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Appl. Environ. Microbiol., Nov 1997, 4292-4297, Vol 63, No. 11
Copyright © 1997, American Society for Microbiology

cis-trans Isomerization of Unsaturated Fatty Acids: Cloning and Sequencing of the cti Gene from Pseudomonas putida P8

R Holtwick, F Meinhardt and H Keweloh
Institut fur Mikrobiologie, Westfalische-Wilhelms-Universitat Munster, 48149 Munster, Germany

Transposon mutants of Pseudomonas putida P8 were generated by applying a mini-Tn5 mutagenesis system. The mutants obtained were checked for their ability to tolerate increased temperatures and elevated phenol concentrations. Approximately 5,800 transposon mutants were used to generate a pool of 600 temperature-sensitive strains; one of these strains was identified as being damaged in its ability to perform cis-trans isomerization of fatty acids. A gene library of P. putida P8 was constructed and screened by using as a probe sequences immediately adjacent to the mini-Tn5 insertion. A DNA fragment that complemented the mutation was isolated and cloned. The corresponding gene, termed cti, is located close to the methionine synthase locus (metH) in P. putida P8. A cti-carrying fragment integrated into a plasmid also conferred the ability for cis-trans isomerization to Escherichia coli; the cti gene was completely sequenced, and the amino acid sequence was deduced.

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