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  • Geraniol and geranial dehydrogenases induced in anaerobic monoterpene degradation by Castellaniella defragrans.

Geraniol and geranial dehydrogenases induced in anaerobic monoterpene degradation by Castellaniella defragrans.

Applied and environmental microbiology (2012-01-31)
Frauke Lüddeke, Annika Wülfing, Markus Timke, Frauke Germer, Johanna Weber, Aytac Dikfidan, Tobias Rahnfeld, Dietmar Linder, Anke Meyerdierks, Jens Harder
ABSTRACT

Castellaniella defragrans is a Betaproteobacterium capable of coupling the oxidation of monoterpenes with denitrification. Geraniol dehydrogenase (GeDH) activity was induced during growth with limonene in comparison to growth with acetate. The N-terminal sequence of the purified enzyme directed the cloning of the corresponding open reading frame (ORF), the first bacterial gene for a GeDH (geoA, for geraniol oxidation pathway). The C. defragrans geraniol dehydrogenase is a homodimeric enzyme that affiliates with the zinc-containing benzyl alcohol dehydrogenases in the superfamily of medium-chain-length dehydrogenases/reductases (MDR). The purified enzyme most efficiently catalyzes the oxidation of perillyl alcohol (k(cat)/K(m) = 2.02 × 10(6) M(-1) s(-1)), followed by geraniol (k(cat)/K(m) = 1.57 × 10(6) M(-1) s(-1)). Apparent K(m) values of <10 μM are consistent with an in vivo toxicity of geraniol above 5 μM. In the genetic vicinity of geoA is a putative aldehyde dehydrogenase that was named geoB and identified as a highly abundant protein during growth with phellandrene. Extracts of Escherichia coli expressing geoB demonstrated in vitro a geranial dehydrogenase (GaDH) activity. GaDH activity was independent of coenzyme A. The irreversible formation of geranic acid allows for a metabolic flux from β-myrcene via linalool, geraniol, and geranial to geranic acid.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
cis-3,7-Dimethyl-2,6-octadien-1-ol, 97%
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Sigma-Aldrich
cis-3,7-Dimethyl-2,6-octadien-1-ol, ≥97%, FCC, FG
Sigma-Aldrich
Citral, 95%
Sigma-Aldrich
Geraniol, natural, ≥97%, FG
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