Position: Home > Articles > Isolation, Purification and Characterization of Malate Dehydrogenase from Duck Heart
FOOD SCIENCE
2013,34
(23)
239-244
鸭心苹果酸脱氢酶的分离纯化及酶学性质
作 者：
孙芳;胡瑞斌;任美凤;唐云明
单 位：
西南大学生命科学学院;西南大学生命科学学院,重庆市甘薯工程研究中心,三峡库区生态环境教育部重点实验室
关键词：
鸭心;苹果酸脱氢酶;分离纯化;性质
摘 要：
取新鲜鸭心,通过匀浆、硫酸铵沉淀、有机溶剂沉淀、DEAE-Sepharose层析以及Superdex-200层析等方法纯化,获得鸭心中所含的两种电泳纯的苹果酸脱氢酶同工酶(c-MDH和m-MDH),并对粗酶液中含量较高的c-MDH进行酶学性质研究。结果表明:c-MDH和m-MDH纯化倍数分别为69.07倍和130.40倍,酶活力回收率分别为25.54%和10.98%,亚基分子质量分别为39.58ku和37.62ku。c-MDH酶学性质研究表明:该酶最适反应温度为55℃,在30℃以下时比较稳定;最适反应pH为7.2,在pH6~9范围内稳定性较好;其对草酰乙酸的K m值为140.93 mol/L;草酸、SDS、Cu2+、Ag+对该酶的抑制作用非常强烈,而Mg2+、K+、Ba2+对该酶表现出明显的激活作用。
译 名：
Isolation, Purification and Characterization of Malate Dehydrogenase from Duck Heart
作 者：
SUN Fang;HU Rui-bin;REN Mei-feng;TANG Yun-ming;Key Laboratory of Eco-environments in Three Gorges Reservoir Region,Ministry of Education,Chongqing Sweetpotato Engineering Research Center,School of Life Science,Southwest University;
关键词：
duck heart;;malate dehydrogenase(MDH);;isolation and purification;;characterization
摘 要：
Two electrophoresis-pure isoenzymes(c-MDH and m-MDH) of malate dehydrogenase from duck heart were obtained through the procedures including homogenization, ammonium sulfate precipitation, organic solvent precipitation, DEAE-sepharose chromatography and Superdex-200 gel filtration. c-MDH present in the crude enzyme solution in a higher amount was characterized. The results showed that the purity of c-MDH and m-MDH w ere increased by 69.07- and 130.40-fold with activity recoveries of 25.54 % and 10.98 %, respectively. The molecular mass of the subunits in c-MDH and m-MDH were 39.58 ku and 37.62 ku, respectively. Enzymatic characterization showed that the optimal reaction temperature and pH for c-MDH were 55 ℃ and 7.2, respectively, and it was stable below 30 ℃ and at pH 6- 9. Meanwhile, its K m using oxobutanedioic acid as the substrate was 140.93 μmol/L. The enzyme activity could be strongly inhibited by oxalic acid, SDS, Cu 2+, Co 2+ and Ag +, while it could be obviously activated by Mg 2+, K +, and Ba 2+.
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