Expanding the promiscuity of a natural-product glycosyltransferase by directed evolution.
Natural products, many of which are decorated with essential sugar residues, continue to serve as a key platform for drug development. Adding or changing sugars attached to such natural products can improve the parent compound's pharmacological properties, specificity at multiple levels, and/or even the molecular mechanism of action. Though some natural-product glycosyltransferases (GTs) are sufficiently promiscuous for use in altering these glycosylation patterns, the stringent specificity of others remains a limiting factor in natural-product diversification and highlights a need for general GT engineering and evolution platforms. Herein we report the use of a simple high-throughput screen based on a fluorescent surrogate acceptor substrate to expand the promiscuity of a natural-product GT via directed evolution. Cumulatively, this study presents variant GTs for the glycorandomization of a range of therapeutically important acceptors, including aminocoumarins, flavonoids and macrolides, and a potential template for engineering other natural-product GTs.
Williams
Gavin J
GJ
Laboratory for Biosynthetic Chemistry, Pharmaceutical Sciences Division, School of Pharmacy, National Cooperative Drug Discovery Program, University of Wisconsin-Madison, 777 Highland Avenue, Madison, Wisconsin 53705, USA.
Zhang
Changsheng
C
Thorson
Jon S
JS
D000595:Amino Acid Sequence; D001688:Biological Products / Q000737:chemistry / Q000494:pharmacology*; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry*; D002384:Catalysis; D005419:Flavonoids / Q000737:chemistry / Q000494:pharmacology; D006031:Glycosylation; D016695:Glycosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018942:Macrolides / Q000737:chemistry / Q000494:pharmacology; D008969:Molecular Sequence Data; D013379:Substrate Specificity