羊心肌乳酸脱氢酶的纯化及性质研究  

Purification and Properties of Lactate Dehydrogenase from Sheep Heart Muscle

作　　者：金黎明[1] 张乐[1] 蒋本国[1] 赵小菁[1] 田文杰[1] 范圣第[1] 

机构地区：[1]大连民族学院生命科学学院,辽宁大连116600

出　　处：《大连民族学院学报》2006年第1期20-23,共4页Journal of Dalian Nationalities University

基　　金：国家自然科学基金资助项目(20472013);大连民族学院博士科研启动基金资助项目(20056106)

摘　　要：建立了一种较为简单、高效的羊心肌乳酸脱氢酶的纯化工艺，经硫酸铵分级沉淀及DEAE—Sepharose离子交换层析纯化，可以得到纯度较高的乳酸脱氢酶，酶的比活力达106．67U／mg，纯化倍数达54．7，酶活力总回收率为48．6％．酶学性质的研究表明：其最适反应温度为50℃，最适反应pH值为7．8，在25～45℃条件下能够保持较高的酶活力，60℃后酶活力下降较快．1．0mmol／L的Mg^2＋、Fe^2＋、Ca^2＋，0．5mmol／L的Ca^2＋对酶活力有促进作用；1．0mmol／L和0．5mmol／L的Cu^2＋和Fe^3＋，1．0mmol／L的Ni^2＋，0．5mmol／L的Mg^2＋和Fe^2＋对酶活力有抑制作用；Co^2＋的作用不太明显．Lactate dehydrogenase （LDH） from sheep heart muscle was purified 54.7 - fold by ammonium sulfate precipitation and DEAE- Sepharose Fast Flow column ion - exchange chromatography. The specific activity of the enzyme was 106.67U/mg. The optimal temperature and pH for the reaction were 50 ℃ and 7.8, respectively.The LDH activity was relatively stable with in 25 - 45 ℃ and decreased quickly over 60 ℃. Cu^2 ＋ and Fe^3 ＋ of 1.0 mmol/L and 0.5 mmol/L were resemble to inhibit the enzyme activity. 1.0 mmol/L Ni^2＋ was an effective inhibitor on LDH activity, whereas the influence was not evident at the concentration of 0.5 mmol/L. 1.0 mmol/L Mg^2＋ showed the ability to enhance the enzyme activity significantly, however, 0.5 mmol/L Mg^2＋ showed the inhibition ability.The effect of Fe^2 ＋ on the enzyme activity was similar to that of Mg^2＋Ca^2＋ enhanced the enzyme activity weakly. The effect of Co^2 ＋ was not evident.

关 键 词：羊心肌 乳酸脱氢酶 纯化 酶学特性 

分 类 号：Q554.9[生物学—生物化学]