Project: Fabrication of hierarchically organized multi-functional heterogeneous biocatalysts for the modular synthesis of ω-amino acids from renewable feedstocks

Acronym	HOMBIOCAT (Reference Number: 61)
Project Topic	Chemical synthesis catalyzed by enzymes is contributing to establish a modern chemistry supported on cleaner, faster and safer chemical reactions. In particular, cell-free metabolic engineering (or systems biocatalysis) in solution is currently emerging as an attractive alternative to synthetic biology using whole cells because isolated enzymes do not present regulation constraints at genomic level and the intensification of the chemical fluxes do negligible effect on the system subsistence. However, it presents major issues in terms of both process- and cost-efficiency because these soluble systems often reach low chemical yields, are notably unstable and their re-usability is rather limited. In order to overcome these limitations, this proposal aims to assemble multi-enzyme systems at the nanoscale of solid and porous materials aided by protein scaffolds that guarantee the hierarchical and spatial organization of the functional modules. This immobilized multi-enzyme cascade will be utilized as heterogeneous multi-functional biocatalyst to transform renewable raw materials into ω-amino acids in one-pot and with in situ cofactor regeneration. The fabrication and exploitation of such heterogeneous biocatalysts will be achieved by i) engineering the proposed artificial pathway for the synthesis of ω-amino acids using bio-oils and fermented diols as raw materials, ii) engineering a modular protein as scaffolding unit to organize the multi-enzyme system at the nanoscale, iii) in vitro assembling of multi-enzyme systems onto the protein scaffolds, iv) immobilizing such multi-enzyme assemblies on solid particles, v) using the hierarchically organized multi-functional heterogeneous biocatalysts for the efficient and sustainable production of long and short ω-amino acids vi) 10 L scaling-up the process under industrially relevant conditions and vii) manufacturing of one modular kit to produce multi-enzyme systems employing common laboratory methods and one modular kit that allows the solid-phase assembly of different functional modules based on scaffolded multi-enzyme systems. These new platforms will open an innovative tool to build sustainable pathways for chemical manufacturing of high added value molecules using renewable raw materials. The rational integration of different enzymes as functional modules with an engineered protein scaffold and a porous material as heterogeneous chassis will be addressed by combining protein engineering, surface chemistry and protein immobilization tools. The research team presents a solid and multidisciplinary background in those areas, which fully qualifies this team to carry out this project at the frontier between the chemistry and the biology. Moreover, the research consortium is hosted in different research institutions that provide a suitable research environment (three academic partners; CIC biomaGUNE, University of Nottingham and Ruhr Universität-Bochum, and one industrial partner: Bioassays) to successfully address the major challenges and meet the main objectives of this project.
Project Results (after finalisation)
Website	visit project website
Network	ERA CoBioTech
Call	ERA CoBioTech “Biotechnology for a sustainable bioeconomy”

Project partner

Number	Name	Role	Country
1	CIC biomaGUNE	Coordinator	Spain
2	Bioassays	Partner	Spain
3	University of Notthingham	Partner	United Kingdom
4	Ruhr-Universität Bochum	Partner	Germany