Highly L and D enantioselective variants of horseradish peroxidase discovered by an ultrahigh-throughput selection method.
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.
Antipov
Eugene
E
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Cho
Art E
AE
Wittrup
K Dane
KD
Klibanov
Alexander M
AM
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
D050505:Mutant Proteins; D006735:Horseradish Peroxidase