响应面法优化葛根蛋白酶解工艺及其体外抗氧化特性分析

庞会娜; 董红影; 肖凤琴; 张红印; 韩荣欣; 王海东; 严铭铭; 邵帅

doi:10.13386/j.issn1002-0306.2021100147

响应面法优化葛根蛋白酶解工艺及其体外抗氧化特性分析

Optimization of Enzymatic Hydrolysis Process of Pueraria Protein by Response Surface Methodology and Its Antioxidant Properties in Vitro

摘要

摘要: 目的：优化葛根蛋白酶解工艺并研究其抗氧化特性。方法：以DPPH自由基清除能力、水解度（DH）为评价指标，结合SDS-PAGE电泳结果，筛选最佳水解蛋白酶；在单因素实验基础上，利用Box-Behnken响应面法优化葛根蛋白酶解工艺，并对最佳葛根蛋白酶解物进行抗氧化特性研究。结果：葛根蛋白酶解最佳工艺条件为：酶解温度55 ℃、pH9、酶底比2%，该条件下制备的葛根蛋白酶解物清除DPPH自由基、ABTS⁺自由基、OH自由基的IC₅₀值分别为0.15、0.38、1.41 mg/mL，还原能力为0.553。结论：该条件下制备的葛根蛋白酶解物具有较好的抗氧化特性。

Abstract: Objective: To optimize the enzymatic hydrolysis process of Pueraria protein and study its antioxidant properties. Methods: DPPH radical scavenging ability and DH were used as the index to screen the best hydrolytic protease. Base on the single factor experiments, the Box-Behnken response surface design was used to optimize the enzymatic hydrolysis process of Pueraria protein, and the antioxidant properties of the optimum hydrolysate in vitro was analyzed. Results: The optimal hydrolysis conditions for alkaline protease were as follows: Temperature 55 ℃, pH9 and enzyme substrate ratio 2%. Under these conditions, the IC₅₀ of DPPH·, ABTS⁺·, and ·OH radicals were 0.15, 0.38, 1.41 mg/mL and a reduction capacity of 0.553. Conclusion: Under these conditions, Pueraria protein hydrolysate had remarkable antioxidant properties.

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