Inhibition of Nitrosomonas europaea by monoterpenes from coastal redwood (Sequoia sempervirens) in whole-cell studies

Inhibition by allelochemicals, including monoterpenes, has been suggested as a factor in the extremely low nitrification rates observed in coastal redwood forests. Similarities between the molecular structure of known nitrification inhibitors and some conifer monoterpenes have been suggested as one reason for the inhibition of autotrophic nitrifiers by conifer monoterpenes. The effect of monoterpenes on nitrification rate and growth of Nitrosomonas europaea was examined in whole-cell pure culture experiments using the five most abundant monoterpenes in coastal redwood needles. These are (in order of decreasing concentration in the needles) limonene, alpha-pinene, sabinene, myrcene, and gamma-terpinene. Four of the five compounds significantly inhibited growth of N. europaea in batch culture experiments. Short-term kinetic studies of the two most inhibitory monoterpenes, limonene and alpha-pinene, were performed on whole cells to evaluate the mode of interaction between these chemicals and nitrification rates. Inhibition constants (K-i) of limonene (38 mu M) and alpha-pinene (95 mu M) were determined. Lineweaver-Burk plots of nitrification in the presence of monoterpenes appear to fit a noncompetitive inhibition model; however, the mechanisms of inhibition may be more complex.