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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (11): 44-49.doi: 10.11924/j.issn.1000-6850.2012-4146

所属专题： 生物技术

• 水产 渔业 • 上一篇    下一篇

文蛤蛋白酶解条件的优化及酶解产物体外抗氧化活性的分析

白仁奥 马英 何传波 魏好程 杨青 王益梅 熊何健   

  • 收稿日期:2012-12-25 修回日期:2013-02-22 出版日期:2013-04-15 发布日期:2013-04-15

Optimization of Enzymatic Hydrolysis of Meretrix Meretrix Linnaeus Protein and Analysis of the Antioxidant Activity of the Hydrolysates in vitro

  • Received:2012-12-25 Revised:2013-02-22 Online:2013-04-15 Published:2013-04-15

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摘要： 为了制备抗氧化活性多肽，利用风味蛋白酶和木瓜蛋白酶双酶复合的方式水解文蛤蛋白，在单因素实验结果的基础上，选取液料比、酶解温度和酶解pH为影响因子，应用Box-Benhnken中心组合设计原理进行三因素三水平实验设计，以文蛤蛋白的水解度(DH%)为响应值，运用响应面(RSM)法对双酶复合水解条件进行优化，并分析了酶解产物的体外抗氧化活性。结果表明：双酶复合酶解的最优酶解条件为水解时间4 h，酶添加量3%，双酶复合比（风味蛋白酶:木瓜蛋白酶）2:1，液固比3.25:1，温度51℃，pH 6.9，在此条件下文蛤蛋白水解度为36.11%，酶解产物清除DPPH和?OH自由基的IC50值分别为2.61、3.47 mg/mL，表现出较强的体外抗氧化活性，因此，可以利用优化后的酶解条件水解文蛤蛋白制备抗氧化多肽。

关键词: 普通克吕格插值法, 普通克吕格插值法

Abstract: In order to get antioxidative peptides, two commercial enzymes, flavorase and papain were used to hydrolyse Meretrix Meretrix Linnaeus. protein. Based on the results of single factor experiments, liquid-solid ratio, hydrolysis temperature and pH were chosed as influence factors. Box-Benhnken Design Center combination method of three factors and three levels experimental design was applied to further optimize hydrolysis conditions and the degree of hydrolysis (DH) was assayed by Response Surface Methodology. The results showed that: the optimum conditions were 4 h reaction time, enzyme dosage was 3.0% (the ratio of flavorase and papain proteases was 2:1), liquid-solid ratio of 3.25:1, temperature of 51℃，and pH 6.9. Under the optimized conditions, the degree of hydrolysis was 36.11%. The hydrolysates showed obvious in vitro antioxidant activity. The values of IC50 of DPPH radical and hydroxyl radical were 2.61 mg/mL and 3.47 mg/mL, respectively. Thus, the optimized conditions could be used for antioxidative peptides preparation.