Flavocytochrome P450BM3: an update on structure and mechanism of a biotechnologically important enzyme
Flavocytochrome P450BM3: an update on structure and mechanism of a biotechnologically important enzyme
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DOI：
10.1042/bst0330747
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发表时间：
2005-08-01
影响因子：
3.9
通讯作者：
Munro, AW
中科院分区：
文献类型：
--
作者：
Warman, AJ;Roitel, O;Munro, AW
自20世纪80年代发现以来，来自巨大芽孢杆菌的脂肪酸羟化酶黄细胞色素P450（细胞色素P450）BM 3（CYP 102 A1）已被用作理解P450超家族酶的结构和机制的范例。P450 BM 3是第一个被发现与其氧化还原伴侣-真核生物样二黄素还原酶融合的P450。这一事实激发了人们对可溶性P450 BM 3作为哺乳动物肝脏P450酶模型的兴趣，该模型使用单独的膜包埋的P450和还原酶来操作类似的电子传递链。P450 BM 3的每个组成结构域的结构现在已经被解析，并且详细的蛋白质工程和分子酶学研究已经确定了几种氨基酸在例如底物结合、辅酶选择性和催化中的作用。P450 BM 3在生物技术应用中的潜力也已得到认可，其变体能够使用合理诱变和强制进化技术产生工业上重要的转化。本文重点介绍了我们对这种重要酶的结构和机制的理解的最新进展，并强调了仍需解决的重要问题。
Since its discovery in the 1980s, the fatty acid hydroxylase flavocytochrome P450 (cytochrome P450) BM3 (CYP102A1) from Bacillus megaterium has been adopted as a paradigm for the understanding of structure and mechanism in the P450 superfamily of enzymes. P450 BM3 was the first P450 discovered as a fusion to its redox partner - a eukaryotic-like diflavin reductase. This fact fuelled the interest in soluble P450 BM3 as a model for the mammalian hepatic P450 enzymes, which operate a similar electron transport chain using separate, membrane-embedded P450 and reductase enzymes. Structures of each of the component domains of P450 BM3 have now been resolved and detailed protein engineering and molecular enzymology studies have established roles for several amino acids in, e.g. substrate binding, coenzyme selectivity and catalysis. The potential of P450 BM3 for biotechnological applications has also been recognized, with variants capable of industrially important transformations generated using rational mutagenesis and forced evolution techniques. This paper focuses on recent developments in our understanding of structure and mechanism of this important enzyme and highlights important problems still to be resolved.