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  Physiology of deletion mutants in the anaerobic beta-myrcene degradation pathway in Castellaniella defragrans

Lueddeke, F., Dikfidan, A., & Harder, J. (2012). Physiology of deletion mutants in the anaerobic beta-myrcene degradation pathway in Castellaniella defragrans. BMC Microbiology, 12: 192.

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 Urheber:
Lueddeke, F.1, Autor           
Dikfidan, A.1, Autor           
Harder, J.1, Autor           
Affiliations:
1Department of Microbiology, Max Planck Institute for Marine Microbiology, Max Planck Society, ou_2481695              

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 Zusammenfassung: Background

Monoterpenes present a large and versatile group of unsaturated hydrocarbons of plant origin with widespread use in the fragrance as well as food industry. The anaerobic β-myrcene degradation pathway in Castellaniella defragrans strain 65Phen differs from well known aerobic, monooxygenase-containing pathways. The initial enzyme linalool dehydratase-isomerase ldi/LDI catalyzes the hydration of β-myrcene to (S)-(+)-linalool and its isomerization to geraniol. A high-affinity geraniol dehydrogenase geoA/GeDH and a geranial dehydrogenase geoB/GaDH contribute to the formation of geranic acid.

A genetic system was for the first time applied for the betaproteobacterium to prove in vivo the relevance of the linalool dehydratase-isomerase and the geraniol dehydrogenase. In-frame deletion cassettes were introduced by conjugation and two homologous recombination events.
Results

Polar effects were absent in the in-frame deletion mutants C. defragrans Δldi and C. defragrans ΔgeoA. The physiological characterization of the strains demonstrated a requirement of the linalool dehydratase-isomerase for growth on acyclic monoterpenes, but not on cyclic monoterpenes. The deletion of geoA resulted in a phenotype with hampered growth rate on monoterpenes as sole carbon and energy source as well as reduced biomass yields. Enzyme assays revealed the presence of a second geraniol dehydrogenase. The deletion mutants were in trans complemented with the broad-host range expression vector pBBR1MCS-4ldi and pBBR1MCS-2geoA, restoring in both cases the wild type phenotype.
Conclusions

In-frame deletion mutants of genes in the anaerobic β-myrcene degradation revealed novel insights in the in vivo function. The deletion of a high-affinity geraniol dehydrogenase hampered, but did not preclude growth on monoterpenes. A second geraniol dehydrogenase activity was present that contributes to the β-myrcene degradation pathway. Growth on cyclic monoterpenes independent of the initial enzyme LDI suggests the presence of a second enzyme system activating unsaturated hydrocarbons.

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Sprache(n): eng - English
 Datum: 2012-09-04
 Publikationsstatus: Erschienen
 Seiten: 11
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: eDoc: 634621
ISI: 000310876600001
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Titel: BMC Microbiology
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: BioMed Central
Seiten: - Band / Heft: 12 Artikelnummer: 192 Start- / Endseite: - Identifikator: ISSN: 1471-2180
CoNE: https://pure.mpg.de/cone/journals/resource/111000136905014