CN110713516A

CN110713516A - Polypeptide VVC with antioxidant function and application thereof

Info

Publication number: CN110713516A
Application number: CN201911109842.XA
Authority: CN
Inventors: 朱作艺; 何建民; 王伟; 张玉; 王君虹; 李雪; 孙素玲
Original assignee: Zhejiang Academy of Agricultural Sciences
Current assignee: Zhejiang Academy of Agricultural Sciences
Priority date: 2019-11-14
Filing date: 2019-11-14
Publication date: 2020-01-21

Abstract

The invention discloses a polypeptide VVC with an antioxidant function, which has an amino acid sequence as follows: Val-Val-Cys. The invention also discloses the application of the polypeptide VVC with the antioxidant function, which comprises the following steps: can be used as antioxidant, especially antioxidant in food, health product, and cosmetic.

Description

Polypeptide VVC with antioxidant function and application thereof

Technical Field

The invention belongs to the technical field of biology, and particularly relates to polypeptide VVC with an antioxidant function and application thereof as an antioxidant in foods, health-care products and cosmetics.

Background

The oxidation of biological molecules is a free radical mediated process, and the oxidative deterioration of food and the free radicals generated by human metabolism can cause serious oxidative damage to cells and tissues, thereby causing body aging, cardiovascular diseases, cancer and other diseases. The antioxidant is applied to food in the form of food additive, which not only can prevent or delay food oxidation, improve food quality and prolong food shelf life, but also can play an active function role in related food products, and then can play an antioxidant effect in human bodies and remove harm of free radicals and the like to human bodies. The antioxidants commonly used in the food industry at present include t-butyl p-hydroxyanisole (BHA), 2, 6-di-t-butyl p-cresol (BHT), t-butyl p-diphenol (TBHQ), etc., and although these antioxidants can effectively inhibit oxidation, they have certain potential risks to human health, and the use limit is set in all countries at present. The search for safe, healthy antioxidants is therefore of increasing interest to researchers.

The polypeptide is a compound with a molecular structure between amino acid and protein, wherein the polypeptide with biological activity is called biological active peptide, and the biological active peptide has various human metabolism and physiological regulation functions, is easier to digest and absorb than raw material protein, and has more excellent physicochemical property, nutritional property and biological activity. The antioxidant peptide refers to polypeptide with antioxidant activity, has the characteristics of eliminating free radicals and inhibiting or slowing down oxidation reaction, can be used for improving the quality of food and prolonging the quality guarantee period of the food, and can relieve the damage of the free radicals to organisms in organisms, so that the antioxidant peptide has good application prospects in the fields of food, health-care products, medicines, cosmetics and the like. Many researchers have prepared antioxidant peptides from animal or plant tissues. In the published scientific and technical literature, the antioxidant peptides are reported to be separated and identified from different animal and plant materials, and the antioxidant peptides obtained by identification have low content in the animal and plant materials, low recovery rate and complicated preparation technology and process such as separation and purification. Therefore, the polypeptide with antioxidant activity, which is separated and identified from animal and plant raw materials, is prepared by a chemical synthesis method according to the identified sequence structure, so that the problems of low content, low recovery rate and the like can be solved, and the polypeptide has important significance for promoting the application of antioxidant peptides in the fields of foods, health care products, cosmetics and the like.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a novel peptide sequence with stronger antioxidant activity and application thereof.

In order to solve the technical problems, the invention provides a polypeptide VVC with an antioxidant function, wherein the amino acid sequence of the polypeptide VVC is as follows: Val-Val-Cys.

The polypeptide VVC is obtained by identifying royal jelly proteolysis products. The polypeptide VVC of the present invention can be prepared by chemical synthesis by a polypeptide synthesis company.

The invention also provides the application of the polypeptide VVC with the antioxidant function, which is characterized in that: as an antioxidant. Especially as antioxidant in food, health product and cosmetics.

The antioxidant contains the antioxidant polypeptide VVC as main ingredient.

The antioxidant peptide provided by the invention has strong antioxidation, is safe, has no toxic or side effect, has small molecular weight, is easy to absorb, can be used as an antioxidant to be applied to food additives, can also be used as a functional component to be applied to functional foods, health-care products and cosmetics.

The antioxidant peptide VVC is used as an index of activity strength aiming at the capability of eliminating ABTS free radicals, hydroxyl free radicals, DPPH free radicals and the reduction capability of iron ions, thereby showing the characteristic of having an antioxidant function.

The antioxidant peptide VVC provided by the invention has strong functional activities of resisting oxidation, removing free radicals, reducing capability and the like. The IC50 value of the antioxidant peptide VVC on ABTS free radical clearance is 0.096mg/mL, the IC50 value of hydroxyl free radical clearance is 0.382mg/mL, the IC50 value of DPPH free radical clearance is 0.124mg/mL, and the reducing capability of the antioxidant peptide VVC on iron ions is equivalent to 3.897mmol/L Trolox when the concentration of the antioxidant peptide VVC is 1.0 mg/mL.

The antioxidant peptide can be mixed with other substances to be used as an antioxidant; the antioxidant peptide or the antioxidant containing the antioxidant peptide as the main component can be applied to the fields of food, health care products, cosmetics and the like.

The tripeptide structure (VVC) has antioxidant activity which is not reported, and belongs to a novel functional peptide with antioxidant activity.

The antioxidant polypeptide can be prepared by enzymatic hydrolysis of royal jelly protein and subsequent separation and purification processes, or prepared by chemical synthesis. The polypeptide has small molecular weight, easy absorption, strong free radical scavenging ability and iron ion reducing ability, and can be used as functional active ingredient or antioxidant in the fields of food, health product and cosmetic.

Drawings

The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.

FIG. 1 is the effect of antioxidant peptides on scavenging ABTS free radicals;

FIG. 2 is the effect of antioxidant peptides on scavenging hydroxyl radicals;

FIG. 3 is a graph showing the effect of antioxidant peptides on scavenging DPPH free radicals;

FIG. 4 is the iron ion reducing ability of antioxidant peptides.

Detailed Description

The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:

example 1: activity measurement of antioxidant peptide

The method for detecting the antioxidant activity comprises the following steps:

1) ABTS free radical scavenging ability: in the experiment, the ABTS free radical scavenging capacity is measured by using a total antioxidant capacity measuring kit (ABTS method, product number A015-2) produced by Nanjing institute of bioengineering, namely, 170 mu L of ABTS working solution and 20 mu L of application solution are added on a 96-well plate, 10 mu L of sample solution (polypeptide VVC solution with the concentration of 0.02-1.0 mg/mL) is added, the mixture is mixed uniformly and reacted for 6min at room temperature, and the absorbance value A at 414nm is measured₁Using distilled water to replace sample as blank control to determine absorbance value A₀. The scavenging ability was calculated as follows and the results are shown in FIG. 1.

Clearance rate ═ a₀-A₁)/A₀×100％。

2) Determination of free hydroxyl scavenging capacity: the experiment uses Nanjing to build a hydroxyl radical test box (product) produced by bioengineering research instituteSequence number A018), the specific operating procedure is as follows: sequentially adding 0.2mL of substrate application solution, 0.2mL of sample solution (polypeptide VVC solution with concentration of 0.02-1.0 mg/mL) and 0.4mL of reagent three application solution, mixing, reacting at 37 ℃ for 1min, immediately adding color developing agent to terminate the reaction from the completion of the reagent three application solution addition to the completion of 1min, standing at room temperature for 20min, and measuring absorbance value A at 550nm₁Using distilled water to replace sample as blank control to determine absorbance value A₀. The scavenging ability was calculated as follows, and the results are shown in FIG. 2.

Clearance rate ═ a₀-A₁)/A₀×100％。

3) Determination of DPPH radical scavenging Capacity: mixing 0.5mL of sample solution (polypeptide VVC solution with concentration of 0.02-1.0 mg/mL) with 2.5mL of DPPH reagent (0.2mmol/L ethanol solution), standing in the dark for 20min, and measuring absorbance value A at 517nm₁Measuring absorbance A by using distilled water as blank control instead of sample₀. The scavenging ability was calculated as follows, and the results are shown in FIG. 3.

Clearance rate ═ a₀-A₁)/A₀×100％

4) And (3) determining the reduction capability of iron ions: the total antioxidant capacity detection kit (product number A015-3-1) produced by Nanjing institute of bioengineering is used for determination in the experiment, and the specific operation steps are as follows: adding 5 mu L of sample solution (polypeptide VVC solution with concentration of 0.02-1.0 mg/mL) on a 96-well plate, adding 180 mu L of FRAP working solution, mixing uniformly, reacting at 37 ℃ for 3-5min, and measuring absorbance value A at 593nm₁Meanwhile, Trolox solutions with different concentrations (concentration of 0.1, 0.2, 0.4, 0.8 and 1.0mmoL/L) are used as standard control solutions, the reduction capacity is defined by corresponding to mmoL/L Trolox solution, and the reduction capacity is expressed by mmoL/L Trolox. The results are shown in FIG. 4.

The antioxidant peptide VVC (serving as a sample) provided by the invention has strong functional activities of resisting oxidation, removing free radicals, reducing capacity and the like. IC of ABTS free radical clearance rate of antioxidant peptide VVC₅₀IC with value of 0.096mg/mL, hydroxyl radical scavenging ratio₅₀IC with value of 0.382mg/mL, DPPH radical clearance rate₅₀The value is 0.124mg/mL, the concentration of the antioxidant peptide VVC is 1.0At mg/mL, the reduction capacity against iron ions is equivalent to 3.897mmol/L Trolox.

Comparative example 1, the polypeptides as shown in table 1 were selected and tested according to the method described in example 1 above, and the results are shown in table 1 below in comparison with the invention.

TABLE 1

Example 2: application of antioxidant peptide

1) And manufacturing a tablet: mixing 30% of antioxidant peptide (VVC), 36% of lactose, 30% of crystalline cellulose and 4% of water, and tabletting with a tabletting machine to obtain tablet with antioxidant effect.

2) And functional beverage preparation: fully mixing the components of the beverage according to the following proportion: 20% of fruit juice, 0.3% of citric acid, 10% of white granulated sugar, 0.2% of spice, 1% of antioxidant peptide (VVC) and 68.5% of water, and the fruit juice beverage with the antioxidant function is prepared.

3) And manufacturing the toning lotion: fully mixing the components of the toning lotion according to the following proportion: 94.98% of water, 2.0% of glycerol, 1% of alcohol, 1.9% of butanediol, 0.02% of preservative and 0.1% of antioxidant peptide (VVC), and the cosmetic lotion with antioxidant function is prepared.

The above percentages are mass%.

Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.

Sequence listing

<110> Zhejiang province academy of agricultural sciences

<120> polypeptide VVC with antioxidant function and application thereof

<160>1

<170>SIPOSequenceListing 1.0

<210>1

<211>3

<212>PRT

<213> Artificial Sequence (Artificial Sequence)

<400>1

Val Val Cys

1

Claims

1. Polypeptide VVC with anti-oxidation function is characterized in that the amino acid sequence is as follows: Val-Val-Cys.

2. Use of the polypeptide VVC having antioxidant function according to claim 1, characterized in that: as an antioxidant.

3. Use of the polypeptide VVC with antioxidant function according to claim 2, characterized in that: can be used as antioxidant in food, health product, and cosmetic.