title: Highly L and D enantioselective variants of horseradish peroxidase discovered by an ultrahigh-throughput selection method.
abstract: A highly efficient selection method for enhanced enzyme enantioselectivity based on yeast surface display and fluorescence-activated cell sorting (FACS) is developed and validated. Its application to horseradish peroxidase has resulted in enzyme variants up to 2 orders of magnitude selective toward either substrate enantiomer at will. These marked improvements in enantioselectivity are demonstrated for the surface-bound and soluble enzymes and rationalized by computational docking studies.
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Eugene
authors.initials: E
authors.lastname: Antipov
authors.forename: Art E
authors.initials: AE
authors.lastname: Cho
authors.forename: K Dane
authors.initials: KD
authors.lastname: Wittrup
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Klibanov
mesh_terms: D005434:Flow Cytometry / Q000379:methods*; D006735:Horseradish Peroxidase / Q000737:chemistry*; D008958:Models, Molecular; D050505:Mutant Proteins / Q000737:chemistry*; D010084:Oxidation-Reduction; D012441:Saccharomyces cerevisiae; D012995:Solubility; D013237:Stereoisomerism; D013379:Substrate Specificity; D013499:Surface Properties
chemical_list: D050505:Mutant Proteins; D006735:Horseradish Peroxidase