CN112522357A - Method for extracting small molecular peptides from fish scales by using complex enzyme and application - Google Patents
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- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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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 or cold insoluble globulin [CIG]
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Abstract
The invention provides a method for extracting small molecular peptides from fish scales by using complex enzyme and application thereof, relating to the technical field of biological extraction, and the method for extracting the small molecular peptides from the fish scales by using the complex enzyme and the application thereof comprise the following steps: selecting fish scales as raw materials, pretreating the purchased materials, recovering the pretreated raw materials, decalcifying and degreasing the raw materials by a selection method, recovering the treated raw materials, detecting the required data by experiments, obtaining the required data, storing and comparing the data, extracting the raw materials by a selected enzyme extraction method by complex enzyme, recovering the extracted raw materials, storing the raw materials in a required environment, storing the temperature of the storage at minus 1 to minus 4 ℃, extracting the fish scales by the complex enzyme, effectively improving the extraction quality compared with single enzyme extraction, obtaining low-molecular-weight collagen peptides by complex enzyme hydrolysis, reducing the extraction difficulty of workers and reducing the cost.
Description
Technical Field
The invention relates to the technical field of biological extraction, in particular to a method for extracting small molecular peptides from fish scales by using complex enzyme and application thereof.
Background
The molecular active peptide is a biochemical substance between amino acid and protein, has a molecular weight smaller than that of the protein and larger than that of the amino acid, and is a fragment of the protein. Two or more amino acids are linked by peptide bonds to form an "amino acid chain" or "amino acid string" called a peptide. Among them, peptides consisting of 10-15 or more amino acids are called polypeptides, peptides consisting of 2 to 9 amino acids are called oligopeptides, peptides consisting of 2 to 15 amino acids are called small peptides or small peptides, the development of world fishery is rising year by year from 2075 million in 1950, FAO statistics, and the total world fishery production in 2010 is about 1.485 million t [1 ]. China has abundant ocean resources and fresh water fish culture conditions, the total yield of aquatic products accounts for 39% of the total yield of the world [2], but the scale of Chinese fresh water fish processing enterprises is small and dispersed. The fish scale is late in research and development, is not well developed and utilized, causes huge resource waste, contains rich type I collagen [3-4] in the fish scale, forms collagen peptide through hydrolysis, has low molecular weight, increased water solubility, improved emulsibility, foamability and frost resistance [5], has oxidation resistance [6-7], is easy to digest and absorb [8], is used as a food ingredient, and is widely applied to industries such as food processing and the like. Because collagen is distributed in the inner layer of fish scales, and the outer layer is adhered by hydroxyapatite, collagen pretreatment is particularly important for the production of collagen peptide, and grass carp is fish of Cyprinus and grass carp. The grass carp is commonly known as Wan fish, grass carp, bai grass carp, grass carp root (northeast), etc. The body length is 3.4-4.0 times of the body height, 3.6-4.3 times of the head length, 7.3-9.5 times of the tail handle length and 6.8-8.8 times of the tail handle height. Long, blunt, 2 rows of teeth in the lower pharynx, in comb shape. The dorsal fin has no spine, the outer edge is straight and is positioned above the ventral fin, and the distance from the starting point to the tail fin base is shorter than that from the osculating end. The gill rake is short and small in number. The body is dark yellow, the abdomen is grey white, the edges of the body side scales are grey black, the pectoral fin and the ventral fin are grey yellow, and other fins are light-colored. Grass carp is a typical herbivorous fish, inhabits rivers and lakes in plain areas, and is generally favored in the middle and lower layers of water and near-shore multi-water grass areas. It is lively, swims rapidly, and often finds food in groups. The young grass carp eats larvae, algae and the like, and the grass carp eats some meat foods, such as earthworm, dragonfly and the like. Wide distribution, and is distributed from Heilongjiang to Yunnan Yuanjiang in China (except Tibet and Xinjiang areas). The grass carp is transplanted to countries such as Asia, Europe, America, non continents and the like, and generally likes to inhabit middle, lower layers and coastal multi-aquatic-grass areas of water areas such as rivers, lakes and the like. The sex mature individuals lay eggs in the flowing water of rivers, and the parent fishes and the juvenile fishes after laying eggs enter branches and rivers and go to the rivers and the lakes, and are generally fed and fattened on the shallow grasslands and the flooded areas submerged by water and the affiliated water bodies of the dry branches (water and grass cluster zones of lakes, rivers, harbor channels and the like). In winter, the river can live through winter in deep water in a dry flow or lake. Grass carp is typical herbivorous fish with lively sexual condition, rapid swimming, frequent foraging in groups and greedy in sex. The fish fry stage ingests zooplankton, the juvenile fish stage also ingests insects, earthworms, algae, duckweed and the like, and when the body length reaches about more than 10 cm, aquatic higher plants are completely ingested, particularly gramineae plants. The species of plants ingested by grass carps vary with the condition of the food base in the living environment. Grass carp and several other domestic fishes reproduce similarly, and cannot lay eggs in still water under natural conditions. The spawning place is selected from river junction of river main stream, deep-trough water area on one side of river curve and river section with both banks suddenly contracted as suitable spawning place. The reproduction season is similar to that of silver carp, and is earlier than that of black carp and bighead carp.
The existing extraction method of the small molecular peptide is difficult, the operation difficulty is increased for workers, most of the small molecular peptides are extracted from marine animals, but due to the fact that the treatment process is complex, the cost is high, the fishy smell of the product is difficult to remove, pollutants such as different types of persistent organic matters and heavy metals can be accumulated in the bodies of the products, the small molecular peptides cannot be effectively extracted, the extraction method is low in efficiency, and the extraction purity is low.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides a method for extracting small molecular peptides from fish scales by using complex enzyme and application thereof, so that the quality of the extracted small molecular peptides is improved, and the anti-aging and beautifying functions of the extracted products are improved.
(II) technical scheme
In order to achieve the purpose, the invention is realized by the following technical scheme: a method for extracting small molecular peptides from fish scales by using complex enzyme and application thereof comprise the following steps:
sp 1: pretreating raw materials: selecting fish scales as raw materials, and preprocessing the selected materials;
sp 2: decalcification treatment: recovering the pretreated raw materials, and decalcifying and degreasing the raw materials by a selection method;
sp3: and (3) component analysis: recovering the processed raw materials, detecting through experiments to obtain required data, and storing and comparing;
sp4: complex enzyme extraction: extracting the raw materials by adopting a complex enzyme extraction method;
sp 5: and (4) recovering and preserving: and recovering the extracted raw materials, and storing the raw materials to a required environment at the temperature of minus 1 to minus 4 ℃.
Preferably, grass carp scales are selected as raw material extraction, the raw material is purchased, stored and moved to a position to be processed, the purchased raw material is pretreated, collected grass carps are subjected to cold water washing for 5min, recovered and stored, the water on the surfaces of the fish scales is wiped off, the grass carps are divided into 20 g/bag and 40 g/bag, the raw material is frozen and stored for later use, the weight of the required raw material is taken out according to requirements, the fish scale raw material is put into 1000mL of 1% sodium chloride boiling water for blanching for 2min for sterilization, the raw material is put into a fish scale cleaning machine for cleaning, and the cleaning liquid is stirred and cleaned for 2h at 40-60 ℃ by adopting warm water with the material-liquid ratio of 1: 15 for one time after the cleaning liquid is changed.
Preferably, 50g of degreased, washed and dried fish scales for later use is placed in a 1000mL beaker according to a feed-liquid ratio of 1: adding 500mL of water into 10, adding a hydrochloric acid solution, heating, stirring, reacting, sampling a decalcifying solution, washing fish scales with clear water for 10 times, drying in the air, and vacuum-drying at 279 ℃ for 24 hours, wherein the hydrochloric acid concentration is 0.37mol/L, the decalcification time is 35min, and the decalcification rate in the decalcification solution reaches 92%.
Preferably, the decalcifying solution is accurately sucked up by 0.5mL into a 250mL volumetric flask, diluted to 50mL with distilled water, 10mL of an ammonia buffer solution with pH of 10 and 3 drops of a chrome black T indicator are added. Titrating with 0.0100mol/L EDTA standard solution until the solution turns from purple to blue, recording the consumption volume of EDTA to obtain the Torr probability, adding 6mol/L hydrochloric acid 9mL into 1mL of centrifuged fish scale decalcification solution by using a chromatograph, hydrolyzing in an oven at 110 ℃ for 12h, and diluting by 200 times to detect the hydroxyproline content of the decalcification solution.
Preferably, the compound enzyme is composed of papain, bromelain and an alkaline compound enzyme, and the mass ratio of the compound enzyme to the papain, the bromelain and the alkaline compound enzyme is 1: 1: 2, the mass of fish scale peptide prepared by hydrolyzing 3% of papain, 3% of bromelain and 6% of alkaline complex enzyme for 5 hours is measured after centrifugation, the mass of collagen peptide with molecular mass of more than 2000u is 6.15%, the mass of collagen peptide with molecular mass of less than 176u is 3.82%, and the contents of oligopeptide with molecular mass distribution of 176-1000 u and 1000-2000 u are 73.3% and 16.6% respectively.
(III) advantageous effects
The invention provides a method for extracting small molecular peptides from fish scales by using complex enzyme and application thereof. The method has the following beneficial effects:
1. according to the invention, the compound enzyme is adopted to extract the fish scales, the extraction quality is effectively improved compared with single enzyme extraction, the low molecular weight collagen peptide is obtained by adopting compound enzyme hydrolysis, the extraction difficulty of workers is reduced, the cost is low, the effective raw materials are cleaned, the peculiar smell is taken out, the extracted small molecular peptide has high purity and high efficiency, the waste fish scale resources are comprehensively utilized under the effects of resisting aging, whitening and the like, the environmental pollution can be reduced, the additional value of fish processing can be obviously improved, and good economic and social benefits are created.
Drawings
FIG. 1 is a schematic flow chart of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The first embodiment is as follows:
as shown in fig. 1, the embodiment of the invention provides a method for extracting small molecular peptides from fish scales by using complex enzyme and an application thereof, comprising the following steps:
sp 1: pretreating raw materials: selecting fish scales as raw materials, and preprocessing the selected materials;
sp 2: decalcification treatment: recovering the pretreated raw materials, and decalcifying and degreasing the raw materials by a selection method;
sp3: and (3) component analysis: recovering the processed raw materials, detecting through experiments to obtain required data, and storing and comparing;
sp4: complex enzyme extraction: extracting the raw materials by adopting a complex enzyme extraction method;
sp 5: and (4) recovering and preserving: and recovering the extracted raw materials, and storing the raw materials to a required environment at the temperature of minus 1 to minus 4 ℃.
Example two:
as shown in figure 1, the embodiment of the invention provides a method for extracting small molecular peptides from fish scales by using complex enzyme and application thereof, selecting grass carp scales as raw material for extraction, purchasing the raw material, storing the raw material, moving the raw material to a position to be processed, pretreating the purchased raw material, removing non-fish scale substances such as gills, fins, viscera and the like of the collected grass carps, washing the grass carps with cold water for 5min, recovering and storing the grass carps, wiping off water on the surfaces of the fish scales, packaging the grass carps into 20 g/bag and 40 g/bag respectively, and freezing and preserving the grass carps for later use, taking out the weight of the required raw materials according to the requirement, blanching the fish scale raw materials in 1000mL of 1% sodium chloride boiling water for 2min for sterilization, putting the raw materials into a fish scale cleaning machine, cleaning the raw materials, adopting warm water with a material-liquid ratio of 1: 15, putting 50g of degreased, washed and dried fish scales for later use into a 1000mL beaker, and mixing the raw materials according to the material-liquid ratio of 1: adding 500mL of water into 10, adding a hydrochloric acid solution, heating, stirring, reacting, sampling a decalcifying solution, washing fish scales with clear water for 10 times, drying in the air, and then placing under 279C for vacuum drying for 24 hours, wherein the hydrochloric acid concentration is 0.37mol/L, the decalcification time is 35min, and the decalcification rate in the decalcification solution reaches 92%.
Example three:
as shown in fig. 1, the embodiment of the invention provides a method for extracting small molecular peptides from fish scales by using complex enzyme and application thereof, wherein 0.5mL of decalcifying solution is accurately absorbed into a volumetric flask with 250mL, the volumetric flask is diluted to 50mL by using distilled water, 10mL of ammonia buffer solution with the pH value of 10 is added, and 3 drops of a chrome black T indicator are added. Titrating by using 0.0100mol/L EDTA standard solution until the solution is changed from purple to blue, recording the EDTA consumption volume to obtain the support probability, using a chromatograph to perform centrifugation on 1mL of fish scale decalcification solution, adding 9mL of 6mo/L hydrochloric acid, putting the fish scale decalcification solution in an oven at 110 ℃, performing hydrolysis for 12h, diluting by 200 times to detect the hydroxyproline content in the decalcification solution, wherein the complex enzyme is composed of papain, bromelain and an alkaline complex enzyme, and the mass ratio of the papain to the bromelain to the alkaline complex enzyme is 1: 1: 2, the papain accounts for 3 percent, the bromelain accounts for 3 percent, the alkaline complex enzyme accounts for 6 percent, the alkaline complex enzyme accounts for 3.82 percent, the fish scale peptide prepared after 5 hours of hydrolysis is measured to obtain molecular weight after centrifugation, the content of the collagen peptide with molecular weight more than 2000u is 6.15 percent, the content of the collagen peptide less than 176u is 3.82 percent, the content of the oligopeptide with molecular weight distribution in 176-1000 u and 1000-2000 u is 73.3 percent and 16.6 percent respectively, the complex enzyme is adopted to extract the fish scales, compared with the single enzyme extraction, the extraction quality is effectively improved, the low molecular weight collagen peptide is obtained by adopting the complex enzyme hydrolysis, the extraction difficulty of workers is reduced, the cost is low, the waste raw materials are cleaned, the peculiar smell is taken out, the extracted small molecular peptide has high purity and high efficiency, the waste fish scale resources are comprehensively utilized under the effects of resisting aging, whitening, the environmental pollution can be reduced, creating good economic and social benefits.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising a reference structure" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (5)
1. A method for extracting small molecular peptides from fish scales by using complex enzyme and application thereof are characterized in that: the method comprises the following steps:
sp 1: pretreating raw materials: selecting fish scales as raw materials, and preprocessing the selected materials;
sp 2: decalcification treatment: recovering the pretreated raw materials, and decalcifying and degreasing the raw materials by a selection method;
sp3 composition analysis: recovering the processed raw materials, detecting through experiments to obtain required data, and storing and comparing;
sp4 complex enzyme extraction: extracting the raw materials by adopting a complex enzyme extraction method;
sp 5: and (4) recovering and preserving: and recovering the extracted raw materials, and storing the raw materials to a required environment at the temperature of minus 1 to minus 4 ℃.
2. The method for extracting small molecular peptides from fish scales by using compound enzymes and the application of the method, as claimed in claim 1, is characterized in that: selecting grass carp scales as an extraction raw material, purchasing the raw material, storing and moving the raw material to a position to be processed, pretreating the purchased raw material, removing non-fish scale substances such as gills, fins, fish viscera and the like of the collected grass carps, washing the raw material with cold water for 5min, recovering and storing the raw material, wiping off water on the surfaces of the fish scales, packaging the raw material into 20 g/bag and 40 g/bag respectively, freezing and preserving the raw material for later use, taking out the component of the required raw material according to requirements, putting the fish scale raw material into 1000mL of 1% sodium chloride boiling water, blanching for 2min for sterilization, putting the raw material into a fish scale cleaning machine, cleaning the raw material, and stirring and cleaning for 2h midway by adopting warm water with the material-liquid ratio of 1: 15 at 40-60 ℃ for one time and replacing cleaning liquid.
3. The method for extracting small molecular peptides from fish scales by using compound enzymes and the application of the method, as claimed in claim 1, is characterized in that: 50g of degreased, washed and dried fish scales for later use is placed in a 1000mL beaker, 500mL of water is added according to the material-liquid ratio of 1:10, a hydrochloric acid solution is added, after heating and stirring reaction are finished, a decalcification solution is sampled, the fish scales are washed with clear water for 10 times, dried and placed under 279C for vacuum drying for 24 hours, the concentration of hydrochloric acid is 0.37mol/L, the decalcification time is 35min, and the decalcification rate in the decalcification solution reaches 92%.
4. The method for extracting small molecular peptides from fish scales by using compound enzymes and the application of the method, as claimed in claim 1, is characterized in that: accurately sucking 0.5mL of decalcifying liquid into a 250mL volumetric flask, diluting to 50mL with distilled water, adding 10mL of ammonia buffer solution with pH of 10 and 3 drops of chrome black T indicator. Titrating with 0.0100mol/L EDTA standard solution until the solution turns from purple to blue, recording the consumption volume of EDTA to obtain the Torr probability, adding 6mol/L hydrochloric acid 9mL into 1mL of centrifuged fish scale decalcification solution by using a chromatograph, hydrolyzing in an oven at 110 ℃ for 12h, and diluting by 200 times to detect the hydroxyproline content of the decalcification solution.
5. The method for extracting small molecular peptides from fish scales by using compound enzymes and the application of the method, as claimed in claim 1, is characterized in that: the compound enzyme is composed of papain, bromelain and an alkaline compound enzyme, and the mass ratio of the compound enzyme to the papain, the bromelain and the alkaline compound enzyme is 1: 1: 2, the mass of fish scale peptide prepared by hydrolyzing 3% of papain, 3% of bromelain and 6% of alkaline complex enzyme for 5 hours is measured after centrifugation, the mass of collagen peptide with molecular mass of more than 2000u is 6.15%, the mass of collagen peptide with molecular mass of less than 176u is 3.82%, and the contents of oligopeptide with molecular mass distribution of 176-1000 u and 1000-2000 u are 73.3% and 16.6% respectively.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115746128A (en) * | 2022-09-30 | 2023-03-07 | 山东杰乐生物科技有限公司 | Preparation method of fish skin and scale collagen for increasing hydroxyproline content |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007326869A (en) * | 2007-07-27 | 2007-12-20 | Seiwa Kasei Co Ltd | Hydrolyzed collagen derived from fish scale |
| JP2008086219A (en) * | 2006-09-29 | 2008-04-17 | Tsuji Seiyu Kk | Collagen peptide powder, method for producing the same and polyphenol-containing product |
| CN101899491A (en) * | 2010-05-18 | 2010-12-01 | 武汉艾诗妮娜生物科技有限公司 | Method for preparing micromolecular collagen peptide from fish scales by utilizing complex enzyme |
| CN103045706A (en) * | 2012-12-27 | 2013-04-17 | 国家海洋局第三海洋研究所 | Method for continuously producing scale collagen peptide chelated calcium salt and scale collagen peptide |
| US20170164638A1 (en) * | 2015-04-30 | 2017-06-15 | China National Research Institute Of Food And Fermentation Industries | Fish protein oligopeptide with low allergenicity and slight fishiness and industrial preparation method and application thereof |
-
2020
- 2020-12-30 CN CN202011601405.2A patent/CN112522357A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008086219A (en) * | 2006-09-29 | 2008-04-17 | Tsuji Seiyu Kk | Collagen peptide powder, method for producing the same and polyphenol-containing product |
| JP2007326869A (en) * | 2007-07-27 | 2007-12-20 | Seiwa Kasei Co Ltd | Hydrolyzed collagen derived from fish scale |
| CN101899491A (en) * | 2010-05-18 | 2010-12-01 | 武汉艾诗妮娜生物科技有限公司 | Method for preparing micromolecular collagen peptide from fish scales by utilizing complex enzyme |
| CN103045706A (en) * | 2012-12-27 | 2013-04-17 | 国家海洋局第三海洋研究所 | Method for continuously producing scale collagen peptide chelated calcium salt and scale collagen peptide |
| US20170164638A1 (en) * | 2015-04-30 | 2017-06-15 | China National Research Institute Of Food And Fermentation Industries | Fish protein oligopeptide with low allergenicity and slight fishiness and industrial preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 朱俊颖等: "复合酶法制备高纯度鱼基质胶原蛋白肽", 《中国食品学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115746128A (en) * | 2022-09-30 | 2023-03-07 | 山东杰乐生物科技有限公司 | Preparation method of fish skin and scale collagen for increasing hydroxyproline content |
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Application publication date: 20210319 |