HIGH-YIELD PURIFICATION OF CYTOCHROME AA(3) AND CYTOCHROME CAA(3) OXIDASES FROM BACILLUS-SUBTILIS PLASMA-MEMBRANES
HIGH-YIELD PURIFICATION OF CYTOCHROME AA(3) AND CYTOCHROME CAA(3) OXIDASES FROM BACILLUS-SUBTILIS PLASMA-MEMBRANES
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DOI：
10.1042/bj3090279
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发表时间：
1995-07-01
影响因子：
4.1
通讯作者：
HILL, BC
中科院分区：
文献类型：
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作者：
HENNING, W;VO, L;HILL, BC
当在通气振荡培养中生长时，枯草芽孢杆菌表达两种不同的含血红素A的末端氧化酶：细胞色素aa（3）-醌醇氧化酶和细胞色素caa（3）氧化酶。本文描述了一种从枯草芽孢杆菌的同一好氧培养物中纯化两种含血红素A氧化酶的高产常规方法。获得了接近总血红素A的40%的产率，并且从4升液体培养物中获得了约6mg的每种纯化的氧化酶。两种纯化的酶都有两个亚基，细胞色素氧化酶的表观分子量分别为71.6 kDa和34.3 kDa，aa（3）-醌醇氧化酶的表观分子量分别为67.6 kDa和37.2 kDa。这些特征与B的aa（3）和caa（3）操纵子中相应结构基因的序列数据一致。枯草芽孢杆菌两个纯化的氧化酶的一些光谱和酶学特征的报道，这是一致的，这两种酶作为细胞色素氧化酶超家族的成员列入。
When grown in aerated shaking culture, Bacillus subtilis expresses two different haem A-containing terminal oxidases: cytochrome aa(3)-quinol oxidase and cytochrome caa(3) oxidase. This paper describes a high-yield conventional procedure for purifying the two haem A-containing oxidases from the same aerobic culture of Bacillus subtilis. Yields of close to 40% of the total haem A are achieved and about 6 mg of each of the purified oxidases is obtained from 4 litres of liquid culture. Both of the purified enzymes have two subunits, with apparent molecular masses of 71.6 kDa and 34.3 kDa for the cytochrome can, oxidase, and 67.6 kDa and 37.2 kDa for aa(3)-quinol oxidase. These features are in agreement with the sequence data for the corresponding structural genes in the aa(3) and caa(3) operons of B. subtilis. Some spectral and enzymic features of the two purified oxidases are reported that are consistent with the inclusion of both of these enzymes as members of the cytochrome oxidase superfamily.