Terpene cyclase catalysis in organic solvent/minimal water media: demonstration and optimization of (+)-alpha-pinene cyclase activity.

A partially purified and lyophilized preparation of (+)-alpha-pinene cyclase from sage (Salvia officinalis) was shown to convert geranyl pyrophosphate to the monoterpene olefins alpha-pinene, camphene, limonene, and myrcene, in hexane with the addition of 0.1 to 10% water. This constitutes the first report of catalysis by a terpene cyclase in "non-aqueous" media. The relative proportions of olefinic products, metal ion requirement, pH optimum, temperature response, and time course of the enzymatic conversion were determined. The cyclase was shown to be stabilized with respect to temperature and time by the use of hydrocarbon solvent, and, in all other characteristics, to exhibit properties closely resembling those observed in aqueous media.