茶叶科学2009，Vol.29Issue(6)：412-418,7.
茶树叶绿素酶提取、分离纯化与性质的初步研究
Preliminary Studies on Extraction, Purification and Characterization of Chlorophyllase from Tea Leaves(Camellia Sinensis)
李远华 1顾玮 1倪德江 1郑芳 1饶睿 1范仕胜1
作者信息
- 1. 华中农业大学园艺林学学院,湖北,武汉,430070
- 折叠
摘要
Abstract
Under the optimized conditions for the enzyme extraction and activity analysis, chlorophyllase was purified and its characterization was studied. The results indicated that the amount of polivinpolypyrrolidone (PVP) had great impact on chlase activity, chlase activity increased to max as pH value rose to 7.5. It was found that the enzyme reaction was stable at 45℃ in 15 min. Using the methods of ammonium sulfate precipitation and column chromatography on Sephadex G-200, Sepharose CL-6B, chlase in the tea leaves was purified. After three steps of Isolation and purification, It was purified 55.96 fold and a yield of 9.45%. there are one chlase stripe by SDS-PAGE. The subunit molecular mass was 37.2 kD. Research on the characterization of the purified chlase proved that the enzyme showed optimal activity at pH 7.5 and 45℃. The survival rate of enzyme activity remained 45% when stored 4℃ for a month. Adding metal ions Ca~(2+)、Mg~(2+)、Zn~(2+)、Fe~(3+) and EDTA into chlase solution could show the promoting role on the activity of chlorophyllase in certain degree.关键词
茶树/叶绿素酶/酶学性质/提取/分离纯化Key words
tea plant/ chlorophyllase/ characterization/ extraction/ isolation and purification分类
轻工纺织引用本文复制引用
李远华,顾玮,倪德江,郑芳,饶睿,范仕胜..茶树叶绿素酶提取、分离纯化与性质的初步研究[J].茶叶科学,2009,29(6):412-418,7.
