CN103539855A - Extraction method of acaudina molpadioidea collagen - Google Patents
Extraction method of acaudina molpadioidea collagen Download PDFInfo
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- CN103539855A CN103539855A CN201310432623.1A CN201310432623A CN103539855A CN 103539855 A CN103539855 A CN 103539855A CN 201310432623 A CN201310432623 A CN 201310432623A CN 103539855 A CN103539855 A CN 103539855A
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- 108010035532 Collagen Proteins 0.000 title claims abstract description 69
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- 238000000605 extraction Methods 0.000 title abstract description 8
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- 238000000034 method Methods 0.000 claims abstract description 32
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- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 claims description 71
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- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 2
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- 229910052753 mercury Inorganic materials 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- 235000019750 Crude protein Nutrition 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 241000258164 Molpadiida Species 0.000 description 2
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- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 1
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- 241000251468 Actinopterygii Species 0.000 description 1
- 241000965254 Apostichopus japonicus Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000258152 Caudinidae Species 0.000 description 1
- 241000555887 Globigerina Species 0.000 description 1
- 241000251511 Holothuroidea Species 0.000 description 1
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- JJWSNOOGIUMOEE-UHFFFAOYSA-N Monomethylmercury Chemical compound [Hg]C JJWSNOOGIUMOEE-UHFFFAOYSA-N 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
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- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
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- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
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- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- -1 highly basic Substances 0.000 description 1
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- 238000009776 industrial production Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
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- 238000004519 manufacturing process Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
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- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 235000011649 selenium Nutrition 0.000 description 1
- 229940091258 selenium supplement Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 235000019158 vitamin B6 Nutrition 0.000 description 1
- 239000011726 vitamin B6 Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Peptides Or Proteins (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses an extraction method of acaudina molpadioidea collagen. The extraction method comprises the following steps: adopting fresh and frozen acaudina molpadioidea or dry acaudina molpadioidea as raw material; cleaning the raw material, decontaminating, combining, peeling, destaining, slicing, pulping, adjusting the liquid-solid ratio, adjusting pH, heating, hydrolyzing, crudely extracting, purifying, drying and the like; and extracting the acaudina molpadioidea collagen. By adopting the method, heavy metal pollution is removed before hydrolyzing; no strong acid or alkali is used in the hydrolyzing process; the operation is simple and convenient; the prepared collagen is high in purity and good in safety; the obtained collagen not only can be applied to food, cosmetics and the like, but also has wide application on the aspect of pharmacological action.
Description
Technical field
The extracting method that the present invention relates to a kind of Haiti melon collagen protein, belongs to technical field of bioengineering.
Background technology
Haiti melon (Acaudina molpadioides Semper), be commonly called as Haiti melon, extra large eggplant, fragrant ginseng etc., Molpadida (Molpadida), Kao Can section (Caudinidae), Acaudina (Acaudina) animal, the long maximum of body can reach 400mm, be generally 100～200mm, be mainly distributed in Area of The East China Sea and ask the globigerina ooze that takes 80 meters of the depth of waters to from tide.
Research shows, Haiti melon nutritive ingredient is very abundant, contain rich in protein, polysaccharide, lower fat and amino acid, calcium, zinc and iron content are high, also contain selenium, vitamin A, vitamin B6, vitamin-E etc., wherein the content of Haiti melon body wall crude protein accounts for 89.16% of Haiti melon dry weight, higher than stichopus japonicus crude protein content, this proteinoid main component is the activated collagen protein of tool, if this low value Haiti melon is extracted to collagen protein purifying, can make full use of Haiti melon resource and improve its economic worth.
Haiti melon resource of China's Area of The East China Sea is very abundant, it is reported standing stock approximately 10
6t, the annual about 3000t of quantity of the catch, Along Zhejiang Coast resident is often edible using Haiti melon as low value sea cucumber, because quality after the heating of Haiti melon is hard, and live in the ooze of shallow sea, body surface has accumulated the harmful heavy metals such as a large amount of lead, cadmium, mercury, makes Haiti melon resource not yet by abundant development and utilization.
At present collagen protein in industrial production mainly from the raw materials such as skin bone of ox, pig, fish, but due to reasons such as epiphytotics generation such as religious, mad cow disease, bird flus, many developed countries have banned use of from ox-hide and ox bone and have extracted collagen protein.Living marine resources are abundant, and pollute less, and in recent years, marine collagen albumen develops an important content that has become each maritimecountry's ocean exploitation of the world.The extracting method of collagen protein is divided into hot water formulation, extraction and zymohydrolysis extracting method etc., wherein soda acid abstraction reaction violent, may residual harmful substance, although zymohydrolysis extracting method high-level efficiency, price is higher, hydrolysis degree is wayward, and can produce bitter peptides; Hot water formulation is extracted collagen protein, and step is simple, easily controls, and cost is low, and product is safer.
In recent years, the report that extracts collagen protein from the fish-skin of ocean is more, but from the melon of Haiti, extract collagen protein report seldom, therefore carrying out Haiti melon collagen protein extracts and fully utilizes, Haiti melon product of exploitation safety, nutrition, health care, for the quality of life and the health care level that improve civilian, significant.
Summary of the invention
In order to overcome the technological deficiencies such as heavy metal content in the melon of Haiti exceeds standard, utilization ratio is low, the present invention to provide a kind of easy and simple to handle, heavy metal free is residual, purity is high, the safe technique of collagen protein of extracting from the melon of Haiti.
The technical solution used in the present invention is:
An extracting method for Haiti melon collagen protein, is characterized in that, comprises the steps:
(1) machinery mill skin: Haiti melon raw material is put into the decorticating mill tool of frosted rotating disk, mill skin 10～20min; If fresh Haiti melon is directly put into decorticating mill tool and is ground skin; If freezing Haiti melon needs first naturally to thaw, then grinds skin; If dry Haiti melon must first soak 1～2d with 4～8 ℃, tap water, after melon water suction in dry Haiti eases back, then grind skin;
(2) enzymolysis decortication: Haiti melon after machinery mill skin, it is that in 1.0～1.5% proteolytic enzyme, solid-liquid ratio (g/mL) is 1:1.0～1.5 that cleaning is placed on concentration, and temperature is 40～45 ℃, enzymolysis 0.5～1h, Haiti melon after enzymolysis scrubs clean with banister brush or hairbrush-cleaning machine;
(3) enzyme that goes out cleans: Haiti melon after decortication is put into 80～90 ℃ of hot water enzyme 5～10min that goes out, and is cooled to after room temperature, removes silt, internal organ, sandspit and tooth in the melon abdomen of Haiti, with tap water, cleans up;
(4) making beating: after above-mentioned decortication decolouring Haiti melon cleans up, be cut into fragment, be placed in tissue mashing machine, add with step (1) in the water of the weight such as Haiti melon raw material, broken homogenate 10～30min, is transferred to extractor;
(5) regulate solid-liquid ratio (g/mL): in extractor, according to the weight of Haiti melon raw material in step (1), itself and tap water volume ratio are 1:10～15;
(6) regulate pH value: in extractor, by pH adjusting agent, regulate pH to weakly alkaline;
(7) constant temperature extracts: control 70～80 ℃ of extractor solution temperatures, constant temperature extracts 10～15h;
(8) slightly carry: said hydrolyzed liquid is placed in whizzer, the centrifugal 30min of 4000 * g, gets supernatant liquor and obtains collagen protein crude extract, or filters extracting solution with the Plate Filtration device in 0.01mm aperture, by liquid, is collagen protein crude extract;
(9) purifying: adopt 0.5 μ m microfiltration membrane under 0.05Pa condition, crude extract in step (8) to be filtered and removes impurity, then, under pressure 0.08～0.1MPa, use 80kDa ultra-filtration membrane to carry out ultrafiltration, collect trapped fluid;
(10) dry: trapped fluid to be carried out to lyophilize or spray drying treatment obtains collagen product.
As the improvement of the technology of the present invention method, between step (3) and step (4), also comprise decolouring step, Haiti melon being about to after decortication decolours with aqueous oxidizing agent solution immersion 2～4h, after decolouring, cleans up and pulls an oar.Preferably, oxygenant is food grade dioxide peroxide, and concentration of aqueous solution is 0.15%～0.20%.
Preferably, in step (2), proteolytic enzyme is food grade papoid, and enzyme is lived as 0.017Kat/g, and enzymolysis time is 0.6h.
Preferably, in step (5), solid-liquid ratio is 1:10.
Preferably, in step (6), pH adjusting agent is food grade NaHCO3, and regulating pH is 7.5～8.0; Most preferred, when pH is 7.6, extraction effect is best.
Preferably, in step (9) purge process, the pressure of ultrafiltration is 0.08MPa.
The invention has the beneficial effects as follows:
1, the present invention be take Haiti melon as raw material extracts collagen protein, Haiti melon resource of low value is fully utilized, and has significantly improved value-added content of product;
2, the present invention adopts machinery and enzymolysis combination peeling method, has effectively removed a difficult problem for Haiti melon epidermis heavy-metal residual;
3, the present invention adopts micro-filtration, ultrafiltration to combine the method for the purifying that dams, and can access the collagen protein of higher degree;
4, the present invention adopts hot water extraction method, and leaching process is not used the chemical reagent such as strong acid, highly basic, and Product Safety is high, and the method meets the requirement of collagen protein green extraction method, has good promotional value.
5, collagen protein extracting method of the present invention is easy and simple to handle, and controllability is strong, is applicable to suitability for industrialized production.
Embodiment
Embodiment 1
(1) bright Haiti melon collagen protein extraction process
(1) machinery mill skin: take the decorticating mill tool that the fresh Haiti of 500g melon is put into frosted rotating disk, mill skin 10min;
(2) enzymolysis decortication: Haiti melon after machinery mill skin, it is that in 1.3% 0.017Kat/g food grade papoid, solid-liquid ratio (g/mL) is 1:1.5 that cleaning is placed on concentration, and temperature is 45 ℃, enzymolysis 0.6h, Haiti melon after enzymolysis scrubs clean with banister brush;
(3) enzyme that goes out cleans: Haiti melon after enzymolysis decortication is put into 90 ℃ of hot water enzyme 5min that goes out, and is cooled to after room temperature, removes silt, internal organ, sandspit and tooth, with tap water, cleans up;
(4) the food grade dioxide peroxide that is 0.15% by concentration soaks 4h and decolours;
(5) making beating: after clean decolouring Haiti melon stripping and slicing, be placed in hollander, add 500g tap water, making beating 10min, after having pulled an oar, is transferred to extractor;
(6) regulate solid-liquid ratio (g/mL): to the tap water that adds 5000g in extractor;
(7) regulate pH value: use food grade NaHCO
3regulate pH to 7.6;
(8) constant temperature extracts: control 70 ℃ of extractor solution temperatures, constant temperature extracts 10h;
(9) slightly carry: said hydrolyzed liquid is placed in whizzer, the centrifugal 30min of 4000 * g, gets supernatant liquor and obtains collagen protein crude extract;
(10) purifying: adopt 0.5 μ m microfiltration membrane under 0.05Pa condition, the crude extract obtaining in (9) to be filtered and removes impurity, then, under pressure 0.08MPa, use 80kDa ultra-filtration membrane to carry out ultrafiltration, collect trapped fluid;
(11) dry: trapped fluid to be carried out to lyophilize and obtain collagen product.
(2) extraction yield of collagen protein, purity and heavy metal content
(1) assay of collagen protein
By measuring the content of oxyproline, be then multiplied by corresponding hydrolysis coefficient, obtain the content of collagen protein.The mensuration of oxyproline is carried out according to GB/T9695.23-2008, and collagen content calculates according to following formula:
Collagen content (%)=hydroxyproline content (%) * 12.6
(2) collagen protein extraction yield
This technique collagen protein extraction yield, is calculated as follows:
(3) mensuration of collagen product purity
The purity of collagen product is calculated as follows:
(4) mensuration of heavy metal content
Inorganic arsenic, total arsenic detects according to the mensuration > > of GB/T5009.11-2003 < < total Arsenic in Food and inorganic arsenic, plumbous (in Pb) detects according to the mensuration > > of GB5009.12-2010 < < Pb in food, cadmium (in Cd) detects according to the mensuration > > of GB/T5009.15-2003 < < cadmium in foods, mercury (in methyl mercury) detects according to total mercury and organomercurial mensuration > > in GB/T5009.17-2003 < < food.
(5) result
According to above detection method, carry out three parallel laboratory tests, the collagen protein extraction yield of survey, purity are in Table 1, and before and after collagen protein extracts, heavy metal content changes in Table 2.As can be seen from the table, present method collagen protein extraction yield, purity are higher, and repeatability good (CV < 5%); Collagen protein heavy metal content is compared remarkable minimizing with the heavy metal content in the melon raw material of Haiti, and meets the limit the quantity of requirement of > > of GB2762-2012 < < pollutants in food.
Table 1 collagen protein extraction yield and product purity
| Sample | Parallel 1(%) | Parallel 2(%) | Parallel 3(%) | X±SD | CV（%） |
| Collagen protein extraction yield | 42.56 | 43.18 | 41.97 | 42.57±0.61 | 1.42 |
| Collagen protein purity | 91.93 | 90.64 | 91.02 | 91.20±0.66 | 0.73 |
Heavy metal content before and after table 2 collagen protein extracts
Embodiment 2:
(1) freezing Haiti melon collagen protein extraction process
(1) machinery mill skin: take the freezing Haiti of 50kg melon, after naturally thawing, put into the decorticating mill tool of frosted rotating disk, mill skin 20min, removes 70～80% of Haiti melon epidermis;
(2) enzymolysis decortication: Haiti melon after machinery mill skin, cleaning and being placed on 50L concentration is, in 1.3% 0.017Kat/g food grade papoid, to keep temperature 45 C, enzymolysis 1h, Haiti melon after enzymolysis scrubs clean with Brush type cleaning machine;
(3) enzyme that goes out cleans: Haiti melon after enzymolysis decortication is put into 90 ℃ of hot water enzyme 10min that goes out, and is cooled to after room temperature, removes silt, internal organ, sandspit and tooth, with tap water, cleans up;
(4) by the concentration of 50L, be that 0.20% food grade dioxide peroxide immersion decortication Haiti melon 4h decolours;
(5) making beating: after above-mentioned decortication decolouring Haiti melon cleans up, dice with dicer, be placed in tissue mashing machine, add 50kg tap water, broken homogenate 30min, is transferred in large-scale extractor;
(6) regulate solid-liquid ratio (g/mL): to the tap water that adds 450kg in extractor;
(7) regulate pH value: use food grade NaHCO
3regulate pH to 7.6;
(8) constant temperature extracts: control 80 ℃ of extractor solution temperatures, constant temperature extracts 10h;
(9) slightly carry: said hydrolyzed liquid filters extracting solution with the Plate Filtration device in 0.01mm aperture, by liquid, is collagen protein crude extract;
(10) purifying: above-mentioned crude extract adopts 0.5 μ m microfiltration membrane to filter under 0.05Pa condition and removes impurity, then, under pressure 0.08MPa, use 80kDa ultra-filtration membrane to carry out ultrafiltration, collects trapped fluid;
(11) dry: trapped fluid is sprayed to be dried obtains collagen product.
(2) collagen protein extraction yield, purity and heavy metal content
According to the measuring method of collagen content, extraction yield, purity and heavy metal content in embodiment 1, collagen protein extraction yield and product purity detection to be met in Table 3, before and after collagen protein extracts, heavy metal content changes in Table 4.As can be seen from the table, the collagen protein extraction yield of present method, purity are higher, and repeatability good (CV < 5%), heavy metal content meets the limit the quantity of requirement of > > of GB2762-2012 < < pollutants in food.
Table 3 collagen protein extraction yield and product purity
| Sample | Parallel 1(%) | Parallel 2(%) | Parallel 3(%) | X±SD | CV（%） |
| Collagen protein extraction yield | 39.56 | 37.55 | 38.89 | 38.67±1.02 | 2.65 |
| Collagen protein purity | 89.65 | 88.42 | 89.02 | 89.03±0.61 | 0.69 |
Heavy metal content before and after table 4 collagen protein extracts
Claims (8)
1. an extracting method for Haiti melon collagen protein, is characterized in that, comprises the steps:
(1) machinery mill skin: Haiti melon raw material is put into the decorticating mill tool of frosted rotating disk, mill skin 10～20min; If fresh Haiti melon is directly put into decorticating mill tool and is ground skin; If freezing Haiti melon needs first naturally to thaw, then grinds skin; If dry Haiti melon must first soak 1～2d with 4～8 ℃, tap water, after melon water suction in dry Haiti eases back, then grind skin;
(2) enzymolysis decortication: Haiti melon after machinery mill skin, it is that in 1.0～1.5% proteolytic enzyme, solid-liquid ratio (g/mL) is 1:1.0～1.5 that cleaning is placed on concentration, and temperature is 40～45 ℃, enzymolysis 0.5～1h, Haiti melon after enzymolysis scrubs clean with banister brush or hairbrush-cleaning machine;
(3) enzyme that goes out cleans: Haiti melon after decortication is put into 80～90 ℃ of hot water enzyme 5～10min that goes out, and is cooled to after room temperature, removes silt, internal organ, sandspit and tooth in the melon abdomen of Haiti, with tap water, cleans up;
(4) making beating: after above-mentioned decortication decolouring Haiti melon cleans up, be cut into fragment, be placed in tissue mashing machine, add with step (1) in the water of the weight such as Haiti melon raw material, broken homogenate 10～30min, is transferred to extractor;
(5) regulate solid-liquid ratio (g/mL): in extractor, according to the weight of Haiti melon raw material in step (1), itself and tap water volume ratio are 1:10～15;
(6) regulate pH value: in extractor, by pH adjusting agent, regulate pH to weakly alkaline;
(7) constant temperature extracts: control 70～80 ℃ of extractor solution temperatures, constant temperature extracts 10～15h;
(8) slightly carry: said hydrolyzed liquid is placed in whizzer, the centrifugal 30min of 4000 * g, gets supernatant liquor and obtains collagen protein crude extract, or filters extracting solution with the Plate Filtration device in 0.01mm aperture, by liquid, is collagen protein crude extract;
(9) purifying: adopt 0.5 μ m microfiltration membrane under 0.05Pa condition, crude extract in step (8) to be filtered and removes impurity, then, under pressure 0.08～0.1MPa, use 80kDa ultra-filtration membrane to carry out ultrafiltration, collect trapped fluid;
(10) dry: trapped fluid to be carried out to lyophilize or spray drying treatment obtains collagen product.
2. according to the extracting method of Haiti melon collagen protein described in claim l, it is characterized in that: between step (3) and step (4), also comprise decolouring step, Haiti melon being about to after decortication decolours with aqueous oxidizing agent solution immersion 2～4h, after decolouring, cleans up and pulls an oar.
3. the extracting method of Haiti according to claim 2 melon collagen protein, is characterized in that: oxygenant is food grade dioxide peroxide, and concentration of aqueous solution is 0.15%～0.20%.
4. according to the extracting method of Haiti melon collagen protein described in claim l, it is characterized in that: in step (2), proteolytic enzyme is food grade papoid, enzyme is lived as 0.017Kat/g, and enzymolysis time is 0.6h.
5. according to the extracting method of Haiti melon collagen protein described in claim l, it is characterized in that: in step (5), solid-liquid ratio is 1:10.
6. according to the extracting method of Haiti melon collagen protein described in claim l, it is characterized in that: in step (6), pH adjusting agent is food grade NaHCO3, regulating pH is 7.5～8.0.
7. the extracting method of Haiti according to claim 6 melon collagen protein, is characterized in that: regulating pH is 7.6.
8. according to the extracting method of Haiti melon collagen protein described in claim l, it is characterized in that: in step (9) purge process, the pressure of ultrafiltration is 0.08MPa.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105595222A (en) * | 2015-12-29 | 2016-05-25 | 福尔康（上海）生物科技股份有限公司 | Acandina molpadioides hydrolysate preparation process |
| CN110981950A (en) * | 2019-12-16 | 2020-04-10 | 温州科技职业学院 | Paddy-potato collagen extraction equipment |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101792382A (en) * | 2010-03-10 | 2010-08-04 | 宋艳 | Method for preparing disinfected-decolorized fish-squamosum organic acid calcium and non-denatured collagen |
| CN102251006A (en) * | 2011-07-29 | 2011-11-23 | 宁波超星海洋生物制品有限公司 | Method for manufacturing collagen from black sea cucumbers from East China Sea |
| CN103027321A (en) * | 2011-09-30 | 2013-04-10 | 温州科技职业学院 | Quick skinning method for Acaudina molpadioides Semper |
-
2013
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101792382A (en) * | 2010-03-10 | 2010-08-04 | 宋艳 | Method for preparing disinfected-decolorized fish-squamosum organic acid calcium and non-denatured collagen |
| CN102251006A (en) * | 2011-07-29 | 2011-11-23 | 宁波超星海洋生物制品有限公司 | Method for manufacturing collagen from black sea cucumbers from East China Sea |
| CN103027321A (en) * | 2011-09-30 | 2013-04-10 | 温州科技职业学院 | Quick skinning method for Acaudina molpadioides Semper |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105595222A (en) * | 2015-12-29 | 2016-05-25 | 福尔康（上海）生物科技股份有限公司 | Acandina molpadioides hydrolysate preparation process |
| CN110981950A (en) * | 2019-12-16 | 2020-04-10 | 温州科技职业学院 | Paddy-potato collagen extraction equipment |
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