Abstract
Alcohol dehydrogenases (ADH) are a family of enzymes that catalyse the interconversion between ketones/aldehydes and alcohols in the presence of NADPH cofactor. It is challenging to desymmetrise the substituted cyclopentane-1,3-dione by engineering an ADH, while the reaction mechanism of the metal independent ADH remains elusive. Here we measured the conversion of a model substrate 2-benzyl-2-methylcyclopentane-1,3-dione by LbADH and found it predominately gave the (2R, 3R) product. Binding mode analysis of the substrate in LbADH from molecular dynamics simulations disclosed the origin of the enantioselectivity of the enzyme; the opening and closing of the loop 191-205 above the substrate are responsible for shaping the binding pocket to orientate the substrate, so as to give different stereoisomer products. Using QM/MM calculations, we elucidated the reaction mechanism of LbADH. Furthermore, we demonstrated the reaction profile corresponding to the production of different stereoisomers, which is in accordance with our experimental observations. This research here will shed a light on the rational engineering of ADH to achieve stereodivergent stereoisomer products.
| Original language | English |
|---|---|
| Pages (from-to) | 31292-31300 |
| Number of pages | 9 |
| Journal | Physical Chemistry Chemical Physics |
| Volume | 25 |
| Issue number | 45 |
| Early online date | 06 Nov 2023 |
| DOIs | |
| Publication status | Published - 07 Dec 2023 |
Bibliographical note
Funding Information:J. Z. acknowledges the financial support of Royal Society International Exchange (IEC\NSFC\191454), Queen's University Belfast (QUB) and Chinese Scholarship Council. The authors acknowledge the financial support from INVEST NI Research and Development Programme, partly financed by the European Regional Development Fund under the Investment for Growth and Jobs Programme 2021–2027. The authors are grateful for the computing resources from QUB high performance computing Tier2 computing resource funded by EPSRC (EP/T022175). We also acknowledge Chenggong Hui for the helpful discussion on IRC analysis.
Funding Information:
J. Z. acknowledges the financial support of Royal Society International Exchange (IEC\NSFC\191454), Queen's University Belfast (QUB) and Chinese Scholarship Council. The authors acknowledge the financial support from INVEST NI Research and Development Programme, partly financed by the European Regional Development Fund under the Investment for Growth and Jobs Programme 2021-2027. The authors are grateful for the computing resources from QUB high performance computing Tier2 computing resource funded by EPSRC (EP/T022175). We also acknowledge Chenggong Hui for the helpful discussion on IRC analysis.
Publisher Copyright:
© 2023 The Royal Society of Chemistry.
ASJC Scopus subject areas
- General Physics and Astronomy
- Physical and Theoretical Chemistry