Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 105, Issue 46, Pages 17694-17699Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0809851105
Keywords
directed evolution; enzyme design; molecular modeling; redox enzymes; stereoselectivity
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Funding
- National Institutes of Health [GM66712]
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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.
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