CN114805549A - Preparation method of fish collagen oligopeptide - Google Patents
Preparation method of fish collagen oligopeptide Download PDFInfo
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- CN114805549A CN114805549A CN202210294750.9A CN202210294750A CN114805549A CN 114805549 A CN114805549 A CN 114805549A CN 202210294750 A CN202210294750 A CN 202210294750A CN 114805549 A CN114805549 A CN 114805549A
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Abstract
The invention discloses a preparation method of fish collagen oligopeptide, which belongs to the technical field of food processing, wherein the fish collagen oligopeptide takes fresh fish skin, fish scales and fish bones as raw materials, a collagen crude extract is prepared, a sodium hydroxide solution is used for adjusting the pH value, then protease and fig juice are added for enzymolysis for 2-6h, after enzyme deactivation, a strain is inoculated for fermentation for 4-16h, after sterilization, activated carbon is used for decoloration and deodorization for 1h, fermentation liquor is collected by filtration, the fermentation liquor is filtered by an ultrafiltration membrane with the molecular weight of 1k, and after nanofiltration, spray drying is carried out to obtain the fish collagen oligopeptide; the fig juice is rich in protease with higher activity, so that the average molecular weight of the obtained fish collagen oligopeptide is smaller, and the prepared fish collagen oligopeptide has the molecular weight of less than 1000 daltons and can be quickly absorbed; after enzyme deactivation, strains are added for fermentation, so that fishy smell and bitter substances in the prepared fish collagen oligopeptide are effectively reduced, the properties of the product are improved, and the fish collagen oligopeptide is more acceptable to consumers.
Description
Technical Field
The invention belongs to the technical field of food processing, and particularly relates to a preparation method of fish collagen oligopeptide.
Background
The fish resources in the ocean and the fresh water are rich. In the process of processing aquatic products, a large amount of processing byproducts such as fish skin, fish bones and fish scales are inevitably generated and are directly discarded, so that environmental pollution and resource waste are easily caused. If the byproducts can be fully utilized, the additional value of fish processing can be provided, the industrial chain is prolonged, the environmental pollution can be reduced, and good economic benefit and social benefit can be obtained.
Collagen is a white, opaque, unbranched fibrous protein, which is mainly present in animal skin, bone, cartilage, teeth, tendons, ligaments and blood vessels, and is an important structural protein of connective tissues, and plays a role in supporting organs and protecting organisms. The fish skin, bone, scale, fin, air bladder, etc. contain collagen, wherein the collagen content in fish skin is the highest, and can be more than 80% of the protein content. The collagen peptide is a product obtained by decomposing and breaking molecular chains of collagen through enzymolysis. The peptide is formed by removing water molecules from 2 or more amino acids, is a structural fragment for forming protein, has small molecular weight, and is easy to absorb. Peptides are the basis of molecular structures of proteins with a variety of physiological effects. Thus, when the amino acid structure is altered, the efficacy of the peptide is also altered. Peptides are of thousands of species and their functions are also diverse.
The fish collagen peptide is widely applied to the fields of food, cosmetics, medicines and the like at present, but the fishy smell and the bitter taste of the fish collagen peptide seriously influence the application of the fish collagen peptide in the food and the cosmetics. Therefore, it is required to prepare a fish collagen oligopeptide with a small average molecular weight and less bitter and fishy smell.
Disclosure of Invention
The invention aims to provide a preparation method of fish collagen oligopeptide to solve the problems in the background technology.
The purpose of the invention can be realized by the following technical scheme: a preparation method of fish collagen oligopeptide comprises the following steps:
the method comprises the following steps: taking fresh fish skin, fish scales and fish bones, adding a sodium hydroxide solution with the mass fraction of 0.5%, soaking for 2-4h, then cleaning for 2-3 times by using clear water, adding softened water, transferring to a grinding machine, and stirring to be pasty to obtain a total protein crude extract;
step two: adjusting pH value of the crude extract to 2-4 with hydrochloric acid, heating to 75-90 deg.C, and extracting gel for 3-8 hr to obtain crude extract of collagen;
step three: adjusting the pH value of the collagen crude extract to 5-8 by using a sodium hydroxide solution, adding protease accounting for 0.1-1% of the mass of the collagen crude extract, adding fig juice accounting for 0.5-5% of the mass of the collagen crude extract, carrying out enzymolysis for 2-6h at 45-60 ℃ to generate fish collagen oligopeptide in an enzymolysis solution, killing enzyme, inoculating a strain, fermenting for 4-16h at 30-42 ℃, debitterizing and deodorizing the fish collagen oligopeptide, adding activated carbon accounting for 2-5% of the mass of the collagen crude extract after sterilization, decoloring and deodorizing for 1h at 60 ℃, filtering and collecting a fermentation liquid;
step four: filtering the fermentation liquor by using an ultrafiltration membrane with the molecular weight of 1k, removing inorganic salt and water-soluble free amino acid from the filtrate by using nanofiltration equipment, and then carrying out spray drying to obtain the fish collagen oligopeptide.
Further, the protease in the third step is one or a mixture of several of alkaline protease, neutral protease, flavor enzyme and bromelain according to any ratio.
Further, the strain in the third step is one or a mixture of several of Lactobacillus curvatus, Pediococcus pentosaceus and Staphylococcus xylosus according to any ratio, and the total inoculation amount of the strain is 2 multiplied by 10 9 -800×10 9 cfu/mL。
Further, in the third step, the protease is prepared by mixing bromelain and flavor enzyme according to the mass ratio of 3: 1.
Further, fig juice is obtained by selecting fig with Chinese national geographical sign of Weihai fig as raw material, cleaning, crushing and squeezing.
The invention has the beneficial effects that:
the molecular weight of the fish collagen oligopeptide prepared by the invention is less than 1000 daltons, and the fish collagen oligopeptide can be rapidly absorbed by organisms; in the preparation process, besides conventional protease, a small amount of fig juice is added, and the fig juice is rich in protease with higher activity, so that the average molecular weight of the obtained fish collagen oligopeptide is smaller, and the national standard requirement can be met; after enzyme deactivation, strains are added for fermentation, so that fishy smell and bitter substances in the prepared fish collagen oligopeptide are effectively reduced, the properties of the product are improved, and the fish collagen oligopeptide is more acceptable to consumers.
Drawings
The invention will be further described with reference to the accompanying drawings.
FIG. 1 is a graph showing the content of hydroxyproline in blood of each group of rats in 24 hours in experiment 2 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 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.
Example 1
The preparation method of the fish collagen oligopeptide comprises the following steps:
the method comprises the following steps: taking fresh fish skin, fish scales and fish bones, adding a sodium hydroxide solution with the mass fraction of 0.5%, soaking for 2 hours, then cleaning for 2 times by using clear water, adding softened water, transferring to a grinding machine, and stirring to be pasty to obtain a total protein crude extract;
step two: adjusting the pH value of the crude extract of the total protein to 2 by using hydrochloric acid, then heating to 75 ℃, and carrying out heat preservation and gel extraction for 3 hours to obtain a crude extract of collagen;
step three: adjusting pH value of 100g of collagen crude extract to 5 with sodium hydroxide solution, and adding 0.1g of protease, wherein the protease is prepared by mixing bromelain and flavourzyme according to the mass ratio of 3: 1; adding 0.5g of fig juice, performing enzymolysis at 45 deg.C for 2 hr to obtain fish collagen oligopeptide, inactivating enzyme, inoculating Lactobacillus curvatus, and inoculating with 2 × 10 strain 9 cfu/mL, fermenting at 30 deg.C for 4h, debitterizing and deodorizing fish collagen oligopeptide, sterilizing, adding 2g active carbon, decolorizing and deodorizing at 60 deg.C for 1h, filtering, and collecting fermentation broth;
step four: filtering the fermentation liquor by using an ultrafiltration membrane with the molecular weight of 1k, removing inorganic salt and water-soluble free amino acid from the filtrate by using nanofiltration equipment, and then carrying out spray drying to obtain the fish collagen oligopeptide.
Example 2
The preparation method of the fish collagen oligopeptide comprises the following steps:
the method comprises the following steps: taking fresh fish skin, fish scales and fish bones, adding a sodium hydroxide solution with the mass fraction of 0.5%, soaking for 3h, then cleaning for 2 times by using clear water, adding softened water, transferring to a grinding machine, and stirring to be pasty to obtain a total protein crude extract;
step two: adjusting the pH value of the crude extract of the total protein to 3 by using hydrochloric acid, then heating to 80 ℃, and carrying out heat preservation and gel extraction for 5 hours to obtain a crude extract of collagen;
step three: taking 100g of collagen crude extract, adjusting the pH value to 7 by using a sodium hydroxide solution, and adding 0.5g of neutral protease; adding 2g of fig juice, performing enzymolysis at 50 deg.C for 4 hr to obtain fish collagen oligopeptide, inactivating enzyme, inoculating Pediococcus pentosaceus with total inoculum size of 800 × 10 9 cfu/mL, fermenting at 35 deg.C for 10h, debitterizing and deodorizing fish collagen oligopeptide, sterilizing, adding 3g active carbon, decolorizing and deodorizing at 60 deg.C for 1h, filtering, and collecting fermentation liquid;
step four: filtering the fermentation liquor by using an ultrafiltration membrane with the molecular weight of 1k, removing inorganic salt and water-soluble free amino acid from the filtrate by using nanofiltration equipment, and then carrying out spray drying to obtain the fish collagen oligopeptide.
Example 3
The preparation method of the fish collagen oligopeptide comprises the following steps:
the method comprises the following steps: taking fresh fish skin, fish scales and fish bones, adding a sodium hydroxide solution with the mass fraction of 0.5%, soaking for 4h, then cleaning for 3 times by using clear water, adding softened water, transferring to a grinding machine, and stirring to be pasty to obtain a total protein crude extract;
step two: adjusting the pH value of the crude extract of the total protein to 4 by using hydrochloric acid, then heating to 90 ℃, and carrying out heat preservation and gel extraction for 8 hours to obtain a crude extract of collagen;
step three: taking 100g of collagen crude extract, adjusting the pH value to 8 by using a sodium hydroxide solution, and adding 1g of alkaline protease; adding 5g of fig juice, performing enzymolysis at 60 deg.C for 6 hr to obtain fish collagen oligopeptide, inactivating enzyme, inoculating Staphylococcus xylosus, and inoculating the strain with a total inoculum size of 400 × 10 9 cfu/mL, fermenting at 42 deg.C for 16h, debitterizing and deodorizing fish collagen oligopeptide, sterilizing, adding 5g active carbon, decolorizing and deodorizing at 60 deg.C for 1h, filtering, and collecting fermentation broth;
step four: filtering the fermentation liquor by using an ultrafiltration membrane with the molecular weight of 1k, removing inorganic salt and water-soluble free amino acid from the filtrate by using nanofiltration equipment, and then carrying out spray drying to obtain the fish collagen oligopeptide.
Example 4
The preparation method of the fish collagen oligopeptide comprises the following steps:
the method comprises the following steps: taking fresh fish skin, fish scales and fish bones, adding a sodium hydroxide solution with the mass fraction of 0.5%, soaking for 4h, then cleaning for 3 times by using clear water, adding softened water, transferring to a grinding machine, and stirring to be pasty to obtain a total protein crude extract;
step two: adjusting the pH value of the crude extract of the total protein to 2.5 by hydrochloric acid, then heating to 90 ℃, and carrying out heat preservation and gel extraction for 6 hours to obtain a crude extract of collagen;
step three: adjusting pH value of 100g of collagen crude extract to 7.8 with sodium hydroxide solution, adding 0.8g of protease, wherein the protease is prepared by mixing bromelain and flavourzyme according to the mass ratio of 3: 1; adding 5g fructus fici juice, and performing enzymolysis at 50 deg.C for 6 hr to obtain fishInactivating enzyme of collagen oligopeptide, and inoculating 200 × 10 collagen oligopeptide 8 cfu/mL Lactobacillus curvatus, 300X 10 8 cfu/mL Pediococcus pentosaceus and 400X 10 8 Fermenting cfu/mL staphylococcus xylosus at 38 deg.C for 6h, debitterizing and deodorizing fish collagen oligopeptide, sterilizing, adding 3g active carbon, decolorizing and deodorizing at 60 deg.C for 1h, filtering, and collecting fermentation liquid;
step four: filtering the fermentation liquor by using an ultrafiltration membrane with the molecular weight of 1k, removing inorganic salt and water-soluble free amino acid from the filtrate by using nanofiltration equipment, and then carrying out spray drying to obtain the fish collagen oligopeptide.
The sources of the species used in examples 1-4 are shown in table 1:
TABLE 1
Comparative example 1
The preparation of fish collagen oligopeptide without adding fig juice and using bacterial fermentation conditions comprises the following steps:
the method comprises the following steps: taking fresh fish skin, fish scales and fish bones, adding a sodium hydroxide solution with the mass fraction of 0.5%, soaking for 4h, then cleaning for 3 times by using clear water, adding softened water, transferring to a grinding machine, and stirring to be pasty to obtain a total protein crude extract;
step two: regulating the pH value of the total protein crude extract to 2.5 by using hydrochloric acid, then heating to 90 ℃, and carrying out heat preservation and gel extraction for 6 hours to obtain a collagen crude extract;
step three: adjusting pH value of 100g of collagen crude extract to 7.8 with sodium hydroxide solution, adding 0.8g of protease, wherein the protease is prepared by mixing bromelain and flavourzyme according to the mass ratio of 3: 1; carrying out enzymolysis for 6h at 50 ℃, adding 3g of activated carbon after enzyme deactivation, decoloring and deodorizing for 1h at 60 ℃, filtering and collecting fermentation liquor;
step four: filtering the fermentation liquor by using an ultrafiltration membrane with the molecular weight of 1k, removing inorganic salt and water-soluble free amino acid from the filtrate by using nanofiltration equipment, and then carrying out spray drying to obtain the fish collagen oligopeptide.
The molecular weight and oligopeptide of example 4 and comparative example 1 were measured according to the national standard of GB22729 ocean fish oligopeptide, and the results are shown in Table 2:
TABLE 2
As can be seen from Table 2, the average molecular weight of the obtained product is lower after the fig juice and the microorganism are added for fermentation, and the national standard requirement can be met.
Test 2
Feeding 24 rats with the same growth condition and health condition for 24h, and averagely dividing the rats into four groups of ABCD; respectively gavage distilled water for a group, respectively gavage the fish collagen oligopeptide (average molecular weight 576 daltons) prepared in the example 4 for a group B, respectively gavage the fish collagen oligopeptide (average molecular weight 900 daltons) prepared in the comparative example 1 for a group C, respectively gavage the oligopeptide (average molecular weight 3364 daltons) provided by the limited biological technology of shengmeno, anhui, respectively, and respectively measuring the hydroxyproline content in the serum of each group of mice when blood samples are taken for 0.5h, 1h, 2h, 4h, 7h and 24h, and calculating the average value of each group, wherein the result is shown in fig. 1.
Since hydroxyproline is a characteristic amino acid of collagen peptide, the content of hydroxyproline in blood is high or low, and the hydroxyproline can reflect the absorption condition of the body. As can be seen from fig. 1, the smaller the molecular weight, the faster the collagen peptide is absorbed and the higher the content in blood.
Test 3
The fish collagen oligopeptides prepared in examples 1 to 4 were labeled as sample 1 to sample 4, respectively, and the fish collagen oligopeptide purchased from Sheng Mei Nuo Biotechnology Ltd, Anhui having an average molecular weight of 582 daltons was labeled as sample 5, and the label was placed on the bottom of a vessel for containing each sample.
The sensory evaluation was performed on samples 1 to 5 by a panel of 5 professional sensory evaluators. The scoring principle is 1-5 points: (1) the mouthfeel is as follows: the bitter taste is extremely heavy for 1 minute; obvious bitter taste is divided into 2 points; bitter taste is divided into 3 points; slightly bitter and astringent taste, and no other peculiar smell of 4 points; has no bitter taste and slight sweet taste of 5 points. (2) Odor: the fishy smell is extremely heavy and is 1 minute; obvious fishy smell is 2 points; 3 points of fishy smell; has slight fishy smell and no other peculiar smell of 4 points; no bitter taste 5 points. The scoring results are shown in table 3:
TABLE 3
| Score the | Sample | 1 | Sample 2 | Sample 3 | Sample No. 4 | Sample No. 5 |
| Taste of the product | 4.31 | 4.29 | 4.30 | 4.33 | 3.24 | |
| Smell(s) | 4.67 | 4.66 | 4.68 | 4.66 | 3.17 |
As can be seen from Table 3, the fish collagen oligopeptides in samples 1 to 4 are superior to the commercial products in taste and smell.
It should be noted that, in this document, terms such as "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.
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 (7)
1. A preparation method of fish collagen oligopeptide is characterized by comprising the following steps:
the method comprises the following steps: adjusting pH of the crude collagen extract to 5-8 with sodium hydroxide solution, adding protease 0.1-1% of the crude collagen extract, adding fig juice 0.5-5% of the crude collagen extract, performing enzymolysis at 45-60 deg.C for 2-6h, inactivating enzyme, inoculating strain, fermenting at 30-42 deg.C for 4-16h, sterilizing, adding active carbon 2-5% of the crude collagen extract, decolorizing at 60 deg.C for 1h, filtering, and collecting the fermentation broth;
step two: filtering the fermentation liquor by using an ultrafiltration membrane with the molecular weight of 1k, performing nanofiltration on the filtrate, and performing spray drying to obtain the fish collagen oligopeptide.
2. The method for preparing fish collagen oligopeptide according to claim 1, wherein the method for preparing the crude collagen extract comprises the following steps: taking fresh fish skin, fish scales and fish bones, adding a sodium hydroxide solution with the mass fraction of 0.5%, soaking for 2-4h, cleaning, adding softened water, and stirring to be pasty to obtain a total protein crude extract; adjusting acid, and extracting gel at 75-90 deg.C for 3-8 hr to obtain collagen crude extract.
3. The method for preparing fish collagen oligopeptide according to claim 1, wherein the protease in the first step is one or more of alkaline protease, neutral protease, flavor enzyme and bromelain, and the mixture is prepared by mixing the alkaline protease, the neutral protease, the flavor enzyme and the bromelain at any ratio.
4. The method for preparing fish collagen oligopeptide according to claim 1, wherein the bacterial strain in the first step is one or more of Lactobacillus curvatus, Pediococcus pentosaceus and Staphylococcus xylosus, and the bacterial strain is mixed in any ratio.
5. The method according to claim 1, wherein the total inoculum size of the bacterial strain in step one is 2X 10 9 -800×10 9 cfu/mL。
6. The method for preparing fish collagen oligopeptide according to claim 2, wherein the acid adjusting method comprises the following steps: adjusting pH of the crude extract to 2-4 with hydrochloric acid.
7. The method of claim 3, wherein the protease is bromelain and flavourzyme mixed together at a mass ratio of 3: 1.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103205480A (en) * | 2013-04-15 | 2013-07-17 | 武汉工业学院 | Method for producing high-quality collagen oligopeptide by using fish skin or fishbone |
| CN103388017A (en) * | 2013-08-06 | 2013-11-13 | 国家海洋局第三海洋研究所 | Preparation technology of fish skin collagen oligopeptide |
| CN109371092A (en) * | 2018-12-26 | 2019-02-22 | 河北肽都生物科技有限公司 | A kind of preparation method of collagen peptide |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103205480A (en) * | 2013-04-15 | 2013-07-17 | 武汉工业学院 | Method for producing high-quality collagen oligopeptide by using fish skin or fishbone |
| CN103388017A (en) * | 2013-08-06 | 2013-11-13 | 国家海洋局第三海洋研究所 | Preparation technology of fish skin collagen oligopeptide |
| CN109371092A (en) * | 2018-12-26 | 2019-02-22 | 河北肽都生物科技有限公司 | A kind of preparation method of collagen peptide |
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