Isotope substitution of promiscuous alcohol dehydrogenase reveals the origin of substrate preference in the transition state

Behiry, Enas M., Ruiz-Pernia, J. Javier, Luk, Louis ORCID: https://orcid.org/0000-0002-7864-6261, Tunon, Inaki, Moliner, Vicent and Allemann, Rudolf K. ORCID: https://orcid.org/0000-0002-1323-8830 2018. Isotope substitution of promiscuous alcohol dehydrogenase reveals the origin of substrate preference in the transition state. Angewandte Chemie International Edition 57 (12) , pp. 3128-3131. 10.1002/anie.201712826

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Abstract

The origin of substrate preference in promiscuous enzymes was investigated by enzyme isotope labeling of the alcohol dehydrogenase from Geobacillus stearothermophilus (BsADH). At physiological temperature, protein dynamic coupling to the reaction coordinate was insignificant. However, the extent of dynamic coupling was highly substrate-dependent at lower temperatures. For benzyl alcohol, an enzyme isotope effect larger than unity was observed, whereas the enzyme isotope effect was close to unity for isopropanol. Frequency motion analysis on the transition states revealed that residues surrounding the active site undergo substantial displacement during catalysis for sterically bulky alcohols. BsADH prefers smaller substrates, which cause less protein friction along the reaction coordinate and reduced frequencies of dynamic recrossing. This hypothesis allows a prediction of the trend of enzyme isotope effects for a wide variety of substrates.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Chemistry
Publisher:	Wiley
ISSN:	1433-7851
Funders:	Biotechnology and Biological Sciences Research Council
Date of First Compliant Deposit:	29 January 2018
Date of Acceptance:	12 January 2018
Last Modified:	08 May 2023 00:01
URI:	https://orca.cardiff.ac.uk/id/eprint/108563

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