| Autor: |
Rade LL; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Generoso WC; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Das S; Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695-7622., Souza AS; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Silveira RL; Institute of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro 21941-594, Brazil., Avila MC; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Vieira PS; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Miyamoto RY; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Lima ABB; Institute of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro 21941-594, Brazil., Aricetti JA; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., de Melo RR; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Milan N; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Persinoti GF; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Bonomi AMFLJ; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Murakami MT; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil., Makris TM; Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695-7622., Zanphorlin LM; Brazilian Biorenewables National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas 13083-100, Brazil. |
| Abstrakt: |
The enzymatic decarboxylation of fatty acids (FAs) represents an advance toward the development 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 OleT JE . Herein, we describe OleTP RN , a poly-unsaturated alkene-producing decarboxylase 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 on high salt concentrations, OleTP RN can also efficiently produce alkenes from unsaturated (oleic and linoleic) acids, the most abundant FAs found in nature. OleTP RN performs carbon-carbon cleavage by a catalytic itinerary that involves hydrogen-atom transfer by the heme-ferryl intermediate Compound I and features a hydrophobic cradle at the distal region of the substrate-binding pocket, not found in OleT JE , 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 OleTP RN 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. |
