Engineering a Non‐Natural Photoenzyme for Improved Photon Efficiency**

Nicholls, Bryce T.; Oblinsky, Daniel G.; Kurtoic, Sarah I.; Grosheva, Daria; Ye, Yuxuan; Scholes, Gregory D.; Hyster, Todd K.

doi:10.1002/ange.202113842

Engineering a Non‐Natural Photoenzyme for Improved Photon Efficiency**

Journal Article · Thu Dec 02 00:00:00 EST 2021 · Angewandte Chemie

DOI:https://doi.org/10.1002/ange.202113842· OSTI ID:1833904

Nicholls, Bryce T. ^[1]; Oblinsky, Daniel G. ^[2]; Kurtoic, Sarah I. ^[2]; Grosheva, Daria ^[2]; Ye, Yuxuan ^[3]; Scholes, Gregory D. ^[2]; ^[3]

Department of Chemistry and Chemical Biology Cornell University Ithaca NY 14853 USA, Department of Chemistry Princeton University Princeton NJ 08544 USA
Department of Chemistry Princeton University Princeton NJ 08544 USA
Department of Chemistry and Chemical Biology Cornell University Ithaca NY 14853 USA

Abstract

Photoenzymes are biological catalysts that use light to convert starting materials into products. These catalysts require photon absorption for each turnover, making quantum efficiency an important optimization parameter. Flavin‐dependent “ene”‐reductases (EREDs) display latent photoenzymatic activity for synthetically valuable hydroalkylations; however, protein engineering has not been used to optimize this non‐natural function. We describe a protein engineering platform for the high throughput optimization of photoenzymes. A single round of engineering results in improved catalytic function toward the synthesis of γ, δ, ϵ‐lactams, and acyclic amides. Mechanistic studies show that key mutations can alter the enzyme's excited state dynamics, enhance its photon efficiency, and ultimately increase catalyst performance. Transient absorption spectroscopy reveals that engineered variants display dramatically decreased radical lifetimes, indicating an evolution toward a concerted mechanism.

View Accepted Manuscript (Publisher)

Sponsoring Organization:: USDOE

Grant/Contract Number:: SC0019370

OSTI ID:: 1833904

Journal Information:: Angewandte Chemie, Journal Name: Angewandte Chemie Journal Issue: 2 Vol. 134; ISSN 0044-8249

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: Germany

Language:: English

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