卡瑞苯西思伯克霍尔德氏菌卤乙酸脱卤酶的分离纯化及性质  

Purification and Properties of Haloactate Dehalogenase from Burkholderia caribensis

作　　者：高玉红[1] 彭谦[1] 李铭刚[1] 崔晓龙[1] 徐丽华[1] 姜成林[1] 

机构地区：[1]云南大学云南省微生物研究所教育部微生物资源开放研究重点实验室,昆明650091

出　　处：《微生物学报》2002年第6期706-712,共7页Acta Microbiologica Sinica

基　　金：云南省科技厅国际合作处资助项目 ( 99C0 1 1 )

摘　　要：从云南西北部土样中分离到一株卡瑞苯西思伯克霍尔德氏菌 (Burkholderiacariben sis) ,它在氯乙酸培养基中能产生较高活性的卤乙酸脱卤酶。经硫酸铵盐析、DEAESephadex A5 0柱层析、羟基磷灰石柱层析、SephadexG 2 0 0凝胶过滤后 ,获得电泳纯酶。用SDS PAGE测定酶分子量为 46kD。水解氯乙酸的Km值为 3 7× 1 0 - 3 mol L。酶反应的最适温度为 40℃ ,最适pH值为 9 5。金属离子及CN- 、EDTA对该酶有不同程度的影响 ,Hg2 + 和CN-Haloacetate dehalogenase from Burkholderia caribensis isolated from soils samples collected from north-west of Yunnan province was pruified. The purification procedures included ammonium sulfate fractionation, and column chromatography with DEAE Sephadex-A50, hydroxyapatite and Sephadex G-200 gel filtration. The purified enzyme with specific activity of 63.52 U/mg was homogeneous by SDS-PAGE. The molecular weight estimated by SDS-polyacrylamide gel electrophoresis was 46kD. K m of the enzyme (with chloroacetic acid as substrate)was 3.7×10 -3mol/L. The metal ion, CN - and EDTA have the different level influence on enzyme. The optimum reaction pH and temperature of the enzyme were 9.5 and 40℃, respectively. Besides, the enzyme was obviously inhibited by Hg 2+ and CN -.

关 键 词：伯克霍尔德氏菌 卤乙酸脱卤酶 纯化 酶学性质 

分 类 号：O629[理学—有机化学]