Abstract
Protein engineering is a growing field with a variety of experimental techniques available for altering protein function. However, creating an enzyme de novo is still in its infancy, so far yielding enzymes of modest catalytic efficiency. In this study, a system of artificial retro-aldolase enzymes found to have chemistry coupled to protein dynamics was examined. The original design was created computationally, and this protein was then subjected to directed evolution to improve the initial low catalytic efficiency. We found that this re-engineering of the enzyme resulted in rapid density fluctuations throughout the enzyme being reshaped via alterations in the hydrogen bonding network. This work also led to the discovery of a second important motion which aids in the release of an intermediate product. These results provide compelling evidence that to engineer efficient protein catalysts, fast protein dynamics need to be considered in the design.
Original language | English (US) |
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Pages (from-to) | 8476-8484 |
Number of pages | 9 |
Journal | ACS Catalysis |
Volume | 10 |
Issue number | 15 |
DOIs | |
State | Published - Aug 7 2020 |
Externally published | Yes |
Keywords
- Markov chain Monte Carlo
- QMMM simulation
- directed evolution
- hydrogen bonding network
- protein dynamics
- transition path sampling
ASJC Scopus subject areas
- Catalysis
- General Chemistry