Origin of the Regioselective Fatty-Acid Hydroxylation versus Decarboxylation by a Cytochrome P450 Peroxygenase: What Drives the Reaction to Biofuel Production?

• Published in: Chemistry : a European journal 2016-04, Vol.22 (16), p.5478-5483
• Main Authors: Faponle, Abayomi S., Quesne, Matthew G., de Visser, Sam P.
• Format: Article
• Language: English
• Subjects: Bifurcations; bioengineering; Biofuels; Biomass; Byproducts; Chemistry; Cytochrome P-450 Enzyme System - chemistry; Cytochrome P-450 Enzyme System - metabolism; cytochromes; Decarboxylation; density functional calculations; Enzymes; Fatty acids; Fatty Acids - chemistry; Fatty Acids - metabolism; heme proteins; Hydroxylation; iron; Kinetics; Origins; Oxidation-Reduction; Pathways; Peroxidases - chemistry; Peroxidases - metabolism
• ISSN: 0947-6539; 1521-3765
• DOI: 10.1002/chem.201600739

The cytochromes P450 are heme‐based mono‐oxygenases or peroxygenases involved in vital reaction processes for human health. A recently described P450 per‐oxygenase, OleTJE, converts long‐chain fatty acids to terminal olefins and as such may have biotechnological relevance in biodiesel production. However, the reaction produces significant amounts of α‐ and β‐hydroxylation by‐products, and their origin are poorly understood. Herein, we elucidate through a QM/MM study on the bifurcation pathways how the three possible products are generated and show how the enzyme can be further engineered for optimum desaturase activity. The studies showed that the polarity and the solvent accessibility of the substrate in the binding pocket destabilize the OH‐rebound pathways and kinetically enable a thermodynamically otherwise unfavorable decarboxylation reaction. The origins of the bifurcation pathways are analyzed with valence‐bond models that highlight the differences in reaction mechanism. A detailed quantum mechanics/molecular mechanics study was performed into the origins of the regioselectivity of aliphatic hydroxylation versus decarboxylation of long‐chain fatty acids. The key variables that determine the bifurcation processes were identified, and suggestions on how to bioengineer this enzyme further for optimum biofuel production are given (see scheme).