Periodic Reporting for period 1 - NUVAPY-BLUE (Nutricosmeceutical valorization of Porphyra/Pyropia spp. (Rhodophyta): biorefinery approach in the frame of Blue Economy)
Periodo di rendicontazione: 2023-06-01 al 2025-05-31
Sintesi del contesto e degli obiettivi generali del progetto
Nutricosmeceutical develops products to improve human health through the use of bioactive compounds (BACs). Algae are organisms that have a wide variety of BACs, high productivity and they can be produced without fresh water or arable land. Following EU Green Deal and Blue Economy, the proposal aims to increase the macroalgae Porphyra/Pyropia productivity in consortium with fish, to produce high value BACs naturally and make nutricosmeceutical more attractive to the market. To reach this objective, the project will focus in: Porphyra production using integrated multitrophic aquaculture with fishpond effluents from Chelon labrosus for alga growing, optimizing natural production of BACs; extract and purify BACs through biorefinery strategy; perform BACs encapsulation and test their sun protection, stability, toxicity, bioavailability; and develop topical and oral sunscreens. Researches show some abiotic factors for BACs development and production in Porphyra, however there is a lack of studies that correlate various abiotic factors along with fish culture for a circular economy and better algae development. The project will produce biofertilizers and biostimulants from residual algal biomass, after extractions, which has not yet been optimized. Natural sunscreens are still scarce on the market, due to the lack of standards in extracts with stability for use in products and low BACs availability. The project’s interdisciplinarity is strong involving aquaculture, biochemistry, biotechnology and nutricosmeceutic.
Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti
During the project ‘NUVAPY-BLUE’, a multidisciplinary biorefinery strategy was applied to maximize the use of biomass from Porphyra species, targeting high-value bioactive compounds for nutraceutical and cosmeceutical applications.
The activities were structured into several technical and scientific workstreams:
Cultivation Optimization: A key focus was the optimization of cultivation conditions to enhance biomass quality and functional compound yield. A novel cultivation setup was tested, simulating tidal cycles to better mimic the natural environment of Porphyra/Pyropia. This simulation led to visibly more robust algae, with improved resistance to environmental stress. Additionally, controlled experiments were conducted using different nitrate concentrations to assess the impact on the endogenous production of mycosporine-like amino acids (MAAs). Parallel trials with varying doses of UV radiation were also performed, aiming to stimulate the biosynthesis of these photoprotective compounds. When testing different nitrate concentrations in combination with varying UV doses, nitrate availability proved to be the most critical factor in enhancing the natural production of MAAs, indicating its predominant role over UV exposure in optimizing the biosynthesis of these compounds. Furthermore, the feasibility of using fish effluent as a natural nitrate source was explored and compared with synthetic nitrate. The results showed that the use of fishpond effluents allowed for comparable MAA production with lower overall nitrate concentrations, indicating a more efficient and sustainable nutrient source that aligns with circular aquaculture and integrated multi-trophic aquaculture systems (IMTA).
Extraction Process Development: Extraction protocols were evaluated using both fresh and freeze-dried biomass of Porphyra. Initial comparisons showed that fresh biomass consistently yielded higher concentrations of bioactive compounds, guiding subsequent extraction trials. Based on this, different extraction strategies were tested, including alkaline and enzymatic hydrolysis. These approaches significantly increased the extraction yield of key antioxidant compounds, particularly polyphenols, phycobiliproteins, and carbohydrates. In contrast, for MAAs (HPLC), the best results were obtained using water-based extraction at elevated temperatures—higher than those typically reported in the literature—suggesting a more efficient release of these thermoresistant molecules under these conditions. Additionally, specific protocols were developed for the extraction and partial purification of the polysaccharide porphyran (GC-MS), a valuable compound with known bioactive and gelling properties.
Bioactivity Screening: The obtained extracts were tested in vitro for antioxidant activity using the ABTS assay and for cellular response in HCT116 human colorectal carcinoma cells. Additionally, the sun protection factors (SPF and UVAPF) of selected extracts were evaluated using PMMA plates, simulating topical application conditions and enabling quantification of photoprotective properties relevant to cosmeceutical applications.
Nanoencapsulation Development: A novel nanoencapsulation strategy is being developed using porphyran, as a natural cross-linking polymer. This approach is designed as a sustainable alternative to traditional encapsulating agents such as sodium tripolyphosphate (TPP). Porphyran-based nanoparticles are being formulated to encapsulate selected bioactive compounds, aiming to enhance their stability, bioavailability, and controlled release. Preliminary characterization of the nanoparticles (e.g. size distribution, zeta potential, encapsulation efficiency) supports the potential of porphyran as a functional biomaterial for drug delivery and cosmeceutical applications.
Bioactive Residue Valorization for Biostimulant Applications: In line with the biorefinery approach, the residual biomass remaining after primary extractions was further valorized through a secondary, simplified water-based extraction under controlled temperature. The resulting extracts were evaluated for potential plant biostimulant activity using bioassays targeting hormone-like effects. Specifically, cytokinin-like activity was assessed via the cucumber cotyledon expansion test, and gibberellin-like activity was evaluated through the germination index assay in watercress seed. The positive responses observed in both tests indicate that the residual biomass still contains bioactive compounds with plant-growth promoting potential, suggesting its applicability as a sustainable input in agricultural or horticultural formulations.
Outcomes of the actions:
• Demonstrated that simulating tidal cycles in cultivation systems improves the physiological robustness and stress resistance of Porphyra/Pyropia biomass.
• Identified nitrate concentration as the key factor in enhancing the endogenous production of mycosporine-like amino acids (MAAs), surpassing the influence of UV exposure.
• Proved the potential of fishpond effluents as an alternative nutrient source, achieving similar MAA yields with lower nitrate concentrations, supporting sustainable aquaculture integration.
• Developed efficient extraction protocols using alkaline and enzymatic hydrolysis, significantly increasing yields of phenolics, phycobiliproteins, and carbohydrates.
• Optimized MAA extraction using high-temperature, water-based methods, achieving superior results to previously reported techniques.
• Successfully extracted and partially purified porphyran, confirming its presence and biofunctional potential through GC-MS analysis.
• Validated the antioxidant potential of extracts through ABTS assay.
• Confirmed photoprotective potential by determining SPF values on PMMA plates, indicating suitability for cosmeceutical applications.
• Initiated development of a natural and sustainable nanoencapsulation system using porphyran as a cross-linking polymer. Replaced synthetic agents like TPP with porphyran, aligning with clean-label and eco-friendly formulation goals.
• Demonstrated that secondary extracts from residual biomass retain biological activity with plant hormone-like effects.
• Supported the zero-waste biorefinery concept by adding value to process residues, contributing to sustainability and circular economy in marine biomass valorization.
The activities were structured into several technical and scientific workstreams:
Cultivation Optimization: A key focus was the optimization of cultivation conditions to enhance biomass quality and functional compound yield. A novel cultivation setup was tested, simulating tidal cycles to better mimic the natural environment of Porphyra/Pyropia. This simulation led to visibly more robust algae, with improved resistance to environmental stress. Additionally, controlled experiments were conducted using different nitrate concentrations to assess the impact on the endogenous production of mycosporine-like amino acids (MAAs). Parallel trials with varying doses of UV radiation were also performed, aiming to stimulate the biosynthesis of these photoprotective compounds. When testing different nitrate concentrations in combination with varying UV doses, nitrate availability proved to be the most critical factor in enhancing the natural production of MAAs, indicating its predominant role over UV exposure in optimizing the biosynthesis of these compounds. Furthermore, the feasibility of using fish effluent as a natural nitrate source was explored and compared with synthetic nitrate. The results showed that the use of fishpond effluents allowed for comparable MAA production with lower overall nitrate concentrations, indicating a more efficient and sustainable nutrient source that aligns with circular aquaculture and integrated multi-trophic aquaculture systems (IMTA).
Extraction Process Development: Extraction protocols were evaluated using both fresh and freeze-dried biomass of Porphyra. Initial comparisons showed that fresh biomass consistently yielded higher concentrations of bioactive compounds, guiding subsequent extraction trials. Based on this, different extraction strategies were tested, including alkaline and enzymatic hydrolysis. These approaches significantly increased the extraction yield of key antioxidant compounds, particularly polyphenols, phycobiliproteins, and carbohydrates. In contrast, for MAAs (HPLC), the best results were obtained using water-based extraction at elevated temperatures—higher than those typically reported in the literature—suggesting a more efficient release of these thermoresistant molecules under these conditions. Additionally, specific protocols were developed for the extraction and partial purification of the polysaccharide porphyran (GC-MS), a valuable compound with known bioactive and gelling properties.
Bioactivity Screening: The obtained extracts were tested in vitro for antioxidant activity using the ABTS assay and for cellular response in HCT116 human colorectal carcinoma cells. Additionally, the sun protection factors (SPF and UVAPF) of selected extracts were evaluated using PMMA plates, simulating topical application conditions and enabling quantification of photoprotective properties relevant to cosmeceutical applications.
Nanoencapsulation Development: A novel nanoencapsulation strategy is being developed using porphyran, as a natural cross-linking polymer. This approach is designed as a sustainable alternative to traditional encapsulating agents such as sodium tripolyphosphate (TPP). Porphyran-based nanoparticles are being formulated to encapsulate selected bioactive compounds, aiming to enhance their stability, bioavailability, and controlled release. Preliminary characterization of the nanoparticles (e.g. size distribution, zeta potential, encapsulation efficiency) supports the potential of porphyran as a functional biomaterial for drug delivery and cosmeceutical applications.
Bioactive Residue Valorization for Biostimulant Applications: In line with the biorefinery approach, the residual biomass remaining after primary extractions was further valorized through a secondary, simplified water-based extraction under controlled temperature. The resulting extracts were evaluated for potential plant biostimulant activity using bioassays targeting hormone-like effects. Specifically, cytokinin-like activity was assessed via the cucumber cotyledon expansion test, and gibberellin-like activity was evaluated through the germination index assay in watercress seed. The positive responses observed in both tests indicate that the residual biomass still contains bioactive compounds with plant-growth promoting potential, suggesting its applicability as a sustainable input in agricultural or horticultural formulations.
Outcomes of the actions:
• Demonstrated that simulating tidal cycles in cultivation systems improves the physiological robustness and stress resistance of Porphyra/Pyropia biomass.
• Identified nitrate concentration as the key factor in enhancing the endogenous production of mycosporine-like amino acids (MAAs), surpassing the influence of UV exposure.
• Proved the potential of fishpond effluents as an alternative nutrient source, achieving similar MAA yields with lower nitrate concentrations, supporting sustainable aquaculture integration.
• Developed efficient extraction protocols using alkaline and enzymatic hydrolysis, significantly increasing yields of phenolics, phycobiliproteins, and carbohydrates.
• Optimized MAA extraction using high-temperature, water-based methods, achieving superior results to previously reported techniques.
• Successfully extracted and partially purified porphyran, confirming its presence and biofunctional potential through GC-MS analysis.
• Validated the antioxidant potential of extracts through ABTS assay.
• Confirmed photoprotective potential by determining SPF values on PMMA plates, indicating suitability for cosmeceutical applications.
• Initiated development of a natural and sustainable nanoencapsulation system using porphyran as a cross-linking polymer. Replaced synthetic agents like TPP with porphyran, aligning with clean-label and eco-friendly formulation goals.
• Demonstrated that secondary extracts from residual biomass retain biological activity with plant hormone-like effects.
• Supported the zero-waste biorefinery concept by adding value to process residues, contributing to sustainability and circular economy in marine biomass valorization.
Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)
Overview of Key Results:
• Optimized cultivation conditions (simulated tidal cycles and nitrate enrichment) significantly enhanced biomass quality and endogenous production of mycosporine-like amino acids (MAAs).
• Fishpond effluents as a natural nitrate source were tested and compared to synthetic nitrate. Results showed that fishpond effluents achieved comparable MAA yields while requiring lower overall nitrate input, highlighting a sustainable and integrated approach aligned with circular aquaculture practices.
• Fresh biomass extraction with alkaline and enzymatic hydrolysis improved yields of antioxidant compounds (phenolics, phycobiliproteins, carbohydrates).
• Efficient MAA extraction using high-temperature water-based methods surpassed literature benchmarks.
• Porphyran extraction was successfully achieved and showed potential both as a bioactive compound and encapsulation agent. Porphyran-based nanoencapsulation is being developed as a natural alternative to synthetic encapsulates.
• In vitro bioactivity testing demonstrated antioxidant activity (ABTS), and SPF properties (PMMA plate assays).
• Biostimulant potential of residual biomass was confirmed through plant hormone-like bioassays.
Potential Impacts:
• Industrial relevance confirmed: As a direct outcome of the project, a follow-up collaboration has been established with Cantabria Labs, a Spanish cosmetics company, to further develop a commercial photoprotector/antioxidant cream. This new project aims to build upon the current findings by incorporating additional plant extracts to boost SPF and antioxidant activity, reinforcing the translational value of the research.
• Blue Bioeconomy alignment: The approach supports zero-waste strategies and marine biomass circularity.
• Nutraceutical and cosmeceutical innovation: Clean-label, natural bioactives with proven efficacy contribute to high-demand product segments.
• Agricultural circularity: Biomass residues with biostimulant activity offer potential for sustainable agriculture inputs.
Key Needs for Further Uptake and Success:
• Scale-up and demonstration activities to validate industrial feasibility of extraction and formulation processes.
• In vivo testing and regulatory compliance for market authorization in cosmetic and food sectors.
• Intellectual Property Strategy development to protect nanoencapsulation and formulation innovations.
• Market alignment and partnerships with companies across cosmetics, nutraceuticals, and sustainable agriculture.
• Optimized cultivation conditions (simulated tidal cycles and nitrate enrichment) significantly enhanced biomass quality and endogenous production of mycosporine-like amino acids (MAAs).
• Fishpond effluents as a natural nitrate source were tested and compared to synthetic nitrate. Results showed that fishpond effluents achieved comparable MAA yields while requiring lower overall nitrate input, highlighting a sustainable and integrated approach aligned with circular aquaculture practices.
• Fresh biomass extraction with alkaline and enzymatic hydrolysis improved yields of antioxidant compounds (phenolics, phycobiliproteins, carbohydrates).
• Efficient MAA extraction using high-temperature water-based methods surpassed literature benchmarks.
• Porphyran extraction was successfully achieved and showed potential both as a bioactive compound and encapsulation agent. Porphyran-based nanoencapsulation is being developed as a natural alternative to synthetic encapsulates.
• In vitro bioactivity testing demonstrated antioxidant activity (ABTS), and SPF properties (PMMA plate assays).
• Biostimulant potential of residual biomass was confirmed through plant hormone-like bioassays.
Potential Impacts:
• Industrial relevance confirmed: As a direct outcome of the project, a follow-up collaboration has been established with Cantabria Labs, a Spanish cosmetics company, to further develop a commercial photoprotector/antioxidant cream. This new project aims to build upon the current findings by incorporating additional plant extracts to boost SPF and antioxidant activity, reinforcing the translational value of the research.
• Blue Bioeconomy alignment: The approach supports zero-waste strategies and marine biomass circularity.
• Nutraceutical and cosmeceutical innovation: Clean-label, natural bioactives with proven efficacy contribute to high-demand product segments.
• Agricultural circularity: Biomass residues with biostimulant activity offer potential for sustainable agriculture inputs.
Key Needs for Further Uptake and Success:
• Scale-up and demonstration activities to validate industrial feasibility of extraction and formulation processes.
• In vivo testing and regulatory compliance for market authorization in cosmetic and food sectors.
• Intellectual Property Strategy development to protect nanoencapsulation and formulation innovations.
• Market alignment and partnerships with companies across cosmetics, nutraceuticals, and sustainable agriculture.