摘要
滁菊在采收后极易因酶促褐变导致其品质下降,故以滁菊鲜花为原材料,针对影响其品质劣变的多酚氧化酶展开研究,明确其三相分离纯化工艺以及酶学特性。考察了硫酸铵浓度、pH值、提取液与叔丁醇体积比、提取时间等工艺参数对滁菊多酚氧化酶纯化效果的影响,结果表明三相分离纯化滁菊多酚氧化酶的较佳工艺参数为:硫酸铵质量浓度0.4 g·mL^(-1),pH值6.0,粗提液与叔丁醇的体积比为1:1.5;按照此工艺条件对纯化后滁菊中多酚氧化酶的酶学特性进行了分析,滁菊多酚氧化酶最适pH值为6.50,最适反应温度为30℃;以邻苯二酚为滁菊多酚氧化酶反应底物时,底物最适浓度为0.35 mol·L^(-1),酶促反应米氏常数为0.1749 mol·L^(-1)。
The quality of Chrysanthemum morifolium Chuju is easy to decline because of enzymatic browning after harvest,therefore,it is necessary to study polyphenol oxidase which affects its quality deterioration.The three phase partitioning process and enzymatic properties of polyphenol oxidase were studied using Chuju flowers as raw materials.The effects of process parameters such as ammonium sulfate concentration,pH value,volume ratio of crude extract to tert-butanol,and extraction time on the purification effect of Chuju polyphenol oxidase were investigated.The results showed that the optimal process parameters for three phase partitioning of Chuju polyphenol oxidase were:ammonium sulfate mass concentration of 0.4 g·mL^(-1),pH 6.0,volume ratio of crude extract to tert-butanol 1:1.5;the enzymatic characteristics of the purified polyphenol oxidase in Chuju were analyzed,and the optimal pH for Chuju polyphenol oxidase was 6.50,the optimal reaction temperature was 30℃;the optimal concentration of the substrate was 0.35 mol·L^(-1) when catechol was used as the substrate of the polyphenol oxidase,and the Michaelis constant of the enzymatic reaction was 0.1749 mol·L^(-1).
作者
苗文娟
陈强
曾妍
朱双杰
董艺凝
MIAO Wenjuan;CHEN Qiang;ZENG Yan;ZHU Shuangjie;DONG Yining(School of Biology Science and Food Engineering,Chuzhou University,Chuzhou 239000)
出处
《安徽农业大学学报》
CAS
CSCD
2021年第2期327-332,共6页
Journal of Anhui Agricultural University
基金
安徽省教育厅自然科学研究一般项目(KJ2018B11)
滁州市第六批“221”产业创新团队项目(特色农产品开发与利用)共同资助。
关键词
滁菊
多酚氧化酶
三相分离
酶学特性
Chrysanthemum morifolium Chuju
polyphenol oxidase
three phase partitioning
enzymatic properties
