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Carbohydrate-based fine chemicals: Development of a glycosylation platform cell factory and optimization of downstream processing for the sustainable production of glycosides.

Periodic Reporting for period 3 - CARBAFIN (Carbohydrate-based fine chemicals: Development of a glycosylation platform cell factory and optimization of downstream processing for the sustainable production of glycosides.)

Reporting period: 2021-01-01 to 2022-06-30

Based on integrated biocatalytic production technologies developed in the project, CARBAFIN has enabled new value chains for the utilization of sucrose from sugar beet biomass in the EU. CARBAFIN has demonstrated bio-based production of a functional glucoside (glucosylglycerol, cellodextrin) that is obtained together with an exploitable fructose stream. CARBAFIN’s glucoside products have large-scale uses as functional ingredients in nutrition and feed, cosmetics and detergents. The fructose streams are feedstocks for the production of 5-hydroxymethylfurfural (HMF), a versatile chemical building block currently considered for making bio-based plastics. CARBAFIN has delivered leading platform technologies in biocatalytic cell factories and downstream processing, both critical for the successful bioprocess development. CARBAFIN production technologies have been optimized for performance efficiency and cost effectiveness under full integration of LCA and economic evaluation. CARBAFIN results demonstrate pilot scale production of sucrose-derived glucosides alongside with conversion of fructose into HMF in the industrial environment. Detailed process evaluation reveals the critical factors of ecological and economic sustainability. CARBAFIN process technologies appear to be competitive for production in today's markets.
Overall, multilevel biocatalyst engineering was applied to boost the synthetic efficiency of cell factories to industrial target values. Protein engineering/screening was used to enhance enzyme selectivity/stability for efficient glucosylglycerol production via direct glucosylation from sucrose to glycerol and for synthesis of defined cello-oligosaccharides. Co-expression strategies in Escherichia coli provide highly active cell factories for cellodextrin production via a multi-step indirect glucosylation that involves glucose 1-phosphate as the key intermediate. The cell factories obtained approach the targeted productivities in synthesis. Advanced kinetic modeling was established as a process engineering tool for flexibly targeted production of glucosylglycerol and cellodextrins. Furthermore, enzyme and reaction engineering were combined to develop biocatalytic synthesis of soluble cellodextrins at controlled degree of polymerization. Immobilized whole cell-derived biocatalysts were developed for continuous glucoside production. Nanofiltration combined with reactive extraction was assessed as process technology for product downstream processing. Methods of process sustainability assessment and economic evaluation were applied to the process technologies developed. Fructose streams resulting from enzymatic conversion were assessed for HMF production. In the last reporting period focus was put on demonstration of production processes in industrial environment as well as on techno-economic and environmental evaluation of the processes.
CARBAFIN’s dissemination activities, in public events (#6) and workshops (#4) involved around 1,200 participants in total. The number of peer-reviewed publications is 23. CARBAFIN has identified 6 key exploitable results (KER) in the category “products and processes”, 4 in category “expertise/skills/data” and 4 in category “learnings and recommendations”. For each KER a lead partner was assigned to follow up on the exploitation strategy jointly defined. The results from category “products and processes” and “expertise/skills/data” are found on the EU innovation radar platform. In addition, business cases for the separate production of glucosylglycerol, cellodextrin and HMF were prepared. CARBAFIN production technologies involve unique selling propositions due to enhanced efficiency, improved eco-credentials or both. They thus enable more sustainable production of CARBAFIN industrial partners in compliance with the Green Deal program.
In addition, CARBAFIN contributes to the establishment of new, fully renewables-based value chains across the European industries, linking the sugar industry sector to the cosmetics, chemicals, polymer and detergents sectors and strengthening the sugar industries' role in food and feed. It supports the efficient cascading use of sugar beet-derived sucrose in combination with biomass-derived base chemicals (e.g. glycerol, glucose). A detailed analysis of industrial stakeholders was performed and strategies to involve these stakeholders in the successful replication of the technology were implemented. The use of CARBAFIN's fructose streams for HMF production promotes the replacement of petroleum-based chemicals with bio-based building blocks. CARBAFIN will thus contribute to the development of a chemical industry that is low-carbon, resource-efficient and sustainable. CARBAFIN helps Europe to maintain global leadership in industrial biotechnology and secure position as innovation leader in sugar-based products. Also beyond the project, CARBAFIN's innovations will involve broadly the engagement of societal actors with particular emphasis on consumer awareness and acceptance. Moreover, CARBAFIN’s public relations activities (e.g. school visits at the partners sites, engagement in European and National Researchers Nights, broadcasting of explanatory videos) underline the relevance of industrial biotechnology for a broader audience.
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