Growth and monoterpene production by transformed shoot cultures of Mentha citrata and Mentha piperita in flasks and fermenters

This paper reports studies on the growth and biosynthesis of monoterpenes by transformed shoot cultures of Mentha citrata and Mentha piperita, originally developed 5 years ago and since maintained by regular subculturing. Throughout this time, the M. citrata culture has stably maintained production of an oil closely resembling that of the parent plant in which linalool and linalyl acetate are the predominant components. However, M. piperita, which initially showed a divergence from the parent plant in producing significant amounts of menthofuran in addition to the characteristic oil components menthol and menthone, has now been found to produce pulegone and menthofuran as the major components. The cultures were subjected to different environmental conditions of varying periods of light and temperature in an attempt to restore menthol and menthone production. Increased illumination reduced the yields of pulegone and menthofuran but did not stimulate the production of either menthol or menthone, which remained only at trace levels (below 0.2 micrograms/g fresh weight). Cultures of M. citrata were, however, stimulated by increased illumination, and produced more linalool and linalyl acetate. Shoot cultures of M. citrata and M. piperita were grown in 14-1 fermenters for up to 60 days during which the biomass increased from approximately 100 g to 2.5 kg and 3.5 kg respectively. Both cultures rapidly consumed sucrose with a concomitant release of glucose, and the uptake of inorganic ions was similar except that M. citrata consumed far less Na+ during the fermentation. The total yields of monoterpenes from the fermentations were 1.16 g (M. piperita) and 0.18 g (M. citrata).