Engineering of a stable whole-cell biocatalyst capable of (S)-styrene oxide formation for continuous two-liquid-phase applications.
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.
Panke
S
S
Institute of Biotechnology, Swiss Federal Institute of Technology Zurich, 8093 Zurich, Switzerland.
de Lorenzo
V
V
Kaiser
A
A
Witholt
B
B
Wubbolts
M G
MG
D001431:Bacteriological Techniques; D002384:Catalysis; D004251:DNA Transposable Elements; D004852:Epoxy Compounds / Q000138:chemical synthesis* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005818:Genetic Engineering; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D016254:Mutagenesis, Insertional; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics*; D011995:Recombination, Genetic; D014169:Transformation, Bacterial
D004251:DNA Transposable Elements; D004852:Epoxy Compounds; C013690:styrene oxide; D006899:Mixed Function Oxygenases; D010105:Oxygenases; C026721:styrene monooxygenase; C026267:4-xylene methylhydroxylase