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Objectives
To provide biomass-derived sugar and lipid substrates for the enzymatic technologies proposed in the subsequent objectives, and evaluate some separation/pre-treatment technologies.

To provide the most adequate enzymatic biocatalysts (new or engineered oxidative enzymes) for development of the bio-chemical technologies proposed in the subsequent objectives.

To develop an enzymatic technology for the production of FDCA as a renewable polymer building block derived from HMF.

To develop new enzymatic technologies for the selective hydroxylation of biomass-derived lipid components in the manufacture of F&F ingredients, APIs and other added value products.

To optimize the above enzymatic (or chemo-enzymatic) technologies in terms of biocatalyst stability, substrate concentration, reaction medium, co-substrate supply (when required) and downstream processing.

To evaluate the technical, economic and environmental feasibility (LCA analysis) of the enzymatic processes, compared with chemical processes for the production of the same or similar compounds.

Project concept

The overall aim of the EnzOx2 project is to develop new bio-chemical technologies based on the use of oxidative enzymes, largely unexplored at the industrial level, to provide innovative solutions in the production of some added value compounds from biomass components to substitute others of petrochemical origin. The potential of oxidative enzymes in such biotransformations has been shown by some of the EnzOx2 partners in previous projects, including several oxidation and oxyfunctionalization reactions catalyzed by different types of fungal oxidoreductases (such as oxidases and peroxygenases).

In this context, EnzOx2 plans to develop a 100% enzymatic conversion of bio-based 5-hydroxymethylfurfural (HMF) into diformylfuran, a platform chemical, and 2,5-furandicarboxylic acid (FDCA), a plastic building-block to be used in substitution of pterphthalic acid. On the other hand, highly selective hydroxylation of plant lipids (such as fatty acids, terpenes and steroids) will be optimized for cost-effective production of flavour and fragrance (F&F) ingredients, active pharmaceutical ingredients (APIs) and others.

Project workflow

To attain these goals, EnzOx2 will take advantage from the largely unexploited diversity of oxidoreductases in sequenced microbial genomes to obtain new enzymes of interest. Moreover, the catalytic performance, selectivity and/or stability of the best candidates will be adapted, when needed, to the required reaction conditions using protein engineering tools. Several concepts such as substrate loading, co-factor addition, biocatalyst stability and downstream processing, among others, will also be considered to further optimize the enzymatic reactions. Finally, life cycle assessment (LCA) analyses of the enzymatic processes, compared with chemical processes for the production of the same or similar compounds will be conducted in order to evaluate their technical, economic and environmental feasibility.