摘要
本实验采取单因素分析法对土豆中酪氨酸粗酶的提取及反应条件进行优化,采用多巴速率氧化法筛选具有酪氨酸酶激活作用的植物精油,并通过酶动力学实验研究其激活机理。结果表明土豆中酪氨酸酶最佳提取条件为:料液比1∶10(g/mL)、磷酸盐缓冲液pH6.8;最佳酪氨酸酶反应条件为:温度35℃、时间13 min、底物L-多巴/粗酶加入量比例5∶5。在最佳提取条件与反应条件下筛选15种植物精油中发现百里香、茶树、依兰精油对酪氨酸酶激活作用显著(P>0.05),IC50值分别为0.488、0.429、0.480 mg/mL,接近阳性对照补骨脂素IC50值0.328 mg/mL,动力学实验发现百里香、茶树、依兰精油激活类型分别为非竞争型、混合型、混合型。
The extraction and eaction of tryosinase from potato,and optimization by single factor experiment were been investigated in the study.The plant essential oil which could increase the catalytic activity of tryosinase was been screened by dopa rate oxidation method.Furthermore,enzyme kinetics model was used to study it’s catalytic mechanism.Results showed that the optimal extraction conditions of tyrosinase in potatoes were as follows:solid-liquid ratio 1∶10(g/mL),phosphate buffer pH value 6.8.The optimum reaction conditions of tryosinase were as follows:reaction temperature 35℃,reaction time 13 min,and the substrate addition L-Dopa/crude enzyme addition ratio at 5∶5.In the screening of 15 plant essential oils under the optimal extraction conditions and reaction conditions,thyme,tea tree and ylang-ylang had significant tryosinase activation(P>0.05),the IC50 values were 0.488,0.429 and 0.480 mg/mL,close to the positive control psoralen IC50 value 0.328 mg/mL.The kinetic experiments found that the activation types of thyme,tea tree and ylang-ylang oil were non-competitive,mixed and mixed.
作者
陈怀庆
廖兴宏
赵慧
杨俊棋
王文君
张岩
CHEN Huai-qing;LIAO Xing-hong;ZHAO Hui;YANG Jun-qi;WANG Wen-jun;ZHANG Yan(Bioengineering Faculty,Zunyi Medical University Zhuhai Campus,Zhuhai 519041,China)
出处
《食品工业科技》
CAS
北大核心
2020年第7期25-29,36,共6页
Science and Technology of Food Industry
基金
贵州省大学生创新创业训练项目(201710661048)
合肥医学院硕士启动基金(F-912)
贵州省科技计划项目(黔科合基础[2019]1345号)。
关键词
土豆
酪氨酸酶
提取
植物精油
激活
动力学
potato
tyrosinase
extraction
plants essential oil
activation
kinetics
