title: Production of alkenes and novel secondary products by P450 OleTJEusing novel H2O2-generating fusion protein systems.
abstract: Jeotgalicoccus sp. 8456 OleTJE(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 OleTJE. AldO oxidizes polyols (including glycerol), generating H2O2as a coproduct and facilitating its use for efficient OleTJE-dependent fatty acid decarboxylation. AldO activity is regulatable by polyol substrate titration, enabling control over H2O2supply to minimize oxidative inactivation of OleTJEand 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 OleTJE.
authors.affiliation: Manchester Institute of Biotechnology, Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), School of Chemistry, The University of Manchester, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Matthews
authors.affiliation: Manchester Institute of Biotechnology, Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), School of Chemistry, The University of Manchester, UK.
authors.forename: Kang Lan
authors.initials: KL
authors.lastname: Tee
authors.affiliation: Manchester Institute of Biotechnology, Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), School of Chemistry, The University of Manchester, UK.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Rattray
authors.affiliation: Manchester Institute of Biotechnology, Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), School of Chemistry, The University of Manchester, UK.
authors.forename: Kirsty J
authors.initials: KJ
authors.lastname: McLean
authors.affiliation: Manchester Institute of Biotechnology, Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), School of Chemistry, The University of Manchester, UK.
authors.forename: David
authors.initials: D
authors.lastname: Leys
authors.affiliation: The Westhollow Technology Center, Houston, TX, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Parker
authors.affiliation: Agilent Technologies UK Ltd., Stockport, Cheshire, UK.
authors.forename: Richard T
authors.initials: RT
authors.lastname: Blankley
authors.affiliation: Manchester Institute of Biotechnology, Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), School of Chemistry, The University of Manchester, UK.
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Munro
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D000475:Alkenes / Q000378:metabolism*; D055162:Biocatalysis; D056804:Biofuels; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D003653:Decarboxylation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D015870:Gene Expression; D006861:Hydrogen Peroxide / Q000378:metabolism*; D007218:Industrial Microbiology; D060847:Metabolic Engineering; D019814:Myristic Acid / Q000378:metabolism; D010084:Oxidation-Reduction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D040141:Staphylococcaceae / Q000201:enzymology / Q000235:genetics; D048372:Streptomyces coelicolor / Q000201:enzymology / Q000235:genetics
chemical_list: D000475:Alkenes; D056804:Biofuels; D005227:Fatty Acids; D011993:Recombinant Fusion Proteins; D019814:Myristic Acid; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide; D000429:Alcohol Oxidoreductases; C515743:alditol oxidase (FAD), Streptomyces coelicolor
keywords: CYP152L1; OleTJE peroxygenase; alkene production; cytochrome P450; fusion enzymes; hydrogen peroxide