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Applied and Environmental Microbiology, December 1999, p. 5619-5623, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Engineering of a Stable Whole-Cell Biocatalyst Capable of (S)-Styrene Oxide Formation for Continuous Two-Liquid-Phase Applications

Sven Panke,1 Víctor de Lorenzo,2 Arnë Kaiser,1 Bernard Witholt,1,* and Marcel G. Wubbolts1,dagger

Institute of Biotechnology, Swiss Federal Institute of Technology Zurich, 8093 Zurich, Switzerland,1 and Centro Nacional de Biotecnologia, Campus de Cantoblanco, 28049 Madrid, Spain2

Received 19 July 1999/Accepted 29 September 1999

Recombinant strains of Pseudomonas putida KT2440 carrying genetic expression cassettes with xylene oxygenase- and styrene monooxygenase-encoding genes on their chromosomes could be induced in shaking-flask experiments to specific activities that rivaled those of multicopy-plasmid-based Escherichia coli recombinants. Such strains maintained the introduced styrene oxidation activity in continuous two-liquid-phase cultures for at least 100 generations, although at a lower level than in the shaking-flask experiments. The data suggest that placement of target genes on the chromosome might be a suitable route for the construction of segregationally stable and highly active whole-cell biocatalysts.

* Corresponding author. Mailing address: Institut für Biotechnologie, ETH Zürich, Hönggerberg HPT, CH-8093 Zürich, Switzerland. Phone: 41-1-633 32 86. Fax: 41-1-633 10 51. E-mail: bw{at}biotech.biol.ethz.ch.

dagger Present address: DSM Biotech GmbH, Jülich, Germany.

Applied and Environmental Microbiology, December 1999, p. 5619-5623, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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