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Applied and Environmental Microbiology, June 1999, p. 2644-2649, Vol. 65, No. 6
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

cis/trans Isomerase of Unsaturated Fatty Acids of Pseudomonas putida P8: Evidence for a Heme Protein of the Cytochrome c Typedagger

Rita Holtwick, Heribert Keweloh, and Friedhelm Meinhardt*

Institut für Mikrobiologie, Westfälische Wilhelms-Universität Münster, 48149 Münster, Germany

Received 14 December 1998/Accepted 29 March 1999

From a pool of 600 temperature-sensitive transposon mutants of Pseudomonas putida P8, 1 strain was isolated that carries a mini-Tn5 insertion within the cytochrome c operon. As a result, genes involved in the attachment of heme to cytochrome c-type proteins are turned off. Accordingly, cytochrome c could not be detected spectrophotometrically. The mutant also exhibited a remarkable reduction of cis-trans isomerization capability for unsaturated fatty acids. Consistent with the genetic and physiological data is the detection of a cytochrome c-type heme-binding motif close to the N terminus of the predicted polypeptide of the cis/trans isomerase (cti) gene (CVACH; conserved amino acids in italics). The functional significance of this motif was proven by site-directed mutagenesis. A possible mechanism of heme-catalyzed cis-trans isomerization of unsaturated fatty acids is discussed.

* Corresponding author. Mailing address: Institut für Mikrobiologie, Westfälische Wilhelms-Universität Münster, Corrensstr. 3, 48149 Münster, Germany. Phone: 49-251-833-9825. Fax: 49-251-833-8388. E-mail: meinhar{at}uni-muenster.de.

dagger This work is dedicated to Karl Esser on the occasion of his 75th birthday.

Applied and Environmental Microbiology, June 1999, p. 2644-2649, Vol. 65, No. 6
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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