Biosynthesis of the Monoterpenes Limonene and Carvone in the Fruit of Caraway¹
I. Demonstration of Enzyme Activities and Their Changes with Development

Harro J. Bouwmeester^*, Jonathan Gershenzon², Maurice C.J.M. Konings, and Rodney Croteau

Research Institute for Agrobiology and Soil Fertility, P.O. Box 14, 6700 AA Wageningen, The Netherlands (H.J.B., M.C.J.M.K.); and Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 (J.G., R.C.)

The biosynthesis of the monoterpenes limonene and carvone in the fruit of caraway (Carum carvi L.) proceeds from geranyl diphosphate via a three-step pathway. First, geranyl diphosphate is cyclized to (+)-limonene by a monoterpene synthase. Second, this intermediate is stored in the essential oil ducts without further metabolism or is converted by limonene-6-hydroxylase to (+)-trans-carveol. Third, (+)-trans-carveol is oxidized by a dehydrogenase to (+)-carvone. To investigate the regulation of monoterpene formation in caraway, we measured the time course of limonene and carvone accumulation during fruit development and compared it with monoterpene biosynthesis from [U-¹⁴C]Suc and the changes in the activities of the three enzymes. The activities of the enzymes explain the profiles of monoterpene accumulation quite well, with limonene-6-hydroxylase playing a pivotal role in controlling the nature of the end product. In the youngest stages, when limonene-6-hydroxylase is undetectable, only limonene was accumulating in appreciable levels. The appearance of limonene-6-hydroxylase correlates closely with the onset of carvone accumulation. At later stages of fruit development, the activities of all three enzymes declined to low levels. Although this correlates closely with a decrease in monoterpene accumulation, the latter may also be the result of competition with other pathways for substrate.

Plant Physiol. (1998) 117: 901-912
Copyright Clearance Center:   0032-0889/98/117/0901/12
© 1998 American Society of Plant Physiologists

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