摘要
首次报道了豆科植物来源的苹果酸脱氢酶——绿豆苹果酸脱氢酶的纯化及表征。前期实验结果已于2005年发表于JournalofFoodBiochemistry,本文对该绿豆中苹果酸脱氢酶的性质及生物进化等做了进一步研究。结果显示:苹果酸脱氢酶对底物草酰乙酸的米氏常数为112μmol/L。酶活性受金属离子效应物影响,其中KCl对酶有激活作用,CaCl2和MgCl2对酶活影响不大,ZnCl2、PbCl2和CuSO4表现出不同程度的抑制酶活作用。锌离子对该酶的作用属于非竞争性抑制。通过Edman降解法测定其N-氨基酸序列为N-ASEPGPERKVAVL-GAAGGIG-20,与其它植物来源的苹果酸脱氢酶的同源性分别达到75%～95%。采用CLUSTALW多序列比对软件产生的进化树结构,与绿豆亲缘关系由近至远依次是大豆、苹果树、西瓜、土豆、芥菜、苜蓿、番茄。
The study was reported for the first time that a novel malate dehydrogenase was purified from leguminous plants. The research was focused on further property and phylogenetic relationship of malate dehydrogenase （MDH） from mung bean （Phaseolus mungo）, in succession with the early results published on Journal of Food Biochemistry. The resuits showed that the enzyme expressed the K, for oxaloacetate was 112μmol/L. The enzyme activity was affected by metal ions. K^＋ activated the enzyme, Ca^2＋ and Mg^2＋ had seldom action, the activity was inhibited in various degree by Zn^2＋, Pb^2＋ and Cu^2＋. The effect of Zn^2＋ was classified as noncompetitive type. The partial N-terminal amino acid sequence data analysis of the first 20 amino acids of the MDH was determined to be 1-ASEPGPERKVAVLGAAGGIG-20 by Edman degeneration, and revealed 75% to 95% homology with other reported plant MDHs. The phylogenetic tree was setted by CLUSTAL W software, demonstrating phylogenetie relationship as Glycine max, Eucalyptus gunnii, CitruUus lanatus, Solanum tuberosum, Arabidopsis thaliana, Medicago sativa, Lycopersicon esculentum.
出处
《中国食品学报》
EI
CAS
CSCD
2006年第1期262-266,共5页
Journal of Chinese Institute Of Food Science and Technology
基金
福建省教育厅资助项目(No.JB05040)
关键词
绿豆
苹果酸脱氢酶
金属离子
抑制
进化
Mung bean Malate dehydrogenase （MDH） Metal ions Inhibition Phylogenetic relationship
