In vivo ESR spin trapping detection of carbon-centered alpha-Farnesene radicals

Lyophilisates of the ascomycetes Penicillium solitum and Aspergillus niger converted alpha-farnesene to 7-hydroxyfarnesene as the major product. The radical mechanism of this bioconversion was proven by electron spin resonance (ESR) and GC-MS using the spin trapping technique. Intermediate carbon-centered radicals of alpha-farnesene were captured using two spin traps, 2-methyl-2-nitrosopropane and alpha-(4-pyridyl-1-oxide)- N- tert-butylnitrone, respectively. The evaluation of the coupling constants and hyperfine couplings of the ESR spectra showed that tertiary carbon radicals were trapped. The radical position at C7 of alpha-farnesene was derived from EI and CI mass spectra of the corresponding MNP spin adduct. The present study demonstrates that the complementary application of ESR and MS spectrometric data allows the detailed evaluation of a radical mechanism of a fungal terpene transformation reaction.