Hydrogen peroxide-independent production of α-alkenes by OleTJE P450 fatty acid decarboxylase

Biotechnology for Biofuels

Yi Liu¹  Cong Wang²  Jinyong Yan²  Wei Zhang²  Wenna Guan²  Xuefeng Lu²  Shengying Li² 

[1] University of Chinese Academy of Sciences, Beijing 100049, China

[2] 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

关键词: Peroxygenase;    P450 fatty acid decarboxylase;    Monooxygenase;    Biofuels;    Alkenes;   

Others  :  793478
DOI  :  10.1186/1754-6834-7-28

 received in 2013-11-21, accepted in 2014-02-10,  发布年份 2014

【 摘 要 】

Background

Cytochrome P450 OleT_JE 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 OleT_JE 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 OleT_JE 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 OleT_JE or OleT_JE-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 OleT_JE 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 OleT_JE.

【 授权许可】

   
2014 Liu et al.; licensee BioMed Central Ltd.

【 图 表 】

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