热带假丝酵母酰基辅酶A氧化酶的纯化及性质研究  

Purification and Characterization of Acyl Coenzyme a Oxidase (ACO) from Candida tropicalis

作　　者：欧阳晶[1] 荣东[1] 郝秀珍[1] 陈远童[1] 

机构地区：[1]中国科学院微生物学研究所微生物资源前期开发国家重点实验室,北京10080

出　　处：《微生物学报》2002年第6期713-719,共7页Acta Microbiologica Sinica

基　　金：九五攻关项目 ( 96 -C -0 3-0 4 )

摘　　要：利用热带假丝酵母由烷烃生产二元酸时 ,二元酸面临被 β氧化降解的代谢途径。酰基辅酶A氧化酶是二元酸 β氧化的限速酶。以热带假丝酵母 1 2 3 0菌株为材料 ,经硫酸铵分级沉淀、阴离子交换柱层析、Blue Sepharose亲和柱层析 ,得到电泳均一的酰基辅酶A氧化酶。该酶有两种亚基 ,分子量分别为 74kD和 78kD。酶作用最适pH和最适温度分别为 8 0和5 0℃。金属离子Ag+ 、Pb2 + 完全抑制酶活性 ,Ba2 + 、Mg2 + 、Ca2 + 对酶活性有明显抑制作用。丙烯酸是酶的反竞争性抑制剂 ,Ki 为 0 6 3 3mmol L ,维生素C是竞争性抑制剂 ,Ki 为 2 0 1× 1 0 - 3mmol L。Acyl Coenzyme A Oxidase (ACO) was rate-limiting enzyme in the β-oxidation of dicarboxylic acid(DC) which can be produced from n-alkane by Candida tropicalis. Electrophoretic purity ACO was purified from Candida tropicalis strain 1230 through ammonium sulfate fractionation, anion-exchange chromatography, affinity chromatography. ACOwas composed of two kinds of subunits with MW 74kD and 78kD, respectively. The optima reactive pH and temperature of ACO were 8.0 and 50℃,respectively. ACO activity can be inhibited completely by Ag +、Pb 2+, and obviously by Ba 2+、Mg 2+、Ca 2+.Acrylic acid was uncompetitive inhibitor of ACO with K i=0.633mmol/L, and Vitamin C was competitive inhibitor with K i=2.01×10 -3mmol/L.

关 键 词：酰基辅酶A氧化酶 热带假丝酵母 纯化 性质 

分 类 号：Q936[生物学—微生物学]