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Microbial degradation of monoterpenes in the absence of molecular oxygen

Anaerobic degradation of natural monoterpenes by microorganisms was evaluated by using Pseudomonas citronellolis DSM 50332 and enrichment cultures containing nitrate as an electron acceptor. P. citronellolis grew anaerobically on 3,7-dimethyl-1-octanol and citronellol but not on geraniol, nerol, and alicyclic monoterpenes. In contrast, several a-, mono-, and bicyclic monoterpenes supported microbial growth and denitrification in enrichment cultures. We found that consumption of linalool, menthol, menth-1-ene, alpha-phellandrene, limonene, 2-carene, alpha-pinene, and fenchone in enrichment cultures depended on the presence of living microorganisms and nitrate. In these experiments, the ratios of number of electrons derived from complete substrate oxidation to number of electrons derived from nitrate reduction ranged from 1.2:1 to 2.9:1. Microbial degradation was accompanied by the formation of small traces of monoterpenes, which were characterized by gas chromatography-mass spectroscopy. The formation of geraniol and geranial from linalool suggested that a 3,1-hydroxyl-delta 1-delta 2-mutase reaction initiates linalool degradation. Seven strains of motile, oval to rod-shaped, facultatively denitrifying bacteria were isolated on agar bottle plates by using linalool, menthol, menth-1-ene, alpha-phellandrene, 2-carene, eucalyptol, and alpha-pinene as sole carbon and energy sources.