黄国文*
湖南科技学院生命科学和化学工程学院，永州425199
摘要:采用磷酸缓冲液浸提法提取羊蹄叶POD,对影响酶提取的主要因素(料液比、浸提液pH,浸提温度和浸提时间)讲行单因素实验和正交实验，用。H沉泞和丙酮沉泞对酶讲行了纯化，并研究了其酶学性质结果表明，羊蹄叶POD的最佳提取条件是料液比为1:40,提取液pH为9,浸提温度是35℃，浸提时间为30 min调节提取液的pH为5以及用1倍和0. 8倍体积的丙酮沉泞提取液时蛋白相对沉泞量多和酶的比活性大羊蹄叶POD以愈}}l木酚和H= 0=为底物的Km分别是0. 12 mmol/L和0. 62 x 10-3mmo1/L} CuS04对羊蹄叶POD活性有激活作用，而ZnS04,MeS04,CaC1=,KC1和NaCI对POD活性有抑制作用研究证明羊蹄叶POD的提取和纯化方法简单有效，其酶学性质为理解羊蹄植物的生长特性和酶学应用提供依据
关键词:羊蹄叶;讨氧化物酶;分离纯化;酶学性质
中图分类号:PS文献标识码:A DOI:10.16333/i.1001-6880.2015.10.017
Purification and Enzymatic
Peroxidases from Rumex
Characteristics of
}aponzcus
Houtt
HUANG Guo-wen‘
rrtment ofBioloeicol Sciences and Ch.em.icol Eneineerine,Hunon Colleee of Science and Tech,noloe}',Yonezh,ou,Hunan 425199，Chino
Abstract:In this stu小，neroxidases(PODS) of Rumex iooonicus Houtt leaves were extracted and purified. Its enzymatic
characteristics were studied. PODS were extracted from leaves of Rumex tooonicus Houtt urine the lixiviatine method with
a phosphate buffer. The main factors affectine extraction,includine ratio of materials to huffer,huffer nH,extraction tem-
nerature and extraction <luration,were evaluated and ontimize<1 by the sinele factor and L9(3")orthoeonal experiment. In
addition，PODS were purified urine pH and acetone precipitation methods for further analysis of its enzymatic properties.
The results showed that the optimal extraction conditions of PODS were as follows:ratio of materials to buffer of 1:40,nH
9 , extraction temperature of 35℃，extraction duration of 35 min. Adiustine pH to 5 and addine 0. 8一1 folds of acetone
in the extraction solutions can eet proteins with relatively' hieh amount and hieh enz}'matic activity'. The obtained PODS
had the Km, of 0. 12 mmol/L and 0. 62 x 10-3 mmol/L for Guaiacol and H, 0,，respectively'. Only' CuS04 enhanced the
PODS activit}',while ZnS04，MeS04，CaCI=，KCl and NaCI inhibited it. The results sueeested that the extraction and puri-
fication methods of PODS were simple and effective for R. }avonicus and laid a foundation for practical application of its
enz}'matic properties and for understandine the erowth features of R. }avonicus.
Kev words:Rum.ex }avonicus Houtt;peroxidase;isolation and purification;enz}'matic characteristics
