7O1D

A de novo Enzyme for the Morita-Baylis-Hillman Reaction BH32.7

7O1D の概要

• エントリーDOI: 10.2210/pdb7o1d/pdb
• 分子名称: BH32.7 protein (2 entities in total)
• 機能のキーワード: de novo enzyme, biosynthetic protein
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 27531.64

構造登録者

Levy, C.W. (登録日: 2021-03-29, 公開日: 2021-11-03, 最終更新日: 2024-05-01)

主引用文献

Crawshaw, R.,Crossley, A.E.,Johannissen, L.,Burke, A.J.,Hay, S.,Levy, C.,Baker, D.,Lovelock, S.L.,Green, A.P.
Engineering an efficient and enantioselective enzyme for the Morita-Baylis-Hillman reaction.
Nat.Chem., 14:313-320, 2022

PubMed Abstract: The combination of computational design and directed evolution could offer a general strategy to create enzymes with new functions. So far, this approach has delivered enzymes for a handful of model reactions. Here we show that new catalytic mechanisms can be engineered into proteins to accelerate more challenging chemical transformations. Evolutionary optimization of a primitive design afforded an efficient and enantioselective enzyme (BH32.14) for the Morita-Baylis-Hillman (MBH) reaction. BH32.14 is suitable for preparative-scale transformations, accepts a broad range of aldehyde and enone coupling partners and is able to promote selective monofunctionalizations of dialdehydes. Crystallographic, biochemical and computational studies reveal that BH32.14 operates via a sophisticated catalytic mechanism comprising a His23 nucleophile paired with a judiciously positioned Arg124. This catalytic arginine shuttles between conformational states to stabilize multiple oxyanion intermediates and serves as a genetically encoded surrogate of privileged bidentate hydrogen-bonding catalysts (for example, thioureas). This study demonstrates that elaborate catalytic devices can be built from scratch to promote demanding multi-step processes not observed in nature.

PubMed: 34916595
DOI: 10.1038/s41557-021-00833-9

実験手法

X-RAY DIFFRACTION (1.8 Å)