Generation of New Glycoanalogues of Polyene Antibiotics by Synthetic Biology—Testing Current Technical Boundaries

Abstract

A number of antifungal drugs are based on polyene macrolides that cause severe side effects. Most of these compounds contain a single aminodeoxysugar, D-mycosamine. Toxicity can be reduced by increasing the extent of glycosylation. The aromatic heptaene 67-121C and two analogues of the degenerate heptaene nystatin have a second sugar attached to the C4′ hydroxyl of mycosamine. Another nystatin analogue has L-digitoxose as a second sugar attached to C35 on the macrolactone ring. The pentaene selvamicin has 4-O-methyl-L-digitoxose at C27, the equivalent position. To assist the production of new antifungals by synthetic biology, we explore further the utility of three classes of polyene glycosyltransferase: extending glycosyltransferases that form disaccharide-containing polyenes, glycosyltransferases that add the L-digitoxose sugars of nystatin A3 and selvamicin, and mycosaminyltransferases that add the primary aminodeoxysugar. In addition, we combine enzymatic hyperglycosylation with a known chemical method for adding sugars to the C3′ amino group of mycosamine. This was used to convert the disaccharide-containing 67-121C heptaene to forms containing branched trisaccharide or tetrasaccharide chains. These analogues are of interest for testing as anti-Leishmania drugs.