7ZP7

Crystal structure of evolved photoenzyme EnT1.3 (truncated) with bound product

Summary for 7ZP7

• Entry DOI: 10.2210/pdb7zp7/pdb
• Descriptor: EnT1.3 C, (1~{R},10~{R},12~{S})-15-oxa-8-azatetracyclo[8.5.0.0^{1,12}.0^{2,7}]pentadeca-2(7),3,5-trien-9-one, 1,2-ETHANEDIOL, ... (4 entities in total)
• Functional Keywords: designed photoenzyme, [2+2]-cyclase, genetic code expansion, engineered enzyme, biosynthetic protein
• Biological source: synthetic construct
• Total number of polymer chains: 2
• Total formula weight: 73961.60

Authors

Hardy, F.J.,Levy, C. (deposition date: 2022-04-26, release date: 2022-09-28, Last modification date: 2024-01-31)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.7 Å)