霍山石斛多酚氧化酶活力测定条件的优化

Optimization of Determination Conditions for Polyphenol Oxidase Activity of Dendrobium Huoshanense

为了对目前霍山石斛多酚氧化酶的测定条件进行优化,以霍山石斛一年生和三年生鲜条为原料,采用邻苯二酚法对霍山石斛中的多酚氧化酶进行提取并测定其活力。从实验中所需缓冲液的pH值、多酚氧化酶的酶浓度、反应温度和生长年限这四个方面对实验提取条件进行优化。结果显示,生长年限为三年生的霍山石斛多酚氧化酶在缓冲溶液pH值为6.5、酶浓度为1.0 ml、反应温度为40℃的条件下测定的酶活力为最高值是39.153 U。得出结论:测定结果较传统方法酶活力显著提高,说明优化后测定条件准确可靠,为霍山石斛多酚氧化酶的活性测定提供了新的条件基础。

[Objectives]Optimization of determination conditions for polyphenol oxidase activity of Dendrobium Huoshanense.[Methods]Fresh twigs of one-year-old and three-year-old Dendrobium huoshanense were prepared,and the catechol method was applied to extract the polyphenol oxidase in Dendrobium huoshanense,with its activity tested.Extraction conditions were optimized from four aspects–the pH value and reaction temperature and enzyme concentration of buffer solution and years of growth.[Results]The tested peroxidase activity of three-year-old Dendrobium huoshanense peaked at 39.153U under the condition where the pH value of the buffer solution is 6.5 and its enzyme concentration is 1.0ml and the reaction temperature of 40℃.[Conclusions]Compared with the traditional method,the enzyme activity of Dendrobium huoshanense was significantly increased,indicating that the optimized conditions were accurate and reliable,which provided a new condition basis for the activity determination of polyphenol oxidase of Dendrobium huoshanense.