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Applied and Environmental Microbiology, July 2006, p. 4819-4828, Vol. 72, No. 7
0099-2240/06/$08.00+0     doi:10.1128/AEM.00853-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of Genes and Proteins Necessary for Catabolism of Acyclic Terpenes and Leucine/Isovalerate in Pseudomonas aeruginosa

Karin Förster-Fromme,^1, Birgit Höschle,^1, Christina Mack,² Michael Bott,² Wolfgang Armbruster,³ and Dieter Jendrossek^1*

Institut für Mikrobiologie, Universität Stuttgart, Stuttgart, Germany,¹ Institut für Biotechnologie 1, Forschungszentrum Jülich, Jülich, Germany;,² Institut für Lebensmittelchemie, Universität Hohenheim, Stuttgart-Hohenheim, Germany³

Received 11 April 2006/ Accepted 16 May 2006

Geranyl-coenzyme A (CoA)-carboxylase (GCase; AtuC/AtuF) and methylcrotonyl-CoA-carboxylase (MCase; LiuB/LiuD) are characteristic enzymes of the catabolic pathway of acyclic terpenes (citronellol and geraniol) and of saturated methyl-branched compounds, such as leucine or isovalerate, respectively. Proteins encoded by two gene clusters (atuABCDEFGH and liuRABCDE) of Pseudomonas aeruginosa PAO1 were essential for acyclic terpene utilization (Atu) and for leucine and isovalerate utilization (Liu), respectively, as revealed by phenotype analysis of 10 insertion mutants, two-dimensional gel electrophoresis, determination of GCase and MCase activities, and Western blot analysis of wild-type and mutant strains. Analysis of the genome sequences of other pseudomonads (P. putida KT2440 and P. fluorescens Pf-5) revealed candidate genes for Liu proteins for both species and candidate genes for Atu proteins in P. fluorescens. This result concurred with the finding that P. fluorescens, but not P. putida, could grow on acyclic terpenes (citronellol and citronellate), while both species were able to utilize leucine and isovalerate. A regulatory gene, atuR, was identified upstream of atuABCDEFGH and negatively regulated expression of the atu gene cluster.

These authors contributed equally to this work.

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