title: Whole-Cell-Catalyzed Multiple Regio- and Stereoselective Functionalizations in Cascade Reactions Enabled by Directed Evolution.
abstract: Biocatalytic cascade reactions using isolated stereoselective enzymes or whole cells in one-pot processes lead to value-added chiral products in a single workup. The concept has been restricted mainly to starting materials and intermediate products that are accepted by the respective wild-type enzymes. In the present study, we exploited directed evolution as a means to create E. coli whole cells for regio- and stereoselective cascade sequences that are not possible using man-made catalysts. The approach is illustrated using P450-BM3 in combination with appropriate alcohol dehydrogenases as catalysts in either two-, three-, or four-step cascade reactions starting from cyclohexane, cyclohexanol, or cyclohexanone, respectively, leading to either (R,R)-, (S,S)-, or meso-cyclohexane-1,2-diol. The one-pot conversion of cyclohexane into (R)- or (S)-2-hydroxycyclohexanone in the absence of ADH is also described.
authors: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470, Mülheim an der Ruhr, Germany.|Fachbereich Chemie der Philipps-Universität, Hans-Meerwein-Strasse, 35032, Marburg, Germany.
authors: Aitao
authors: A
authors: Li
authors: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470, Mülheim an der Ruhr, Germany.|Fachbereich Chemie der Philipps-Universität, Hans-Meerwein-Strasse, 35032, Marburg, Germany.
authors: Adriana
authors: A
authors: Ilie
authors: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470, Mülheim an der Ruhr, Germany.|Fachbereich Chemie der Philipps-Universität, Hans-Meerwein-Strasse, 35032, Marburg, Germany.
authors: Zhoutong
authors: Z
authors: Sun
authors: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470, Mülheim an der Ruhr, Germany.|Fachbereich Chemie der Philipps-Universität, Hans-Meerwein-Strasse, 35032, Marburg, Germany.
authors: Richard
authors: R
authors: Lonsdale
authors: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
authors: Jian-He
authors: JH
authors: Xu
authors: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470, Mülheim an der Ruhr, Germany. reetz@mpi-muelheim.mpg.de.|Fachbereich Chemie der Philipps-Universität, Hans-Meerwein-Strasse, 35032, Marburg, Germany. reetz@mpi-muelheim.mpg.de.
authors: Manfred T
authors: MT
authors: Reetz
keywords: C−H activation; P450 monooxygenase; cascade reactions; directed evolution; enantioselectivity