The oxygen-independent metabolism of cyclic monoterpenes in Castellaniella defragrans 65Phen

BMC Microbiology

Jens Harder¹  Richard Reinhardt⁴  Bruno Hüttel⁴  Thomas Schweder³  Dörte Becher²  Stephanie Markert³  Eva-Maria Disch¹  Jan Petasch¹ 

[1] Department of Microbiology, Max Planck Institute for Marine Microbiology, Celsiusstraße 1, Bremen D-28359, Germany;Department of Microbiology, Ernst-Moritz-Arndt-University, Greifswald, Germany;Department of Pharmaceutical Biotechnology, Ernst-Moritz-Arndt-University, Greifswald, Germany;Max Planck Genome Centre Cologne, Cologne, Germany

关键词: Phellandrene;    Limonene;    Biodegradation;    Isoprenoids;    Monoterpene;   

Others  :  1140938
DOI  :  10.1186/1471-2180-14-164

 received in 2014-02-14, accepted in 2014-06-12,  发布年份 2014

【 摘 要 】

Background

The facultatively anaerobic betaproteobacterium Castellaniella defragrans 65Phen utilizes acyclic, monocyclic and bicyclic monoterpenes as sole carbon source under oxic as well as anoxic conditions. A biotransformation pathway of the acyclic β-myrcene required linalool dehydratase-isomerase as initial enzyme acting on the hydrocarbon. An in-frame deletion mutant did not use myrcene, but was able to grow on monocyclic monoterpenes. The genome sequence and a comparative proteome analysis together with a random transposon mutagenesis were conducted to identify genes involved in the monocyclic monoterpene metabolism. Metabolites accumulating in cultures of transposon and in-frame deletion mutants disclosed the degradation pathway.

Results

Castellaniella defragrans 65Phen oxidizes the monocyclic monoterpene limonene at the primary methyl group forming perillyl alcohol. The genome of 3.95 Mb contained a 70 kb genome island coding for over 50 proteins involved in the monoterpene metabolism. This island showed higher homology to genes of another monoterpene-mineralizing betaproteobacterium, Thauera terpenica 58Eu^T, than to genomes of the family Alcaligenaceae, which harbors the genus Castellaniella. A collection of 72 transposon mutants unable to grow on limonene contained 17 inactivated genes, with 46 mutants located in the two genes ctmAB (cyclic terpene metabolism). CtmA and ctmB were annotated as FAD-dependent oxidoreductases and clustered together with ctmE, a 2Fe-2S ferredoxin gene, and ctmF, coding for a NADH:ferredoxin oxidoreductase. Transposon mutants of ctmA, B or E did not grow aerobically or anaerobically on limonene, but on perillyl alcohol. The next steps in the pathway are catalyzed by the geraniol dehydrogenase GeoA and the geranial dehydrogenase GeoB, yielding perillic acid. Two transposon mutants had inactivated genes of the monoterpene ring cleavage (mrc) pathway. 2-Methylcitrate synthase and 2-methylcitrate dehydratase were also essential for the monoterpene metabolism but not for growth on acetate.

Conclusions

The genome of Castellaniella defragrans 65Phen is related to other genomes of Alcaligenaceae, but contains a genomic island with genes of the monoterpene metabolism. Castellaniella defragrans 65Phen degrades limonene via a limonene dehydrogenase and the oxidation of perillyl alcohol. The initial oxidation at the primary methyl group is independent of molecular oxygen.

【 授权许可】

   
2014 Petasch et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

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