Production of alkenes and novel secondary products by P450 Ole<scp>T_JE</scp> using novel H₂O₂‐generating fusion protein systems

Jeotgalicoccus sp. 8456 OleT_JE (CYP152L1) is a fatty acid decarboxylase cytochrome P450 that uses hydrogen peroxide (H₂ O₂ ) 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 H₂ O₂ 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 H₂ O₂ 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 .