6TQ5

Alcohol dehydrogenase from Candida magnoliae DSMZ 70638 (ADHA): complex with NADP+

Summary for 6TQ5

• Entry DOI: 10.2210/pdb6tq5/pdb
• Descriptor: Enzyme subunit, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, ISOPROPYL ALCOHOL, ... (5 entities in total)
• Functional Keywords: alcohol dehydrogenase, thermal stability, protein engineering, oxidoreductase
• Biological source: Starmerella magnoliae
• Total number of polymer chains: 4
• Total formula weight: 105970.66

Authors

Rovida, S.,Aalbers, F.S.,Fraaije, M.W.,Mattevi, A. (deposition date: 2019-12-16, release date: 2020-04-08, Last modification date: 2024-01-24)

Primary citation

Aalbers, F.S.,Furst, M.J.,Rovida, S.,Trajkovic, M.,Gomez Castellanos, J.R.,Bartsch, S.,Vogel, A.,Mattevi, A.,Fraaije, M.W.
Approaching boiling point stability of an alcohol dehydrogenase through computationally-guided enzyme engineering.
Elife, 9:-, 2020

PubMed Abstract: Enzyme instability is an important limitation for the investigation and application of enzymes. Therefore, methods to rapidly and effectively improve enzyme stability are highly appealing. In this study we applied a computational method (FRESCO) to guide the engineering of an alcohol dehydrogenase. Of the 177 selected mutations, 25 mutations brought about a significant increase in apparent melting temperature (Δ ≥ +3 °C). By combining mutations, a 10-fold mutant was generated with a of 94 °C (+51 °C relative to wild type), almost reaching water's boiling point, and the highest increase with FRESCO to date. The 10-fold mutant's structure was elucidated, which enabled the identification of an activity-impairing mutation. After reverting this mutation, the enzyme showed no loss in activity compared to wild type, while displaying a of 88 °C (+45 °C relative to wild type). This work demonstrates the value of enzyme stabilization through computational library design.

PubMed: 32228861
DOI: 10.7554/eLife.54639

Experimental method

X-RAY DIFFRACTION (1.6 Å)