6tq3

Publication Abstract from PubMed

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 (DeltaTm >/= +3 degrees C). By combining mutations, a 10-fold mutant was generated with a Tm of 94 degrees C (+51 degrees C relative to wildtype), 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 wildtype, while displaying a Tm of 88 degrees C (+45 degrees C relative to wildtype). This work demonstrates the value of enzyme stabilization through computational library design.

Approaching boiling point stability of an alcohol dehydrogenase through computationally-guided enzyme engineering.,Aalbers FS, Furst MJ, Rovida S, Trajkovic M, Gomez Castellanos JR, Bartsch S, Vogel A, Mattevi A, Fraaije MW Elife. 2020 Mar 31;9. pii: 54639. doi: 10.7554/eLife.54639. PMID:32228861^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.