1,2-Hydrogenation and Transhydrogenation Catalyzed by 3-Ketosteroid Δ1-Dehydrogenase from Sterolibacterium denitrificans—Kinetics, Isotope Labelling and QM:MM Modelling Studies
Abstract
:1. Introduction
2. Results and Discussion
2.1. 1,2-Hydrogenation
2.1.1. pH Dependency
2.1.2. Synthesis of 1α,2β-Deuterated-AD
2.2. Transhydrogenation
3. Material and Methods
3.1. Materials
3.2. Protein Expression and Purification
3.3. Spectrophotometric Assays
3.4. 1,2-Hydrogenation
3.5. Transhydrogenation
3.6. HPLC and LC-MS Detection
3.7. Product Purification and NMR Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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∆(E + ZPE) (kcal/mol) | ||||
---|---|---|---|---|
1α,2β-d2-AD | AD | |||
2β Attack | 2α Attack | 2β Attack | 2α Attack | |
E:S | 0 | 0 | 0 | 0 |
TS1 | 10.85 | 14.19 | 9.94 | 14.05 |
E:I | 3.13 | 3.13 | 2.92 | 2.92 |
TS2 | 13.39 | 13.50 | 12.59 | 12.59 |
E:P | −9.63 | −9.46 | −9.45 | −9.45 |
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Wojtkiewicz, A.M.; Glanowski, M.; Waligórski, P.; Janeczko, T.; Szaleniec, M. 1,2-Hydrogenation and Transhydrogenation Catalyzed by 3-Ketosteroid Δ1-Dehydrogenase from Sterolibacterium denitrificans—Kinetics, Isotope Labelling and QM:MM Modelling Studies. Int. J. Mol. Sci. 2022, 23, 14660. https://doi.org/10.3390/ijms232314660
Wojtkiewicz AM, Glanowski M, Waligórski P, Janeczko T, Szaleniec M. 1,2-Hydrogenation and Transhydrogenation Catalyzed by 3-Ketosteroid Δ1-Dehydrogenase from Sterolibacterium denitrificans—Kinetics, Isotope Labelling and QM:MM Modelling Studies. International Journal of Molecular Sciences. 2022; 23(23):14660. https://doi.org/10.3390/ijms232314660
Chicago/Turabian StyleWojtkiewicz, Agnieszka M., Michał Glanowski, Piotr Waligórski, Tomasz Janeczko, and Maciej Szaleniec. 2022. "1,2-Hydrogenation and Transhydrogenation Catalyzed by 3-Ketosteroid Δ1-Dehydrogenase from Sterolibacterium denitrificans—Kinetics, Isotope Labelling and QM:MM Modelling Studies" International Journal of Molecular Sciences 23, no. 23: 14660. https://doi.org/10.3390/ijms232314660