榛子粕抗氧化肽制备条件的优化及体外模拟消化的研究

Research in Preparing Condition of Antioxidant Peptide Released from Defatted Hazelnut Meal and the Activity of Gastrointestinal Digestion

为选出最佳的水解蛋白酶,选用碱性蛋白酶、中性蛋白酶、木瓜蛋白酶3种蛋白酶水解榛子粕蛋白,研究单一蛋白酶和复合蛋白酶水解产物的水解度、DPPH自由基清除能力和氨基酸含量,对榛子粕抗氧化肽制备工艺进行优化,分析体外模拟消化前后榛子粕抗氧化肽DPPH自由基清除能力和氨基酸含量的变化。结果表明:木瓜蛋白酶、碱性蛋白酶和中性蛋白酶水解产物水解度分别为2.89%,6.26%,9.50%。1N复合蛋白酶水解产物、1A复合蛋白酶水解产物、2NA复合蛋白酶水解产物和2AN复合蛋白酶水解产物水解度分别为6.31%,7.20%,6.25%,4.99%。木瓜蛋白酶、碱性蛋白酶和中性蛋白酶水解产物清除DPPH自由基的IC50分别为12.0,4.60和2.53mg·mL^(-1)。1N复合蛋白酶水解产物、1A复合蛋白酶水解产物、2NA复合蛋白酶水解产物和2AN复合蛋白酶水解产物清除DPPH自由基的IC50分别为3.05,3.50,3.25和2.81mg·mL^(-1)。木瓜蛋白酶、碱性蛋白酶和中性蛋白酶水解产物氨基酸总量分别为22.08,34.13和41.36g·100g-1。1N复合蛋白酶水解产物、1A复合蛋白酶水解产物、2NA复合蛋白酶水解产物和2AN复合蛋白酶水解产物的氨基酸总量分别为18.18,18.47,26.70和24.49g·100g-1。因此,选用中性蛋白酶为水解榛子粕蛋白的最佳酶源;通过响应面分析,确定制备榛子粕抗氧化肽的最佳工艺条件为:温度43.7℃、时间1.7h、加酶量17000U·g-1,中性蛋白酶水解产物水解度为11.57%、DPPH清除率80.38%、氨基酸含量46.07g·100g-1;体外模拟胃肠消化后,榛子粕抗氧化肽的清除DPPH自由基的IC50升高3.27mg·mL^(-1)、氨基酸总量降低l0.05g·100g-1。

Researching the degree of hydrolysis, DPPH free redical scavenging activity and the composition of ammonia acid from alkaline protease, neutral protease, papain protease hydrolyzing protein of defatted hazelnut meal. Protease screening was employed to prepare antioxidant peptide released from defatted hazelnut meal with enzymolysis method. Changes in DPPH free redical scavenging activity and ammonia acid composition of antioxidant peptide released from defatted hazelnut meal before and after in vitro digestion were investigated. The results indicated that neutral protease was screened out from the following three kinds of protease. The results respectively showed that the degree of hydrolysis from papain protease, alkaline protease, neutral protease hydrolyzing protein of defatted hazelnut meal is 2.89%, 6.26% and 9.50%. The degree of 1 N compound enzymatic hydrolysates,1 A compound enzymatic hydrolysates, 2 NA compound enzymatic hydrolysates, 2 AN compound enzymatic hydrolysates is 6.31%,7.20%, 6.25%, 4.99%. IC50 of scavenging DPPH free redical of hydrolysis from papain protease, alkaline protease, neutral protease hydrolyzing protein of defatted hazelnut meal is 12.0, 4.60 and 2.53 mg·mL^-1. IC50 of scavenging DPPH free redical of 1 N compound enzymatic hydrolysates, 1 A compound enzymatic hydrolysates, 2 NA compound enzymatic hydrolysates, 2 AN compound enzymatic hydrolysates is 3.05, 3.50, 3.25 and 2.81 mg·mL^-1. The composition of total Amion acid from papain protease, alkaline protease, neutral protease hydrolyzing protein of defatted hazelnut meal is 22.08, 34.13 and 41.36 g·100 g^-1. The composition of total Amion acid of 1 N compound enzymatic hydrolysates, 1 A compound enzymatic hydrolysates, 2 NA compound enzymatichydrolysates. 2 AN compound enzymatic hydrolysates was 18.18, 18.47, 26.70 and 24.49 g·100 g^-1. The optimum hydrolysis conditions obtained from the experiments were as the following： hydrolysis temperature of 43.7 ℃,enzyme dosage 17000 U·g^-1,hydrolysis time 1.7 h, the degree of hydrolysis 11.57%, DPPH free redical scavenging activity 80.38%,the content of amino acid in neutral protease hydrolysates 46.07 g·100 g^-1. IC50 of scavenging DPPH free redical apparently decreased by 3.27 mg·mL^-1 and the composition of total Amion acid decreased by 10.05 g·100 g^-1 after in vitro digestion.