Thermostable Flavin Reductase That Couples with Dibenzothiophene Monooxygenase, from Thermophilic Bacillus sp. DSM411: Purification, Characterization, and Gene Cloning
Thermostable Flavin Reductase That Couples with Dibenzothiophene Monooxygenase, from Thermophilic Bacillus sp. DSM411: Purification, Characterization, and Gene Cloning
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DOI：
10.1271/bbb.68.1712
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发表时间：
2004-01
期刊：
影响因子：
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通讯作者：
T. Ohshiro;H. Yamada;T. Shimoda;Toshiyuki Matsubara;Y. Izumi
中科院分区：
文献类型：
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作者：
T. Ohshiro;H. Yamada;T. Shimoda;Toshiyuki Matsubara;Y. Izumi
黄素还原酶是微生物二苯并噻吩(DBT)脱硫过程中所需的加氧酶。嗜热性芽孢杆菌的一种酶。选择DSM411与红球菌D-1的DBT单加氧酶(DszC)偶联。从芽孢杆菌中分离纯化黄素还原酶，得到了均一的黄素还原酶。DSM411，天然酶为分子量为16 kDa的单体。虽然最好的底物是黄素单核苷酸和NADH，但该酶还使用了其他黄素化合物，对硝基芳香化合物和NADPH的作用较小。纯化的酶与DszC偶联，具有铁还原酶活性。在迄今鉴定的黄素还原酶中，目前的酶是最嗜热和最耐热的。该基因编码155个氨基酸的蛋白质，计算质量为17,325 Da。该酶在大肠杆菌中得到高效表达，其粗提物的比活力约为野生型芽孢杆菌的440倍。DSM411。
Flavin reductase is essential for the oxygenases involved in microbial dibenzothiophene (DBT) desulfurization. An enzyme of the thermophilic strain, Bacillus sp. DSM411, was selected to couple with DBT monooxygenase (DszC) from Rhodococcus erythropolis D-1. The flavin reductase was purified to homogeneity from Bacillus sp. DSM411, and the native enzyme was a monomer of Mr 16 kDa. Although the best substrates were flavin mononucleotide and NADH, the enzyme also used other flavin compounds and acted slightly on nitroaromatic compounds and NADPH. The purified enzyme coupled with DszC and had a ferric reductase activity. Among the flavin reductases so far characterized, the present enzyme is the most thermophilic and thermostable. The gene coded for a protein of 155 amino acids with a calculated mass of 17,325 Da. The enzyme was overproduced in Escherichia coli, and the specific activity in the crude extracts was about 440-fold higher than that of the wild-type strain, Bacillus sp. DSM411.