title: Hydrogen peroxide-independent production of α-alkenes by OleTJE P450 fatty acid decarboxylase.
abstract: BACKGROUND
  Cytochrome P450 OleTJE from Jeotgalicoccus sp. ATCC 8456, a new member of the CYP152 peroxygenase family, was recently found to catalyze the unusual decarboxylation of long-chain fatty acids to form α-alkenes using H2O2 as the sole electron and oxygen donor. Because aliphatic α-alkenes are important chemicals that can be used as biofuels to replace fossil fuels, or for making lubricants, polymers and detergents, studies on OleTJE fatty acid decarboxylase are significant and may lead to commercial production of biogenic α-alkenes in the future, which are renewable and more environmentally friendly than petroleum-derived equivalents.
  
  
  RESULTS
  We report the H2O2-independent activity of OleTJE for the first time. In the presence of NADPH and O2, this P450 enzyme efficiently decarboxylates long-chain fatty acids (C12 to C20) in vitro when partnering with either the fused P450 reductase domain RhFRED from Rhodococcus sp. or the separate flavodoxin/flavodoxin reductase from Escherichia coli. In vivo, expression of OleTJE or OleTJE-RhFRED in different E. coli strains overproducing free fatty acids resulted in production of variant levels of multiple α-alkenes, with a highest total hydrocarbon titer of 97.6 mg·l-1.
  
  
  CONCLUSIONS
  The discovery of the H2O2-independent activity of OleTJE not only raises a number of fundamental questions on the monooxygenase-like mechanism of this peroxygenase, but also will direct the future metabolic engineering work toward improvement of O2/redox partner(s)/NADPH for overproduction of α-alkenes by OleTJE.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Cong
authors.initials: C
authors.lastname: Wang
authors.forename: Jinyong
authors.initials: J
authors.lastname: Yan
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Wenna
authors.initials: W
authors.lastname: Guan
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Lu
authors.affiliation: Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No, 189 Songling Road, Qingdao, Shandong 266101, China. lishengying@qibebt.ac.cn.
authors.forename: Shengying
authors.initials: S
authors.lastname: Li