A bioactive peptide compound beverage and its preparation method

Abstract

The present invention provides a bioactive peptide composite beverage and a preparation method thereof. For the first time, the present invention isolates antioxidant active short peptides from crocodile meat. The active short peptides prepared by the isolation method have a high purity. Moreover, through activity analysis, it can be known that the active short peptides have good DPPH free radical scavenging rate and superoxide anion scavenging rate, and can be used to prepare a beverage product with nutritional and health functions.

Description

Technical Field

[0001] The present invention relates to the field of compound beverages, and more specifically, to a bioactive peptide compound beverage and a preparation method thereof. Background Art

[0002] Crocodile meat is a kind of meat with low fat and low cholesterol. Healthy and high-quality crocodile meat is not only delicious, but also has the effects of nourishing the heart and lungs, enriching blood and strengthening bones, tonifying the kidney and consolidating essence, and expelling pathogenic factors and removing dampness. Regular consumption can have effects such as tonifying qi and nourishing blood, relieving asthma and coughing, and beautifying and prolonging life.

[0003] Antioxidant peptides in bioactive peptides play an important role in scavenging free radicals, inhibiting lipid peroxidation, and improving the body's resistance to aging and diseases, and have extremely high application value in the fields of food, cosmetics, medicine, etc. However, the current research on crocodile bioactive peptides is not yet systematic, and there is little research at home and abroad, especially on crocodile antioxidant active short peptides. Summary of the Invention

[0004] In view of the technical problems existing in the prior art, the present invention provides a bioactive peptide compound beverage and a preparation method thereof. The present invention first isolates antioxidant active short peptides from crocodile meat. The active short peptides prepared by the isolation method have a high purity, and through activity analysis, it can be known that the active short peptides have good DPPH free radical scavenging rate and superoxide anion scavenging rate, and can be used to prepare a beverage product with nutritional and health functions.

[0005] Specifically, the present invention first provides a bioactive peptide compound beverage, which, by mass, includes: 10-25 parts by mass of crocodile antioxidant active peptide, 0.2-0.6 part by mass of table salt, 0.003-0.007 part by mass of sucralose, 0.005-0.01 part by mass of citric acid, 0.05-0.1 part by mass of sodium benzoate, 0.02-0.05 part by mass of potassium sorbate, and 100-200 parts by mass of sterile water.

[0006] Preferably, the bioactive peptide compound beverage, by mass, includes: 10 parts by mass of crocodile antioxidant active peptide, 0.2 part by mass of table salt, 0.003 part by mass of sucralose, 0.005 part by mass of citric acid, 0.05 part by mass of sodium benzoate, 0.02 part by mass of potassium sorbate, and 100 parts by mass of sterile water.

[0007] Preferably, the bioactive peptide compound beverage, by mass, includes: 20 parts by mass of crocodile antioxidant active peptide, 0.4 part by mass of table salt, 0.005 part by mass of sucralose, 0.007 part by mass of citric acid, 0.08 part by mass of sodium benzoate, 0.04 part by mass of potassium sorbate, and 150 parts by mass of sterile water.

[0008] Preferably, the bioactive peptide compound beverage, by mass, comprises: 25 parts by mass of crocodile antioxidant active peptide, 0.6 part by mass of table salt, 0.007 part by mass of sucralose, 0.01 part by mass of citric acid, 0.1 part by mass of sodium benzoate, 0.05 part by mass of potassium sorbate and 200 parts by mass of sterile water.

[0009] Preferably, the sequence of the crocodile antioxidant active peptide is as shown in SEQ ID NO.1.

[0010] Another aspect of the present invention is to provide a preparation method of the bioactive peptide compound beverage, comprising the following steps:

[0011] 1) Weigh the crocodile antioxidant active peptide, sodium benzoate, and potassium sorbate by parts, dissolve them in sterile water and mix well, heat to 60 - 80 °C, and stir thoroughly to dissolve them;

[0012] 2) Weigh the table salt, sucralose, and citric acid by parts, and stir and mix them evenly for standby;

[0013] 3) Mix the mixture obtained in step 1) with the mixture obtained in step 2), stir evenly and then homogenize to obtain the bioactive peptide compound beverage.

[0014] Preferably, the preparation method of the bioactive peptide compound beverage comprises the following steps:

[0015] 1) Weigh the crocodile antioxidant active peptide, sodium benzoate, and potassium sorbate by parts, dissolve them in sterile water and mix well, heat to 60 °C, and stir thoroughly to dissolve them;

[0016] 2) Weigh the table salt, sucralose, and citric acid by parts, and stir and mix them evenly for standby;

[0017] 3) Mix the mixture obtained in step 1) with the mixture obtained in step 2), stir evenly and then homogenize to obtain the bioactive peptide compound beverage.

[0018] Preferably, the preparation method of the bioactive peptide compound beverage comprises the following steps:

[0019] 1) Weigh the crocodile antioxidant active peptide, sodium benzoate, and potassium sorbate by parts, dissolve them in sterile water and mix well, heat to 75 °C, and stir thoroughly to dissolve them;

[0020] 2) Weigh the table salt, sucralose, and citric acid by parts, and stir and mix them evenly for standby;

[0021] 3) Mix the mixture obtained in step 1) with the mixture obtained in step 2), stir evenly and then homogenize to obtain the bioactive peptide compound beverage.

[0022] Preferably, the preparation method of the bioactive peptide compound beverage comprises the following steps:

[0023] 1) Weigh crocodile antioxidant active peptide, sodium benzoate, and potassium sorbate by parts, dissolve them in sterile water and mix well, heat to 80 °C and stir thoroughly to dissolve them;

[0024] 2) Weigh table salt, sucralose, and citric acid by parts, and stir and mix them evenly for standby;

[0025] 3) Mix the mixture obtained in step 1) with the mixture obtained in step 2), stir evenly and then homogenize to obtain the bioactive peptide compound beverage.

[0026] Furthermore, the present invention also provides the use of crocodile antioxidant active peptide in the preparation of bioactive peptide compound beverage.

[0027] The advantages of the invention are as follows: The present invention first isolates antioxidant active short peptides from crocodile meat. The active short peptides prepared by the isolation method have a high purity. And through activity analysis, it can be known that the active short peptides have good DPPH free radical scavenging rate and superoxide anion scavenging rate, and can be used to prepare a beverage product with nutritional and health functions. Specific Embodiments

[0028] The following combines specific embodiments to further elaborate on the present invention, so that those skilled in the art can understand the present invention more clearly.

[0029] The following embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention. Based on the specific embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work belong to the protection scope of the present invention.

[0030] In the embodiments of the present invention, unless otherwise specified, all raw material components are commercially available products well-known to those skilled in the art; in the embodiments of the present invention, unless specifically specified, the technical means used are all conventional means well-known to those skilled in the art.

[0031] Example 1

[0032] An extraction method of crocodile antioxidant active peptide comprises the following steps:

[0033] (1) Take 1 kg of crocodile meat, remove the remaining fat, and use a pulverizer to obtain minced crocodile meat;

[0034] (2) Add 4 times the mass of water to the obtained minced crocodile meat and adjust its pH to 7.0;

[0035] (3) Add 100 U of complex protease and continuously stir and enzymatically hydrolyze at 65 °C for 6 h, then centrifuge to separate the supernatant to obtain crocodile protein solution; wherein, the complex protease is composed of papain and alkaline protease, and the dosage ratio of papain to alkaline protease is 1:2;

[0036] (4) After enzymatic hydrolysis, perform enzyme inactivation treatment at 100 °C and cool it to room temperature;

[0037] (5) Centrifuge the cooled enzymatic hydrolysate at 1500 g for 5 min, take the supernatant; and filter the obtained supernatant with an ultrafiltration membrane to retain polypeptide molecules with a molecular weight cut-off between 4000 - 6000 Da;

[0038] (7) Sequence the retained polypeptide molecules with an Obitrap Fusion Lumos mass spectrometer. After identification and analysis, a total of 1 main active peptide is obtained, and the sequence is shown in SEQ ID NO.1 in sequence, and it is reserved for use after spray drying or vacuum freeze-drying at low temperature. After identification by high-performance liquid chromatography, the purity of the active peptide prepared by the method of the present invention reaches 95.26%.

[0039] Example 2

[0040] A bioactive peptide compound beverage, by mass, includes: 10 parts by mass of crocodile antioxidant active peptide (Example 1), 0.2 part by mass of table salt, 0.003 part by mass of sucralose, 0.005 part by mass of citric acid, 0.05 part by mass of sodium benzoate, 0.02 part by mass of potassium sorbate, and 100 parts by mass of sterile water.

[0041] Among them, the preparation method of the bioactive peptide compound beverage includes the following steps:

[0042] 1) Weigh the crocodile antioxidant active peptide, sodium benzoate, and potassium sorbate by parts and dissolve them in sterile water and mix well. Heat to 60 °C and stir well to dissolve them;

[0043] 2) Weigh the table salt, sucralose, and citric acid by parts, and stir and mix them evenly for standby;

[0044] 3) Mix the mixture obtained in step 1) with the mixture obtained in step 2), stir well and homogenize to obtain the bioactive peptide compound beverage.

[0045] Example 3

[0046] A bioactive peptide compound beverage, by mass, includes: 20 parts by mass of crocodile antioxidant active peptide (Example 1), 0.4 part by mass of table salt, 0.005 part by mass of sucralose, 0.007 part by mass of citric acid, 0.08 part by mass of sodium benzoate, 0.04 part by mass of potassium sorbate, and 150 parts by mass of sterile water.

[0047] Among them, the preparation method of the bioactive peptide compound beverage comprises the following steps:

[0048] 1) Weigh crocodile antioxidant active peptide, sodium benzoate, and potassium sorbate by parts, dissolve them in sterile water and mix well, heat to 75 °C and stir thoroughly to dissolve them;

[0049] 2) Weigh table salt, sucralose, and citric acid by parts, and stir and mix them evenly for standby;

[0050] 3) Mix the mixture obtained in step 1) with the mixture obtained in step 2), stir evenly and then homogenize to obtain the bioactive peptide compound beverage.

[0051] Example 4

[0052] A bioactive peptide compound beverage, by mass parts, comprises: 25 parts by mass of crocodile antioxidant active peptide (Example 1), 0.6 part by mass of table salt, 0.007 part by mass of sucralose, 0.01 part by mass of citric acid, 0.1 part by mass of sodium benzoate, 0.05 part by mass of potassium sorbate, and 200 parts by mass of sterile water.

[0053] Among them, the preparation method of the bioactive peptide compound beverage comprises the following steps:

[0054] 1) Weigh crocodile antioxidant active peptide, sodium benzoate, and potassium sorbate by parts, dissolve them in sterile water and mix well, heat to 80 °C and stir thoroughly to dissolve them;

[0055] 2) Weigh table salt, sucralose, and citric acid by parts, and stir and mix them evenly for standby;

[0056] 3) Mix the mixture obtained in step 1) with the mixture obtained in step 2), stir evenly and then homogenize to obtain the bioactive peptide compound beverage.

[0057] Comparative Example 1

[0058] A compound beverage, with the same formula and preparation method as in Example 1, except that it does not contain crocodile antioxidant active peptide.

[0059] Verification Example

[0060] 1. DPPH free radical scavenging assay

[0061] Treatment group: Take 1 mL of each of the compound beverages in Examples 2-4 and Comparative Example 1, add 1 mL of 0.5 mol / L DPPH solution, mix well, react in the dark at room temperature for 30 min, centrifuge at 5000 revolutions per minute for 5 minutes, and measure the absorbance A1 at a wavelength of 517 nm. Blank group: 1 mL of deionized water is added to 1 mL of 0.5 mol / L DPPH solution. After mixing well, react in the dark at room temperature for 30 min, centrifuge at 5000 revolutions per minute for 5 minutes, and measure the absorbance A0 at a wavelength of 517 nm. The calculation formula is as shown in Formula I: DPPH free radical scavenging rate (%) = (1 - (A1÷A0)) × 100% (Formula I).

[0062] 2. Determination of superoxide anion radical scavenging

[0063] Treatment group: Take 5 mL of 0.05 mol / L Tris-HCl buffer solution (pH 8.2), preheat it in a water bath at 25 °C for 20 min, add 4 mL of the compound beverages in Examples 2-4 and Comparative Example 1 respectively, then add 1 mL of 3 mmol / L pyrogallol solution, mix well, place in a water bath at 25 °C for 5 min, add 1 mL of 10 mol / L HCl to terminate the reaction, and measure the absorbance B1 at 320 nm; Blank group: Replace the compound beverage with the same volume of distilled water, after the same treatment, measure the absorbance B0 at 320 nm. The calculation formula is as shown in Formula II: Superoxide anion radical scavenging (%) = (1 - (B1÷B0)) × 100% (Formula II).

[0064] Table 1. Activity analysis of compound beverage

[0065] DPPH free radical scavenging rate (%) Superoxide anion scavenging rate (%) Example 2 82.52±1.21 35.64±0.89 Example 3 95.43±1.05 50.85±1.66 Example 4 90.86±2.13 48.25±2.54 Comparative Example 1 5.48±0.56 1.31±0.36

[0066] As shown in Table 1, the compound beverage containing active short peptides of the present invention has excellent antioxidant activity, with a high DPPH free radical scavenging rate and superoxide anion scavenging rate, and is a beverage product with nutritional and health functions.

[0067] It is necessary to point out here that the above embodiments are only for further elaboration and explanation of the technical solution of the present invention, and are not a further limitation of the technical solution of the present invention. The method of the present invention is only a preferred implementation scheme and is not used to limit the protection scope of the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A bioactive peptide compound beverage, characterized in that: In parts by mass, it includes: 10-25 parts by mass of crocodile antioxidant active peptide, 0.2-0.6 parts by mass of salt, 0.003-0.007 parts by mass of sucralose, 0.005-0.01 parts by mass of citric acid, 0.05-0.1 parts by mass of sodium benzoate, 0.02-0.05 parts by mass of potassium sorbate and 100-200 parts by mass of sterile water; wherein the sequence of the crocodile antioxidant active peptide is shown in SEQ ID NO.

1.

2. The compound beverage according to claim 1, characterized in that In parts by mass, the composition comprises: 10 parts by mass of crocodile antioxidant active peptide, 0.2 parts by mass of salt, 0.003 parts by mass of sucralose, 0.005 parts by mass of citric acid, 0.05 parts by mass of sodium benzoate, 0.02 parts by mass of potassium sorbate and 100 parts by mass of sterile water.

3. The compound beverage according to claim 1, characterized in that In parts by mass, the composition comprises: 20 parts by mass of crocodile antioxidant active peptide, 0.4 parts by mass of salt, 0.005 parts by mass of sucralose, 0.007 parts by mass of citric acid, 0.08 parts by mass of sodium benzoate, 0.04 parts by mass of potassium sorbate and 150 parts by mass of sterile water.

4. The compound beverage according to claim 1, characterized in that In terms of mass parts, the composition includes: 25 mass parts of crocodile antioxidant active peptide, 0.6 mass parts of salt, 0.007 mass parts of sucralose, 0.01 mass parts of citric acid, 0.1 mass parts of sodium benzoate, 0.05 mass parts of potassium sorbate and 200 mass parts of sterile water.

5. The method for preparing the bioactive peptide compound beverage according to any one of claims 1 to 4, characterized in that: The steps include: 1) Weigh crocodile antioxidant active peptide, sodium benzoate and potassium sorbate according to the number of parts, dissolve them in sterile water, mix well, heat to 60-80℃ and stir thoroughly to dissolve; 2) Weigh salt, sucralose and citric acid in sterile water, mix well, and stir to mix well for later use; 3) The mixed solution obtained in step 1) is mixed with the mixed solution obtained in step 2), stirred evenly and homogenized to obtain a bioactive peptide compound beverage.

6. The method according to claim 5, characterized in that The method comprises the following steps: 1) Weigh crocodile antioxidant active peptide, sodium benzoate and potassium sorbate according to the number of parts, dissolve them in sterile water, mix well, heat to 60℃ and stir thoroughly to dissolve; 2) Weigh salt, sucralose and citric acid in sterile water, mix well, and stir to mix well for later use; 3) The mixed solution obtained in step 1) is mixed with the mixed solution obtained in step 2), stirred evenly and homogenized to obtain a bioactive peptide compound beverage.

7. The method according to claim 5, characterized in that The method comprises the following steps: 1) Weigh crocodile antioxidant active peptide, sodium benzoate and potassium sorbate according to the number of parts, dissolve them in sterile water, mix well, heat to 75℃ and stir thoroughly to dissolve; 2) Weigh salt, sucralose and citric acid in sterile water, mix well, and stir to mix well for later use; 3) The mixed solution obtained in step 1) is mixed with the mixed solution obtained in step 2), stirred evenly and homogenized to obtain a bioactive peptide compound beverage.

8. The method according to claim 5, characterized in that The method comprises the following steps: 1) Weigh crocodile antioxidant active peptide, sodium benzoate and potassium sorbate according to the number of parts, dissolve them in sterile water, mix well, heat to 80℃ and stir thoroughly to dissolve; 2) Weigh salt, sucralose and citric acid in sterile water, mix well, and stir to mix well for later use; 3) The mixed solution obtained in step 1) is mixed with the mixed solution obtained in step 2), stirred evenly and homogenized to obtain a bioactive peptide compound beverage.

9. Use of crocodile antioxidant active peptide in preparing a bioactive peptide compound beverage, characterized in that: The sequence of the crocodile antioxidant active peptide is shown in SEQ ID NO.1.