Manchester Institute of Biotechnology, Department of Chemistry, University of Manchester, Manchester, UK.
Innovation/Biodomain, Shell International Exploration and Production Inc., Westhollow Technology Center, Houston, TX, USA.
Nat Chem Biol. 2020 Nov;16(11):1255-1260. doi: 10.1038/s41589-020-0603-0. Epub 2020 Jul 27.
The direct C-H carboxylation of aromatic compounds is an attractive route to the corresponding carboxylic acids, but remains challenging under mild conditions. It has been proposed that the first step in anaerobic microbial degradation of recalcitrant aromatic compounds is a UbiD-mediated carboxylation. In this study, we use the UbiD enzyme ferulic acid decarboxylase (Fdc) in combination with a carboxylic acid reductase to create aromatic degradation-inspired cascade reactions, leading to efficient functionalization of styrene through CO fixation. We reveal that rational structure-guided laboratory evolution can expand the substrate scope of Fdc, resulting in activity on a range of mono- and bicyclic aromatic compounds through a single mutation. Selected variants demonstrated 150-fold improvement in the conversion of coumarillic acid to benzofuran + CO and unlocked reactivity towards naphthoic acid. Our data demonstrate that UbiD-mediated C-H activation is a versatile tool for the transformation of aryl/alkene compounds and CO into commodity chemicals.
芳香族化合物的直接 C-H 羧化是一种将相应羧酸转化的有吸引力的途径，但在温和条件下仍然具有挑战性。据提议，在厌氧微生物对难降解芳香族化合物的降解中，第一步是 ubiD 介导的羧化作用。在这项研究中，我们使用 ubiD 酶阿魏酸脱羧酶（Fdc）与羧酸还原酶相结合，创建了受芳香族化合物降解启发的级联反应，通过 CO 固定有效地对苯乙烯进行功能化。我们揭示了合理的基于结构的实验室进化可以扩展 Fdc 的底物范围，通过单个突变使其对一系列单环和双环芳香族化合物具有活性。选定的变体使阿魏酸转化为苯并呋喃+CO 的转化率提高了 150 倍，并对萘酸表现出反应性。我们的数据表明， ubiD 介导的 C-H 活化是一种将芳基/烯烃化合物和 CO 转化为商品化学品的通用工具。
