Replacement of natural cofactors by selected hydrogen peroxide donors or organic peroxides results in improved activity for CYP3A4 and CYP2D6
Replacement of natural cofactors by selected hydrogen peroxide donors or organic peroxides results in improved activity for CYP3A4 and CYP2D6
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DOI：
10.1002/cbic.200600006
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发表时间：
2006-06-01
期刊：
影响因子：
3.2
通讯作者：
Auclair, Karine
中科院分区：
文献类型：
--
作者：
Chefson, Amandine;Zhao, Jin;Auclair, Karine
细胞色素P450酶（P450或CYP）形成一个普遍存在的血红素蛋白家族，能够催化多种底物的单加氧作用。P450在合成有机化学中引起了相当大的兴趣，因为它们具有催化氧插入非活化的C3 OH键的能力。这种有机化学中有用的反应在几十年来受到了广泛关注，但仍然是一个重大的挑战。一些金属催化剂已成功使用[1]，仿生非血红素铁催化剂也已开发出来[2]，但区域和/或立体选择性通常仍然较差。生物催化剂如P450酶代表了一种有前途的替代品。[3]在合成中使用P450的一个限制是需要辅因子的复杂系统，包括NADPH和氧化还原配偶体，如细胞色素P450还原酶（CPR）或铁氧还蛋白/铁氧还蛋白还原酶系统。一些团体试图克服这一缺点。电化学方法，[4]二茂钴，[5]和钴（III）sepulchrate，[6]都被用来ACHTUNGTRENNUNG替代辅因子，尽管成功或适用性有限。虽然已知许多P450酶也接受过氧化物或过氧化氢水溶液作为氧源（分流途径），[7]但这种途径通常效率不高。P450 BM-3的一些突变体已经通过定向进化工程化，以在不存在辅因子的情况下有效地利用过氧化氢。[8]P450 BM-3的血红素结构域突变体已被改造为在过氧化氢而不是其天然辅因子的存在下催化区域和立体选择性氧化。[9]然而，初始反应速率显著低于在自然条件下用野生型酶（NADPH）观察到的那些。此外，这种酶天然地对脂肪酸非常特异，并且必须突变以接受任何新的底物。[10个国家]
The cytochrome P450 enzymes (P450s or CYPs) form a ubiquitous family of heme proteins able to catalyze the monooxygenation of a wide range of substrates. P450s are of considerable interest in synthetic organic chemistry because of their impressive ability to catalyze the insertion of oxygen into nonactivated CÀH bonds. This useful reaction in organic chemistry has received much attention over several decades, but still remains a significant challenge. Some metal catalysts have been successfully used [1] and biomimetic non-heme iron catalysts have been developed,[2] but the regio-and/or stereoselectivity usually remains poor. Biocatalysts such as P450 enzymes represent a promising alternative.[3] One limitation to the use of P450s in synthesis is the need for a complex system of cofactors including NADPH and a redox partner such as cytochrome P450 reductase (CPR) or a ferrodoxin/ferredoxin reductase system. A number of groups have attempted to overcome this drawback. Electrochemical methods,[4] cobaltocene,[5] and cobaltACHTUNGTRENNUNG (III) sepulchrate,[6] have all been used to ACHTUNGTRENNUNGreplace the cofactors, albeit with limited success or applicability. Although many P450 enzymes are also known to accept peroxides or aqueous hydrogen peroxide as a source of oxygen (shunt pathway),[7] this pathway is generally not efficient. Some mutants of P450BM-3 have been engineered by directed evolution to efficiently use hydrogen peroxide in the absence of cofactors.[8] A heme-domain mutant of P450BM-3 has been engineered to catalyze regio-and stereoselective oxidations in the presence of hydrogen peroxide instead of its natural cofactors.[9] The initial reaction rates were, however, significantly lower than those observed with the wild-type enzyme under natural conditions (NADPH). Moreover, this enzyme is naturally very specific for fatty acids and must be mutated to accept any new substrates.[10]