ultra-filtration membrane retains fractional dose.
In step (5), described resin is the resin of specificity absorption dermatan sulfate.
In step (6), described drying under reduced pressure temperature is 65-80 DEG C.
In step (8), described NaOH solution concentration is 0.1M.
The invention has the advantages that:
1, the present invention utilizes enzyme process preparing product, and technique is simple, and mild condition, energy consumption is low.
2, the present invention utilizes black shark skin can prepare multiple product simultaneously, and raw material availability is high.
3, the molecular weight of collagen peptide that prepared by the present invention is mainly distributed in 300-2000 dalton, is easily absorbed by the body.
4, the present invention realizes its comprehensive utilization be worth, and considerably reduces waste pollution problem in fish-skin production process.
Accompanying drawing explanation
Fig. 1 is the graph of molecular weight distribution of shark collagen peptide;
Fig. 2 is the melanic infrared spectra of shark;
Fig. 3 is the infrared spectra of shark dermatan sulfate.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described.
Embodiment 1
100kg black shark dry hide powder is broken to 25 orders, add 400kg purified water, heat up 90 DEG C, keep 1 hour, be cooled to 53 DEG C, adjust ph 8.3, add the compound protease (0.5kg Sumizyme MP and 0.5kg pancreatin) of 1kg, enzymolysis is stirred 10 hours at 53 DEG C, adjust ph is 4.5, be warming up to 90 DEG C of heating to go out enzyme 20min, be cooled to 50 DEG C, carry out centrifugation, isolated clear liquid carries out ultrafiltration through 10kDa film, permeate for ultrafiltration carries out nanofiltration (interception is 300 dalton) desalination, large aperture adsorption resin decolours, de-raw meat obtains collagen peptide 42.79kg.In ultrafiltration and concentration liquid, add the alcohol of 4 times 95%, leave standstill 2 hours, obtain throw out, throw out is dissolved in water, and then regulates pH to 11, adds 1% hydrogen peroxide of solution weight, leave standstill 6 hours, filter, by the pH regulator to 6 of filtrate, add the alcohol of its volume 5 times 95%, leave standstill 2-4 hour, collecting precipitation, through dehydrated alcohol dehydration, obtains dermatan sulfate 1.35kg in 4 hours through 70 DEG C of drying under reduced pressure.The NaOH process of the solid 0.1M that centrifugation goes out, after 2 hours, is carried out centrifugal, isolated solid again, obtains hydroxyapatite 12.21kg through washing, high-temperature calcination.Isolated alkaline liquid, regulates ph value to be 1 with hydrochloric acid, and leave standstill 4 hours, throw out obtains melanochrome 1.17kg through washing and drying.
Embodiment 2
100kg black shark dry hide powder is broken to 30 orders, add 600kg purified water, heat up 85 DEG C, keep 2 hours, be cooled to 55 DEG C, adjust ph 8.0, add the compound protease (1kg Sumizyme MP and 1kg collagenase) of 2kg, enzymolysis is stirred 11 hours at 55 DEG C, adjust ph is 4.0, be warming up to 80 DEG C of heating to go out enzyme 30min, be cooled to 60 DEG C, carry out centrifugation, isolated clear liquid carries out ultrafiltration through 15kDa film, permeate for ultrafiltration carries out nanofiltration (interception is 300 dalton) desalination, large aperture adsorption resin decolours, de-raw meat obtains collagen peptide 42.65kg.In ultrafiltration and concentration liquid, add the alcohol of 5 times 92%, leave standstill 3 hours, obtain throw out, throw out is dissolved in water, and then regulates pH to 11.5, adds 1.5% hydrogen peroxide of solution weight, leave standstill 7 hours, filter, by the pH regulator to 6.5 of filtrate, add the alcohol of its volume 6 times 92%, leave standstill 2 hours, collecting precipitation, through dehydrated alcohol dehydration, obtains dermatan sulfate 1.42kg in 3 hours through 80 DEG C of drying under reduced pressure.The NaOH process of the solid 0.1M that centrifugation goes out, after 2 hours, is carried out centrifugal, isolated solid again, obtains hydroxyapatite 11.83kg through washing, high-temperature calcination.Isolated alkaline liquid, regulates ph value to be 3 with hydrochloric acid, and leave standstill 5 hours, throw out obtains melanochrome 1.04kg through washing and drying.
Embodiment 3
100kg black shark dry hide powder is broken to 30 orders, add 500kg purified water, heat up 90 DEG C, keep 1.5 hours, be cooled to 50 DEG C, adjust ph 7.5, add the compound protease (1.0kg papoid and 1kg collagenase) of 2kg, enzymolysis 12h is stirred at 50 DEG C, adjust ph is 5.0, be warming up to 85 DEG C of heating to go out enzyme 25min, be cooled to 55 DEG C, carry out centrifugation, isolated clear liquid is through dermatan sulfate resin absorption, nanofiltration (interception is 300 dalton) desalination is carried out for the permeate after absorption, large aperture adsorption resin decolours, de-raw meat obtains collagen peptide 42.56kg.To de-wash in concentrated solution add 5 times 92% alcohol, leave standstill 4 hours, obtain throw out, throw out is dissolved in water, and then regulates pH to 10.5, adds 1.0% hydrogen peroxide of solution weight, leave standstill 8 hours, filter, by the pH regulator to 6.3 of filtrate, add the alcohol of its volume 5.5 times 94%, leave standstill 4 hours, collecting precipitation, through dehydrated alcohol dehydration, obtains dermatan sulfate 1.37kg in 3.5 hours through 78 DEG C of drying under reduced pressure.The NaOH process of the solid 0.1M that centrifugation goes out, after 3 hours, is carried out centrifugal, isolated solid again, obtains hydroxyapatite 11.94kg through washing, high-temperature calcination.Isolated alkaline liquid, regulates ph value to be 2 with hydrochloric acid, and leave standstill 6 hours, throw out obtains melanochrome 1.09kg through washing and drying.
Embodiment result shows, the pre-treatment of black shark skin, enzymolysis, centrifugation are gone out clear liquid and solid sediment by the present invention; Clear liquid obtains collagen peptide and dermatan sulfate respectively through ultrafiltration or resin absorption.Solid sediment obtains hydroxyapatite and melanochrome through NaOH process, centrifugation.This invented technology feature obtains collagen peptide, dermatan sulfate, hydroxyapatite and melanochrome continuously, achieves the comprehensive utilization of shark skin, reduces the discharge of extracting waste in preparation process, save production cost, and improves economical efficiency; This technique is simple in addition, can realize industrialization.
Above are only specific embodiments of the invention, but design concept of the present invention is not limited thereto, all changes utilizing this design the present invention to be carried out to unsubstantiality, all should belong to the behavior of invading scope.
Table 1 shark skin collagen peptide amino acid forms
Claims (9)
1. extract collagen peptide, dermatan sulfate, hydroxyapatite and a melanic technique from black shark skin, it is characterized in that: it comprises the following steps:
(1) the black shark skin after cleaning being dried is pulverized, and adds purified water, is warmed up to 65-90 DEG C, pre-treatment 0.5-2.5 hour;
(2) by the pretreatment fluid cooling in step (1), regulate pH to 7.5-9.0 with alkali, add compound protease and carry out enzymolysis, hydrolysis temperature is 50-58 DEG C, and enzymolysis time is 4-12 hour;
(3) the enzymolysis solution pH value in step (2) is adjusted to 3.0-6.0, is warming up to 75-90 DEG C, be incubated the 10-30 minute enzyme that goes out;
(4) material in step (3) is carried out solid-liquid separation, obtain clear liquid and solid sediment respectively;
(5) by the clear liquid in step (4), utilize Ultra filtration membrane or resin absorption to be separated, obtain concentrated solution and permeate, concentrated solution is the aqueous solution of sulfur acid dermatan, and permeate is the aqueous solution containing collagen peptide;
(6) concentrated solution in step (5) is added the alcohol of its weight 3-6 times more than 90%, leave standstill 2-4 hour, obtain throw out, throw out is dissolved in water, and then regulates pH to 10.5-11.5, adds the 0.5-3% hydrogen peroxide of solution weight, leave standstill 6-8 hour, filter, by the pH regulator of filtrate to 6-7, add its volume 4-7 doubly more than 90% alcohol, leave standstill 2-4 hour, collecting precipitation, through dehydrated alcohol dehydration, obtains dermatan sulfate through drying under reduced pressure;
(7) by the permeate in step (5), through nanofiltration desalination, large aperture adsorption resin decolouring, porn star, spraying dry obtains collagen peptide;
(8) the solid sediment NaOH solution process 1-4 hour in step (4), carries out solid-liquid separation, isolated solid, through washing, high-temperature calcination, obtains hydroxyapatite; Isolated liquid, add hydrochloric acid and regulate ph value for 1-3, leave standstill 2-6 hour, throw out, through washing, is dried and is obtained melanochrome.
2. one according to claim 1 extracts collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique from black shark skin, it is characterized in that in step (1), and the amount adding purified water is 2-6 times of shark dry hide weight.
3. one according to claim 1 extracts collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique from black shark skin, it is characterized in that in step (2), described compound protease is any one or combination of pancreatin, Sumizyme MP, Collagenase and papoid, and the addition of often kind of enzyme is the 0.5%-2.0% of fish-skin dry weight.
4. one according to claim 1 extracts collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique from black shark skin, it is characterized in that in step (3), the pH value of enzymolysis solution, is adjusted to 3.0-6.0 before the enzyme that goes out.
5. one according to claim 1 extracts collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique from black shark skin, it is characterized in that, in step (4), described solid-liquid separation is centrifugation.
6. one according to claim 1 extracts collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique from black shark skin, it is characterized in that, in step (5), it is 6000-15000 dalton that described ultra-filtration membrane retains fractional dose.
7. one according to claim 1 extracts collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique from black shark skin, it is characterized in that in step (5), and described resin is the resin of specificity absorption dermatan sulfate.
8. one according to claim 1 extracts collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique from black shark skin, it is characterized in that, in step (6), described drying under reduced pressure temperature is 65-80 DEG C.
9. one according to claim 1 extracts collagen peptide, dermatan sulfate, hydroxyapatite and melanic technique from black shark skin, it is characterized in that, in step (8), described NaOH solution concentration is 0.1M.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510340198.2A CN104862366B (en) | 2015-06-18 | 2015-06-18 | A kind of technique from black sharkskin extraction collagen peptide, dermatan sulfate, hydroxyapatite and melanin |
Applications Claiming Priority (1)
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| CN108079382A (en) * | 2017-12-29 | 2018-05-29 | 连云港碳谷材料科技有限公司 | A kind of preparation method of porous hydroxyapatite |
| CN109078164A (en) * | 2018-08-22 | 2018-12-25 | 厦门肽王基因科技有限公司 | Application of the grey gummy shark fish-skin mixed type protein peptides as skeleton development promotor and micro element supplement agent |
| WO2020000536A1 (en) * | 2018-06-26 | 2020-01-02 | 福州大学 | Preparation method for antioxidant peptide |
| JP2020054262A (en) * | 2018-10-01 | 2020-04-09 | ユーハ味覚糖株式会社 | Mucus membrane protection-related gene expression enhancer and use of the same |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105348383A (en) * | 2015-12-15 | 2016-02-24 | 中国海洋大学 | Odourless fishskin collagen and preparation method thereof |
| CN108079382A (en) * | 2017-12-29 | 2018-05-29 | 连云港碳谷材料科技有限公司 | A kind of preparation method of porous hydroxyapatite |
| CN108079382B (en) * | 2017-12-29 | 2021-01-26 | 连云港碳谷材料科技有限公司 | Preparation method of porous hydroxyapatite |
| WO2020000536A1 (en) * | 2018-06-26 | 2020-01-02 | 福州大学 | Preparation method for antioxidant peptide |
| CN109078164A (en) * | 2018-08-22 | 2018-12-25 | 厦门肽王基因科技有限公司 | Application of the grey gummy shark fish-skin mixed type protein peptides as skeleton development promotor and micro element supplement agent |
| CN109078164B (en) * | 2018-08-22 | 2022-04-01 | 厦门肽王基因科技有限公司 | Application of shark skin mixed protein peptide of Astrocaryum platypomum as skeletal development promoter and microelement supplement |
| JP2020054262A (en) * | 2018-10-01 | 2020-04-09 | ユーハ味覚糖株式会社 | Mucus membrane protection-related gene expression enhancer and use of the same |
| JP7095536B2 (en) | 2018-10-01 | 2022-07-05 | ユーハ味覚糖株式会社 | Mucosal protection-related gene expression promoter and its uses |
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