Preparation method and application of marine fish oligopeptide powder

Abstract

The invention provides a preparation method and application of marine fish oligopeptide powder, and belongs to the technical field of fish collagen oligopeptide.The preparation method comprises the following steps that S1, frozen cod fish skin is unfrozen and cleaned, then water is added for soaking, the temperature and pH are adjusted, and a fish skin mixed solution is obtained; s2, adding neutral protease and flavor enzyme into the fish skin mixed liquid for enzymolysis; and S3, performing enzyme deactivation and filtration on the fish skin mixed liquid after enzymolysis to obtain filtrate, performing low-temperature concentration on the filtrate, performing sterilization, performing drying, and performing sub-packaging to obtain the marine fish oligopeptide powder. The invention provides a preparation method and application of marine fish oligopeptide powder. The purpose that the prepared marine fish oligopeptide powder is stable in quality is achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of fish collagen oligopeptides, and in particular to a preparation method of marine fish oligopeptide powder and application thereof. Background Art

[0002] Fish collagen oligopeptides are a collagen hydrolyzate derived from deep-sea fish skin, produced through low-temperature, bio-directed enzymatic hydrolysis. Their average molecular weight is less than 1,000 Daltons, and they exhibit high solubility and absorption rates exceeding 90%. By removing non-target components like fat and purines while retaining core nutrients like methionine, they are widely used in health foods, cosmetics, and specialty nutritional supplements.

[0003] Publication No. CN105063153B, a Chinese patent application titled "A Method for Preparing a Food-Grade Low-Salt Marine Fish Oligopeptide Powder," discloses a method for preparing a food-grade low-salt marine fish oligopeptide powder. The powder is prepared using food-grade marine low-fat fish meal as raw material through water washing and pre-desalting, colloid mill grinding, enzymatic hydrolysis, membrane separation, concentration, and spray drying. The raw fish meal is washed and pre-desalted to remove most of the salt from the raw fish meal, reducing the pressure on subsequent nanofiltration membrane desalination. The enzymatic hydrolysis is performed using a food-grade industrial protease preparation, and the hydrolyzate is separated using a 1,000-10,000 Da ultrafiltration membrane to remove pigments and peptides with larger molecular weight. A 150-500 Da nanofiltration membrane is then used to remove free amino acids and residual salt from the hydrolyzate. The resulting food-grade low-salt marine fish oligopeptide powder has a high oligopeptide content and low salt content, making it widely applicable to the food industry and suitable for large-scale industrial production.

[0004] Publication No. CN113755550B, entitled "A Marine Fish Oligopeptide Prepared from Deep-Sea Cod Skin as Raw Material and Preparation Method Thereof," provides a marine fish oligopeptide prepared from deep-sea cod skin as raw material and a preparation method thereof, comprising the following steps: (1) pretreatment: taking deep-sea cod skin, cleaning it, and then steam-exploding it to obtain steam-exploded deep-sea cod skin; (2) fermentation: placing the steam-exploded deep-sea cod skin in a fermentation tank, inoculating it with a bacterial mixture, fermenting it, filtering it, and collecting the fermentation liquid; the bacterial mixture is prepared from Bacillus licheniformis, Lactobacillus casei, brewer's yeast, and kelp polysaccharide liquid; (3) enzymatic hydrolysis: mixing the fermentation liquid with water, then adding a complex enzyme and Ganoderma lucidum spore powder, hydrolyzing it, inactivating the enzyme, filtering it, collecting the hydrolyzed liquid, vacuum concentrating it, and spray drying it to obtain the marine fish oligopeptide. The marine fish oligopeptide prepared by the above technical solution has excellent wrinkle-removing and moisturizing effects.

[0005] However, the enzymatic hydrolysis in the above two technical solutions relies on alkaline protease hydrolysis, but is limited by the high and low sensitivity of enzyme activity to temperature and pH, resulting in poor product quality stability of oligopeptides. Summary of the Invention

[0006] In view of this, the present invention provides a preparation method of marine fish oligopeptide powder and application thereof, so as to achieve the purpose of stable quality of the prepared marine fish oligopeptide powder.

[0007] To achieve the above object, the present invention provides a method for preparing marine fish oligopeptide powder, comprising the following steps: S1, frozen cod fish skin is thawed, soaked in water after cleaning, and adjusted temperature and pH to obtain fish skin mixed solution; S2, adding neutral protease and flavor enzyme to the fish skin mixed solution for enzymolysis; S3, the fish skin mixed solution after enzymolysis is inactivated, filtered to obtain a filtrate, the filtrate is low-temperature concentrated, sterilized and dried, and the marine fish oligopeptide powder is obtained after subpackaging.

[0008] In the technical solution of the present invention, enzymatic hydrolysis adopts the combined use of neutral protease and flavor enzyme, wherein the flavor enzyme contains aminopeptidase and carboxypeptidase, which can hydrolyze amino acids one by one from the end of the peptide chain, and further degrade the medium peptide chain produced by the neutral protease into oligopeptides and a small amount of free amino acids. Through the action of the flavor enzyme, the proportion of small molecular peptides in the product can be increased to more than 80%, thereby making the product easier to be absorbed by the human body and enhancing the functional activity of the product in food and health products.

[0009] Flavor enzyme and neutral protease have a synergistic effect. Neutral protease destroys the fiber structure of fish skin, exposing more enzymatic sites, and flavor enzyme takes over the hydrolysis. The combination of the two can increase the protein utilization rate to more than 90%, reducing raw material waste, replacing trypsin to reduce industrial production costs, and ensuring the stability of product quality.

[0010] Optionally, the amount of the neutral protease added is 0.066%~0.1% of the mass of the cod skin, the enzyme activity concentration of the neutral protease is 1000~1500U / g, the amount of the flavor enzyme added is 0.2%~0.33% of the mass of the cod skin, the enzyme activity concentration of the flavor enzyme is 300~500U / g, and the enzymatic hydrolysis time is 2~4h.

[0011] Optionally, in step S1, the mass ratio of fish skin to water when soaking in water is 1:9-11.

[0012] Optionally, the temperature and pH are adjusted to 40-50° C. and 6-7.

[0013] Optionally, the enzyme inactivation temperature is 115-120°C.

[0014] Optionally, the filtration is performed using a 0.1 UM microfiltration membrane.

[0015] Optionally, the low-temperature concentration is to concentrate the filtrate to a concentration of 35-40%.

[0016] Optionally, the sterilization temperature is 130-135° C., and the sterilization time is 2-4 seconds.

[0017] Optionally, the drying is spray drying, and the inlet temperature of the spray drying is 155-170°C, and the outlet temperature is 80-120°C.

[0018] In order to achieve the above-mentioned purpose, the present invention also provides a marine fish oligopeptide powder for use in health care products.

[0019] The above technical solution of the present invention includes at least the following beneficial effects: The flavor enzyme in the technical solution provided by the present invention can improve the flavor and taste of the product and remove undesirable substances. The cod skin enzymatic hydrolysate may produce bitterness due to residual hydrophobic peptide chains. The flavor enzyme can specifically hydrolyze these bitter peptides and simultaneously release flavor amino acids such as glutamic acid and aspartic acid, making the product taste softer.

[0020] Neutral proteases preferentially hydrolyze peptide bonds formed by hydrophobic amino acids in proteins, rapidly breaking down macromolecules like collagen and elastin in cod skin into medium-length peptide chains with molecular weights of approximately 1,000-5,000 Da. This paves the way for further hydrolysis by flavor enzymes. Neutral proteases have a relatively mild hydrolytic ability and are less likely to over-degrade peptide chains into free amino acids, thereby retaining more bioactive oligopeptides and enhancing the product's functional properties. Furthermore, in a neutral environment, impurities like fat and pigments in the fish skin are less susceptible to chemical changes, facilitating subsequent separation and purification, resulting in a high-purity oligopeptide powder with a light color and minimal impurities.

[0021] Neutral protease has good temperature tolerance and stable activity within the range of 40-60°C. It can flexibly adapt to the temperature control requirements in industrial production while reducing energy consumption. It is responsible for quickly destroying the components of fish skin tissue, releasing proteins such as collagen and forming medium peptide chains. Flavor enzyme deep hydrolysis is responsible for accurately cutting medium peptide chains into oligopeptides while improving the flavor. The two are used together, and the process is stable and the product quality is stable.

[0022] The test results of the marine fish oligopeptide powder prepared according to the present invention show that the marine fish oligopeptide powder has obvious ROS scavenging effect on the zebrafish aging model induced by hydrogen peroxide; has the effect of inhibiting β-galactosidase activity in the zebrafish aging model induced by hydrogen peroxide; and has the effect of promoting the enhancement of telomerase activity in the zebrafish aging model induced by hydrogen peroxide; that is, the marine fish oligopeptide powder prepared according to the present invention has anti-aging effects and can be used in health care products, especially in health care products with anti-aging effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1The chromatograms are of ten consecutive batches of marine fish oligopeptide powder prepared in Example 1 of the present invention; Figure 2 is the ROS fluorescence value of zebrafish after sample treatment in Example 1 of the present invention; Figure 3 is the zebrafish β-galactosidase staining intensity after sample treatment in Example 1 of the present invention; Figure 4 A typical graph of zebrafish β-galactosidase staining intensity after sample treatment; Figure 5 It is the zebrafish β-galactosidase staining intensity after sample treatment in Example 1 of the present invention. DETAILED DESCRIPTION

[0024] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the following will be combined with the appended drawings of the embodiments of the present invention. Figures 1 to 5 , clearly and completely describing the technical solutions of the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the described embodiments of the present invention, all other embodiments derived by ordinary technicians in this field fall within the scope of protection of the present invention.

[0025] Example 1 The present invention provides a method for preparing marine fish oligopeptide powder, comprising the following steps: 3 tons of frozen cod skin were thawed, washed with deionized water for 3 times, and deionized water was added in an amount of 10 times that of the fish skin. The temperature was adjusted to 45° C. and the pH was adjusted to 7.0 to obtain a fish skin mixed solution.

[0026] Neutral protease and flavor enzyme were added to the fish skin mixture, wherein the amount of neutral protease added was 0.066% of the mass of the cod skin, i.e., 2 kg of neutral protease was added; the amount of flavor enzyme added was 0.25% of the mass of the cod skin, i.e., 7.5 kg of flavor enzyme was added; the enzyme activity concentration of the neutral protease was 1200 U / g, the enzyme activity concentration of the flavor enzyme was 400 U / g, and the enzymatic hydrolysis time was 3 h.

[0027] The enzymatically hydrolyzed fish skin mixture is inactivated at 115°C, and then filtered through a .1UM microfiltration membrane to obtain a filtrate. The filtrate is concentrated at low temperature to a concentration of 35%. The concentrated filtrate is sterilized and then dried. The sterilization temperature is 130°C and the sterilization time is 3s. The drying is spray drying. The inlet temperature of the spray drying is 160°C and the outlet temperature is 100°C. After packaging, the marine fish oligopeptide powder is obtained.

[0028] Example 2 The present invention provides a method for preparing marine fish oligopeptide powder, comprising the following steps: 3 tons of frozen cod skin were thawed, washed with deionized water for 3 times, and deionized water was added in an amount of 9 times that of the fish skin. The temperature was adjusted to 50° C. and the pH was adjusted to 6.5 to obtain a fish skin mixed solution.

[0029] Neutral protease and flavor enzyme were added to the fish skin mixture, wherein the amount of neutral protease added was 0.1% of the mass of the cod skin, i.e., 3 kg of neutral protease was added; the amount of flavor enzyme added was 0.33% of the mass of the cod skin, i.e., 9.9 kg of flavor enzyme was added; the enzyme activity concentration of the neutral protease was 1000 U / g, the enzyme activity concentration of the flavor enzyme was 300 U / g, and the enzymatic hydrolysis time was 4 h.

[0030] The enzymatically hydrolyzed fish skin mixture is inactivated at 120°C to inactivate the enzyme, and then filtered using a 0.1UM microfiltration membrane to obtain a filtrate, which is then concentrated at low temperature to a concentration of 38%. The concentrated filtrate is sterilized and then dried at a temperature of 135°C for 2 seconds. The drying is performed by spray drying at an inlet temperature of 170°C and an outlet temperature of 80°C. The marine fish oligopeptide powder is obtained after packaging.

[0031] Example 3 The present invention provides a method for preparing marine fish oligopeptide powder, comprising the following steps: 3 tons of frozen cod skin were thawed, washed with deionized water for 3 times, and deionized water was added in an amount of 11 times that of the fish skin. The temperature was adjusted to 40° C. and the pH was adjusted to 6 to obtain a fish skin mixed solution.

[0032] 1500U of neutral protease and 300U of flavor enzyme were added to the fish skin mixture, and the enzymatic hydrolysis time was 2h.

[0033] Neutral protease and flavor enzyme were added to the fish skin mixture, wherein the amount of neutral protease added was 0.08% of the mass of the cod skin, i.e., 2.4 kg of neutral protease was added; the amount of flavor enzyme added was 0.2% of the mass of the cod skin, i.e., 6 kg of flavor enzyme was added; the enzyme activity concentration of the neutral protease was 1500 U / g, the enzyme activity concentration of the flavor enzyme was 500 U / g, and the enzymolysis time was 4 h.

[0034] The enzymatically hydrolyzed fish skin mixture is inactivated at 118°C to inactivate the enzyme, and then filtered using a 0.1UM microfiltration membrane to obtain a filtrate. The filtrate is concentrated at low temperature to a concentration of 40%. The concentrated filtrate is sterilized and then dried. The sterilization temperature is 133°C and the sterilization time is 4s. The drying is spray drying. The inlet temperature of the spray drying is 155°C and the outlet temperature is 120°C. After packaging, the marine fish oligopeptide powder is obtained.

[0035] The preparation method of Example 1 was used to repeatedly produce 10 batches, and the ten batches of marine fish oligopeptides were detected using LC-Q-TOF-MS liquid chromatography-quadrupole-time of flight mass spectrometry system to obtain chromatograms, namely, Figure 1 .

[0036] Depend on Figure 1 As can be seen, the retention time, peak area, and signal intensity of the corresponding peaks in the chromatograms of the ten batches are consistent, indicating that the production process of the marine fish oligopeptide is stable, that is, the preparation method of the present invention ensures stable product quality. In addition, a comparison of the overlap of the liquid phase chromatograms under different temperature conditions reveals that it has good thermal stability.

[0037] The marine fish oligopeptide obtained in Example 1 was tested for its anti-aging properties.

[0038] 1. Evaluation of ROS scavenging efficacy: Sample preparation information: Marine fish oligopeptide powder is prepared into a 20.0 mg / mL stock solution with standard dilution water and prepared immediately before use.

[0039] Positive control: catalase, prepared into a 200 mg / mL stock solution with ultrapure water and stored at -20°C in the dark.

[0040] Experimental Animals: Zebrafish were maintained in aquaculture water at 28°C (water quality: 200 mg of instant sea salt per liter of reverse osmosis water, conductivity 450–550 µS / cm, pH 6.5–8.5, and hardness 50–100 mg / L CaCO₃). The laboratory animal use license number is SYXK (Zhejiang) 2012-0171. Animal husbandry and management were in compliance with AAALAC accreditation (certification number: 001458).

[0041] Wild-type AB zebrafish were bred in natural pairs. Zebrafish at 6 hours post-fertilization (hpf) were used for maximum detectable concentration (MTC) determination and ROS scavenging efficacy evaluation.

[0042] Detection method: 1. MTC determination: Wild-type AB strain zebrafish at 6 hpf were randomly selected and plated in a 6-well plate. Thirty zebrafish were treated in each well (experimental group). Marine fish oligopeptide powder (concentrations shown in Table 1-1) was administered in water. A normal control group and a model control group were also established, with a volume of 3 mL per well. Except for the normal control group, all other experimental groups were administered hydrogen peroxide in water to establish a zebrafish aging model. During sample treatment, the number of zebrafish deaths in each experimental group was counted daily and promptly removed. After 5 days of treatment at 28°C, the mean temperature (MTC) of the samples in the aging model zebrafish was determined.

[0043] 2. Evaluation of ROS Scavenging Efficacy Wild-type AB zebrafish (6 hpf) were randomly selected and plated in a 6-well plate. Thirty zebrafish were treated in each well (experimental group). Marine fish oligopeptide powder (concentrations shown in Table 1) and a positive control, catalase, at a concentration of 2000 μg / mL were administered in water. A normal control group and a model control group were also established. The volume per well was 3 mL. Except for the normal control group, all other experimental groups were treated with hydrogen peroxide in water to establish a zebrafish aging model. After co-treatment with hydrogen peroxide until 4 dpf, the reactive oxygen species (ROS) detection reagent CM-H2DCFDA was added to each experimental group. The zebrafish were then transferred to a 96-well microplate plate, with one zebrafish per well in a 100 μL volume. The zebrafish were incubated in a 28°C incubator until 5 dpf. At the end of the experiment, ROS fluorescence was measured in each experimental group using a multi-function microplate reader. The results are shown in Table 2. Statistical analysis of this indicator was used to evaluate the ROS-scavenging efficacy of the samples in aging zebrafish. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software, and p < 0.05 indicated that the difference was statistically significant. Figure 1 .

[0044] Test results: 1. MTC Table 1. Results of the concentration exploration experiment for evaluating the anti-aging efficacy of samples (n = 30)

[0045] It can be seen that under the experimental conditions, the MTC of marine fish oligopeptide powder for model zebrafish is 2000.

[0046] 2. Evaluation of ROS Scavenging Efficacy Table 2. Results of the ROS scavenging efficacy evaluation experiment (n = 10)

[0047] Compared with the model control group, **p<0.01, ***p<0.001 According to the ROS scavenging efficacy evaluation results (Table 2 and Figure 1 ) showed that the ROS fluorescence value of zebrafish in the model control group (5041) was compared with that in the normal control group (1107), with a p < 0.001, indicating that the model was successfully established. The ROS fluorescence value of zebrafish in the positive control catalase 2000 μg / mL concentration group was 2892, with a p < 0.01 compared with the model control group, indicating that catalase has a significant ROS scavenging effect on zebrafish.

[0048] The ROS fluorescence values ​​of zebrafish in the 500, 1000 and 2000 μg / mL concentration groups of marine fish oligopeptide powder were 1034, 3895 and 3556, respectively, and the ROS scavenging efficacy was 102%, 29% and 38%, respectively. Compared with the model control group, p<0.001&p>0.05&p>0.05, indicating that marine fish oligopeptide powder has a significant ROS scavenging effect on the hydrogen peroxide-induced zebrafish aging model under the experimental concentration conditions.

[0049] It can be seen that under the experimental conditions, marine fish oligopeptide powder has the effect of scavenging ROS.

[0050] 2. Evaluation of β-galactosidase activity inhibition efficacy The samples, positive controls, and experimental animals were the same as above.

[0051] Detection Methods: Wild-type AB zebrafish (6 hpf) were randomly selected and plated in 6-well plates, with 30 zebrafish treated in each well (experimental group). Marine fish oligopeptide powder (concentrations shown in Table 3) was administered in water, along with a positive control of catalase at a concentration of 2000 μg / mL. A normal control group and a model control group were also established, with a volume of 3 mL per well. All experimental groups, except the normal control group, were treated with hydrogen peroxide in water to establish a zebrafish aging model. After co-treatment with hydrogen peroxide until 5 dpf, the zebrafish were fixed overnight with 4% tissue cell fixative and stained using a β-galactosidase staining kit. Following staining, 10 zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. Images were then analyzed and data collected using NIS-Elements D 3.20 advanced image processing software. The overall β-galactosidase staining intensity of the zebrafish was statistically analyzed, and the inhibitory efficacy of the samples on β-galactosidase activity in the aging model zebrafish was evaluated using statistical analysis of this indicator. Statistical analysis was performed using SPSS 26.0 software. P < 0.05 indicated a statistically significant difference. Figure 3 Typical images of zebrafish β-galactosidase staining intensity after sample treatment are shown in Figure 4 , blue indicates the staining intensity of β-galactosidase, and the darker the color, the stronger the activity.

[0052] Test results: Table 3 Experimental results of zebrafish β-galactosidase staining intensity after sample treatment (n = 10)

[0053] Compared with the model control group, *p<0.05, **p<0.01, ***p<0.001.

[0054] According to the evaluation results of β-galactosidase activity inhibition efficacy (Table 3, Figure 3 ) showed that the zebrafish β-galactosidase staining intensity in the model control group (51,822 pixels) compared with the normal control group (47,204 pixels) was p < 0.01, indicating that the model was successfully established. The positive control group, catalase at a concentration of 2000 μg / mL, had a β-galactosidase staining intensity of 46,256 pixels, with a p < 0.05 compared with the model control group. This inhibitory efficacy against β-galactosidase activity in zebrafish was 121%, demonstrating that catalase has the ability to inhibit β-galactosidase activity.

[0055] The zebrafish β-galactosidase staining intensity in the 500, 1000 and 2000 μg / mL marine fish oligopeptide powder groups were 45809, 51491 and 53973 pixels, respectively, and the β-galactosidase activity inhibition efficacy was 130%, 7% and -47%, respectively. Compared with the model control group, p<0.05&p>0.05&p>0.05, indicating that marine fish oligopeptide powder has the effect of inhibiting β-galactosidase activity in the hydrogen peroxide-induced zebrafish aging model under the experimental concentration conditions.

[0056] 3. Evaluation of the efficacy of increasing telomerase activity The samples and positive controls were the same as above.

[0057] Experimental Animals: Zebrafish were maintained in aquaculture water at 28°C (water quality: 200 mg of instant sea salt per liter of reverse osmosis water, conductivity 450–550 µS / cm, pH 6.5–8.5, and hardness 50–100 mg / L CaCO₃). The laboratory animal use license number is SYXK (Zhejiang) 2012-0171. Animal husbandry and management were in compliance with AAALAC accreditation (certification number: 001458).

[0058] Wild-type AB zebrafish, bred in natural pairs, were used for evaluation of the efficacy of increasing telomerase activity.

[0059] Detection method: 6 hpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). Marine fish oligopeptide powder was administered in water (concentrations are shown in Table 4), and the positive control catalase concentration was 2000 μg / mL. A normal control group and a model control group were also set up, with a capacity of 3 mL per well. Except for the normal control group, hydrogen peroxide was administered in water to establish a zebrafish aging model in all other experimental groups. After the samples were co-treated with hydrogen peroxide until 5 dpf, the supernatant of the zebrafish homogenate was obtained, and the reaction was performed using a telomerase ELISA kit. The telomerase activity of each experimental group was determined using a multifunctional microplate reader. The statistical analysis results of this indicator were used to evaluate the efficacy of the sample in increasing the telomerase activity of the aging model zebrafish. The statistical processing results were expressed as mean ± SE. SPSS26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant. The results are shown in Table 4. Figure 5 .

[0060] Test results: Table 4 Results of zebrafish telomerase activity experiments after sample treatment (n = 3)

[0061] Compared with the model control group, *p<0.05, **p<0.01, ***p<0.001 According to the evaluation results of the efficacy of increasing telomerase activity (Table 4, Figure 5 ) showed that the telomerase activity in the model control group (2.54 IU / gprot) was compared with that in the normal control group (2.82 IU / gprot), with a p < 0.05, indicating successful model establishment. The telomerase activity in the positive control group, catalase at a concentration of 2000 μg / mL, was 3.26 IU / gprot, with a p < 0.001 compared with the model control group. This demonstrated a 28% enhancement of telomerase activity in zebrafish, demonstrating that catalase significantly enhances telomerase activity in the hydrogen peroxide-induced aging model.

[0062] The telomerase activities of zebrafish in the 500, 1000 and 2000 μg / mL concentration groups of marine fish oligopeptide powder were 3.69, 3.64 and 2.95 IU / gprot, respectively, and the telomerase activity enhancement efficacy was 45%, 43% and 16%, respectively. Compared with the model control group, p<0.01&p<0.01&p>0.05, indicating that marine fish oligopeptide powder has the effect of promoting the enhancement of telomerase activity in the hydrogen peroxide-induced zebrafish aging model under the experimental concentration conditions.

[0063] That is, under the conditions of this experiment, marine fish oligopeptide powder has the effect of promoting the increase of telomerase activity.

[0064] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing marine fish oligopeptide powder, characterized in that: The following steps are involved: S1, the frozen cod fish skin is thawed, soaked in water after cleaning, and the temperature and pH are adjusted to obtain a cod fish skin mixed solution; S2, adding neutral protease and flavor enzyme to the fish skin mixed solution for enzymolysis; S3, the fish skin mixed solution after enzymolysis is inactivated, filtered to obtain a filtrate, the filtrate is low-temperature concentrated, sterilized and dried, and the marine fish oligopeptide powder is obtained after subpackaging.

2. The preparation method of marine fish oligopeptide powder according to claim 1, wherein The amount of the neutral protease added is 0.066% to 0.1% of the mass of the cod skin, the enzyme activity concentration of the neutral protease is 1000 to 1500 U / g, the amount of the flavor enzyme added is 0.2% to 0.33% of the mass of the cod skin, the enzyme activity concentration of the flavor enzyme is 300 to 500 U / g, and the enzymatic hydrolysis time is 2 to 4 hours.

3. The preparation method of marine fish oligopeptide powder according to claim 1, wherein In the step S1, the mass ratio of fish skin to water during soaking is 1:9-11.

4. The preparation method of marine fish oligopeptide powder according to claim 1, wherein The temperature and pH are adjusted to 40-50° C. and 6-7.

5. The preparation method of marine fish oligopeptide powder according to claim 1, wherein The temperature for inactivating the enzyme is 115-120°C.

6. The preparation method of marine fish oligopeptide powder according to claim 1, wherein The filtration is performed using a 0.1 UM microfiltration membrane.

7. The preparation method of marine fish oligopeptide powder according to claim 1, wherein The low-temperature concentration is to concentrate the filtrate to a concentration of 35-40%.

8. The preparation method of marine fish oligopeptide powder according to claim 1, wherein The sterilization temperature is 130-135° C., and the sterilization time is 2-4 seconds.

9. The preparation method of marine fish oligopeptide powder according to claim 1, wherein The drying is spray drying, the inlet temperature of the spray drying is 155-170°C, and the outlet temperature is 80-120°C.

10. A marine fish oligopeptide powder is used in health care products, characterized in that: The marine fish oligopeptide powder is obtained by the preparation method of the marine fish oligopeptide powder according to any one of claims 1 to 9.

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