Month: July 2018

Evolutionary repurposing of a sulfatase: A new Michaelis complex leads to efficient transition state charge offset

Charlotte M. Miton, Stefanie Jonas, Gerhard Fischer, Fernanda Duarte, Mark F. Mohamed, Bert van Loo, Bálint Kintses, Shina C. L. Kamerlin, Nobuhiko Tokuriki, Marko Hyvönen, and Florian Hollfelder

Proceedings of the National Academy of Sciences of the USA (2017)

DOI: 10.1073/pnas.1607817115
Pubmed: 30012610

PDB coordinates:

4CYR (3D view), 4CXS (3D view), 4CXU (3D view), 4CYS (3D view)
5AJ9 (3D view), 4CXK (3D view)


The recruitment and evolutionary optimization of promiscuous enzymes is key to the rapid adaptation of organisms to changing environments. Our understanding of the precise mechanisms underlying enzyme repurposing is, however, limited: What are the active-site features that enable the molecular recognition of multiple substrates with contrasting catalytic requirements? To gain insights into the molecular determinants of adaptation in promiscuous enzymes, we performed the laboratory evolution of an arylsulfatase to improve its initially weak phenylphosphonate hydrolase activity. The evolutionary trajectory led to a 100,000-fold enhancement of phenylphosphonate hydrolysis, while the native sulfate and promiscuous phosphate mono- and diester hydrolyses were only marginally affected (≤50-fold). Continue reading →

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