An expanded genetic code for probing the role of electrostatics in enzyme catalysis by vibrational Stark spectroscopy

To find experimental validation for electrostatic interactions essential for catalytic reactions represents a challenge due to practical limitations in assessing electric fields within protein structures. This review examines the applications of non-canonical amino acids (ncAAs) as genetically encoded probes for studying the role of electrostatic interactions in enzyme catalysis. ncAAs constitute sensitive spectroscopic probes to detect local electric fields by exploiting the vibrational Stark effect (VSE) and thus have the potential to map the protein electrostatics. Mapping the electrostatics in proteins will improve our understanding of natural catalytic processes and, in beyond, will be helpful for biocatalyst engineering. This article is part of a Special Issue entitled “Biochemistry of Synthetic Biology - Recent Developments” Guest Editor: Dr. Ilka Heinemann and Dr. Patrick O'Donoghue. [Display omitted] •Enzyme electrostatics have a great impact on catalytic reactions.•Strong evidence for this concept is provided on the basis of VSS.•Genetically encoded VSE-active ncAAs substantially broaden the potential of VSS.•VSS with ncAAs enables studying the role of global electrostatics in biocatalysis.