Cofactor Regeneration of NAD+ from NADH: Novel Water-Forming NADH Oxidases
Cofactor Regeneration of NAD+ from NADH: Novel Water-Forming NADH Oxidases
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DOI：
10.1002/1615-4169(200212)344:10
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发表时间：
2002-12
影响因子：
5.4
通讯作者：
B. Riebel;P. R. Gibbs;William B. Wellborn;A. Bommarius
中科院分区：
文献类型：
--
作者：
B. Riebel;P. R. Gibbs;William B. Wellborn;A. Bommarius
脱氢酶具有极好的对映体选择性，可用于制备对映体纯醇、羟基酸和氨基酸。然而，对于经济合成，脱氢酶的共底物，NAD(P)(H)辅因子，必须再生。虽然从NADH中再生NADH的问题可以认为已经解决，但从NADH中再生NAD的逆问题仍然需要一个明确和实用的解决方案。一种可能的解决方法是NADH氧化酶催化NADH氧化为NAD，同时伴有氧的还原。-)可以将O2还原为不需要的H2O2或无害的H2O。我们从伯氏疏螺旋体和圣弗朗西斯乳杆菌中发现并克隆了两个迄今为止只有机器注释的NADH氧化酶功能的新基因。我们已经过表达了相应的蛋白，可以证明标注的功能是正确的。这两种新型NADH氧化酶可以将O2还原为H2O。
Dehydrogenases with their superb enantio- selectivity can be employed advantageously to pre- pare enantiomerically pure alcohols, hydroxy acids, and amino acids. For economic syntheses, however, the co-substrate of dehydrogenases, the NAD(P)(H) cofactor, has to be regenerated. Whereas the problem of regenerating NADH from NAD can be consid- ered solved, the inverse problem of regenerating NAD from NADH still awaits a definitive and practical solution. A possible solution is the oxidation of NADH to NAD with concomitant reduction of oxygen catalyzed by NADH oxidase (E.C. 1.6.-.-) which can reduce O2 either to undesirable H2O2 or to innocuous H2O. We have found and cloned two novel genes from Borrelia burgdorferi and Lactobacillus sanfranciscensis with hitherto only machine-annotat- ed NADH oxidase function. We have overexpressed the corresponding proteins and could prove the annotated function to be correct. As demonstrated with a more sensitive assay than employed previously, the two novel NADH oxidases reduce O2 to H2O.