Discovery and development of oxidative enzymes for the in vivo biosynthesis of hydrocarbons

• Grant number: 21/14726-0
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: July 01, 2022
• End date: June 30, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Leticia Maria Zanphorlin
• Grantee: Vandierly Sampaio de Melo
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated scholarship(s): 24/18327-0 - Enzyme Cascades for Sustainable Chemical Synthesis, BE.EP.DR

Abstract

The disadvantages of high dependence on fossil sources for energy generation and for obtaining various products have been the subject of global discussions with a focus on the transition to sustainable development models. The global agenda for sustainability encompasses action in all sectors of the economy with the aim of adopting renewable alternatives to replace or complement fossil raw materials already in use, reduce atmospheric emissions and mitigate adverse socio-environmental impacts. In this scenario, Brazil has a high potential for the production of biofuels and green chemicals from biomass, considering the concept of biorefinery. In this line of action to obtain renewable products, hydrocarbons have been one of the best for the development of alternative routes to first generation biofuels, ethanol and fatty acid methyl esters (biodiesel), and petrochemicals such as polymers, solvents and surfactants of fossil origin. Biohydrocarbons are drop-in molecules that have physicochemical properties similar to those of petroleum. In general, these compounds can be produced from the fatty acid biosynthesis pathway. An attractive way to produce olefin-type hydrocarbons (alkenes) is by enzymatic decarboxylation of these fatty acids by decarboxylase enzymes. However, this is a very recent area of study and rare enzymes have been characterized. In this sense, our research group has been dedicating efforts for some years in an attempt to discover and elucidate the mechanisms of new fatty acid decarboxylases for obtaining terminal alkenes. As a result of these efforts, three patent applications have already been generated (BR10202001600, BR10202002353 and BR1020210246723). The literature describes two main types of fatty acid decarboxylases, the CYP152 enzymes (belonging to the P450 superfamily) and the oxidative UndA enzymes, which are capable of decarboxylating fatty acids, producing 1-alkenes. In particular, the enzyme UndA, is a non-heme iron-dependent decarboxylase/oxidase, which has a preference for C8-C14 medium chains. It is known that it is an extremely conserved enzyme in the genus Pseudomonas and homologous in the genus Burkholderia were also identified, however, only one with a structure with different ligands was determined. Therefore, knowledge about the molecular mechanisms of these decarboxylases is still very scarce, mainly due to the lack of structural information. In order to discover new UndA-type decarboxylases, our research group carried out a microbial consortium in the presence of different sources of fatty acids and together with bioinformatics methodologies, we identified five new Unds-like. Therefore, this doctoral project aims at the molecular study of these enzymes through a multidisciplinary approach that involves biochemical and biophysical tools. Through this project, we intend to elucidate the mechanisms used by decarboxylases for the production of alkenes, as well as the specificity of each enzyme against different sizes of fatty acid. Finally, this project has enormous potential to contribute to the development of biological routes for the production of olefins, an important renewable chemical that can be used in a range of industrial applications. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DE MELO, VANDIERLY SAMPAIO; DE MELO, RICARDO RODRIGUES; RADE, LETICIA LEANDRO; MIYAMOTO, RENAN YUJI; MILAN, NATALIA; DE SOUZA, CLAUDIA MARIA; DE OLIVEIRA, VINICIUS MARTINS; SIMOES, ISABELLE TAIRA; LIMA, EVANDRO ANTONIO DE; GUILHERME, EDERSON PAULO XAVIER; et al. Thermoascus aurantiacus harbors an esterase/lipase that is highly activated by anionic surfactant. Biochemical and Biophysical Research Communications, v. 733, p. 9-pg., 2024-08-26. (17/14253-9, 18/04897-9, 21/14410-2, 21/14726-0, 18/02865-2, 19/08855-1)