CAJ | 학술논문

在优化茶树叶绿素酶提取及活性分析条件的基础上,对茶树叶绿素酶的分离纯化及其性质进行了研究,结果表明:加入与鲜叶等量PVP,用pH7.5的磷酸二氢钠-磷酸氢二钠缓冲液提取的叶绿素酶活性最高,叶绿素酶的活性分析最佳条件为在45℃条件下反应15 min;经过分离纯化后,叶绿素酶的纯度提高了55.96倍,酶的得率为9.45%,SDS-PAGE结果显示有一条蛋白质的条带,酶的亚基分子量为37.2 kD;对纯化后的叶绿素酶的性质研究表明,该酶最适pH为7.5,45℃时候反应活性最高,在4℃保存条件下,随着存放时间的延长,酶活性逐渐下降,保存一个月后叶绿素酶活性剩余45%,用2 mmol/L和7 mmol/L的金属离子(Ca~(2+)、Mg~(2+)、Zn~(2+)、Fe~(3+))、EDTA处理,都对叶绿素酶活性有不同程度的促进作用.
재우화다수협록소매제취급활성분석조건적기출상,대다수협록소매적분리순화급기성질진행료연구,결과표명:가입여선협등량PVP,용pH7.5적린산이경납-린산경이납완충액제취적협록소매활성최고,협록소매적활성분석최가조건위재45℃조건하반응15 min;경과분리순화후,협록소매적순도제고료55.96배,매적득솔위9.45%,SDS-PAGE결과현시유일조단백질적조대,매적아기분자량위37.2 kD;대순화후적협록소매적성질연구표명,해매최괄pH위7.5,45℃시후반응활성최고,재4℃보존조건하,수착존방시간적연장,매활성축점하강,보존일개월후협록소매활성잉여45%,용2 mmol/L화7 mmol/L적금속리자(Ca~(2+)、Mg~(2+)、Zn~(2+)、Fe~(3+))、EDTA처리,도대협록소매활성유불동정도적촉진작용.
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.