阅读说明：本技术 一种金枪鱼蛋白水解物和芍药苷的协同抗氧化组合物及其应用 (Synergistic antioxidant composition of tuna protein hydrolysate and paeoniflorin and application thereof ) 是由 黄雅钦 杜博玮 邓桂雅 李雪娟 于 2019-08-29 设计创作，主要内容包括：一种金枪鱼蛋白水解物和芍药苷的协同抗氧化组合物及其应用,涉及生物技术领域。含有金枪鱼蛋白水解物和芍药苷,其中金枪鱼蛋白水解物与芍药苷质量比范围为1：7-7：1。金枪鱼蛋白水解物与芍药苷不同比例混合后具有协同作用,有效地提高金枪鱼蛋白水解物的抗氧化活性,可以制备用于预防和治疗与氧化应激有关的疾病,如银屑病的药物。(A synergistic antioxidant composition of tuna protein hydrolysate and paeoniflorin and application thereof relate to the technical field of biology. Contains tuna protein hydrolysate and paeoniflorin, wherein the mass ratio of the tuna protein hydrolysate to the paeoniflorin is 1: 7-7: 1. the tuna protein hydrolysate and paeoniflorin in different proportions have synergistic effect, effectively improve antioxidant activity of the tuna protein hydrolysate, and can be used for preparing medicines for preventing and treating diseases related to oxidative stress, such as psoriasis.)

1. An antioxidant composition containing tuna protein hydrolysate and paeoniflorin, which is characterized by comprising the tuna protein hydrolysate and the paeoniflorin, wherein the mass ratio of the tuna protein hydrolysate to the paeoniflorin is 1: 7-7: 1.

2. the antioxidant composition comprising tuna protein hydrolysate and paeoniflorin according to claim 1, wherein the ratio by mass of tuna protein hydrolysate: paeoniflorin 1-7: 1.

3. the antioxidant composition comprising tuna protein hydrolysate and paeoniflorin according to claim 1, wherein the ratio by mass of tuna protein hydrolysate: paeoniflorin ═ 1: 1. 2: 1. 3: 1. 5: 1. 7: 1.

4. the antioxidant composition as claimed in claim 1, wherein the antioxidant composition is prepared by mixing tuna protein hydrolysate with paeoniflorin according to a ratio, and the tuna protein hydrolysate is hydrolyzed under conditions of enzymolysis by adding trypsin 0.5% (g/g) and chymotrypsin 0.1% (g/g) at the same time, at a pH of 8 and 40 ℃ for 2 hours.

5. Use of an antioxidant composition as claimed in any one of claims 1-4 for the preparation of a medicament for the prevention and treatment of diseases triggered by free radicals and diseases associated with oxidative stress.

Technical Field

The invention relates to the technical field of biology, and particularly relates to a synergistic antioxidant composition of tuna protein hydrolysate and paeoniflorin and application thereof.

Background

Active oxygen is generated by various enzymatic reactions and chemical processes, which are necessary for many physiological functions in addition to functioning as an auxiliary messenger in the human body, but may damage neuronal functions when they are excessively accumulated in the human body. Many neurodegenerative diseases, including alzheimer's disease, parkinson's disease and huntington's disease, are characterized by severe or prolonged oxidative stress. The main result of oxidative stress is irreversible damage to macromolecules by reactive oxygen species. Thus, in theory, antioxidant therapy can delay the spread of neuronal damage and improve neurological prognosis.

Peony is a well-known traditional Chinese medicine and is widely used as a traditional Chinese medicine prescription for treating certain neurodegenerative diseases, traumatic injuries and inflammations. Paeoniflorin is a main active ingredient extracted from Paeonia lactiflora pall, and has neuroprotective, anti-ischemic, antioxidant, antiinflammatory and anticancer effects. The neuroprotective potential of paeoniflorin has been demonstrated in animal models of various neurological diseases. However, paeoniflorin is very expensive, and tests show that the in vitro oxidation resistance of paeoniflorin needs to be improved, so that a compound which is low in cost, good in biocompatibility, safe, high in oxidation resistance and capable of generating a synergistic oxidation resistance effect with paeoniflorin needs to be found.

Tuna is one of the most important commercial fish species. The fishery processing of tuna produces a large amount of by-products such as bones, scales, viscera and skin. It is well known that these by-products contain a large amount of collagen. On the other hand, most of commercially available collagens are derived from pigs and cows. However, tuna collagen has become a good choice due to religious problems. Studies have shown that tuna-derived protein hydrolysate has antioxidant activity, and thus, the tuna protein hydrolysate is likely to be a potential source of neuroprotection, antihypertensive, antitumor and anti-inflammatory peptides. If the tuna protein hydrolysate can generate a synergistic antioxidant effect with paeoniflorin, the treatment and health care effects of the composition can be greatly enhanced.

Disclosure of Invention

In order to solve the technical problems in the prior art, the invention provides a composition with high antioxidation and containing tuna protein hydrolysate and paeoniflorin and application thereof.

The technical scheme of the invention is as follows:

an antioxidant composition containing tuna protein hydrolysate and paeoniflorin, which is characterized by comprising the tuna protein hydrolysate and the paeoniflorin, wherein the mass ratio of the tuna protein hydrolysate to the paeoniflorin is 1: 7-7: 1, preferably 1 to 7: 1.

in a preferred embodiment of the invention, the tuna protein hydrolysate: paeoniflorin ═ 1: 1.

in a preferred embodiment of the invention, the tuna protein hydrolysate: paeoniflorin 2: 1.

in a preferred embodiment of the invention, the tuna protein hydrolysate: paeoniflorin ═ 3: 1.

in a preferred embodiment of the invention, the tuna protein hydrolysate: paeoniflorin-5: 1.

in a preferred embodiment of the invention, the tuna protein hydrolysate: paeoniflorin-7: 1.

mixing tuna protein hydrolysate and paeoniflorin according to a ratio to prepare the antioxidant composition, wherein the tuna protein hydrolysate is subjected to enzymolysis under the conditions that 0.5% (g/g) of trypsin and 0.1% (g/g) of chymotrypsin are added simultaneously, the pH value is 8, and the hydrolysis is carried out for 2 hours at 40 ℃.

Compared with the prior art, the invention has the following advantages:

the composition of the tuna protein hydrolysate and paeoniflorin has strong oxidation resistance and can effectively remove ABTS free radicals. According to the composition product obtained by the preparation method, after the tuna protein hydrolysate and the paeoniflorin are mixed in different proportions, experiments prove that the tuna protein hydrolysate and the paeoniflorin have a synergistic antioxidant effect, so that the antioxidant activity of the tuna protein hydrolysate is improved, and therefore, the composition product can be used for preparing medicines for preventing and treating diseases caused by free radicals and diseases related to oxidative stress.

Drawings

FIG. 1 is a graph of synergy as a function of inhibition rate.

Detailed Description

The invention will be further illustrated and understood by the following non-limiting examples.

In the invention, the antioxidant activity and synergistic effect of the sample are evaluated by measuring the scavenging capacity of the single tuna protein hydrolysate, the single paeoniflorin and the compositions with different proportions on ABTS free radicals. Preparing each sample into solution with gradient concentration, measuring the clearance rate of ABTS free radicals, and calculating IC₅₀The value is obtained. Synergy is characterized by the calculation of CI values.

The determination method of ABTS free radical clearance rate comprises the following steps: mixing ABTS and potassium persulfate according to a volume ratio of 1: 1, and is protected from light for 12-14 hours, thereby exciting ABTS free radicals. ABTS free radicals are diluted by PBS solution until the light absorption value is 0.7 +/-0.02 to prepare ABTS working solution. Preparing sample into solutions with different concentrations, adding 0.09mL of each solution with different concentrations into 0.11mL of working solution, mixing, reacting at 37 deg.C in dark place for 10min, and measuring absorbance A at 734nm_i(ii) a Respectively taking 0.09mL of the solution with each concentration, adding 0.11mLPBS, and measuring the light absorption value A after reaction_j(ii) a The absorbance A is measured by the reaction of 0.11mLABTS working solution and 0.09mL deionized water₀As a reference. The clearance calculation formula is:

EABTS＝[1-(A_i-A_j)/A₀]×100

in the formula: EABTS is the clearance rate of ABTS free radicals,%; a. the_iAdding 0.11mL of ABTS solution into 0.09mL of sample solution to obtain a light absorption value; a. the_jAdding 0.11mL LPBS light absorption value for 0.09mL sample solution; a. the₀Absorbance of 0.09mL deionized water was added for 0.11mL of ABTS solution.

IC₅₀The calculation of (2): IC (integrated circuit)₅₀Is the concentration at which the individual samples achieve 50% clearance of ABTS. Firstly, diluting the sample into a series of concentrations, measuring the clearance rate of the sample to ABTS at each concentration, drawing a relation curve of the clearance rate and the concentration, and obtaining the concentration of the ABTSIC of the sample was plotted₅₀The value is obtained.

Combination Index (CI) is used for data analysis of the degree of synergy of the combination subjects, and is formulated as follows:

(D)₁and (D)₂Is the sample concentration at which the test substance 1 and the test substance 2 act in combination and the effect is X, (Dx)₁And (Dx)₂The sample concentrations were measured when test substance 1 and test substance 2 were applied alone with the effect of X. CI<1. CI is 1 and CI>1 is expressed as synergy, additivity or antagonism, respectively.

The invention will be further illustrated with reference to specific examples. The tuna protein hydrolysate of the following examples of the present invention was prepared by adding trypsin 0.5% (g/g, based on the weight of the tuna protein, the same applies hereinafter) and chymotrypsin 0.1% (g/g) simultaneously, at a pH of 8, and hydrolyzing at 40 ℃ for 2 hours.