Redox modulation of transmembrane fatty acid desaturases for production of medium-chain hydrocarbons using biofactories

• Grant number: 24/08184-8
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: September 01, 2024
• End date: August 31, 2028
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Leticia Maria Zanphorlin
• Grantee: Isabelle Taira Simões
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil

Abstract

Metalloenzymes, especially those with redox function, play a crucial role in nature, and the last decade has witnessed the discovery of oxidative enzymes that catalyze the biosynthesis of hydrocarbons from fatty acids, employing non-canonical mechanisms that lead to the cleavage of the C-C bond in route. There is substantial diversity in the structure, mode of action, cofactor requirement, and specificity among these metalloenzymes, which are generally divided into heme monooxygenases and iron-dependent mono- and dinuclear oxygenases. Aiming to comprehend such diversity, our group has contributed to the discovery and understanding of a new decarboxylase from the heme-protein family, which exhibits improved functional aspects, mainly regarding substrate scope, and is driven by mechanisms entirely distinct from those reported so far. P450 class decarboxylases prefer long chains of fatty acids and also exhibit branched catalytic activity, producing hydroxylated products as well. In an attempt to reduce these undesired coproducts and also customize the production of medium-chain hydrocarbons, which have been of great interest for application in aviation fuel and renewable olefin-based chemicals, this project aims to explore iron enzymes (UndB) that exhibit decarboxylase activity, similar to P450, but are not heme proteins. These enzymes are initially dinuclear with transmembrane characteristics, therefore, few have been characterized. In parallel, this project will also focus on constructing a microbial chassis for the production of medium-chain fatty acids. Thus, by the end of the project, we will offer a biotechnological platform for the biological production of 1-undecene, an olefin considered an important building block for various sectors, and thus, we will be able to contribute to a circular economy. It is important to emphasize that, knowing the challenge of studying transmembrane proteins, we have already begun the project, and after several tests, we have established the production of some desired proteins in the soluble fraction exhibiting decarboxylase activity.