沙棘籽蛋白ADH激活肽的酶解工艺优化及稳定性研究Optimization of Enzymatic Hydrolysis Process of ADH Activating Peptides of Sea Buckthorn Seed Protein and Study on Stability
罕璐,孙宇轩,黄梅桂,毕秀芳,郝刚
摘要(Abstract):
该研究以沙棘籽蛋白为原料,探究酶的种类、底物浓度、pH值、酶底比、酶解时间等因素对水解度、酶解产物乙醇脱氢酶(alcohol dehydrogenase, ADH)激活率、蛋白回收率和TCA可溶性氮得率的影响。结果显示,木瓜蛋白酶酶解产物ADH激活率最高,为21.81%。根据单因素试验结果选取酶解温度、pH值、酶解时间3个关键因素,以ADH激活率为响应值进行响应面分析,最佳酶解工艺为酶解时间2.3 h、pH值6.52、酶解温度56.4℃、底物浓度5%、酶底比4 000 U/g。在此条件下,酶解产物ADH激活率可达29.74%,表明单因素试验和响应面法有效优化了沙棘籽ADH激活肽的制备工艺。稳定性研究结果表明,在酶解温度37~121℃和pH值2~10的条件下,多肽的ADH激活活性并未发生显著变化,并且在不同浓度的K~+、Na~+、Ca~(2+)处理后仍保持较高活性,Fe~(2+)和Fe~(3+)对其活性具有抑制作用且呈浓度依赖性。该研究为沙棘资源的综合利用和产品的开发提供了理论依据。
关键词(KeyWords): 沙棘籽;乙醇脱氢酶;ADH激活肽;酶解;响应面法
基金项目(Foundation):
作者(Author): 罕璐,孙宇轩,黄梅桂,毕秀芳,郝刚
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