title: Glycosyltransferase engineering for carbohydrate synthesis.
abstract: Glycosyltransferases (GTs) are powerful tools for the synthesis of complex and biologically-important carbohydrates. Wild-type GTs may not have all the properties and functions that are desired for large-scale production of carbohydrates that exist in nature and those with non-natural modifications. With the increasing availability of crystal structures of GTs, especially those in the presence of donor and acceptor analogues, crystal structure-guided rational design has been quite successful in obtaining mutants with desired functionalities. With current limited understanding of the structure-activity relationship of GTs, directed evolution continues to be a useful approach for generating additional mutants with functionality that can be screened for in a high-throughput format. Mutating the amino acid residues constituting or close to the substrate-binding sites of GTs by structure-guided directed evolution (SGDE) further explores the biotechnological potential of GTs that can only be realized through enzyme engineering. This mini-review discusses the progress made towards GT engineering and the lessons learned for future engineering efforts and assay development.
authors: Department of Chemistry, University of California, Davis, CA 95616, U.S.A.
authors: John B
authors: JB
authors: McArthur
authors: Department of Chemistry, University of California, Davis, CA 95616, U.S.A. xiichen@ucdavis.edu.
authors: Xi
authors: X
authors: Chen
mesh_terms: D002241:Carbohydrates / Q000096:biosynthesis* / Q000737:chemistry; D019020:Directed Molecular Evolution; D057075:Enzyme Assays; D016695:Glycosyltransferases / Q000737:chemistry / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D013329:Structure-Activity Relationship
chemical_list: D002241:Carbohydrates; D016695:Glycosyltransferases
keywords: carbohydrate; directed evolution; enzymatic synthesis; glycosyltransferase; mutagenesis; protein engineering