Biosynthesis of menthofuran in Mentha x piperita: stereoselective and mechanistic studies.

Mentha x piperita shoot tips and first leaf pair were fed with aqueous solutions of [(2)H(2)]- and [(2)H(2)]/[(18)O]-labeled pulegone. The essential oil was analyzed by solid phase microextraction and enantioselective multidimensional gas chromatography/mass spectrometry. After feeding experiments with labeled pulegone racemate, both labeled (S)-menthofuran and (R)-menthofuran were detectable simultaneously together with genuine (R)-menthofuran. It could be shown that both labeled pulegone enantiomers are converted by Mentha x piperita to the corresponding labeled menthofuran enantiomers, favoring the labeled analogue of the nongenuine (S)-pulegone. The oxygen in menthofuran is introduced by enzymatic oxidation of pulegone, as concluded from feeding experiments with mixed labeled [(2)H(2)]/[(18)O]pulegone.