Efficient Lewis acid catalysis of an abiological reaction in a de novo protein scaffold

• Published in: Nature chemistry Vol. 13; no. 3; pp. 231 - 235
• Main Authors: Basler, Sophie, Studer, Sabine, Zou, Yike, Mori, Takahiro, Ota, Yusuke, Camus, Anna, Bunzel, H. Adrian, Helgeson, Roger C., Houk, K. N., Jiménez-Osés, Gonzalo, Hilvert, Donald
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-03-2021; Nature Publishing Group
• Subjects: 631/92/469; 639/638/77/603; 639/638/92/607; Active control; Analytical Chemistry; Biochemistry; Catalysis; Catalysts; Catalytic Domain; Chemical reactions; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Cycloaddition Reaction; Density Functional Theory; Design; Directed evolution; Directed Molecular Evolution; Enzymes; Evolution; Hydrogen Bonding; Inorganic Chemistry; Kinetics; Lewis acid; Lewis Acids - chemistry; Metal ions; Metalloproteins - chemistry; Metalloproteins - metabolism; Molecular Docking Simulation; Organic Chemistry; Physical Chemistry; Proteins; Scaffolds; Stereochemistry; Stereoselectivity; Substrate Specificity; Zinc; Zinc-binding protein
• ISSN: 1755-4330; 1755-4349
• DOI: 10.1038/s41557-020-00628-4

New enzyme catalysts are usually engineered by repurposing the active sites of natural proteins. Here we show that design and directed evolution can be used to transform a non-natural, functionally naive zinc-binding protein into a highly active catalyst for an abiological hetero-Diels–Alder reaction. The artificial metalloenzyme achieves >10
4
turnovers per active site, exerts absolute control over reaction pathway and product stereochemistry, and displays a catalytic proficiency (1/
K
TS
 = 2.9 × 10
10
 M
−1
) that exceeds all previously characterized Diels–Alderases. These properties capitalize on effective Lewis acid catalysis, a chemical strategy for accelerating Diels–Alder reactions common in the laboratory but so far unknown in nature. Extension of this approach to other metal ions and other de novo scaffolds may propel the design field in exciting new directions.
A de novo designed zinc-binding protein has been converted into a highly active, stereoselective catalyst for a hetero-Diels–Alder reaction. Design and directed evolution were used to effectively harness Lewis acid catalysis and create an enzyme more proficient than other reported Diels–Alderases.