羟基阿特拉津脱乙胺基水解酶(AtzB)典型酶学性质的研究  

Typical characterization of hydroxyatrazine N-ethylaminohydrolase (AtzB)

作　　者：张颖[1] 梁海晶[1] 马广鹏[2] 姜昭[1] 王志刚[3] 南超[1] 王溪[1] 葛世杰[1] 

机构地区：[1]东北农业大学资源与环境学院,哈尔滨150030 [2]中国农村技术开发中心,北京100045 [3]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006

出　　处：《东北农业大学学报》2013年第5期18-23,共6页Journal of Northeast Agricultural University

基　　金：国家"十二五"科技计划项目(2011BAD04B02-1)

摘　　要：采用Ni-NTA亲和层析法对基因重组菌中的羟基阿特拉津脱乙胺基水解酶(AtzB)进行分离纯化,并对该酶的典型酶学性质进行研究。纯化处理后,AtzB的纯度提高15倍,酶活回收率高达15.2%。酶学性质研究表明,AtzB的最适反应温度为40℃;最适反应pH为9.0。在最适反应条件下,AtzB对底物最大反应速率Vmax为3.17μmol.L-1.min-1,米氏常数Km为0.45 mmol.L-1。反应缓冲液中Cu(Ⅱ)对酶活力有相对较强的抑制作用(P<0.05),Zn(Ⅱ)对酶活力影响相对较小(P<0.05),而Co(Ⅱ)、Cr(Ⅲ)、Ca(Ⅱ)和Ni(Ⅱ)则对酶活力基本没有影响。盐度对AtzB活力影响较大,当缓冲液中NaCl浓度达到1 mol.L-1时,AtzB完全失活。The purification of Hydroxyatrazine N-Ethylaminohydrolase was achieved by combination method of Ni-NTA affinity chromatography, and the properties were subsequently measured. AtzB was purified 15-fold to apparent homogeneity with 15.2% overall recovery. The properties of purified AtzB showed that the optimum temperature and pH was 40 ℃ and 9.0, respectively. Kinetic studies of the AtzB showed that the Vmax and Km were 3.17 pmol. L-1 min-1 and 0.45 mmol. L-1, respectively. The activity of pudfied AtzB was strongly inhibited by Cu（ll） （P〈0.05）, and moderately inhibited by Zn（ll） （P〈0.05）, However, Co （11）, Cr （111）, Ca （11） and Ni （11） have little effect on the activity of purified AtzB. In addition, the influence of NaCI on enzyme activity is strong, when its concentration reached 1 tool . L-1, the purified AtzB was a reflection of complete inactivity.

关 键 词：羟基阿特拉津脱乙胺基水解酶 羟基阿特拉津 蛋白纯化 酶学性质 

分 类 号：S592[农业科学—作物学]