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Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - ALGAE4A-B (Development of Microalgae-based novel high added-value products for the Cosmetic and Aquaculture industry)

Teaser

Summary

Algae4A-B seeks to combine both basic and applied biotechnology research with state-of-the-art biomass production technologies to develop novel microalgae based products aimed towards the aquaculture and cosmetic Industries. The main objective is the characterization and the production of application customised microalgae biomass, as a source for high-added-value cosmeceuticals and nutraceuticals. Despite their enormous potential and biodiversity, a surprisingly small number of microalgae have been analysed and only a handful of strains and derived biomolecules are currently used in health and cosmetic applications. The diversification of the production of microalgae biomass towards 2 independent applications will allow the microalgae industry to gain access to alternative markets in an uncertain, highly competitive and fast changing market.

Work performed

WP1-Development and optimization of low input application-based culture systems
New facilities for microalgae culture under different conditions have been set up. Trials to optimize the facilities were performed with the 2 target species. Results obtained revealed good performance of the system, which allows for more research at lab-scale for the evaluation of culture conditions on microalgae growth and productivity of target compounds. Up-scaling of T. chuii from indoor cultures to 650 L outdoor photobioreactors was implemented and comparative analyses performed in terms of cell density and nutrient consumption. The results demonstrated an optimal behaviour of T. chuii, which will serve as reference for the up-scale optimization in other species.
WP2-Development of â€œ-omicâ€ resources
We developed an RT-qPCR based platform for the transcriptomic analysis of T. chuii. RNAseq transcriptomic data were recorded for human primary fibroblasts incubated with microalgae exopolysaccharides to study the molecular effects of these molecules in skin physiology. Approach for sampling for the microbiome and transcriptome was established for sole. Target molecular resources were set for the selected key innate immune tissue (skin and gut). Reference panels to study the innate immune response system and compare expression data between cosmetics and aquaculture were established. Methods for the catabolomic analysis and purification of target enzymes from microalgae biomass were developed, and data on enzymes activities of interest obtained.
WP3-Screening and bioactivity assays for functional analysis of microalgae derived products
Bioinformatics analysis for the transcription analysis and updated list of genes for the transcriptional studies was performed, to study the transcriptional effects of microalgae extracts on human primary skin cells. Gene expression results were obtained, using both the RT-qPCR platform and RNAseq on the in vitro effects of microalgae extracts and exopolysaccharides on human skin cells, under normal and/or oxidative stress and wounding. The methodology for primary cell culture of skin cells was optimized and preliminary assays to assess the impact of micro-algal extracts on proliferation and apoptosis were done. The RT-qPCR assays developed for human skin are transferred to fish skin. Cytotoxicity analysis of micro-algal extracts on human skin was established and a candidate protein from marine organism was tested. Screening capabilities for bioactive compounds using biosensors and protocols for the application of a biosensor in wound healing scenarios were developed. A screening approach for the action of micro-algal extracts on fish pathogens was established.
WP4-Downstream processing of high value added products
Evaluation of the efficiency of micro-algal cell lysis procedures was performed. The impact of total input content was evaluated in relation to protein recovery and SOD activity in fractions using standardized conditions. Trials were performed with Nannochloropsis for protease purification using different ATPS systems and optimal pH for system performance was determined. Attempts to identify cyanobacteria producing polysaccharides was carried out, with a final selection of 15 strains. Beta-glucan from Phaeodactylum sp. biomass was isolated to examine bioactivity. Trials to isolate and characterize theca of Tetraselmis sp. were performed. Assays aiming at characterizing the hatching enzymes (HE) in sole were carried out. 3 putative HE-coding genes were identified, which reveal to be expressed in the hatching gland by in situ hybridization technics.
WP5-Microalgae for aquaculture
The toxicity effects after exposure of embryos and larvae at different concentrations were measured. Biological performance tests indicated a clear association with toxicity effects of microalgae on larval performance. A trial confirmed that the calculated doses were safe and modified the expression profiles of DNMT methyltran

Final results

The predicted direct impact on R&D activities and product development of both the academic and commercial partners are still relevant and will be likely reached at the end of the project. The results dealing with new strategies for microalgae growing conditions in large-scale photobioreactors allows envisioning improved production of selected bioactive compounds with interest in the cosmetic and aquaculture industry (WP1). Sequencing transcriptomes (WP2) of micro-algae and human and fish cells treated with micro-algal extracts figure out biological mechanisms induced by the micro-algal compounds highlighted in the project. â€œOmicsâ€ tools implemented were correlated with biological assays dedicated to cosmeceutical (WP3) and aquaculture applications (WP5). The identification of promising novel molecules and process to obtain them (WP4) were followed by formulation of the molecules, notably by encapsulation strategies, in order to release the bioactive compound in the most appropriate conditions in vivo.