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Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - ABACUS (Algae for a biomass applied to the production of added value compounds)

Teaser

Summary

Algae production is currently limited to a few small industries, mainly for the feed, nutrition and cosmetic sectors. This is ripe for expansion. However, creating an economically viable and sustainable method of growing large quantities of algae and converting them into commercial products that the markets accept remains a challenge. The commercialisation of high value compounds sourced from microalgae could grow the existing industry considerably, while product innovation based on new molecular targets and biorefinery schemes could open new markets.
ABACUS unfolds a business-oriented and technology-driven development of a new algal biorefinery, thereby bringing to the market innovative algae-based ingredients for high-end applications, spanning from algal terpenes for fragrances to long-chain terpenoids (carotenoids) for nutraceuticals and cosmetic actives. One key objective of ABACUS is to obtain more than 10% photosynthates of targeted terpenoids in algal biomass. For this purpose, ABACUS selects and optimizes unique proprietary strains from 4 large culture collections owned by projectâ€™s partners. Moreover, ABACUS focuses on optimizing the cultivation steps and mastering the production of target products by process optimization, online monitoring and automated control of photobioreactors. Specific sensors for terpenoids and for the parameters relevant to their production (light, PO2, PCO2, nutrients) are tested and/or developed. ABACUS also investigates the fractionation steps to provide green low-cost downstream processing with a view to reduce operational expenses of the whole production line. Life cycle analysis and techno-economic analysis are fundamental guidelines to ensure that technologies and products are economically and environmentally sustainable. Applicability of targeted ingredients is assessed by the industrial partners (SMEs and large industries with established access to markets), considering cosmetic and nutraceutical applications as lead targets for commercialization. ABACUS aims to demonstrate biorefinery processes allowing valorizing up to 95% of the algal biomass into high value ingredients and by-products. ABACUS key advantage lies in its business-oriented workplan, gathering key players along the whole product development chain and incorporating most-advanced technologies for efficient growth and fractionation of microalgae.

Work performed

Two market studies and a field market survey were performed to identify relevant market trends. This led to the identification of the major product specifications in the field of cosmetics, food, nutraceuticals and fragrances. With all this information, the initial roadmap of the project was drawn, with main strains, products of interest and target markets defined.
31 candidate micro-algae/cyanobacteria species were included into the screening process at lab scale. This exceeds the initial target defined in the proposal (only 10 strains). A suite of standard methods, for algal cultivation, biochemical analysis and genetical engineering was developed and applied. The main technical outputs are the completion of the initial screening and the construction and validation of GM-cyanobacteria with induced capacity to produce light terpenes with long-term phylogenetic stability.
Pilot-scale cultivation of 7 algae strains was performed, focussing on scale-up of cultures, media optimization, water treatment and recycling, and PBR improvements in order to reduce maintenance and operational costs. New tools (probes and sensors) for process monitoring and automation were developed.
Most promising scenarii for large-scale cultivation were defined between the several partners operating industrial PBRs, and trial industrial runs were performed. First large scale cultures delivered hundreds to thousands grams of algal biomasses for downstream processing activities.
Green extraction and fractionation/purification procedures employing innovative compressed fluids were developed. Wet biomass extraction was applied to obtain increased yield and purity. An integrated processes with multiple product outputs was considered and the fractions/extracts produced characterized.
Applicability activities centered on the definition of compounds to quantify in algae main extracts and physico-chemical features to analyze in the by-products. Desirable storage and transfer conditions were set for the samples to be exchanged between the partners. Special focus was the development of a documentation system allowing traceability and centralization of the data generated.
An in-depth literature review on carotenoid extraction techniques and on LCA studies on carotenoid extraction from microalgae iwas performed. A preliminary LCA model including all process steps and based on secondary data extracted from literature was implemented as well as the collection of real lab-scale data related to the extraction processes.
Core tools for dissemination and communication were set up. Proactive involvement of all project partners in scientific conferences and business fairs increased the outreach of the project, with a sustained frequency of public appearance over the lapsed period. Project management structure is operational, with an internal collaborative platform for sharing results between consortium partners, and with frequent technical, executive and advisory meetings to secure quality control and risk management. Since part of the research implies the creation and use of GMOs, ethics and safety requirements were assessed for all partners involved.

Final results

31 candidate species were accurately screened at lab scale with assessment of criteria such as biomass productivity, comprehensive biochemical proximate analyses and genotyping. This will lead to at least one peer-reviewed publication.
Two cyanobacteria were genetically engineered for the production of light terpenes. Their genetic stability was verified during more than a year. In addition, unprecedented plasmid vectors for cyanobacterial production of a rare sesquiterpene (highly relevant to fragrance industry) were constructed. This synthetic biology work on photosynthetic cyanobacteria will lead to at least two peer-reviewed publications.
So far, two proprietary strains (wild type) belonging to ABACUS partners\'s culture collections were found superior to publicly available strains for astaxanthin production at industrial scale. This ensures freedom to operate in wo value chains for future industrial developments.
Several carotenoid producing strains were grown at pilot scale at very high productivity (exceeding 1g/L/day) and at high concentrations (approaching 10 g/L). Algal biomass with target terpenoid contents exceeding 5% was obtained. Supply of light was optimized allowing productivity increase of +110%. Optimization of culture media (less ingredients) resulted in elevated growth at reduced costs. All these achievements will significatively improve profitability of targeted algae-based ingredients.
Several new probes and sensors were designed to facilitate monitoring and automation of industrial PBRs, corresponding to a crucial need of the industry. These new sensors and probes still remain to be qualified in industrially-representative environment.