Design and optimization of enzymatic activity in a de novo β-barrel scaffold
Abstract
While native scaffolds offer a large diversity of shapes and topologies for enzyme engineering, their often unpredictable behavior in response to sequence modification makes de novo generated scaffolds an exciting alternative. Here we explore the customization of the backbone and sequence of a de novo designed eight stranded beta-barrel protein to create catalysts for a retro-aldolase model reaction. We show that active and specific catalysts can be designed in this fold and use directed evolution to further optimize activity and stereoselectivity. Our results support previous suggestions that different folds have different inherent amenability to evolution and this property could account, in part, for the distribution of natural enzymes among different folds. Show more
Permanent link
https://doi.org/10.3929/ethz-b-000579643Publication status
publishedExternal links
Journal / series
Protein ScienceVolume
Pages / Article No.
Publisher
WileySubject
biocatalysis; computational modeling; enzyme design; enzyme mechanism; protein designOrganisational unit
03492 - Hilvert, Donald (emeritus) / Hilvert, Donald (emeritus)
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