Production of alkenes and novel secondary products by P450 OleT JE using novel H 2 O 2 -generating fusion protein systems

Jeotgalicoccus sp. 8456 OleT (CYP152L1) is a fatty acid decarboxylase cytochrome P450 that uses hydrogen peroxide (H O ) to catalyse production of terminal alkenes, which are industrially important chemicals with biofuel applications. We report enzyme fusion systems in which Streptomyces coelicolor alditol oxidase (AldO) is linked to OleT . AldO oxidizes polyols (including glycerol), generating H O as a coproduct and facilitating its use for efficient OleT -dependent fatty acid decarboxylation. AldO activity is regulatable by polyol substrate titration, enabling control over H O supply to minimize oxidative inactivation of OleT and prolong activity for increased alkene production. We also use these fusion systems to generate novel products from secondary turnover of 2-OH and 3-OH myristic acid primary products, expanding the catalytic repertoire of OleT .