Regioselective Hydroxylation of Stilbenes by Engineered Cytochrome P450 from<i>Thermobifida fusca</i>YX
作者:Ansgar Rühlmann、Dragutin Antovic、Thomas J. J. Müller、Vlada B. Urlacher
DOI:10.1002/adsc.201601168
日期:2017.3.20
synthesis of (E)‐4,4′‐dihydroxystilbene via direct double hydroxylation of (E)‐stilbene. The construction of a triple mutant led to a more than six‐fold increased catalytic efficiency compared to the wild type enzyme. CYP154E1 and variants thereof accepted not only (E)‐stilbene but also possessed remarkable activity towards ortho‐ and meta‐substituted hydroxystilbenes leading to resveratrol, (E)‐2,4′‐dihydroxystilbene
自从过去的几十年以来,植物中的白藜芦醇白藜芦醇因其广泛的药用特性而受到了公众和研究界的广泛关注。除白藜芦醇外,由于其他潜在的生物活性不同,对其他植物类芪也有越来越高的兴趣。为了满足对类胡萝卜素的不断增长的需求,需要替代和可持续的生产方法。我们确定了细胞色素P450单加氧酶154E1从嗜热裂孢菌YX(CYP154E1),使(合成ë)-4,4'- dihydroxystilbene通过(的直管型双羟化ësti二苯乙烯。与野生型酶相比,三重突变体的构建使催化效率提高了六倍以上。CYP154E1及其变体不仅接受(E)-二苯乙烯,而且对邻位和间位取代的羟基苯乙烯有明显的活性,导致白藜芦醇,(E)-2,4'-二羟基二苯乙烯,(E)-2,4',5 -三羟基sti和(E)-3,4'-二羟基sti。蛋白质工程和甲基-β-环糊精作为底物增溶剂的结合,使产品效价高达4.2 g L -1,酶总周转数(TTN)高达20