Purification and properties of pyrazon dioxygenase from pyrazon-degrading bacteria.
Purification and properties of pyrazon dioxygenase from pyrazon-degrading bacteria.
复制标题
来自吡嗪降解菌的吡嗪双加氧酶的纯化和性质。
DOI：
10.1111/j.1432-1033.1977.tb11370.x
复制
发表时间：
1977
期刊：
影响因子：
--
通讯作者：
F. Lingens
中科院分区：
文献类型：
--
作者：
K. Sauber;C. Fröhner;G. Rosenberg;J. Eberspächer;F. Lingens
deae -纤维素层析和Sephadex凝胶过滤表明，吡嗪降解菌的吡嗪双加氧酶由三种不同的酶组分组成。没有一种组分单独氧化吡嗪的苯基部分，只有三种组分结合才能检测到氧化。通过电子顺磁共振和紫外线测量，确定了三种组分的蛋白质性质：组分A1（分子量约为180000，颜色为红棕色）是一种铁硫蛋白。结果表明，每摩尔蛋白质中约含有2摩尔铁和2摩尔无机硫。该酶组分经圆盘电泳纯化达到均匀性。组分A2是一种黄色蛋白，分子量约为67000。FAD被证明是该蛋白的假体基。组分B（分子量约12000，呈棕色）为铁氧还蛋白型蛋白，经圆盘电泳纯化后均质。提出了一种三组分合作的假设方案：组分A2接受NADH作为共底物并发挥铁氧还蛋白还原酶的作用。铁氧还蛋白，组分B，具有电子载体的功能。底物的转化受到组分A1，即末端双加氧酶的影响。
Chromatography on DEAE-cellulose and gel filtration on Sephadex revealed that pyrazon dioxygenase from pyrazon-degrading bacteria consists of three different enzyme components. No component alone oxidizes the phenyl moiety of pyrazon, only when the three components are combined can oxidation be detected. Following electron paramagnetic resonance and ultraviolet measurements the protein nature of the three components was determined: component A1 (molecular weight about 180000,red-brown in colour) is an iron-sulphur protein. The existence of approximately two moles of iron and two moles of inorganic sulphur per mole of protein was demonstrated. This enzyme component was purified to homogeneity in disc electrophoresis. Component A2 is a yellow protein of a molecular weight of about 67000. FAD was shown to be the prosthetic group of this protein. Component B (molecular weight about 12000, brown in colour) is a protein of the ferredoxin type, which was purified to homogeneity, as demonstrated by disc electrophoresis. A hypothetical scheme for the cooperation of the three components is proposed: component A2 accepts as cosubstrate NADH and functions as a ferredoxin reductase. The ferredoxin, component B, has the function of an electron carrier. The conversion of the substrates is effected by component A1, the terminal dioxygenase.