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The Enigmatic P450 Decarboxylase OleT Is Capable of, but Evolved To Frustrate, Oxygen Rebound Chemistry

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Department of Chemistry and Biochemistry, University of South Carolina, Columbia, South Carolina 29208, United States
Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States
§ Department of Chemistry, Barnard College, Columbia University, New York, New York 10027, United States
*Department of Chemistry and Biochemistry, University of South Carolina, 631 Sumter St., Columbia, SC 29208. Telephone: 803-777-6626. Fax: 803-777-9521. E-mail: [email protected]
Cite this: Biochemistry 2017, 56, 26, 3347–3357
Publication Date (Web):June 12, 2017
Copyright © 2017 American Chemical Society

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    OleT is a cytochrome P450 enzyme that catalyzes the removal of carbon dioxide from variable chain length fatty acids to form 1-alkenes. In this work, we examine the binding and metabolic profile of OleT with shorter chain length (n ≤ 12) fatty acids that can form liquid transportation fuels. Transient kinetics and product analyses confirm that OleT capably activates hydrogen peroxide with shorter substrates to form the high-valent intermediate Compound I and largely performs C–C bond scission. However, the enzyme also produces fatty alcohol side products using the high-valent iron oxo chemistry commonly associated with insertion of oxygen into hydrocarbons. When presented with a short chain fatty acid that can initiate the formation of Compound I, OleT oxidizes the diagnostic probe molecules norcarane and methylcyclopropane in a manner that is reminiscent of reactions of many CYP hydroxylases with radical clock substrates. These data are consistent with a decarboxylation mechanism in which Compound I abstracts a substrate hydrogen atom in the initial step. Positioning of the incipient substrate radical is a crucial element in controlling the efficiency of activated OH rebound.

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.biochem.7b00338.

    • Additional results, including UV–vis binding titrations of C6, C8, C10, and C12 FAs (Figure S1), gas chromatogram of the reaction headspace from OleT:DA reactions (Figure S2), gas chromatograms of the products of OleT reactions of C8, C10, and C12 FAs (Figure S3), time course and fitting for Ole-I decay with DA-d19 (Figure S4), decoy CL screening using guaiacol oxidation as a probe (Figure S5), gas chromatograms and reaction products from MCP oxidations (Figure S6), and MS of ring-opened norcarane reaction products (Figure S7) (PDF)

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    16. Olivia M. Manley, Ruixi Fan, Yisong Guo, Thomas M. Makris. Oxidative Decarboxylase UndA Utilizes a Dinuclear Iron Cofactor. Journal of the American Chemical Society 2019, 141 (22) , 8684-8688.
    17. Kaustuv Mittra, Michael T. Green. Reduction Potentials of P450 Compounds I and II: Insight into the Thermodynamics of C–H Bond Activation. Journal of the American Chemical Society 2019, 141 (13) , 5504-5510.
    18. Mathias Pickl, Sara Kurakin, Fabián G. Cantú Reinhard, Philipp Schmid, Alexander Pöcheim, Christoph K. Winkler, Wolfgang Kroutil, Sam P. de Visser, Kurt Faber. Mechanistic Studies of Fatty Acid Activation by CYP152 Peroxygenases Reveal Unexpected Desaturase Activity. ACS Catalysis 2019, 9 (1) , 565-577.
    19. Courtney E. Wise, Chun H. Hsieh, Nathan L. Poplin, Thomas M. Makris. Dioxygen Activation by the Biofuel-Generating Cytochrome P450 OleT. ACS Catalysis 2018, 8 (10) , 9342-9352.
    20. F. Peter Guengerich, Francis K. Yoshimoto. Formation and Cleavage of C–C Bonds by Enzymatic Oxidation–Reduction Reactions. Chemical Reviews 2018, 118 (14) , 6573-6655.
    21. Chen Lu, Fenglin Shen, Shuaibo Wang, Yuyang Wang, Juan Liu, Wen-Ju Bai, Xiqing Wang. An Engineered Self-Sufficient Biocatalyst Enables Scalable Production of Linear α-Olefins from Carboxylic Acids. ACS Catalysis 2018, 8 (7) , 5794-5798.
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    23. Xiongyi Huang, John T. Groves. Oxygen Activation and Radical Transformations in Heme Proteins and Metalloporphyrins. Chemical Reviews 2018, 118 (5) , 2491-2553.
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    27. Osami Shoji, Yuichiro Aiba, Shinya Ariyasu, Hiroki Onoda. Heme-containing proteins: Structures, functions, and engineering. 2023, 194-214.
    28. Yuanyuan Jiang, Shengying Li. P450 fatty acid decarboxylase. 2023, 339-374.
    29. Rodolpho R.C. Monteiro, Silvia S.O. da Silva, Célio L. Cavalcante, F. Murilo T. de Luna, Juan M. Bolivar, Rodrigo S. Vieira, Roberto Fernandez-Lafuente. Biosynthesis of alkanes/alkenes from fatty acids or derivatives (triacylglycerols or fatty aldehydes). Biotechnology Advances 2022, 61 , 108045.
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    36. Yuanyuan Jiang, Wei Peng, Zhong Li, Cai You, Yue Zhao, Dandan Tang, Binju Wang, Shengying Li. Unexpected Reactions of α,β‐Unsaturated Fatty Acids Provide Insight into the Mechanisms of CYP152 Peroxygenases. Angewandte Chemie International Edition 2021, 60 (46) , 24694-24701.
    37. Libo Zhang, Dumei Ma, Yingwu Yin, Qian Wang. Using Small Molecules to Enhance P450 OleT Enzyme Activity in Situ. Chemistry – A European Journal 2021, 27 (35) , 8940-8945.
    38. Dumei Ma, Libo Zhang, Yingwu Yin, Qian Wang. Structure-based design, synthesis of novel probes for cytochrome P450 OleT. Chinese Chemical Letters 2021, 32 (4) , 1466-1469.
    39. Bekir Engin Eser, Yan Zhang, Li Zong, Zheng Guo. Self-sufficient Cytochrome P450s and their potential applications in biotechnology. Chinese Journal of Chemical Engineering 2021, 30 , 121-135.
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    41. Olivia M. Manley, Thomas M. Makris. Cytochrome P450 Enzyme Mechanisms. 2021, 254-268.
    42. Julia Armbruster, Mathilde Steinmassl, Christina A. Müller Bogotá, Gabriele Berg, Bernd Nidetzky, Alexander Dennig. P450 Jα : A New, Robust and α‐Selective Fatty Acid Hydroxylase Displaying Unexpected 1‐Alkene Formation. Chemistry – A European Journal 2020, 26 (68) , 15910-15921.
    43. Daniel Bauer, Ioannis Zachos, Volker Sieber. Production of Propene from n ‐Butanol: A Three‐Step Cascade Utilizing the Cytochrome P450 Fatty Acid Decarboxylase OleT JE . ChemBioChem 2020, 21 (22) , 3273-3281.
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    45. F. Peter Guengerich. Cytochrome P450 Catalysis in the Biosynthesis of Natural Products. 2020, 96-113.
    46. Yuanyuan Jiang, Zhong Li, Cong Wang, Yongjin J. Zhou, Huifang Xu, Shengying Li. Biochemical characterization of three new α-olefin-producing P450 fatty acid decarboxylases with a halophilic property. Biotechnology for Biofuels 2019, 12 (1)
    47. Vivek S. Bharadwaj, Seonah Kim, Michael T. Guarnieri, Michael F. Crowley. Different Behaviors of a Substrate in P450 Decarboxylase and Hydroxylase Reveal Reactivity-Enabling Actors. Scientific Reports 2018, 8 (1)
    48. Hazel M. Girvan, Harshwardhan Poddar, Kirsty J. McLean, David R. Nelson, Katherine A. Hollywood, Colin W. Levy, David Leys, Andrew W. Munro. Structural and catalytic properties of the peroxygenase P450 enzyme CYP152K6 from Bacillus methanolicus. Journal of Inorganic Biochemistry 2018, 188 , 18-28.
    49. Thomas M. Makris. Cytochrome P450 Decarboxylases. 2018, 127-143.
    50. Jia Wang, Zunwei Chen, Hui Chen, Yuezhong Wen. Effect of hydrogen peroxide on Microcystic aeruginosa: Role of cytochromes P450. Science of The Total Environment 2018, 626 , 211-218.
    51. Samiro Bojarra, Dennis Reichert, Marius Grote, Álvaro Gómez Baraibar, Alexander Dennig, Bernd Nidetzky, Carolin Mügge, Robert Kourist. Bio‐based α,ω‐Functionalized Hydrocarbons from Multi‐step Reaction Sequences with Bio‐ and Metallo‐catalysts Based on the Fatty Acid Decarboxylase OleT JE. ChemCatChem 2018, 10 (5) , 1192-1201.
    52. Robert Kourist, Anna Schweiger, Hanna Büchsenschütz. Enzymatic Decarboxylation as a Tool for the Enzymatic Defunctionalization of Hydrophobic Bio-based Organic Acids. 2018, 89-118.
    53. Laura N. Jeffreys, Hazel M. Girvan, Kirsty J. McLean, Andrew W. Munro. Characterization of Cytochrome P450 Enzymes and Their Applications in Synthetic Biology. 2018, 189-261.
    54. Hiroki Onoda, Osami Shoji, Kazuto Suzuki, Hiroshi Sugimoto, Yoshitsugu Shiro, Yoshihito Watanabe. α-Oxidative decarboxylation of fatty acids catalysed by cytochrome P450 peroxygenases yielding shorter-alkyl-chain fatty acids. Catalysis Science & Technology 2018, 8 (2) , 434-442.
    55. Anamitra Chatterjee, Sondre H. Hopen Eliasson, Vidar R. Jensen. Selective production of linear α-olefins via catalytic deoxygenation of fatty acids and derivatives. Catalysis Science & Technology 2018, 8 (6) , 1487-1499.
    56. Xu Zhang, Frank Jordan, Michal Szostak. Transition-metal-catalyzed decarbonylation of carboxylic acids to olefins: exploiting acyl C–O activation for the production of high value products. Organic Chemistry Frontiers 2018, 5 (16) , 2515-2521.
    57. Huriye Erdogan, An Vandemeulebroucke, Thomas Nauser, Patricia L. Bounds, Willem H. Koppenol. Jumpstarting the cytochrome P450 catalytic cycle with a hydrated electron. Journal of Biological Chemistry 2017, 292 (52) , 21481-21489.
    58. Huifang Xu, Linlin Ning, Wenxia Yang, Bo Fang, Cong Wang, Yun Wang, Jian Xu, Severine Collin, Frederic Laeuffer, Laurent Fourage, Shengying Li. In vitro oxidative decarboxylation of free fatty acids to terminal alkenes by two new P450 peroxygenases. Biotechnology for Biofuels 2017, 10 (1)

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