响应面法优化林蛙油抗氧化肽提取工艺的研究.doc
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响应面法优化林蛙油抗氧化肽提取工艺的研究
摘要
为制备高活性的抗氧化肽，本实验以林蛙油为底物，以羟自由基(·OH)的清除率为指标，在料液比固定为1:50的情况下，用中性蛋白酶进行酶解，通过单因素实验初步研究了pH、酶解温度、酶解时间、加酶量（E/S）四个因素对羟自由基清除率的变化的影响，进而通过Box-Behnken实验设计和响应面分析法综合优化了林蛙油抗氧化肽的提取条件。
结果发现在酶解时间为10.06 h、pH为7.47、加酶量为2.03%、酶解温度为51.58 ℃时，所得酶解液对羟自由基(·OH)的清除率最大，达到了41.38%。且四个因素中，温度对酶解液清除羟自由基的能力影响最大，其次依次为pH、时间、加酶量。
关键词：林蛙油，抗氧化肽，提取，响应面，最优条件
Study on extracting antioxidant peptide of Oviductus Ranae by response surface methodology
Abstract
For the preparation of highly active antioxidant peptide, this experiment to Oviductus Ranae as the substrate, hydroxyl radicals (·OH)of the clearance for index, liquid materials in than the fixed for 1:50 with neutral protease for enzyme solution, through the single factor experiment preliminary studies the pH, enzymatic hydrolysis temperature, enzyme solution, and time of enzyme (E/S) four factors on the hydroxyl radicals of the influence of the clearance changes, and then through the Box-Behnken experimental design and the response surface method integrated optimizing the Oviductus Ranae antioxidant peptide extraction conditions.
It is found that the enzyme solution time of 10.06 h, pH 7.47, add enzyme content is 2.03%, enzymatic hydrolysis temperature was 51.58 ℃, the enzyme obtained solution of hydroxyl radicals liquid (·OH) of the largest clearance, to 41.38%. And four factors, the temperature of enzyme secretion clearance solution of free radicals the biggest influence ability, followed by pH, time, and enzyme quantity.
Key words: Oviductus Ranae , antioxidant peptide, extraction, response surface，the optimal conditions
目录
前言 1
1目的意义 1
2研究现状 2
2.1 林蛙油 2
2.2 抗氧化肽 4
3 实验流程 6
2材料与方法 7
2.1实验材料 7
2.2仪器与试剂 7
2.2.1仪器设备 7
2.2.2 实验试剂 7
2.3方法 7
2.3.1蛋白质酶解机理 7
2.3.2响应面法 8
2.3.3邻二氮菲法 8
2.4 实验过程 9
2.4.1林蛙油的预处理 9
2.4.2 实验步骤 9
2.4.3 单因素实验 9
2.4.4 响应曲面优化实验法 9
3 结果及讨论 11
3.1 单因素实验的结果 11
3.1.1 温度对林蛙油抗氧化肽提取的影响 11
3.1.2 pH对林蛙油抗氧化肽提取的影响 11
3.1.3 加酶量对林蛙油抗氧化肽提取的影响 12
3.1.4 时间对林蛙油抗氧化肽提取的影响 12
3.2 响应曲面优化实验结果 13
3.3 讨论 19
4
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