Biological Chemistry

Carbohydrate-containing molecules are widespread throughout nature and have a multitude of biological roles. Their chemical synthesis is challenging and, therefore, novel enzymatic methods for the synthesis of carbohydrate-containing molecules offer stereoselective and regioselective advantages. The proposed research will advance methods to produce novel carbohydrate-containing molecules by altering the substrate specificity of glycosynthases involved in the regio- and stereoselective synthesis of glycosidic linkages. It will use a novel, positive selection strategy, based on bacterial cell death to select mutant glycosynthases with modified substrate specificity. There exists a strong need to develop such methods to fully realize the potential of enzymes to produce carbohydrate-containing molecules, and the significance of developing methods to alter enzyme substrate specificity for enzyme systems that couple synthetic fragments cannot be understated. The proposed research will: (a) make fundamental discoveries in evolving novel enzyme function; (b) develop the ability to generate a positive selection method for enzymatic bond formation and (c) result in the characterization of novel glycosidases and glycosynthases. Funds are requested to enable a graduate student and PDF to conduct the research under the supervision of Dr. Jakeman.