Method for preparing fish collagen peptides by enzymatic hydrolysis after biological fermentation, products and applications

Abstract

The present invention discloses a method for preparing fish collagen peptides by enzymatic hydrolysis after biological fermentation, and relates to the field of biotechnology. The preparation method of the fish collagen peptide comprises the following steps: (1) adding an extractant to fish tissue for extraction, filtering, and obtaining a filtrate; (2) adding a carbon source to the filtrate to obtain a fermentation medium; (3) inoculating a plant lactobacillus strain on a seed culture medium, culturing on a shaking table, and obtaining a seed liquid; (4) inoculating the seed liquid into a fermentation medium, and fermenting and culturing; (5) first adding an enzyme activator, then adding an enzymolysis agent for enzymatic hydrolysis, inactivating the enzyme, centrifuging, and drying to obtain fish collagen peptide powder; the enzyme activator described in step (5) is a fungus polysaccharide, oligomannose, and raffinose. The preparation method of the present invention ensures that the prepared fish collagen peptide has high quality and good antibacterial effect, can effectively improve symptoms such as skin sagging, dull color, greasy, and freckles, and has good application prospects.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a method for preparing fish collagen peptides by enzymatic hydrolysis after biological fermentation, as well as products and applications thereof. Background Art

[0002] Collagen, also known as collagen, is a fibrous protein with a helical structure formed by three intertwined peptide chains. It is the most abundant structural protein in animals. Collagen peptides (CPs) are denatured hydrolyzates of collagen obtained from animal tissue through pretreatment, physical and chemical separation, or biological modification (enzymatic cleavage or fermentation). Collagen peptides are a class of small molecule amino acid sequences with multiple biological functions, primarily obtained through collagen hydrolysis. Fish-derived collagen peptides come from various sources, including fish skin, scales, and bones. These are byproducts of the fish processing industry. Extracting collagen peptides from these sources can help reduce the added value of fishery byproducts, improve their utilization, promote the development of related industries, and reduce resource waste.

[0003] The biological activity of collagen peptides refers to peptide compounds that are beneficial to the body's life activities and have physiological functions. At present, the biological activity functions of collagen peptides are widely recognized. The biological activities of collagen peptides include antioxidant, anti-fatigue, hypoglycemic, hypolipidemic and antibacterial.

[0004] There are three main methods for producing collagen peptides: 1. Enzymatic hydrolysis of collagen to obtain bioactive peptides with multiple functions. 2. Preparation of collagen peptides requires the use of extracellular enzymes produced by microbial fermentation. 3. Chemical hydrolysis: Collagen peptides are produced through acid-base catalytic decomposition of peptide bonds. This reaction is drastic, difficult to control, and results in a high degree of protein hydrolysis.

[0005] Chinese patent CN108866137A discloses a method for preparing fish skin / scale collagen peptides, comprising the following steps: using at least one of fish skin and fish scales as raw material; washing and removing impurities from the raw material, and then steam-exploding it; adding water to the steam-exploded raw material for aqueous extraction; adjusting the pH of the extract to 6.0-8.5, then adding a protease preparation and subjecting it to an enzymatic hydrolysis reaction at 40-65°C for 0.5-2 hours; inactivating the enzymes and filtering the hydrolyzed product to obtain a crude hydrolyzate; ultrafiltration of the crude hydrolyzate using an ultrafiltration membrane, with the resulting permeate serving as a refined solution; vacuum concentrating the refined solution to a solids content of 25-40%, and then spray drying it. The resulting fish skin / scale collagen peptide has an average relative molecular weight of less than 3 kDa and a protein content exceeding 95%. This invention addresses the issue of utilizing fish skin and scales in fish processing, thereby increasing the added value of aquatic product processing waste, but it does not further improve the antibacterial effect of the fish collagen peptides.

[0006] Chinese patent CN113088550A discloses a fish collagen peptide and a preparation method thereof, which comprises the following steps: (1) taking fish glue scraps and soaking them through a deodorizing and soaking process; (2) cleaning the soaked fish glue and removing impurities; (3) adding purified water to the fish glue after removing impurities to a fixed volume, steaming under high pressure, then cooling and reducing the pressure to obtain a colloid; (4) crushing the colloid and placing it in a reaction tank, and then adding protease and flavor enzyme for enzymatic hydrolysis; (5) heating the reaction tank in step (4) to kill the activity of the protease and flavor enzyme, then filtering and removing waste residue, collecting the filtrate, and obtaining the fish collagen peptide. The preparation method provided by the invention uses fish glue scraps as raw materials, solves the problem of waste in processing costs and environmental pollution of fish glue products, and the enzymatic hydrolysis product can be fully absorbed by the human body. However, the quality and antibacterial effect of the fish collagen peptide prepared by the invention are not improved.

[0007] The current preparation method of fish collagen results in the prepared fish collagen having low quality and poor antibacterial properties. Summary of the Invention

[0008] The purpose of the present invention is to provide a method, product and application of preparing fish collagen peptides by enzymatic hydrolysis after biological fermentation, so that the prepared fish collagen peptides have higher quality and better antibacterial effect, can effectively improve skin condition, and have good application prospects.

[0009] In order to achieve the above-mentioned object of the invention, the technical solution of the present invention is as follows:

[0010] In one aspect, the present invention provides a method for preparing fish collagen peptide, comprising the following steps:

[0011] (1) Adding fish tissue to an extractant for extraction, filtering, and obtaining a filtrate;

[0012] (2) Adding a carbon source to the filtrate to obtain a fermentation medium;

[0013] (3) inoculating the Lactobacillus plantarum strain on the seed culture medium and culturing on a shaking table to obtain the seed liquid;

[0014] (4) inoculating the seed liquid into the fermentation medium and fermenting and culturing;

[0015] (5) First add enzyme activator, then add enzymolysis agent for enzymolysis, inactivate enzyme, centrifuge, and dry to obtain fish collagen peptide powder;

[0016] The enzyme activators described in step (5) are fungus polysaccharide, oligomannose and raffinose.

[0017] Specifically, the fish tissue in step (1) is selected from one or more of fish skin, fish bones, fish meat, and fish scales; further, the fish tissue is fish scales.

[0018] According to some embodiments of the present invention, the fish tissue includes but is not limited to fish tissue of marine fish and fish tissue of freshwater fish.

[0019] Specifically, the fish tissue further includes a pretreatment step.

[0020] Furthermore, the pretreatment is as follows: soaking the fish scales in 1-5 mol / L baking soda for 20-40 minutes to remove the odor, rinsing with water until the pH is neutral, and then sterilizing at 121° C. for 20 minutes.

[0021] Furthermore, the sterilization condition is 121° C. for 20 minutes.

[0022] Specifically, the mass ratio of fish tissue to extractant in step (1) is 1:(1-8).

[0023] Furthermore, in step (1), the mass ratio of fish tissue to extractant is 1:5.

[0024] Specifically, the extractant is water.

[0025] Specifically, the extraction time in step (1) is 40-60 min; further, the extraction time in step (1) is 50 min.

[0026] Specifically, in step (1), stirring is performed every 15 minutes during the extraction process.

[0027] Specifically, the carbon source in step (2) is selected from one or more of glucose, glycerol, sucrose, oligofructose, crystalline fructose, white sugar, xylose, mannose, soluble starch, dextrin, sorbitol, and lactose.

[0028] Furthermore, the carbon source in step (2) is glucose.

[0029] Furthermore, the amount of glucose added is 1-3%; furthermore, the amount of glucose added is 1%.

[0030] Furthermore, after adding glucose, the mixture was kept at 100° C. for 20 minutes and cooled to obtain a fermentation medium.

[0031] Specifically, the inoculation amount described in step (3) is 5-10%.

[0032] Furthermore, the inoculation amount described in step (3) is 5%.

[0033] Specifically, the shaking culture conditions in step (3) are: 35-40°C, a rotation speed of 220-260 rpm, and shaking culture for 12-24 hours.

[0034] Furthermore, the shaking culture conditions in step (3) are: 37° C., 220-260 rpm, and 12 h of shaking culture.

[0035] Specifically, the inoculation amount of the seed liquid in step (4) is 1-5%.

[0036] Furthermore, the inoculation amount of the seed solution in step (4) is 3%.

[0037] Specifically, the fermentation culture conditions in step (4) are 35-40° C., and the fermentation time is 150-300 min.

[0038] Furthermore, the fermentation culture condition in step (4) is 38° C. for 210 min.

[0039] Specifically, the pH is adjusted to 6.5-7.5 before adding the enzyme preservative in step (5).

[0040] Furthermore, the pH is adjusted to 7.0 before adding the enzyme preservative in step (5).

[0041] Furthermore, the mass ratio of the wood ear polysaccharide, oligomannose and raffinose is (1-5): (0.1-2): (0.5-1.5).

[0042] Furthermore, the mass ratio of the wood ear polysaccharide, oligomannose and raffinose is 1:0.1:0.5.

[0043] Specifically, the enzymatic hydrolyzing agents in step (5) are papain and flavor protease, and the mass ratio of papain to flavor protease is (1-5):1.

[0044] Furthermore, the mass ratio of papain to flavor protease is 2:1.

[0045] Specifically, the enzymatic hydrolysis temperature in step (5) is 35-40°C, and the enzymatic hydrolysis time is 10-20 hours.

[0046] Furthermore, in step (5), the enzymatic hydrolysis temperature is 38° C., and the enzymatic hydrolysis time is 15 h.

[0047] Specifically, the enzyme inactivation conditions in step (5) are: inactivation at 100°C for 15 minutes.

[0048] Specifically, the centrifugal conditions in step (5) are 3000-5000 r / min and centrifugation for 15-30 min.

[0049] In another aspect, the present invention provides fish collagen peptide prepared by the above preparation method.

[0050] In another aspect, the present invention provides the fish collagen peptide prepared by the above preparation method or the use of the above fish collagen peptide in preparing products for repairing skin condition.

[0051] Specifically, the products include but are not limited to nutritional products, medicines and cosmetics.

[0052] Furthermore, the product is taken orally.

[0053] More specifically, the cosmetics can be conventional preparations such as lotion, toner, toner, essence water, face cream, face mask, cleansing cream, facial cleanser, shampoo and conditioner, shaving cream, hair milk, hair spray, lipstick, sunscreen, hand cream, etc.

[0054] More specifically, the cosmetics may further include matrix materials and conventional auxiliary materials.

[0055] More specifically, the matrix raw materials include but are not limited to colloid raw materials, powdery raw materials, oil raw materials, wax raw materials, coagulants, and surfactants.

[0056] More specifically, the conventional excipients include but are not limited to antioxidants, adhesives, lubricants, moisturizers, preservatives, whitening agents, film agents, emulsifiers, and cosmetic nutritional additives.

[0057] Furthermore, the medicine may also contain a pharmaceutically acceptable carrier.

[0058] Furthermore, the pharmaceutically acceptable carrier is selected from one or more of an excipient, a stabilizer, a diluent, a binder, a preservative, and a lubricant.

[0059] Specifically, the dosage form of the drug is drops, mixtures, tinctures, injections, tablets, powders, oral liquids, capsules, granules, ointments, suspensions, powders, emulsions, solutions, pellets, pills, buccal tablets, freeze-dried powder injections, gels, suppositories or aerosols.

[0060] The beneficial effects of the present invention are:

[0061] The present invention provides a method for preparing fish collagen peptides, which improves the preparation process by adding an enzyme activator and a specific enzymolytic agent, and further restricting the types and ratios of the enzyme activator and enzymolytic agent. This results in the prepared fish collagen peptides having high hydroxyproline and total nitrogen contents, low ash content, high quality, and good antibacterial effects. The collagen peptides prepared by the present invention can effectively improve symptoms such as sagging skin, dull skin color, oiliness, and freckles, and have good application prospects. DETAILED DESCRIPTION

[0062] In order to make the technical means, creative features, purpose and efficacy of the present invention easy to understand, the present invention is further illustrated below in conjunction with specific examples, but the following examples are only preferred embodiments of the present invention, not all. Based on the examples in the implementation manner, other embodiments obtained by those skilled in the art without making creative work are all within the scope of protection of the present invention. In the following examples, unless otherwise specified, the operating methods used are all conventional operating methods, the equipment used are all conventional equipment, and the equipment and materials used in each embodiment are all the same.

[0063] The fish scales used in the following Examples 1-2 are grass carp scales, and those used in Examples 3-4 are silver carp scales.

[0064] The Lactobacillus plantarum used in the following examples and comparative examples was purchased from ATCC and was Lactobacillus plantarum ATCC14917.

[0065] Example 1 Preparation of fish collagen peptide

[0066] (1) Soak the fish scales in 2 mol / L baking soda for 30 min to remove the odor, rinse with water until the pH is neutral, sterilize at 121°C for 20 min, add water for extraction, and extract for 50 min. Stir every 15 min and filter to obtain the filtrate.

[0067] (2) Add 1% glucose to the filtrate, maintain at 100°C for 20 minutes, and cool to obtain a fermentation medium;

[0068] (3) Take the activated Lactobacillus plantarum strain and inoculate the strain into a seed culture medium at a 5% inoculum volume. Incubate the culture medium in a shaking incubator at 37°C and 220 rpm for 12 h to obtain a seed solution.

[0069] (4) The seed liquid was inoculated into the fermentation medium at an inoculum size of 3%, and fermented at 38°C for 3 hours and 30 minutes, and the pH was adjusted to 7.0;

[0070] (5) First, add enzyme activator and keep at room temperature for 10 minutes. Then, add enzymatic agent and perform enzymatic hydrolysis at 38°C for 15 hours. Stir 3 times during the enzymatic hydrolysis. Inactivate the enzyme at 100°C for 15 minutes. Centrifuge (3000r / min, 15 minutes). After the supernatant is concentrated under reduced pressure, freeze-dried to obtain fish collagen peptide powder.

[0071] In step (1), the mass volume ratio of fish scales to water is 1:5;

[0072] The enzyme activator in step (5) is fungus polysaccharide, oligomannosaccharide and raffinose, and the mass ratio of fungus polysaccharide, oligomannosaccharide and raffinose is 1:0.1:0.5;

[0073] The enzymatic hydrolyzing agents in step (5) are papain and flavor protease, and the mass ratio of papain to flavor protease is 2:1.

[0074] Example 2 Preparation of fish collagen peptide

[0075] (1) Soak the fish scales in 2 mol / L baking soda for 30 min to remove the odor, rinse with water until the pH is neutral, sterilize at 121°C for 20 min, add water for extraction, and extract for 40 min. Stir every 15 min and filter to obtain the filtrate.

[0076] (2) Add 2% glucose to the filtrate, maintain at 100°C for 20 minutes, and cool to obtain a fermentation medium;

[0077] (3) Take the activated Lactobacillus plantarum strain and inoculate it into a seed culture medium (MRS culture medium) at a 5% inoculum volume. Incubate it in a shaking incubator at 37°C and 220 rpm for 12 h to obtain a seed solution.

[0078] (4) The seed liquid was inoculated into the fermentation medium at an inoculum size of 3%, fermented at 38°C for 200 min, and the pH was adjusted to 7.0;

[0079] (5) First, add enzyme activator and keep at room temperature for 10 minutes. Then, add enzymatic agent and perform enzymatic hydrolysis at 38°C for 16 hours. Stir 3 times during the enzymatic hydrolysis. Inactivate the enzyme at 100°C for 15 minutes. Centrifuge (3000r / min, 15 minutes). After the supernatant is concentrated under reduced pressure, freeze-dried to obtain fish collagen peptide powder.

[0080] In step (1), the mass volume ratio of fish scales to water is 1:6;

[0081] The enzyme activator in step (5) is fungus polysaccharide, oligomannosaccharide and raffinose, and the mass ratio of fungus polysaccharide, oligomannosaccharide and raffinose is 2:0.5:1;

[0082] The enzymatic hydrolyzing agents in step (5) are papain and flavor protease, and the mass ratio of papain to flavor protease is 2.5:1.

[0083] Example 3 Preparation of fish collagen peptide

[0084] (1) Soak the fish scales in 2 mol / L baking soda for 30 min to remove the odor, rinse with water until the pH is neutral, sterilize at 121°C for 20 min, add water for extraction, and extract for 50 min. Stir every 15 min and filter to obtain the filtrate.

[0085] (2) Add 1% glucose to the filtrate, maintain at 100°C for 20 minutes, and cool to obtain a fermentation medium;

[0086] (3) Take the activated Lactobacillus plantarum strain and inoculate it into the seed culture medium at a 5% inoculum volume. Incubate it in a shaking incubator at 37°C and 220 rpm for 12 h to obtain the seed solution.

[0087] (4) The seed liquid was inoculated into the fermentation medium at a 5% inoculum size, fermented at 38°C for 220 min, and the pH was adjusted to 7.0;

[0088] (5) First, add enzyme activator and keep at room temperature for 10 minutes. Then, add enzymatic agent and perform enzymatic hydrolysis at 38°C for 15 hours. Stir 3 times during the enzymatic hydrolysis. Inactivate the enzyme at 100°C for 15 minutes. Centrifuge (3000r / min, 15 minutes). After the supernatant is concentrated under reduced pressure, freeze-dried to obtain fish collagen peptide powder.

[0089] In step (1), the mass volume ratio of fish scales to water is 1:8;

[0090] The enzyme activator in step (5) is fungus polysaccharide, oligomannosaccharide and raffinose, and the mass ratio of fungus polysaccharide, oligomannosaccharide and raffinose is 2:1:1;

[0091] The enzymatic hydrolyzing agents in step (5) are papain and flavor protease, and the mass ratio of papain to flavor protease is 3:1.

[0092] Example 4 Preparation of fish collagen peptide

[0093] (1) Soak the fish scales in 2 mol / L baking soda for 30 min to remove the odor, rinse with water until the pH is neutral, sterilize at 121°C for 20 min, add water for extraction, and extract for 60 min. Stir every 15 min and filter to obtain the filtrate.

[0094] (2) Add 1% glucose to the filtrate, maintain at 100°C for 20 minutes, and cool to obtain a fermentation medium;

[0095] (3) Take the activated Lactobacillus plantarum strain and inoculate it into the seed culture medium at a 5% inoculum volume. Incubate it in a shaking incubator at 37°C and 220 rpm for 12 h to obtain the seed solution.

[0096] (4) The seed liquid was inoculated into the fermentation medium at an inoculum rate of 3%, fermented at 38°C for 240 minutes, and the pH was adjusted to 7.5;

[0097] (5) First, add enzyme activator and keep at room temperature for 10 minutes. Then, add enzymatic agent and perform enzymatic hydrolysis at 38°C for 16 hours. Stir 3 times during the enzymatic hydrolysis. Inactivate the enzyme at 100°C for 15 minutes. Centrifuge (3000r / min, 15 minutes). After the supernatant is concentrated under reduced pressure, freeze-dried to obtain fish collagen peptide powder.

[0098] In step (1), the mass volume ratio of fish scales to water is 1:8;

[0099] The enzyme activator in step (5) is fungus polysaccharide, oligomannosaccharide and raffinose, and the mass ratio of fungus polysaccharide, oligomannosaccharide and raffinose is 3:2:1.5;

[0100] The enzymatic hydrolyzing agents in step (5) are papain and flavor protease, and the mass ratio of papain to flavor protease is 4:1.

[0101] Comparative Example 1 Preparation of fish collagen peptide

[0102] The only difference between Comparative Example 1 and Example 1 is that the mass ratio of fish scales to water in step (1) is 1:10, and the rest are the same.

[0103] Comparative Example 2 Preparation of fish collagen peptide

[0104] The only difference between Comparative Example 2 and Example 1 is that the enzyme activator described in step (5) is fungus polysaccharide, oligomannosaccharide and chitosan, and the mass ratio of fungus polysaccharide, oligomannosaccharide and chitosan is 1:0.1:0.5. The rest are the same.

[0105] Comparative Example 3 Preparation of fish collagen peptide

[0106] The only difference between Comparative Example 3 and Example 1 is that the enzyme activator described in step (5) is fungus polysaccharide, oligomannosaccharide and raffinose, and the mass ratio of fungus polysaccharide, oligomannosaccharide and raffinose is 6:0.5:1. The rest are the same.

[0107] Comparative Example 4 Preparation of fish collagen peptide

[0108] The only difference between Comparative Example 4 and Example 1 is that the enzymatic hydrolyzing agents in step (5) are trypsin and bromelain, and the rest are the same.

[0109] Comparative Example 5 Preparation of fish collagen peptide

[0110] The only difference between Comparative Example 5 and Example 1 is that the mass ratio of papain to flavor protease in step (5) is 6:1, and the rest are the same.

[0111] Experimental Example 1

[0112] The finished collagen peptide powders prepared in Examples 1-4 and Comparative Examples 1-5 were tested for physical and chemical indicators. The proline detection method was carried out in accordance with GB / T 9695.23, the total nitrogen detection method was carried out in accordance with GB 5009.5, and the ash detection method was carried out in accordance with GB 5009.4.

[0113] The test results are shown in Table 1 below.

[0114] Table 1.

[0115]

[0116] As can be seen from the above table, the hydroxyproline and total nitrogen contents of the fish collagen peptides prepared in Examples 1-4 of the present invention are significantly better than the requirements of the "National Food Safety Standard Collagen Peptide GB 31645-2018", and the ash content is significantly lower than the required ≤7.0g / 100g. The above shows that the fish collagen peptides prepared by the present invention are of high quality. The hydroxyproline and total nitrogen contents of the fish collagen peptides prepared in Comparative Examples 1-5 are significantly lower than those in Examples 1-4, and the ash content is higher than that in Examples 1-4. The fish collagen peptides prepared in Comparative Examples 1-5 are of poor quality.

[0117] Experimental Example 2

[0118] The collagen peptides prepared in Examples 1-4 and Comparative Examples 1-5 were stored at 25°C and 85% relative humidity for 6 months, and the total colony count test was performed using GB / T 4789.2-2010 Food Hygiene Microbiology Test - Determination of Total Colony Count. The results are shown in Table 2 below.

[0119] Table 2.

[0120]

[0121] As can be seen from the above table, the colony count of the fish collagen peptides prepared in Examples 1-4 was significantly reduced, and they had good antibacterial properties, and their antibacterial effects were significantly better than those of the fish collagen peptides prepared in Comparative Examples 1-5.

[0122] Experimental Example 3

[0123] The collagen peptides prepared in Examples 1-4 and Comparative Examples 1-5 were subjected to skin nourishing and moisturizing tests.

[0124] Experimental subjects: The experimental population had symptoms such as loose skin, dull color, oily skin, and freckles. They were aged 28-60 years old, with 10 subjects in each group, 5 males and 5 females in each group, and the ages of the groups were similar.

[0125] Experimental process: Drink 5g of collagen peptide powder prepared in Examples 1-4 and Comparative Examples 1-5 at 9 o'clock every morning, dissolve it in warm water and take it. The test time is 30 days. During the test period, products with similar efficacy should not be taken.

[0126] Efficacy evaluation criteria:

[0127] (1) Significant: Skin is more hydrated, oil production is significantly reduced, skin whiteness is significantly improved, and fine lines on the skin are reduced.

[0128] (2) Effective: The skin becomes moisturized, oil production is reduced, and whiteness is improved.

[0129] (3) Ineffective: There was no obvious improvement in skin symptoms before and after taking the drug.

[0130] The test results are shown in Table 3 below.

[0131] Table 3.

[0132]

[0133] As can be seen from the above table, the collagen peptides prepared in Examples 1-4 of the present invention can effectively improve symptoms such as skin sagging, dull color, oiliness, and freckles. The collagen peptides prepared in Comparative Examples 1-5 have significantly worse improvement effects on skin conditions (skin sagging, dull color, oiliness, and freckles) than those in Examples 1-4.

[0134] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing fish collagen peptide, characterized in that: The following steps are involved: (1) Add fish tissue to water for extraction, filter, and obtain filtrate; (2) Adding a carbon source to the filtrate to obtain a fermentation medium; (3) inoculating the Lactobacillus plantarum strain on the seed culture medium and culturing on a shaking table to obtain the seed liquid; (4) inoculating the seed liquid into the fermentation medium and fermenting and culturing; (5) First add enzyme activator, then add enzymolysis agent for enzymolysis, inactivate enzyme, centrifuge, and dry to obtain fish collagen peptide powder; The fish tissue in step (1) is fish scales, and the fish is grass carp or silver carp; the mass ratio of fish scales to water in step (1) is 1:(5-8); The enzyme activator in step (5) is fungus polysaccharide, oligomannosaccharide and raffinose, and the mass ratio of the fungus polysaccharide, oligomannosaccharide and raffinose is 3: (0.3-2): 1.5; The enzymatic hydrolyzing agents in step (5) are papain and flavor protease, and the mass ratio of papain to flavor protease is (2-4):

1.

2. The preparation method according to claim 1, characterized in that The carbon source in step (2) is selected from one or more of glucose, glycerol, sucrose, oligofructose, crystalline fructose, white sugar, xylose, mannose, soluble starch, dextrin, sorbitol, and lactose.

3. The preparation method according to claim 1, characterized in that The inoculation amount described in step (3) is 5-10%; the shaking culture conditions are: 35-40°C, a rotation speed of 220-260 rpm, and a shaking culture for 12-24 hours.

4. The preparation method according to claim 1, characterized in that The inoculation amount of the seed liquid in step (4) is 1-5%; the fermentation culture condition in step (4) is 35-40° C., and the fermentation time is 150-300 min.

5. The preparation method according to claim 1, characterized in that The enzymatic hydrolysis temperature in step (5) is 35-40°C, and the enzymatic hydrolysis time is 10-20 hours.

6. Fish collagen peptide prepared by the preparation method according to any one of claims 1 to 5.

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