Robust ω-Transaminases by Computational Stabilization of the Subunit Interface
Qinglong Meng, Nikolas Capra, Cyntia M. Palacio, Elisa Lanfranchi, Marleen Otzen, Luc Z. van Schie, H.J. Rozeboom, A.M.W.H. Thunnissen, Hein J. Wijma, Dick B. Janssen
Abstract
)-1-phenylethylamine (>99% enantiomeric excess) with high yield (92%, in comparison to 24% with the wild-type transaminase). Crystal structures mostly confirmed the expected structural changes and revealed that the most stabilizing mutation, I154V, featured a rarely described stabilization mechanism: namely, removal of steric strain. The results show that computational interface redesign can be a rapid and powerful strategy for transaminase stabilization.
Topics & Concepts
IsopropylamineChemistryEnantiopure drugBiocatalysisProtein subunitTransaminaseProtein engineeringPoint mutationStereochemistryDirected evolutionSteric effectsEnzymeCatalysisMutationBiochemistryReaction mechanismMutantEnantioselective synthesisGeneEnzyme Catalysis and ImmobilizationEnzyme Structure and FunctionMicrobial Metabolic Engineering and Bioproduction