Preparation method of oyster peptide for improving male sexual function as well as product and application thereof

Abstract

The invention provides a preparation method of oyster peptide for improving male sexual function, and the preparation method comprises the following steps: obtaining a raw material which is a product obtained by crushing and degreasing oyster meat or dried oyster meat; adjusting the pH value of the raw materials to 6-8, inoculating 0.5-5% by volume of leuconostoc mesenteroides subsp. Cremoris with the initial concentration of 1-8 * 10 < 9 > cfu / mL and 0.5-8% by weight of Lactobacillus kefiranofaciens subsp. Kefiranofaciens with the initial concentration of 1-8 * 10 < 9 > cfu / g, and fermenting at 30-42 DEG C for 2-6 hours to obtain fermentation liquor; adjusting the pH value of the fermentation liquor to 1-3, adding 0.5-5% of pepsase, carrying out enzymolysis for 2-8 hours at the enzymolysis temperature of 30-40 DEG C, then adjusting the pH value to 6-8, adding 0.1-5% of ficin protease and flavourzyme according to the ratio of the ficin protease to the flavourzyme being 1: (1-6), and carrying out enzymolysis for 2-8 hours at the temperature of 50-60 DEG C; raising the temperature to 85-95 DEG C, and keeping the temperature for 20-40 minutes to obtain enzymatic hydrolysate; and clarifying the enzymatic hydrolysate, decolorizing the clarified solution, and refining the decolorized solution to obtain the oyster peptide. According to the preparation method disclosed by the invention, the obtained oyster peptide has the effect of obviously promoting male sexual health.

Description

Technical Field

[0001] The present application relates to the technical field of oyster peptides, and in particular to a preparation method of oyster peptides for improving male sexual function, peptide-containing products, and applications thereof. Background Art

[0002] Male sexual dysfunction is a common condition, affecting approximately 10% of adult men and severely impacting their quality of life and health. Therefore, developing nutritional supplements with clear active ingredients to improve male sexual function is of great significance. Summary of the Invention

[0003] To at least partially solve the above problems, according to a first aspect of the present application, the present application provides a method for preparing oyster peptide for improving male sexual function, comprising:

[0004] Step S1, obtaining raw materials, wherein the raw materials are oyster meat or dried oyster meat that has been crushed and defatted;

[0005] Step S2: adjust the pH value of the raw material to 6-8, inoculate 0.5-5% by volume, and the initial concentration is 1-8×10 9 cfu / mL of Leuconostoc mesenteroides subsp. cremoris and an initial concentration of 0.5-8 wt% was 1-8×10 9 cfu / g of Lactobacillus kumiss-like subspecies, and fermenting at 30-42°C for 2-6h to obtain a fermentation broth;

[0006] Step S3, adjusting the pH value of the fermentation liquid to 1-3, adding 0.5-5% pepsin, enzymolysis temperature 30-40° C., enzymolysis for 2-8 hours, then adjusting the pH value to 6-8, adding 0.1%-5% ficin and flavor protease, the ratio of ficin to flavor protease is 1:1-6, the temperature is 50-60° C., enzymolysis for 2-8 hours; raising the temperature to 85-95° C., keeping the temperature for 20-40 minutes, and obtaining an enzymolysis solution;

[0007] Step S4, clarifying the enzymatic hydrolyzate to obtain a clarified solution;

[0008] Step S5, decolorizing the clarified liquid to obtain a decolorized liquid;

[0009] Step S6: purify the decolorized liquid to obtain oyster peptide.

[0010] Preferably, in step S2, the pH value is adjusted to 6-7 after defatting, and the initial concentration of 1-3 volume % inoculated is 4-8×10 9 cfu / mL of Leuconostoc mesenteroides subsp. cremoris and an initial concentration of 3-5 wt% was 4-8×10 9cfu / g of Lactobacillus kumiss-like subspecies, and fermented at 30-37°C for 3-5h to obtain a fermentation liquid.

[0011] Preferably, in step S3, the pH value of the fermentation broth is adjusted to 1.5-2.3, 1-3% pepsin is added, the enzymatic hydrolysis temperature is 35-37° C., and the enzymatic hydrolysis is carried out for 2-6 hours. The pH value is then adjusted to between 6.5 and 7, 1-3% ficin and flavor protease are added, the ratio of ficin to flavor protease is 1:2-4, the temperature is 53-57° C., and the enzymatic hydrolysis is carried out for 4-6 hours. The temperature is raised to 88-92° C. and kept warm for 25-35 minutes to obtain an enzymatic hydrolyzate.

[0012] Preferably, in step S1, obtaining raw materials includes:

[0013] Obtain shucked oyster meat or dried oyster meat, sterilize it, and then crush it;

[0014] After crushing, add 2-10 times of water, add 0.1%-1% NaOH, ultrasonically treat at 15kHz to 40kHz for 1-4h, drain, add 3-10 times of water, adjust the pH value to between 6-7, and inoculate 1-8% by volume at a temperature of 25-35℃ with an initial concentration of 1-8×10 6 cfu / mL of Aspergillus oryzae, ferment for 1-5 days to perform defatting. More preferably, after crushing, add 3-6 times of water, add 0.1-0.5% NaOH, ultrasonically treat at 15kHz to 40kHz for 2-4h, drain, add 3-10 times of water, adjust the pH value to between 6.5-6.8, and inoculate 4-8% by volume at a temperature of 28-30°C to an initial concentration of 3-8×10 6 cfu / mL of Aspergillus oryzae, fermented for 2-4 days for defatting.

[0015] Preferably, in step S4, clarifying the enzymatic hydrolysate to obtain a clarified solution comprises:

[0016] 0.1-5% of tamarind gum, linseed gum, and fenugreek gum are added to the enzymatic hydrolyzate in a ratio of 1:1-6:2-6, and the mixture is allowed to stand at room temperature for 1-8 hours, and the supernatant is collected as a clarified solution. More preferably, 1-4% of tamarind gum, linseed gum, and fenugreek gum are added to the enzymatic hydrolyzate in a ratio of 1:2-4:2-5, and the mixture is allowed to stand at room temperature for 2-6 hours, and the supernatant is collected as a clarified solution.

[0017] Preferably, in step S5, decolorizing the clarified liquid to obtain a decolorized liquid comprises:

[0018] 0.5-5% of activated carbon from nut shells is added to the clarified liquid, the temperature is 55-65°C, decolorization is carried out for 30-90 minutes, and plate-and-frame filtration is performed to obtain a decolorized liquid. More preferably, 2-4% of activated carbon from nut shells is added to the clarified liquid, the temperature is 58-62°C, decolorization is carried out for 40-60 minutes, and plate-and-frame filtration is performed to obtain a decolorized liquid.

[0019] Preferably, in step S6, the decolorizing solution is purified to obtain oyster peptide, comprising:

[0020] Adjust the pH of the decolorizing solution to 8.5-12, add the decolorizing solution to the cation exchange resin at a flow rate of 5-20 BV / h, and collect the effluent between 5-50 minutes;

[0021] The effluent is nanofiltered to remove inorganic salts and water-soluble free amino acids, and spray-dried to obtain oyster peptides. More preferably, the pH of the decolorizing solution is adjusted to 9-11, the decolorizing solution is added to the cation exchange resin at a flow rate of 10-15 BV / h, and the effluent is collected between 10-40 minutes.

[0022] According to the second aspect of the present application, the present application provides a peptide-containing product containing the oyster peptide prepared by the above-mentioned preparation method.

[0023] Preferably, the peptide-containing product includes food, health care product or medicine, and the peptide-containing product is in the form of tablets, powder or oral liquid.

[0024] According to the third aspect of the present application, the present application provides the use of the above-mentioned peptide-containing preparation in improving male sexual function.

[0025] Oysters were first recorded in the Shennong Bencao Jing, which listed them as top-grade products. Traditional Chinese Medicine believes that oysters belong to the liver and kidney meridians, and have the effects of calming the liver and suppressing yang, softening and dispersing lumps, and astringing and consolidating. Modern scientific research has found that oysters are rich in amino acids and polysaccharides, as well as a variety of minerals such as zinc and selenium, which can relieve fatigue and promote sperm motility. The preparation method of oyster peptides for improving male sexual function provided in this application has a high peptide content, a low average molecular weight, and high taurine and zinc content, which has a significant effect on promoting male sexual health. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the molecular weight spectrum of the oyster peptide prepared by the method of Example 1;

[0027] Figure 2 This is the molecular weight spectrum of the oyster peptide prepared by the method of Example 2;

[0028] Figure 3 This is the molecular weight spectrum of the oyster peptide prepared by the method of Example 3;

[0029] Figure 4This is the molecular weight spectrum of the oyster peptide prepared by the method of Comparative Example 1;

[0030] Figure 5 This is the molecular weight spectrum of the oyster peptide prepared by the method of Comparative Example 2;

[0031] Figure 6 This is the molecular weight spectrum of the oyster peptide prepared by the method of Comparative Example 3;

[0032] Figure 7 This is the molecular weight spectrum of the oyster peptide prepared by the method of Comparative Example 4.

[0033] in, Figures 1 to 7 In the chart, the horizontal axis is time, the unit is minutes (min); the vertical axis is absorbance, the unit is milliabsorptivity (mAU) DETAILED DESCRIPTION

[0034] The technical solutions of the preferred embodiments of the present application will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present application, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection claimed in this application. In the following embodiments and comparative examples, unless otherwise specified, the operating methods used are all conventional operating methods, the equipment used are all conventional equipment, the materials used are common commercially available products, and the equipment and materials used are all the same.

[0035] Example 1

[0036] Sterilize and crush the raw materials: weigh 100g of dried oyster meat, add 10 times the amount of water, wash three times, heat to 90°C, and keep warm for 30 minutes; use a meat grinder to grind the sterilized oyster meat into 3-5mm particles;

[0037] Degreasing: add 5 times water, add 0.5% NaOH, ultrasonicate at 20kHz for 2h, drain, add 5 times water again, adjust pH to 6.5, inoculate 6% Aspergillus oryzae (5×10 6 cfu / mL), fermented for 4 days;

[0038] Preparation of fermentation broth: After defatting, adjust the pH to 7 and inoculate 1% Leuconostoc mesenteroides subsp. cremoris (5×10 9 cfu / mL), 3% Lactobacillus kumiss subsp. (6×10 9 cfu / g), and fermented at 37°C for 5 h to obtain the fermentation liquid;

[0039] Preparation of enzymatic hydrolysate: Adjust the pH of the fermentation broth to 2, add 2% pepsin, hydrolyze at 37°C for 4 hours, adjust the pH to 6.5, add 3% ficin and flavor protease in a ratio of 1:2, hydrolyze at 55°C for 4 hours; then raise the temperature to 90°C and keep warm for 30 minutes to obtain enzymatic hydrolysate;

[0040] Clarification of the enzymatic hydrolysate: Add 2.1% of a mixture of tamarind gum, linseed gum, and fenugreek gum (ratio 1:2:4) to the enzymatic hydrolysate, let it stand at room temperature for 4 hours, and take the supernatant as the clarified solution;

[0041] Decolorization of the clarified liquid: add 2% activated carbon from fruit shells to the clarified liquid, decolorize at 60°C for 60 minutes, and filter on a plate and frame to obtain a decolorized liquid;

[0042] Purification of decolorant: adjust the pH to 10, add decolorant to the cation exchange resin at a flow rate of 15 BV / h, and collect the effluent between 10 and 40 minutes as the purified solution;

[0043] Spray drying of the refined liquid: The refined liquid is nanofiltered to remove inorganic salts and water-soluble free amino acids, and oyster peptides are obtained after spray drying.

[0044] Example 2

[0045] Sterilize and crush the raw materials: weigh 100g of dried oyster meat, add 10 times the amount of water, wash three times, heat to 90°C, and keep warm for 30 minutes; use a meat grinder to grind the sterilized oyster meat into 3-5mm particles;

[0046] Degreasing: add 5 times water, add 0.5% NaOH, ultrasonicate at 20kHz for 2h, drain, add 5 times water again, adjust pH to 6.5, inoculate 6% Aspergillus oryzae (5×10 6 cfu / mL), fermented for 4 days;

[0047] Preparation of fermentation broth: After defatting, adjust the pH to 6 and inoculate 0.5% Leuconostoc mesenteroides subsp. cremoris (1×10 9 cfu / mL), 0.5% Lactobacillus kumiss subsp. (1×10 9 cfu / g), 30℃, and fermented for 2 h to obtain the fermentation liquid;

[0048] Preparation of enzymatic hydrolysate: Adjust the pH of the fermentation broth to 1.5, add 1% pepsin, hydrolyze at 35°C for 2 h, adjust the pH to 6, add 0.1% ficin and flavor protease in a ratio of 1:1, hydrolyze at 50°C for 2 h, raise the temperature to 90°C, and keep warm for 30 min to obtain enzymatic hydrolysate;

[0049] Clarification of the enzymatic hydrolysate: add 0.1% of a mixture of tamarind gum, linseed gum, and fenugreek gum (ratio 1:1:2) to the enzymatic hydrolysate, let it stand at room temperature for 1 hour, and take the supernatant as the clarified solution;

[0050] Decolorization of the clarified liquid: add 0.5% activated carbon from fruit shells to the clarified liquid, decolorize at 60°C for 30 minutes, and filter on a plate and frame to obtain a decolorized liquid;

[0051] Purification of decolorant: adjust the pH to 8.5, add decolorant to the cation exchange resin at a flow rate of 5 BV / h, and collect the effluent between 5 and 50 minutes as the purified solution;

[0052] Spray drying of the refined liquid: The refined liquid is nanofiltered to remove inorganic salts and water-soluble free amino acids, and oyster peptides are obtained after spray drying.

[0053] Example 3

[0054] Sterilize and crush the raw materials: weigh 100g of dried oyster meat, add 10 times the amount of water, wash three times, heat to 90°C, and keep warm for 30 minutes; use a meat grinder to grind the sterilized oyster meat into 3-5mm particles;

[0055] Degreasing: add 5 times water, add 0.5% NaOH, ultrasonicate at 20kHz for 2h, drain, add 5 times water again, adjust pH to 6.5, inoculate 6% Aspergillus oryzae (5×10 6 cfu / mL), fermented for 4 days;

[0056] Preparation of fermentation broth: After defatting, adjust the pH value to 8 and inoculate 5% Leuconostoc mesenteroides subsp. cremoris (8×10 9 cfu / mL), 8% Lactobacillus kumiss subsp. (8×10 9 cfu / g), 42℃, and fermented for 6 h to obtain the fermentation liquid;

[0057] Preparation of enzymatic hydrolysate: Adjust the pH of the fermentation broth to 3, add 5% pepsin, hydrolyze at 40°C for 8 hours, adjust the pH to 8, add 5% ficin and flavor protease in a ratio of 1:6, hydrolyze at 60°C for 8 hours; raise the temperature to 90°C and keep warm for 30 minutes to obtain enzymatic hydrolysate;

[0058] Clarification of the enzymatic hydrolysate: Add 5% of a mixture of tamarind gum, linseed gum, and fennel gum (ratio 1:6:6) to the enzymatic hydrolysate, let it stand at room temperature for 8 hours, and take the supernatant as the clarified solution;

[0059] Decolorization of the clarified liquid: add 5% activated carbon from fruit shells to the clarified liquid, decolorize at 60°C for 90 minutes, and filter on a plate and frame to obtain a decolorized liquid;

[0060] Purification of decolorant: adjust the pH to 12, add decolorant to the cation exchange resin at a flow rate of 20 BV / h, and collect the effluent between 5 and 50 minutes as the purified solution;

[0061] Spray drying of the refined liquid: The refined liquid is nanofiltered to remove inorganic salts and water-soluble free amino acids, and oyster peptides are obtained after spray drying.

[0062] Comparative Example 1

[0063] Sterilize and crush the raw materials: weigh 100g of dried oyster meat, add 10 times the amount of water, wash three times, heat to 90°C, and keep warm for 30 minutes; use a meat grinder to grind the sterilized oyster meat into 3-5mm particles;

[0064] Degreasing: add 5 times water, add 0.5% NaOH, ultrasonicate at 20kHz for 2h, drain, add 5 times water again, adjust pH to 6.5, inoculate 6% Aspergillus oryzae (5×10 6 cfu / mL), fermented for 4 days;

[0065] Preparation of enzymatic hydrolysate: Adjust the pH to 2, add 2% pepsin, hydrolyze at 37°C for 4 hours, adjust the pH to 6.5, add 3% ficin and flavor protease in a ratio of 1:2, hydrolyze at 55°C for 4 hours; raise the temperature to 90°C and keep warm for 30 minutes to obtain enzymatic hydrolysate;

[0066] Clarification of the enzymatic hydrolysate: Add 2.1% of a mixture of tamarind gum, linseed gum, and fenugreek gum (ratio 1:2:4) to the enzymatic hydrolysate, let it stand at room temperature for 4 hours, and take the supernatant as the clarified solution;

[0067] Decolorization of the clarified liquid: add 2% activated carbon from fruit shells to the clarified liquid, decolorize at 60°C for 60 minutes, and filter on a plate and frame to obtain a decolorized liquid;

[0068] Purification of decolorant: adjust the pH to 10, add decolorant to the cation exchange resin at a flow rate of 15 BV / h, and collect the effluent between 10 and 40 minutes as the purified solution;

[0069] Spray drying of the refined liquid: The refined liquid is nanofiltered to remove inorganic salts and water-soluble free amino acids, and oyster peptides are obtained after spray drying.

[0070] Comparative Example 2

[0071] Sterilize and crush the raw materials: weigh 100g of dried oyster meat, add 10 times the amount of water, wash three times, heat to 90°C, and keep warm for 30 minutes; use a meat grinder to grind the sterilized oyster meat into 3-5mm particles;

[0072] Degreasing: add 5 times water, add 0.5% NaOH, ultrasonicate at 20kHz for 2h, drain, add 5 times water again, adjust pH to 6.5, inoculate 6% Aspergillus oryzae (5×10 6 cfu / mL), fermented for 4 days;

[0073] Preparation of fermentation broth: After defatting, adjust the pH to 7 and inoculate 1% Leuconostoc mesenteroides subsp. cremoris (5×10 9 cfu / mL), 3% Lactobacillus kumiss subsp. (6×10 9 cfu / g), and fermented at 37°C for 5 h to obtain the fermentation liquid;

[0074] Clarification of the fermentation broth: Add 2.1% of a mixture of tamarind gum, linseed gum, and fenugreek gum (ratio 1:2:4) to the fermentation broth, let it stand at room temperature for 4 hours, and take the supernatant as the clarified liquid;

[0075] Decolorization of the clarified liquid: add 2% activated carbon from fruit shells to the clarified liquid, decolorize at 60°C for 60 minutes, and filter on a plate and frame to obtain a decolorized liquid;

[0076] Purification of decolorant: adjust the pH to 10, add decolorant to the cation exchange resin at a flow rate of 15 BV / h, and collect the effluent between 10 and 40 minutes as the purified solution;

[0077] Spray drying of the refined liquid: The refined liquid is nanofiltered to remove inorganic salts and water-soluble free amino acids, and oyster peptides are obtained after spray drying.

[0078] Comparative Example 3

[0079] Sterilize and crush the raw materials: weigh 100g of dried oyster meat, add 10 times the amount of water, wash three times, heat to 90°C, and keep warm for 30 minutes; use a meat grinder to grind the sterilized oyster meat into 3-5mm particles;

[0080] Degreasing: add 5 times water, add 0.5% NaOH, ultrasonicate at 20kHz for 2h, drain, add 5 times water again, adjust pH to 6.5, inoculate 6% Aspergillus oryzae (5×10 6 cfu / mL), fermented for 4 days;

[0081] Preparation of fermentation broth: After defatting, adjust the pH to 7 and inoculate 1% Leuconostoc mesenteroides subsp. cremoris (5×10 9 cfu / mL), 3% Lactobacillus kumiss subsp. (6×10 9 cfu / g), and fermented at 37°C for 5 h to obtain the fermentation liquid;

[0082] Preparation of enzymatic hydrolysate: Adjust the pH of the fermentation broth to 2, add 2% pepsin, hydrolyze at 37°C for 4 hours, adjust the pH to 6.5, add 3% ficin and flavor protease in a ratio of 1:2, hydrolyze at 55°C for 4 hours; then raise the temperature to 90°C and keep warm for 30 minutes to obtain enzymatic hydrolysate;

[0083] Decolorization of the enzymatic hydrolyzate: add 2% activated carbon from fruit shells to the enzymatic hydrolyzate, decolorize at 60°C for 60 minutes, and filter on a plate and frame to obtain a decolorized solution;

[0084] Purification of decolorant: adjust the pH to 10, add decolorant to the cation exchange resin at a flow rate of 15 BV / h, and collect the effluent between 10 and 40 minutes as the purified solution;

[0085] Spray drying of the refined liquid: The refined liquid is nanofiltered to remove inorganic salts and water-soluble free amino acids, and oyster peptides are obtained after spray drying.

[0086] Comparative Example 4

[0087] Sterilize and crush the raw materials: weigh 100g of dried oyster meat, add 10 times the amount of water, wash three times, heat to 90°C, and keep warm for 30 minutes; use a meat grinder to grind the sterilized oyster meat into 3-5mm particles;

[0088] Degreasing: add 5 times water, add 0.5% NaOH, ultrasonicate at 20kHz for 2h, drain, add 5 times water again, adjust pH to 6.5, inoculate 6% Aspergillus oryzae (5×10 6 cfu / mL), fermented for 4 days;

[0089] Preparation of fermentation broth: After defatting, adjust the pH to 7 and inoculate 1% Leuconostoc mesenteroides subsp. cremoris (5×10 9 cfu / mL), 3% Lactobacillus kumiss subsp. (6×10 9 cfu / g), and fermented at 37°C for 5 h to obtain the fermentation liquid;

[0090] Preparation of enzymatic hydrolysate: Adjust the pH of the fermentation broth to 2, add 2% pepsin, hydrolyze at 37°C for 4 hours, adjust the pH to 6.5, add 3% ficin and flavor protease in a ratio of 1:2, hydrolyze at 55°C for 4 hours; then raise the temperature to 90°C and keep warm for 30 minutes to obtain enzymatic hydrolysate;

[0091] Clarification of the enzymatic hydrolysate: Add 2.1% of a mixture of tamarind gum, linseed gum, and fenugreek gum (ratio 1:2:4) to the enzymatic hydrolysate, let it stand at room temperature for 4 hours, and take the supernatant as the clarified solution;

[0092] Decolorization of the clarified liquid: add 2% activated carbon from fruit shells to the clarified liquid, decolorize at 60°C for 60 minutes, and filter on a plate and frame to obtain a decolorized liquid;

[0093] Spray drying of the decolorized liquid: The refined liquid is nanofiltered to remove inorganic salts and water-soluble free amino acids, and oyster peptides are obtained after spray drying.

[0094] Oyster peptide detection: The oyster peptide samples obtained in Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4 were tested for peptide content, average molecular weight, taurine content, and zinc content. The peptide content and average molecular weight were determined according to the method specified in GB 22729, the taurine content was determined according to the second method of GB 5009.169, and the zinc content was determined according to the first method of GB 5009.14. The test results are shown in Table 1.

[0095] Table 1

[0096] Group Peptide content (%) Average molecular weight Taurine content (%) Zinc content (mg / kg) Example 1 64.8 378 2.3 60.3 Example 2 63.5 441 1.9 58.7 Example 3 61.1 316 2.2 64.2 Comparative Example 1 32.7 839 1.2 21.3 Comparative Example 2 21.0 1065 0.7 24.8 Comparative Example 3 45.1 519 1.4 33.7 Comparative Example 4 36.5 739 0.2 12.6

[0097] It can be concluded from Table 1 that the oyster peptide prepared by the method of Example 1 has a peptide content significantly higher than that of Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4, and slightly higher than that of Example 2 and Example 3; the oyster peptide prepared by the method of Example 1 has an average molecular weight significantly lower than that of Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4, slightly lower than that of Example 2, and slightly higher than that of Example 3; the oyster peptide prepared by the method of Example 1 has a taurine content significantly higher than that of Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4, and slightly higher than that of Example 2 and Example 3; the oyster peptide prepared by the method of Example 1 has a zinc content significantly higher than that of Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4, slightly higher than that of Example 2, and slightly lower than that of Example 3.

[0098] Verification of the efficacy of oyster peptides: After 3 days of adaptation, male Wistar rats were divided into a normal group, a model group and 7 experimental groups, with 20 rats in each group.

[0099] Testicles were removed from the rats in the model group and the seven experimental groups. The normal and model groups were fed a normal diet. The experimental groups were additionally gavaged with oyster peptide samples prepared in Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4 at a dose of 100 mg / kg BW, in addition to the normal diet.

[0100] After four consecutive weeks, serum testosterone levels in each rat group were measured using an enzyme-linked immunosorbent assay (ELISA), and nitric oxide levels were measured using a nitrate reductase assay. Animals were purchased from Liaoning Changsheng Biotechnology Co., Ltd., and a nitric oxide (NO) assay kit was purchased from Nanjing Jiancheng Bioengineering Research Institute. A rat testosterone (T) ELISA kit was purchased from Shanghai ELISA Biotechnology Co., Ltd. The experimental results are shown in Table 2.

[0101] Table 2

[0102] Group Testosterone (pg / mL) NO (μmol / L) Normal group 213.32±50.41*** 25.73±8.63** Model Group 45.84±33.95 10.29±6.59 Example 1 196.73±62.40*** 23.64±7.27** Example 2 179.48±68.47*** 21.05±6.44** Example 3 188.91±77.52*** 24.33±8.31** Comparative Example 1 94.59±46.77* 13.55±4.09 Comparative Example 2 60.01±54.83 12.96±5.43 Comparative Example 3 110.34±72.58* 17.05±9.43* Comparative Example 4 76.27±49.24 13.31±5.82

[0103] Compared with the model group, *** represents P < 0.001, ** represents P < 0.01, and * represents P < 0.05.

[0104] As can be seen from Table 2, the oyster peptides prepared in Examples 1, 2, 3, Comparative Examples 1, 2, 3, and 4 all increased testosterone and NO levels in rats with removed testicles. However, the oyster peptides prepared using the methods of Examples 1, 2, and 3 showed the most significant effects, reaching over 80% of the testosterone and NO levels in normal rats. While the oyster peptides prepared in Comparative Examples 2 and 4 showed a trend toward increasing testosterone and NO levels in rats with removed testicles, there were no statistically significant differences.

[0105] Based on the above-mentioned embodiments of the present application, in the absence of explicit negation or conflict, the technical features of one embodiment may be beneficially combined with one or more other embodiments.

[0106] Although some specific embodiments of the present application have been described in detail through examples, those skilled in the art will understand that the above examples are for illustration only and are not intended to limit the scope of the present application. Those skilled in the art will understand that the above embodiments may be modified or some technical features may be replaced with equivalents without departing from the scope and spirit of the present application. The scope of the present application is defined by the appended claims.

Claims

1. A method for preparing oyster peptide for improving male sexual function, characterized in that: include: Step S1, obtaining raw materials, wherein the raw materials are oyster meat or dried oyster meat that has been crushed and defatted; Step S2: adjust the pH value of the raw material to 6-8, inoculate 0.5-5 volume% of the initial concentration to 1-8×10 9 cfu / mL of Leuconostoc mesenteroides subsp. cremoris and an initial concentration of 0.5-8 wt% was 1-8×10 9 cfu / g of Lactobacillus kumiss-like subspecies, and fermenting at 30-42°C for 2-6h to obtain a fermentation broth; Step S3, adjusting the pH value of the fermentation liquid to 1-3, adding 0.5-5% pepsin, enzymolysis temperature 30-40° C., enzymolysis for 2-8 hours, then adjusting the pH value to 6-8, adding 0.1%-5% ficin and flavor protease, the ratio of ficin to flavor protease is 1:1-6, the temperature is 50-60° C., enzymolysis for 2-8 hours; raising the temperature to 85-95° C., keeping the temperature for 20-40 minutes, and obtaining an enzymolysis solution; Step S4, clarifying the enzymatic hydrolyzate to obtain a clarified solution; Step S5, decolorizing the clarified liquid to obtain a decolorized liquid; Step S6: purify the decolorized liquid to obtain oyster peptide.

2. The method according to claim 1, characterized in that In step S2, after degreasing, the pH value is adjusted to 6-7, and the initial concentration of 1-3 volume % is inoculated to 4-8×10 9 cfu / mL of Leuconostoc mesenteroides subsp. cremoris and an initial concentration of 3-5 wt% was 4-8×10 9 cfu / g of Lactobacillus kumiss-like subspecies, and fermented at 30-37°C for 3-5h to obtain a fermentation liquid.

3. The method according to claim 1, characterized in that In step S3, the pH value of the fermentation liquid is adjusted to 1.5-2.3, 1-3% pepsin is added, the enzymatic hydrolysis temperature is 35-37° C., and the enzymatic hydrolysis is carried out for 2-6 hours. The pH value is then adjusted to between 6.5 and 7, 1-3% ficin and flavor protease are added, the ratio of ficin to flavor protease is 1:2-4, the temperature is 53-57° C., and the enzymatic hydrolysis is carried out for 4-6 hours. The temperature is raised to 88-92° C. and kept warm for 25-35 minutes to obtain an enzymatic hydrolyzate.

4. The method according to claim 1, wherein In step S1, obtaining raw materials includes: Obtain shucked oyster meat or dried oyster meat, sterilize it, and then crush it; After crushing, add 2-10 times of water, add 0.1%-1% NaOH, ultrasonically treat at 15kHz to 40kHz for 1-4h, drain, add 3-10 times of water, adjust the pH value to 6-7, and inoculate 1-8% by volume at a temperature of 25-35℃. The initial concentration is 1-8×10 6 cfu / mL of Aspergillus oryzae, fermented for 1-5 days for defatting.

5. The method according to claim 1, wherein In step S4, the enzymatic hydrolyzate is clarified to obtain a clarified solution, comprising: Add 0.1-5% of tamarind gum, linseed gum and fennel gum to the enzymatic hydrolysate in a ratio of 1:1-6:2-6, let it stand at room temperature for 1-8 hours, and take the supernatant as the clarified liquid.

6. The method according to claim 1, characterized in that In step S5, the clarified liquid is decolorized to obtain a decolorized liquid, comprising: Add 0.5-5% of activated carbon from fruit shells to the clarified liquid, decolorize at 55-65°C for 30-90 minutes, and filter on a plate and frame to obtain a decolorized liquid.

7. The method according to claim 1, characterized in that In step S6, the decolorized liquid is purified to obtain oyster peptide, comprising: Adjust the pH of the decolorizing solution to 8.5-12, add the decolorizing solution to the cation exchange resin at a flow rate of 5-20 BV / h, and collect the effluent between 5-50 minutes; The effluent was nanofiltered to remove inorganic salts and water-soluble free amino acids, and then spray-dried to obtain oyster peptides.

8. A peptide-containing product comprising an oyster peptide prepared by the preparation method according to any one of claims 1 to 7.

9. The peptide-containing product according to claim 8, characterized in that The peptide-containing products include foods, health products or medicines, and the peptide-containing products are in the form of tablets, powders or oral liquids.

10. Use of the peptide-containing preparation according to claim 8 in improving male sexual function.

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