7ZP5

Crystal structure of designed photoenzyme EnT1.0

7ZP5 の概要

• エントリーDOI: 10.2210/pdb7zp5/pdb
• 分子名称: Diisopropyl-fluorophosphatase, PHOSPHATE ION (3 entities in total)
• 機能のキーワード: designed photoenzyme, [2+2]-cyclase, genetic code expansion, engineered enzyme, biosynthetic protein
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 72670.21

構造登録者

Hardy, F.J.,Levy, C. (登録日: 2022-04-26, 公開日: 2022-09-28, 最終更新日: 2024-01-31)

主引用文献

Trimble, J.S.,Crawshaw, R.,Hardy, F.J.,Levy, C.W.,Brown, M.J.B.,Fuerst, D.E.,Heyes, D.J.,Obexer, R.,Green, A.P.
A designed photoenzyme for enantioselective [2+2] cycloadditions.
Nature, 611:709-714, 2022

PubMed Abstract: The ability to program new modes of catalysis into proteins would allow the development of enzyme families with functions beyond those found in nature. To this end, genetic code expansion methodology holds particular promise, as it allows the site-selective introduction of new functional elements into proteins as noncanonical amino acid side chains. Here we exploit an expanded genetic code to develop a photoenzyme that operates by means of triplet energy transfer (EnT) catalysis, a versatile mode of reactivity in organic synthesis that is not accessible to biocatalysis at present. Installation of a genetically encoded photosensitizer into the beta-propeller scaffold of DA_20_00 (ref. ) converts a de novo Diels-Alderase into a photoenzyme for [2+2] cycloadditions (EnT1.0). Subsequent development and implementation of a platform for photoenzyme evolution afforded an efficient and enantioselective enzyme (EnT1.3, up to 99% enantiomeric excess (e.e.)) that can promote intramolecular and bimolecular cycloadditions, including transformations that have proved challenging to achieve selectively with small-molecule catalysts. EnT1.3 performs >300 turnovers and, in contrast to small-molecule photocatalysts, can operate effectively under aerobic conditions and at ambient temperatures. An X-ray crystal structure of an EnT1.3-product complex shows how multiple functional components work in synergy to promote efficient and selective photocatalysis. This study opens up a wealth of new excited-state chemistry in protein active sites and establishes the framework for developing a new generation of enantioselective photocatalysts.

PubMed: 36130727
DOI: 10.1038/s41586-022-05335-3

実験手法

X-RAY DIFFRACTION (1.54 Å)