酶解公犊奶牛肉制备抗氧化活性肽口服液的工艺优化

为提高公犊奶牛肉的附加值，研究酶解公犊奶牛肉制备抗氧化性活性肽的最佳工艺。采用中性蛋白酶和碱性蛋白酶等5 种酶对公犊奶牛肉进行酶解，以1,1-二苯基-2-三硝基苯肼自由基清除率为评价指标，筛选出酶解产物抗氧化性最高的最适蛋白酶。用最适蛋白酶对公犊奶牛肉进行酶解，通过单因素试验，研究不同酶解时间、酶解温度、酶解pH值和酶添加量对酶解产物抗氧化性的影响，然后再用响应面分析法对酶解条件进行优化。结果表明：中性蛋白酶为最佳的蛋白酶，最优酶解条件为加酶量0.61 g/100 mL、pH值6.80、酶解温度54 ℃、酶解时间2.92 h。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法对酶解产物进行分析，结果表明，公犊奶牛肉中的蛋白质分子被成功水解为分子质量小于7 kDa的小分子多肽和游离氨基酸；通过感官评价方法研制出一款澄清透明、淡黄色、具有牛肉风味的抗氧化活性肽口服液。

Optimization of the Preparation Process for Enzymatic Hydrolysate with Antioxidant Activity from Veal Protein and Formulation of an Active Peptide Oral Liquid

For value-added utilization of veal,the meat from male dairy calves,the preparation of enzymatic hydrolysis with antioxidant activity from veal protein was optimized.Enzymatic hydrolysis was carried out by using five proteases,such as neutral protease and alcalase.To determine the optimal protease for the maximum antioxidant activity,the resulting hydrolysates were screened for 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical scavenging activity.Furthermore,the optimization of hydrolysis conditions including hydrolysis time,pH value,temperature and enzyme dose for improved DPPH radical scavenging activity was investigated by one-factor-at-a-time method and response surface methodology(RSM).The results showed that neutral protease was the best enzyme for the hydrolysis of beef protein.The optimal hydrolysis conditions were as follows:enzyme dose 0.61 g/100 mL,pH 6.80,temperature 54℃and hydrolysis time 2.92 h.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)showed that protein in meat samples was successfully hydrolyzed into small peptides with a molecular mass less than 7 kDa and free amino acids.The hydrolysate prepared under the optimized conditions was used as an ingredient for formulation of an antioxidant peptide oral liquid.According to sensory evaluation,the oral liquid was clear and yellowish in color with a beef flavor.