Production of alkenes and novel secondary products by P450 OleTJE using novel H2O2-generating fusion protein systems

Jeotgalicoccus sp. 8456 OleT(JE) (CYP152L1) is a fatty acid decarboxylase cytochrome P450 that uses hydrogen peroxide (H2O2) 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(JE). AldO oxidizes polyols (including glycerol), generating H2O2 as a coproduct and facilitating its use for efficient OleT(JE)-dependent fatty acid decarboxylation. AldO activity is regulatable by polyol substrate titration, enabling control over H2O2 supply to minimize oxidative inactivation of OleT(JE) 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(JE).