CN108707641A - The method that collagen polypeptide is extracted from fish scale - Google Patents
The method that collagen polypeptide is extracted from fish scale Download PDFInfo
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- CN108707641A CN108707641A CN201810380095.2A CN201810380095A CN108707641A CN 108707641 A CN108707641 A CN 108707641A CN 201810380095 A CN201810380095 A CN 201810380095A CN 108707641 A CN108707641 A CN 108707641A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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- 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, cold insoluble globulin [CIG]
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
Abstract
The invention discloses the methods that collagen polypeptide is extracted from fish scale：Clean fish scale is put into hydrochloric acid solution first and saturated sodium bicarbonate solution impregnates, it is dried after immersion for use, then it will pass through pretreated fish scale and impregnate solvent swell using acid, it is washed to neutrality, it is crushed after draining, distilled water is added later and carries out hot-water extraction, removal of impurities is filtered by vacuum after extracting, freeze-drying obtains fish scale collagen powder, finally by the lactobacillus inoculum after activation culture to carrying out cultivation and fermentation in the zymotic fluid containing fish scale collagen powder, after filtering, centrifugation, take supernatant to be concentrated by evaporation, obtain scale collagen polypeptide.The present invention preparation method is simple, reaction condition is mild, extraction efficiency is high, it is at low cost, convenient for commercialization, prepared scale collagen polypeptide yield and purity are high, molecular weight is small, bioactivity is good, hydrophily is strong, can be widely used in the fields such as cosmetics, health food and pharmaceutical products.
Description
Technical field
The present invention relates to technical field of bioengineering, more particularly to the method that collagen polypeptide is extracted from fish scale.
Background technology
Collagen is a kind of native biopolymer of zooblast synthesis, as the important component in extracellular matrix
Almost be present in a organized way in, account for about the 30% of animal total protein, there is collagen high resistance to extend ability so that skin
Equal connective tissues are maintained certain structure and with special mechanical properties to realize support and protection cell, tissue
And body function, it is very crucial to the structure and performance etc. of cell and tissue.
Isin glue collagen polypeptide is a kind of functional food or health product raw material, many studies have shown that in Isin glue collagen polypeptide
There are a large amount of biological active components, including immune with anti-oxidant, inhibition ACE activity, antiulcer, inhibition arthritis, raising
Power, the polypeptide for enhancing the bioactivity such as bone density and pre- preventing bone rarefaction.
In recent years, as the improvement of people's living standards, people to the diversity of aquatic resources demand and practicability by
It gradually improves, to promote the rapid development of China's fish processing industry.But it will produce while fish processing industry high speed development
A large amount of fish pomace, such as fish-skin, fish scale, internal organ, fish head etc., these leftover bits and pieces account for about the 40-50% of raw material fish, wherein about
5% is fish scale, a variety of nutriments containing needed by human body in fish scale, except containing a large amount of calcareous, also containing brain vegetative activity not
Substance needed for Phospholipids, phosphoprotein and the other vital movements that can lack is accounted for as contained abundant collagen in fish scale
20-40% as novel collagen resource there is very high development and utilization to be worth.
The prior art such as Authorization Notice No. is the Chinese invention patent of 103045706 B of CN, discloses a kind of continuous production
The method that fish scale collagen peptide chelates calcium salt and fish scale collagen peptide, includes the following steps：Pre-process fish scale, Enzymatic Extraction scale collagen
Polypeptide and calcium solution, purifying scale collagen polypeptide and calcium solution, peptide calcium chelating technology prepare fish scale collagen peptide chelating calcium salt soln,
Separation fish scale collagen peptide chelating calcium salt soln obtain sediment and supernatant, sediment water dissolution it is dry fish scale collagen peptide chelating
Calcium salt powder finished product, supernatant concentration removal ethyl alcohol isolate and purify again scale collagen peptide solution, scale collagen peptide solution are dry
To scale collagen Gly-His-Lys finished product.The fish scale collagen peptide chelating calcium salt product bioavilability of this method production is high, safe, and
Peptide calcium giving young employees remedial-courses in general knowledge and vocational skills, the scale collagen peptide product average relative molecular mass that is produced is low, purity is high.But the shortcoming of this method
It is preparation process complexity, and yield is relatively low.
Invention content
The purpose of the present invention is to provide it is a kind of it is simple and practicable, reaction condition is mild, extraction efficiency is high, it is at low cost, be convenient for
The method that collagen polypeptide is extracted in the slave fish scale commercially produced, prepared scale collagen polypeptide yield and purity
Height, molecular weight is small, and bioactivity is good, and hydrophily is strong.
The present invention in view of the above technology in the problem of mentioning, the technical solution taken is：
The method that collagen polypeptide is extracted from fish scale, including fish scale pretreatment, the extraction of fish scale collagen, zymotic fluid system
Standby, scale collagen polypeptide preparation, is as follows：
Fish scale pre-processes：By the dope on fish scale surface and other impurities cleaning, drying, fish scale is then put into a concentration of 3-5%
Hydrochloric acid solution in impregnate 20-24h, slough the calcium on fish scale, then the fish scale after decalcification is put into saturated sodium bicarbonate solution
12-16h is impregnated, fat and foreign protein in fish scale is removed, is finally dried for standby the fish scale handled well at 35-40 DEG C, it is right
Fish scale, which carries out pretreatment, can will be wrapped in the non-collagenic structure proteins such as the fat around collagen fabric and globulin, keratin
Matter also has the calcareous removal in fish scale, is convenient for the extraction mask work of follow-up collagen polypeptide, is conducive to improve fish phosphorus collagen
The purity of polypeptide；
The extraction of fish scale collagen：It is 1 according to weight proportion that pretreated fish scale, which will be passed through, with acid soaking agent:4-6 is swollen
30-50min is cleaned repeatedly with water to neutrality, pulverization process after draining away the water, and adds pearl white weight 8- in pearl white later
10 times of distilled water extracts 6-8h in 45-50 DEG C of shaking bath, and removal of impurities is filtered by vacuum after extracting, and freeze-drying obtains
Fish scale collagen powder impregnates the collagen that can be effectively extracted with the method that hot-water extraction is combined in fish scale by acid,
Collagenous fibres are fully swollen in extraction process, are conducive to the dissolving of collagenous fibres so that the recovery rate of collagen carries significantly
It is high；
The preparation of zymotic fluid：By fish scale collagen powder, yeast extract, diammonium hydrogen phosphate, citric acid, agar, glucose, adenosine two
Phosphate, magnesium sulfate, manganese sulfate and distilled water are uniformly mixed according to proportioning, and in 120-125 DEG C of high pressure sterilization 20-30min, hair
The composition collocation of zymotic fluid is reasonable, and nutritional sufficiency is suitble to the growth metabolism of strain；
The preparation of scale collagen polypeptide：Lactic acid bacteria is subjected to activation culture in degreasing milk medium, activation culture is followed by
Kind in zymotic fluid, while the sucrose of the 5-10% cultivation and fermentation 18-24h at 40-50 DEG C is added, and the pH for controlling zymotic fluid is
6.0-6.5 is filtered after fermentation, and filtrate is centrifuged 20-30min in 3500-4000rpm, and supernatant is taken to be placed in rotation
Turn to concentrate in evaporimeter, temperature is controlled at 50-60 DEG C, is concentrated into the sticky wall built-up of liquid, is obtained scale collagen polypeptide, is led to
The scale collagen polypeptide yield and purity height prepared by lactobacillus-fermented are crossed, molecular weight is small, and bioactivity is good, hydrophily
By force, have a wide range of application, and the flavor substances such as the acetaldehyde generated during the fermentation and biacetyl can reach the purpose that deodorization removes raw meat,
So that collagen polypeptide carries out defishying without follow-up, resource is greatly saved, reduces energy consumption, reduces cost.
Preferably, it is 1 that the acid soaking agent used in the extraction process of fish scale collagen, which is weight proportion,:0.3-
0.6 citric acid and the mixture of uridine triphosphate, uridine triphosphate destroy the hydroxyl in fish scale first under the cooperation of citric acid
Base apatite, then permeates into fish scale rapidly, is fully swollen collagenous fibres, and the broken schiff bases and salt changed between fiber molecule
Key so that the collagen in collagenous fibres is separated rapidly after interaction with the target molecules out, and then improves the extraction efficiency of collagen, shortens
Extracting cycle reduces cost.
Preferably, the material composition and its parts by weight of zymotic fluid are：45-60 parts of fish scale collagen powder, yeast extract 5-8
Part, 2-4 parts of diammonium hydrogen phosphate, 2-4 parts of citric acid, 5-8 parts of agar, 15-20 parts of glucose, 0.8-1.0 parts of adenosine diphosphate ester,
0.2-0.5 parts of magnesium sulfate, 0.05-0.1 parts of manganese sulfate, 80-100 parts of distilled water, wherein adenosine diphosphate ester and fermentation culture
Middle other components play synergistic effect, on the one hand can promote the synthesis and activation of protease during the growth metabolism of lactic acid bacteria,
The quantity for increasing protease on lactobacillus cell wall may advantageously facilitate enzymolysis efficiency, and then improve scale collagen polypeptide
On the other hand yield can make collagen triple helix space structure untwist by influencing the active group on hydroxyproline,
Molecular structure becomes loose, and the enzyme binding site on polypeptide chain increases, and increases the combination probability of enzyme-to-substrate, not only carries significantly
High degree of hydrolysis, and obtained polypeptide molecular weight is small, and bioactivity is high.
Preferably, lactic acid bacteria is lactobacillus bulgaricus, the mixture of lactobacillus buchneri, streptococcus thermophilus, weight
Proportioning is 1:1:0.5-0.8, additive amount are the lactic acid bacteria after inoculation 0.8-1.0% activation cultures in every liter of zymotic fluid.
Further preferably, the specific method of lactic acid bacteria activation culture is：30-40min is stood after fresh milk is boiled, from
It is so cooling, after fat deposit to appear, a layer lotion is removed, sterilize 30-50min at 100-105 DEG C, cooling up to skimmed milk culture
Then lactobacillus bulgaricus, lactobacillus buchneri, streptococcus thermophilus are respectively connected in degreasing milk medium by base, 40-45 DEG C of temperature
6-10h is cultivated in case, continuous activation 3-4 times, the lactic acid bacteria metabolic capability through overactivation is strong, quantity is sufficient, and can effectively ferment water
Solve the triple helix polypeptide chain of fish scale collagen so that peptide chain shortens, and molecular weight reduces, and generates more biologically active
Collagen polypeptide.
Compared with the prior art, the advantages of the present invention are as follows：
1）Method that the present invention extracts collagen polypeptide from fish scale is simple and practicable, reaction condition is mild, extraction efficiency is high, at
This is low, comprehensively utilizes and discards fish scale resource, can not only reduce environmental pollution, and can also significantly improve the additional of fish processing
Value, creates good economic and social profit；
2）The flavor substances such as the acetaldehyde and biacetyl that are generated in lactic acid bacteria fermentation process of the present invention can reach the purpose that deodorization removes raw meat,
Without subsequently carrying out deodorant operation, resource is greatly saved, reduces energy consumption；
3）The present invention extracts collagen polypeptide yield and pure height from fish scale, and molecular weight is small, and bioactivity is good, and hydrophily is strong,
It can be widely used in the fields such as cosmetics, health food and pharmaceutical products.
Specific implementation mode
The present invention program is described further below by embodiment：
Embodiment 1：
The method that collagen polypeptide is extracted from fish scale, including fish scale pretreatment, the extraction of fish scale collagen, zymotic fluid
It prepares, the preparation of scale collagen polypeptide, is as follows：
1）Fish scale pre-processes：By the dope on fish scale surface and other impurities cleaning, drying, fish scale is then put into a concentration of 3%
Hydrochloric acid solution in impregnate 20h, slough the calcium on fish scale, then the fish scale after decalcification is put into saturated sodium bicarbonate solution and is impregnated
12h removes fat and foreign protein in fish scale, is finally dried for standby the fish scale handled well at 40 DEG C, is carried out to fish scale pre-
The nencollagenous proteins such as the fat being wrapped in around collagen fabric and globulin, keratin can be also had fish scale by processing
In calcareous removal, be convenient for the extraction mask work of follow-up collagen polypeptide, be conducive to improve fish phosphorus collagen polypeptide
Purity；
2）The extraction of fish scale collagen：It is 1 according to weight proportion that pretreated fish scale, which will be passed through, with acid soaking agent:4 swellings
50min is cleaned repeatedly with water to neutrality, pulverization process after draining away the water, and 10 times of pearl white weight is then added in pearl white
Distilled water, 6h is extracted in 45 DEG C of shaking baths, after extracting be filtered by vacuum removal of impurities, freeze-drying obtain scale collagen egg
White powder impregnates the collagen that can be effectively extracted with the method that hot-water extraction is combined in fish scale, in extraction process by acid
Collagenous fibres are fully swollen, and the dissolving of collagenous fibres is conducive to so that the recovery rate of collagen greatly improves；
3）The preparation of zymotic fluid：By fish scale collagen powder, yeast extract, diammonium hydrogen phosphate, citric acid, agar, glucose, adenosine
Bisphosphate, magnesium sulfate, manganese sulfate and distilled water are uniformly mixed according to proportioning, and in 120 DEG C of high pressure sterilization 20min, zymotic fluid
Composition collocation it is reasonable, nutritional sufficiency is suitble to the growth metabolism of strain；
4）The preparation of scale collagen polypeptide：Lactic acid bacteria is subjected to activation culture in degreasing milk medium, after activation culture
It is inoculated into zymotic fluid, while the sucrose of addition 5% cultivation and fermentation 18h at 40 DEG C, and the pH for controlling zymotic fluid is 6.0-6.5,
It is filtered after fermentation, and filtrate is centrifuged into 20min in 4000rpm, take supernatant to be placed in Rotary Evaporators and concentrate, temperature
Degree control is concentrated into the sticky wall built-up of liquid, scale collagen polypeptide is obtained, prepared by lactobacillus-fermented at 60 DEG C
Scale collagen polypeptide yield and purity are high, and molecular weight is small, and bioactivity is good, and hydrophily is strong, has a wide range of application, and is fermenting
The flavor substances such as the acetaldehyde and biacetyl that generate in the process can reach the purpose that deodorization removes raw meat so that after collagen polypeptide is not necessarily to
It is continuous to carry out defishying, resource is greatly saved, reduces energy consumption, reduces cost.
It is 1 that the acid soaking agent used in the extraction process of above-mentioned fish scale collagen, which is weight proportion,:0.3 citric acid
With the mixture of uridine triphosphate, uridine triphosphate destroys the hydroxyapatite in fish scale, so first under the cooperation of citric acid
It is permeated afterwards into fish scale rapidly, is fully swollen collagenous fibres, and the broken schiff bases and sat linkage changed between fiber molecule so that collagen
Collagen in fiber is separated rapidly after interaction with the target molecules out, improves the extraction efficiency of collagen, substantially reduces extracting cycle, drop
Low cost.
The material composition and its parts by weight of above-mentioned zymotic fluid be：45 parts of fish scale collagen powder, 6 parts of yeast extract, phosphoric acid hydrogen
4 parts of diammonium, 2 parts of citric acid, 5 parts of agar, 15 parts of glucose, 1.0 parts of adenosine diphosphate ester, 0.2 part of magnesium sulfate, manganese sulfate 0.05
On the one hand part, 90 parts of distilled water can promote the synthesis and activation of protease during the growth metabolism of lactic acid bacteria, increase lactic acid bacteria
The quantity of protease, may advantageously facilitate enzymolysis efficiency on cell wall, improve the yield of scale collagen polypeptide, on the other hand logical
The active group that crossing influences on hydroxyproline can make collagen triple helix space structure untwist, and molecular structure becomes pine
It dissipating, the enzyme binding site on polypeptide chain increases, and increases the combination probability of enzyme-to-substrate, not only substantially increases degree of hydrolysis, and
And obtained polypeptide molecular weight is small, bioactivity is high.
Above-mentioned lactic acid bacteria be lactobacillus bulgaricus, lactobacillus buchneri, streptococcus thermophilus mixture, weight proportion is
1:1:0.5, additive amount is to be inoculated with the lactic acid bacteria after 0.8% activation culture in every liter of zymotic fluid.
The specific method of above-mentioned lactic acid bacteria activation culture is：30min is stood after fresh milk is boiled, natural cooling waits for
After there is fat deposit, a layer lotion is removed, sterilize 30min at 102 DEG C, cooling up to degreasing milk medium, then will be Bulgarian
Lactobacillus, lactobacillus buchneri, streptococcus thermophilus are respectively connected in degreasing milk medium, and 6h is cultivated in 40 DEG C of incubators, continuously activates 3
Secondary, the lactic acid bacteria metabolic capability through overactivation is strong, quantity is sufficient, can effectively fermentation hydrolysis fish scale collagen triple helix it is more
Peptide chain so that peptide chain shortens, and molecular weight reduces, and generates more biologically active collagen polypeptides.
Embodiment 2：
The method that collagen polypeptide is extracted from fish scale, including fish scale pretreatment, the extraction of fish scale collagen, zymotic fluid
It prepares, the preparation of scale collagen polypeptide, is as follows：
1）Fish scale pre-processes：By the dope on fish scale surface and other impurities cleaning, drying, fish scale is then put into a concentration of 5%
Hydrochloric acid solution in impregnate the calcium sloughed for 24 hours on fish scale, then the fish scale after decalcification is put into saturated sodium bicarbonate solution and is impregnated
16h removes fat and foreign protein in fish scale, is finally dried for standby the fish scale handled well at 40 DEG C；
2）The extraction of fish scale collagen：It is 1 according to weight proportion that pretreated fish scale, which will be passed through, with acid soaking agent:6 swellings
50min is cleaned repeatedly with water to neutrality, pulverization process after draining away the water, and 8 times of pearl white weight is then added in pearl white
Distilled water extracts 8h in 50 DEG C of shaking baths, and removal of impurities is filtered by vacuum after extracting, and freeze-drying obtains fish scale collagen
Powder；
3）The preparation of zymotic fluid：By 60 parts of fish scale collagen powder, 8 parts of yeast extract, 4 parts of diammonium hydrogen phosphate, 3 parts of citric acid, fine jade
80 parts of 8 parts of fat, 20 parts of glucose, 1.0 parts of adenosine diphosphate ester, 0.5 part of magnesium sulfate, 0.06 part of manganese sulfate, distilled water mixing are equal
It is even, and in 125 DEG C of high pressure sterilization 30min；
4）The preparation of scale collagen polypeptide：Lactic acid bacteria is subjected to activation culture in degreasing milk medium, after activation culture
Be inoculated into zymotic fluid, at the same be added 10% sucrose at 45 DEG C cultivation and fermentation for 24 hours, and control zymotic fluid pH be 6.0-
6.5, it is filtered after fermentation, and filtrate is centrifuged into 30min in 3500rpm, takes supernatant to be placed in dense in Rotary Evaporators
Contracting, temperature control at 60 DEG C, are concentrated into the sticky wall built-up of liquid, obtain scale collagen polypeptide.
It is 1 that the acid soaking agent used in the extraction process of above-mentioned fish scale collagen, which is weight proportion,:0.6 citric acid
With the mixture of uridine triphosphate.
Above-mentioned lactic acid bacteria be lactobacillus bulgaricus, lactobacillus buchneri, streptococcus thermophilus mixture, weight proportion is
1:1:0.8, additive amount is to be inoculated with the lactic acid bacteria after 1.0% activation culture in every liter of zymotic fluid.
The specific method of above-mentioned lactic acid bacteria activation culture is：30min is stood after fresh milk is boiled, natural cooling waits for
After there is fat deposit, a layer lotion is removed, sterilize 30min at 105 DEG C, cooling up to degreasing milk medium, then will be Bulgarian
Lactobacillus, lactobacillus buchneri, streptococcus thermophilus are respectively connected in degreasing milk medium, and 8h is cultivated in 42 DEG C of incubators, continuously activates 4
It is secondary.
Embodiment 3：
The method that collagen polypeptide is extracted from fish scale, including fish scale pretreatment, the extraction of fish scale collagen, zymotic fluid
It prepares, the preparation of scale collagen polypeptide, is as follows：
1）Fish scale pre-processes：By the dope on fish scale surface and other impurities cleaning, drying, fish scale is then put into a concentration of 4%
Hydrochloric acid solution in impregnate 20h, then the fish scale after decalcification is put into saturated sodium bicarbonate solution and impregnates 16h, finally will processing
Good fish scale is dried for standby at 40 DEG C；
2）The extraction of fish scale collagen：It is 1 according to weight proportion that pretreated fish scale, which will be passed through, with acid soaking agent:5 swellings
50min is cleaned repeatedly with water to neutrality, pulverization process after draining away the water, and 10 times of pearl white weight is then added in pearl white
Distilled water, 7h is extracted in 45 DEG C of shaking baths, after extracting be filtered by vacuum removal of impurities, freeze-drying obtain scale collagen egg
White powder；
3）The preparation of zymotic fluid：By 50 parts of fish scale collagen powder, 6 parts of yeast extract, 3 parts of diammonium hydrogen phosphate, 3 parts of citric acid, fine jade
100 parts of 8 parts of fat, 15 parts of glucose, 0.9 part of adenosine diphosphate ester, 0.5 part of magnesium sulfate, 0.05 part of manganese sulfate, distilled water mixing are equal
It is even, and in 120 DEG C of high pressure sterilization 30min；
4）The preparation of scale collagen polypeptide：Lactic acid bacteria is subjected to activation culture in degreasing milk medium, after activation culture
Be inoculated into zymotic fluid, at the same be added 8% sucrose at 50 DEG C cultivation and fermentation for 24 hours, and control zymotic fluid pH be 6.0-6.5,
It is filtered after fermentation, and filtrate is centrifuged into 20min in 4000rpm, take supernatant to be placed in Rotary Evaporators and concentrate, temperature
Degree control is concentrated into the sticky wall built-up of liquid, obtains scale collagen polypeptide at 60 DEG C.
It is 1 that the acid soaking agent used in the extraction process of above-mentioned fish scale collagen, which is weight proportion,:0.5 citric acid
With the mixture of uridine triphosphate.
Above-mentioned lactic acid bacteria be lactobacillus bulgaricus, lactobacillus buchneri, streptococcus thermophilus mixture, weight proportion is
1:1:0.6, additive amount is to be inoculated with the lactic acid bacteria after 1.0% activation culture in every liter of zymotic fluid.
The specific method of above-mentioned lactic acid bacteria activation culture is：40min is stood after fresh milk is boiled, natural cooling waits for
After there is fat deposit, a layer lotion is removed, sterilize 30min at 105 DEG C, cooling up to degreasing milk medium, then will be Bulgarian
Lactobacillus, lactobacillus buchneri, streptococcus thermophilus are respectively connected in degreasing milk medium, and 7h is cultivated in 45 DEG C of incubators, continuously activates 4
It is secondary.
Comparative example 1：
Compared with Example 1, the material composition of zymotic fluid and its parts by weight are comparative example 1 in step 3：Fish scale collagen
45 parts of powder, 6 parts of yeast extract, 4 parts of diammonium hydrogen phosphate, 2 parts of citric acid, 5 parts of agar, 15 parts of glucose, 0.2 part of magnesium sulfate, sulfuric acid
0.05 part of manganese, 90 parts of water, method and step all same in addition to this.
In order to compare effect of the present invention, respectively prepared by testing example 1, embodiment 2, embodiment 3 and comparative example 1
Scale collagen polypeptide in relative molecular mass in the content of the collagen peptide of 500-1000, concrete outcome such as table 1.
Content table of the relative molecular mass of 1 scale collagen polypeptide of table in 500-1000
| Content/% of the relative molecular mass in 500-1000 | |
| Embodiment 1 | 85 |
| Embodiment 2 | 92 |
| Embodiment 3 | 89 |
| Comparative example 1 | 70 |
As it can be seen from table 1 compared with comparative example 1, the average molecular prepared by embodiment 1, embodiment 2 and embodiment 3
Quality is higher in the content of the collagen polypeptide of 500-1000, it is seen then that the collagen polypeptide that the present invention is extracted from fish scale
Molecular weight is small, and bioactivity is good, can be widely used in the fields such as cosmetics, health food and pharmaceutical products.
Routine operation in the operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (8)
1. the method for extracting collagen polypeptide from fish scale, including fish scale pretreatment, the extraction of fish scale collagen, fish glue from scale
The preparation of former polypeptide, it is characterised in that：It is described to be as follows：
1）Fish scale pre-processes：Clean fish scale is successively put into hydrochloric acid solution and saturated sodium bicarbonate solution and is impregnated, knot is impregnated
It is dried after beam for use；
2）The extraction of fish scale collagen：Pretreated fish scale will be passed through and impregnate solvent swell using acid, neutrality is washed to, drains
After crush, then be added distilled water carry out hot-water extraction, after extracting be filtered by vacuum removal of impurities, freeze-drying obtain scale collagen
Albumen powder, the acidity soaking agent are the mixture of citric acid and uridine triphosphate；
3）It is prepared by zymotic fluid；
4）The preparation of scale collagen polypeptide：It is carried out sucrose in the lactobacillus inoculum to zymotic fluid after activation culture, is added
Culture, and controls temperature, pH and the incubation time of zymotic fluid, after filtering, centrifugation, take supernatant to be concentrated by evaporation, that is, make
Obtain scale collagen polypeptide.
2. the method according to claim 1 for extracting collagen polypeptide from fish scale, it is characterised in that：The step 1
A concentration of 3-5% of hydrochloric acid solution.
3. the method according to claim 1 for extracting collagen polypeptide from fish scale, it is characterised in that：The step 2
The weight proportion of citric acid and uridine triphosphate is 1 in acid soaking agent:0.3-0.6.
4. the method according to claim 1 for extracting collagen polypeptide from fish scale, it is characterised in that：The step 2
Weight ratio by pretreated fish scale and acid soaking agent is 1:4-6.
5. the method according to claim 1 for extracting collagen polypeptide from fish scale, it is characterised in that：The step 3
The material composition and its parts by weight of zymotic fluid be：45-60 parts of fish scale collagen powder, 5-8 parts of yeast powder, diammonium hydrogen phosphate 2-4
Part, 2-4 parts of citric acid, 5-8 parts of potassium acetate, 15-20 parts of glucose, 0.8-1.0 parts of adenosine diphosphate ester, magnesium sulfate 0.2-0.5
Part, 0.05-0.1 parts of manganese sulfate, 80-100 parts of water.
6. the method according to claim 1 for extracting collagen polypeptide from fish scale, it is characterised in that：The step 4
Lactic acid bacteria be lactobacillus bulgaricus, lactobacillus buchneri, streptococcus thermophilus mixture, weight proportion 1:1:0.5-0.8,
Additive amount is the lactic acid bacteria after inoculation 0.8-1.0% activation cultures in every liter of zymotic fluid.
7. the method according to claim 1 for extracting collagen polypeptide from fish scale, it is characterised in that：The step 4
The specific method of lactic acid bacteria activation culture is：30-40min, natural cooling, fat deposit to appear are stood after fresh milk is boiled
Afterwards, a layer lotion is removed, sterilize 30-50min at 100-105 DEG C, cooling up to degreasing milk medium, then by bulgarian milk bar
Bacterium, lactobacillus buchneri, streptococcus thermophilus are respectively connected in degreasing milk medium, and 6-10h is cultivated in 40-45 DEG C of incubator, continuous living
Change 3-4 times.
8. the method according to claim 1 for extracting collagen polypeptide from fish scale, it is characterised in that：The step 4
Control broth temperature is 40-50 DEG C, pH is 6.0-6.5 and incubation time 18-24h.
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| CN115487355A (en) * | 2022-09-30 | 2022-12-20 | 中国海洋大学 | Preparation method and application of fish scale matrix bone repair material |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102318843A (en) * | 2011-07-01 | 2012-01-18 | 中国科学院海洋研究所 | A kind of fish scale collagen polypeptide and preparation method thereof |
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| CN115487355A (en) * | 2022-09-30 | 2022-12-20 | 中国海洋大学 | Preparation method and application of fish scale matrix bone repair material |
| CN115487355B (en) * | 2022-09-30 | 2023-09-22 | 中国海洋大学 | Preparation method and application of fish scale matrix bone repair material |
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