IMMOBILIZATION OF CYCLASE ENZYMES FOR THE PRODUCTION OF MONOTERPENES AND SESQUITERPENES

The first steps in the biosynthesis of most monoterpenes and sesquiterpenes are catalysed by a novel class of enzymes termed cyclases. Several techniques were tested for the immobilization of a partially purified monoterpene cyclase from Foeniculum vulgare [geranyl pyrophosphate:(-)-endo-fenchol cyclase] and a partially purified sesquiterpene cylase from Pogostemon cablin [farnesyl pyrophosphate:(-)-patchoulol cyclase]. Although both enzymes could be immobilized by entrapment and adsorption methods, the most efficient immobilization, in terms of activity recovery, was achieved by covalent attachment involving coupling of the protein amino groups to N-hydroxysuccinimide-activated 6-aminohexanoic acid Sepharose 4B. The immobilized forms of these cyclases were markedly more stable in storage than the free forms. © 1990.