Development of Genetically Encoded Biosensors for Reporting the Methyltransferase-Dependent Biosynthesis of Semisynthetic Macrolide Antibiotics
Yiwei Li, Megan R. Reed, H.T. Wright, T. Ashton Cropp, Gavin J. Williams
Abstract
Clarithromycin is an improved semisynthetic analogue of the naturally occurring macrolide, erythromycin. The subtle modification of a methyl group on the C-6 hydroxyl group endows the molecule with improved acid stability and results in a clinically useful antibiotic. Here, we show that the effector specificity of the biosensor protein, MphR, can be evolved to selectively recognize clarithromycin and therefore report on the production of this molecule in vivo. In addition, a crystal structure of the evolved variant reveals the molecular basis for selectivity and provides a guide for the evolution of a new metabolic function using this biosensor.
Topics & Concepts
MethyltransferaseEffectorClarithromycinBiosensorMacrolide AntibioticsBiochemistryChemistryComputational biologyAntibioticsDirected Molecular EvolutionBiosynthesisErythromycinFunction (biology)Combinatorial chemistryBiologyEnzymeMethylationDirected evolutionGeneGeneticsMutantRNA and protein synthesis mechanismsBiochemical and Molecular ResearchMicrobial Natural Products and Biosynthesis