Flavoproteins in Directed Evolution

Kille, Sabrina

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Flavoproteins in Directed Evolution

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Kille, Sabrina
Research Department Reetz, Max-Planck-Institut für Kohlenforschung, Max Planck Society;

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Citation

Kille, S. (2010). Flavoproteins in Directed Evolution. PhD Thesis, Ruhr-Universität Bochum, Bochum.

Cite as: https://hdl.handle.net/11858/00-001M-0000-000F-8D0D-4

Abstract

Two enzymes, an enoate reductase (YqjM) and a monooxygenase (P450_BM3), were
successfully altered towards acceptance of new substrates applying directed evolution. Using iterative CASTing (combinatorial active site saturation test) highly regio- and stereoselective mutants were obtained in both cases. Additionally it was possible to control the regioselectivity (P450_BM3) and enantioselectivity (YqjM) of the catalyzed reaction.
Moreover this work was used to shed more light on the iterative CASTing methodology and its application and the gained insights contribute to a more efficient application of the method.