6TF8

Structure of the engineered artificial aldolase I133F RA95.5-8F with a bound substrate, pentan-2-one

Summary for 6TF8

• Entry DOI: 10.2210/pdb6tf8/pdb
• Descriptor: RA95.5-8F_133F (2 entities in total)
• Functional Keywords: artificial enzyme, aldolase, lyase
• Biological source: Saccharolobus solfataricus P2
• Total number of polymer chains: 4
• Total formula weight: 116245.95

Authors

Mori, T.,Macdonald, D.S. (deposition date: 2019-11-13, release date: 2020-06-03, Last modification date: 2024-10-23)

Primary citation

Macdonald, D.S.,Garrabou, X.,Klaus, C.,Verez, R.,Mori, T.,Hilvert, D.
Engineered Artificial Carboligases Facilitate Regioselective Preparation of Enantioenriched Aldol Adducts.
J.Am.Chem.Soc., 142:10250-10254, 2020

PubMed Abstract: Controlling regio- and stereoselectivity of aldol additions is generally challenging. Here we show that an artificial aldolase with high specificity for acetone as the aldol donor can be reengineered via single active site mutations to accept linear and cyclic aliphatic ketones with notable efficiency, regioselectivity, and stereocontrol. Biochemical and crystallographic data show how the mutated residues modulate the binding and activation of specific aldol donors, as well as their subsequent reaction with diverse aldehyde acceptors. Broadening the substrate scope of this evolutionarily naïve catalyst proved much easier than previous attempts to redesign natural aldolases, suggesting that such proteins may be excellent starting points for the development of customized biocatalysts for diverse practical applications.

PubMed: 32427470
DOI: 10.1021/jacs.0c02351

Experimental method

X-RAY DIFFRACTION (1.903 Å)