A direct electrode-driven P450 cycle for biocatalysis.
The large potential of redox enzymes to carry out formation of high value organic compounds motivates the search for innovative strategies to regenerate the cofactors needed by their biocatalytic cycles. Here, we describe a bioreactor where the reducing power to the cycle is supplied directly to purified cytochrome CYP101 (P450cam; EC 1.14.15.1) through its natural redox partner (putidaredoxin) using an antimony-doped tin oxide working electrode. Required oxygen was produced at a Pt counter electrode by water electrolysis. A continuous catalytic cycle was sustained for more than 5 h and 2,600 enzyme turnovers. The maximum product formation rate was 36 nmol of 5-exo-hydroxycamphor/nmol of CYP101 per min.
Reipa
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Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.
Mayhew
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Vilker
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D019149:Bioreactors; D002164:Camphor; D019475:Camphor 5-Monooxygenase / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D004566:Electrodes; D004579:Electron Transport; D004867:Equipment Design; D004926:Escherichia coli / Q000235:genetics; D010084:Oxidation-Reduction; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017971:Tin Compounds
D011994:Recombinant Proteins; D017971:Tin Compounds; D002164:Camphor; D019475:Camphor 5-Monooxygenase; C045358:stannic oxide