Dimer-assisted mechanism of (un)saturated fatty acid decarboxylation for alkene production

Author(s):Show less -

Rade, Leticia L. ; Generoso, Wesley C. ; Das, Suman ; Souza, Amanda S. ; Silveira, Rodrigo L. ; Avila, Mayara C. ; Vieira, Plinio S. ; Miyamoto, Renan Y. ; Lima, Ana B. B. ; Aricetti, Juliana A. ; de Melo, Ricardo R. ; Milan, Natalia ; Persinoti, Gabriela F. ; Bonomi, Antonio M. F. L. J. ; Murakami, Mario T. ; Makris, Thomas M. ; Zanphorlin, Leticia M.

• Document type: Journal article
• Source: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA; v. 120, n. 22, p. 10-pg., 2023-05-22.

Abstract

The enzymatic decarboxylation of fatty acids (FAs) represents an advance toward the develop-ment of biological routes to produce drop-in hydrocarbons. The current mechanism for the P450-catalyzed decarboxylation has been largely established from the bacterial cytochrome P450 OleTJE. Herein, we describe OleTPRN, a poly-unsaturated alkene-producing decar-boxylase that outrivals the functional properties of the model enzyme and exploits a distinct molecular mechanism for substrate binding and chemoselectivity. In addition to the high conversion rates into alkenes from a broad range of saturated FAs without dependence high salt concentrations, OleTPRN can also efficiently produce alkenes from unsaturated (oleic and linoleic) acids, the most abundant FAs found in nature. OleTPRN performs carbon-carbon cleavage by a catalytic itinerary that involves hydrogen-atom transfer the heme-ferryl intermediate Compound I and features a hydrophobic cradle at the distal region of the substrate-binding pocket, not found in OleTJE, which is proposed to play a role in the productive binding of long-chain FAs and favors the rapid release of products from the metabolism of short-chain FAs. Moreover, it is shown that the dimeric configuration of OleTPRN is involved in the stabilization of the A-A' helical motif, a second-coordination sphere of the substrate, which contributes to the proper accommodation of the aliphatic tail in the distal and medial active-site pocket. These findings provide an alternative molecular mechanism for alkene production by P450 peroxygenases, creating new opportunities for biological production of renewable hydrocarbons. (AU)

• FAPESP's process: 19/12599-0 - Elucidation of the mechanisms of non-conventional peroxygenases with application in the production of advanced biofuels
• Grantee: Mayara Chagas de Ávila
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 19/08855-1 - New mechanisms of P450: an enzymatic strategy for renewable hydrocarbons
• Grantee: Leticia Maria Zanphorlin
• Support Opportunities: Regular Research Grants
• FAPESP's process: 20/01967-6 - Decarboxylation enzymatic mechanisms of fatty acids into terminal olefins to obtain drop-in biofuels
• Grantee: Natalia Milan
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 18/04897-9 - Production of biodiesel and renewable hydrocarbons via enzymatic route: from the enzyme synthesis to the economic evaluation of the integrated process
• Grantee: Letícia Leandro Rade
• Support Opportunities: Scholarships in Brazil - Post-Doctoral