{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T23:15:48Z","timestamp":1769469348585,"version":"3.49.0"},"reference-count":49,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,1,25]],"date-time":"2026-01-25T00:00:00Z","timestamp":1769299200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Iceland, Liechtenstein, and Norway through the project MICROBOOST","award":["PT-INNOVATION-0102\u2014EEA.BG.CALL2.030.2021"],"award-info":[{"award-number":["PT-INNOVATION-0102\u2014EEA.BG.CALL2.030.2021"]}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["2021.08142.BD"],"award-info":[{"award-number":["2021.08142.BD"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["UIDB\/04326\/2020"],"award-info":[{"award-number":["UIDB\/04326\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["LA\/P\/0101\/2020"],"award-info":[{"award-number":["LA\/P\/0101\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Processes"],"abstract":"<jats:p>Multi-product biorefineries, which transform biomass feedstocks into multiple valuable bio-based products, are pivotal for transitioning from a fossil-based economy to a sustainable circular bioeconomy. This work proposes a processing pipeline for fractionating the macrocomponents of Nannochloropsis oceanica, which can serve as a basis for multi-product microalgae biorefineries. It consists of high-pressure homogenization (1200 bar, 1 cycle) to permeabilize the cells, and sequential membrane processing (0.2 \u00b5m dia-microfiltration followed by 100 kDa ultrafiltration) and ethanolic extraction (60 mL ethanol\/g dry weight, 1 h) to fractionate the disrupted biomass. This biorefinery resulted in four final fractions: (1) enriched in water-soluble proteins (39.0 \u00b1 2.8% w\/w proteins; 10.7 \u00b1 0.8% w\/w carbohydrates); (2) remaining soluble components (5.7 \u00b1 0.4% w\/w proteins; 4.3 \u00b1 0.9% w\/w carbohydrates); (3) lipid-rich extract (62.4 \u00b1 5.8% w\/w lipids); and (4) non-extracted components (11.8 \u00b1 4.5% w\/w lipids), with mass recovery yields of 23.2 \u00b1 2.1%, 6.9 \u00b1 1.0%, 10.6 \u00b1 1.9%, and 60.4 \u00b1 4.1%, respectively. The ultrafiltration protein selectivity was not optimal, despite yielding a 2.6 times more concentrated fraction. Lipid extraction yield (35\u201360%) and purity (56\u201368%) were highly affected by the water content of the microfiltration retentate. Overall, 10.0 \u00b1 0.9% of the proteins, 9.7 \u00b1 1.8% of the carbohydrates, and 42.4 \u00b1 13.4% of the lipids of N. oceanica were recovered in fractions 1, 2, and 3, respectively.<\/jats:p>","DOI":"10.3390\/pr14030420","type":"journal-article","created":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T12:41:16Z","timestamp":1769431276000},"page":"420","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Fractioning Macrocomponents of Nannochloropsis oceanica by High-Pressure Homogenization, Membrane Processing, and Ethanolic Extraction"],"prefix":"10.3390","volume":"14","author":[{"given":"Pedro","family":"Cunha","sequence":"first","affiliation":[{"name":"GreenCoLab\u2014Associa\u00e7\u00e3o Oceano Verde, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"},{"name":"Allmicroalgae\u2014Natural Products, Rua 25 de Abril, 2445-413 Alcoba\u00e7a, Portugal"},{"name":"Center of Marine Sciences, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"given":"Bernardo","family":"Carvalho","sequence":"additional","affiliation":[{"name":"GreenCoLab\u2014Associa\u00e7\u00e3o Oceano Verde, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"},{"name":"Center of Marine Sciences, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"given":"Mariam","family":"Kholany","sequence":"additional","affiliation":[{"name":"GreenCoLab\u2014Associa\u00e7\u00e3o Oceano Verde, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"},{"name":"Center of Marine Sciences, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3634-1128","authenticated-orcid":false,"given":"Helena","family":"Cardoso","sequence":"additional","affiliation":[{"name":"Allmicroalgae\u2014Natural Products, Rua 25 de Abril, 2445-413 Alcoba\u00e7a, Portugal"}]},{"given":"Hugo","family":"Pereira","sequence":"additional","affiliation":[{"name":"GreenCoLab\u2014Associa\u00e7\u00e3o Oceano Verde, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"},{"name":"Center of Marine Sciences, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3101-693X","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Varela","sequence":"additional","affiliation":[{"name":"GreenCoLab\u2014Associa\u00e7\u00e3o Oceano Verde, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"},{"name":"Center of Marine Sciences, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Basso, T., Basso, T., and Basso, L. (2021). Recent Advances in Algal Biomass Production. Biotechnological Applications of Biomass, IntechOpen.","DOI":"10.5772\/intechopen.89320"},{"key":"ref_2","unstructured":"Enzing, C., Ploeg, M., Barbosa, M., and Sijtsma, L. (2017). Microalgae-Based Products for the Food and Feed Sector: An Outlook for Europe, European Commission. JRC Scientific and Policy Reports."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2205","DOI":"10.1007\/978-3-319-32886-7_38","article-title":"Microalgal Biorefinery for Bulk and High-Value Products: Product Extraction within Cell Disintegration","volume":"Volume 3","author":"Postma","year":"2017","journal-title":"Handbook of Electroporation"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1002\/bbb.2187","article-title":"Recent Advances in the Integrated Biorefinery Concept for the Valorization of Algal Biomass through Sustainable Routes","volume":"15","author":"Katiyar","year":"2021","journal-title":"Biofuels Bioprod. Biorefin."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1016\/j.nbt.2015.02.001","article-title":"Biological Potential of Microalgae in China for Biorefinery-Based Production of Biofuels and High Value Compounds","volume":"32","author":"Li","year":"2015","journal-title":"New Biotechnol."},{"key":"ref_6","first-page":"99","article-title":"From Current Algae Products to Future Biorefinery Practices: A Review","volume":"166","author":"Eppink","year":"2017","journal-title":"Adv. Biochem. Eng. Biotechnol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Slegers, P.M., Olivieri, G., Breitmayer, E., Sijtsma, L., Eppink, M.H.M., Wijffels, R.H., and Reith, J.H. (2020). Design of Value Chains for Microalgal Biorefinery at Industrial Scale: Process Integration and Techno-Economic Analysis. Front. Bioeng. Biotechnol., 8.","DOI":"10.3389\/fbioe.2020.550758"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"151124","DOI":"10.1016\/j.cej.2024.151124","article-title":"Unlocking the Potential of Microalgae as Sustainable Bioresources from up to Downstream Processing: A Critical Review","volume":"488","author":"Diankristanti","year":"2024","journal-title":"Chem. Eng. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"114962","DOI":"10.1016\/j.carbpol.2019.06.001","article-title":"Structural Analysis and Potential Immunostimulatory Activity of Nannochloropsis oculata Polysaccharides","volume":"222","author":"Pandeirada","year":"2019","journal-title":"Carbohydr. Polym."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Ribeiro, C., Santos, E.T., Costa, L., Brazinha, C., Saraiva, P., and Crespo, J.G. (2022). Nannochloropsis sp. Biorefinery: Recovery of Soluble Protein by Membrane Ultrafiltration\/Diafiltration. Membranes, 12.","DOI":"10.3390\/membranes12040401"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.algal.2013.02.001","article-title":"Optimizing Protein Isolation from Defatted and Non-Defatted Nannochloropsis Microalgae Biomass","volume":"2","author":"Gerde","year":"2013","journal-title":"Algal Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1416","DOI":"10.1016\/j.biortech.2017.05.124","article-title":"A Biorefinery for Nannochloropsis: Induction, Harvesting, and Extraction of EPA-Rich Oil and High-Value Protein","volume":"244","author":"Chua","year":"2017","journal-title":"Bioresour. Technol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"102248","DOI":"10.1016\/j.algal.2021.102248","article-title":"Process Development of Enzymatically-Generated Algal Protein Hydrolysates for Specialty Food Applications","volume":"55","author":"Wilken","year":"2021","journal-title":"Algal Res."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Soto-Sierra, L., and Nikolov, Z.L. (2022). Feasibility of Membrane Ultrafiltration as a Single-Step Clarification and Fractionation of Microalgal Protein Hydrolysates. Front. Bioeng. Biotechnol., 10.","DOI":"10.3389\/fbioe.2022.957268"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.1007\/s10811-020-02037-z","article-title":"Simultaneous Extraction and Fractionation of Lipids from the Microalga Nannochloropsis sp. for the Production of EPA-Rich Polar Lipid Concentrates","volume":"32","year":"2020","journal-title":"J. Appl. Phycol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"101729","DOI":"10.1016\/j.algal.2019.101729","article-title":"Obtaining Highly Pure EPA-Rich Lipids from Dry and Wet Nannochloropsis gaditana Microalgal Biomass Using Ethanol, Hexane and Acetone","volume":"45","author":"Grima","year":"2020","journal-title":"Algal Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.biortech.2016.04.025","article-title":"Enhanced Lipid Recovery from Nannochloropsis Microalgae by Treatment with Optimized Cell Wall Degrading Enzyme Mixtures","volume":"212","author":"Zuorro","year":"2016","journal-title":"Bioresour. Technol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.algal.2018.09.011","article-title":"Production of Protein-Rich Extracts from Disrupted Microalgae Cells: Impact of Solvent Treatment and Lyophilization","volume":"36","author":"Grossmann","year":"2018","journal-title":"Algal Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1016\/j.biortech.2017.10.062","article-title":"Effect of an Enzymatic Treatment with Cellulase and Mannanase on the Structural Properties of Nannochloropsis Microalgae","volume":"249","author":"Maffei","year":"2018","journal-title":"Bioresour. Technol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.algal.2013.12.004","article-title":"Aqueous Extraction of Proteins from Microalgae: Effect of Different Cell Disruption Methods","volume":"3","author":"Safi","year":"2014","journal-title":"Algal Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.algal.2013.11.017","article-title":"Release of Hydro-Soluble Microalgal Proteins Using Mechanical and Chemical Treatments","volume":"3","author":"Safi","year":"2014","journal-title":"Algal Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.biortech.2016.11.068","article-title":"Biorefinery of Microalgal Soluble Proteins by Sequential Processing and Membrane Filtration","volume":"225","author":"Safi","year":"2017","journal-title":"Bioresour. Technol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.biortech.2017.05.012","article-title":"Energy Consumption and Water-Soluble Protein Release by Cell Wall Disruption of Nannochloropsis gaditana","volume":"239","author":"Safi","year":"2017","journal-title":"Bioresour. Technol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.memsci.2014.04.010","article-title":"Integration of Membrane Technology in Microalgae Biorefineries","volume":"464","author":"Gerardo","year":"2014","journal-title":"J. Memb. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.biortech.2012.11.084","article-title":"A Biorefinery from Nannochloropsis sp. Microalga\u2014Extraction of Oils and Pigments. Production of Biohydrogen from the Leftover Biomass","volume":"135","author":"Nobre","year":"2013","journal-title":"Bioresour. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"42513","DOI":"10.1039\/D5RA02952J","article-title":"Integrated Biodiesel and Biopolymer Production from Nannochloropsis Biomass: A Closed-Loop Biorefinery Approach","volume":"15","author":"Nirmala","year":"2025","journal-title":"RSC Adv."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Blanco-Llamero, C., Garc\u00eda-Garc\u00eda, P., and Se\u00f1or\u00e1ns, F.J. (2025). An Integrated Biorefinery Process to Revalorize Marine Biomass from the Microalga Nannochloropsis gaditana Using Pressurized Green Solvents. Mar. Drugs, 23.","DOI":"10.3390\/md23070263"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1016\/j.biortech.2014.11.049","article-title":"Energy Evaluation of Algal Cell Disruption by High Pressure Homogenisation","volume":"184","author":"Yap","year":"2015","journal-title":"Bioresour. Technol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.biortech.2013.04.074","article-title":"Quantitative Evaluation of the Ease of Rupture of Industrially Promising Microalgae by High Pressure Homogenization","volume":"140","author":"Spiden","year":"2013","journal-title":"Bioresour. Technol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1021\/ac60111a017","article-title":"Colorimetric Method for Determination of Sugars and Related Substances","volume":"28","author":"Dubois","year":"1956","journal-title":"Anal. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1016\/j.algal.2015.07.013","article-title":"Nitrogen-to-Protein Conversion Factors Revisited for Applications of Microalgal Biomass Conversion to Food, Feed and Fuel","volume":"11","author":"Templeton","year":"2015","journal-title":"Algal Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1139\/y59-099","article-title":"A Rapid Method of Total Lipid Extraction and Purification","volume":"37","author":"Bligh","year":"1959","journal-title":"Can. J. Biochem. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.bej.2012.10.008","article-title":"Critical Analysis of Quantitative Indicators of Cell Disruption Applied to Saccharomyces cerevisiae Processed with an Industrial High Pressure Homogenizer","volume":"70","author":"Spiden","year":"2013","journal-title":"Biochem. Eng. J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"101616","DOI":"10.1016\/j.algal.2019.101616","article-title":"Evaluating Microalgal Cell Disruption upon Ultra High Pressure Homogenization","volume":"42","author":"Bernaerts","year":"2019","journal-title":"Algal Res."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Singh, R. (2015). Introduction to Membrane Technology. Membrane Technology and Engineering for Water Purification, Elsevier.","DOI":"10.1016\/B978-0-444-63362-0.00001-X"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"102082","DOI":"10.1016\/j.algal.2020.102082","article-title":"Recovery of Microalgal Protein by Three-Steps Membrane Filtration: Advancements and Feasibility","volume":"51","author":"Gifuni","year":"2020","journal-title":"Algal Res."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Jafari, S., and Castro-Mu\u00f1oz, R. (2021). Recovery of High Added Value Compounds from Microalgae Cultivation Using Membrane Technology. Membrane Separation of Food Bioactive Ingredients, Springer.","DOI":"10.1007\/978-3-030-84643-5"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1016\/j.memsci.2013.12.027","article-title":"Comparison of Membrane Fouling at Constant Flux and Constant Transmembrane Pressure Conditions","volume":"454","author":"Miller","year":"2014","journal-title":"J. Memb. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"101556","DOI":"10.1016\/j.algal.2019.101556","article-title":"The Effects of Different Extraction Methods of Lipids from Nannochloropsis oceanica on the Contents of Omega-3 Fatty Acids","volume":"41","author":"Figueiredo","year":"2019","journal-title":"Algal Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1501","DOI":"10.1007\/s10811-013-0189-y","article-title":"Influence of Extraction Solvent System on the Extractability of Lipid Components from the Biomass of Nannochloropsis gaditana","volume":"26","author":"Ryckebosch","year":"2014","journal-title":"J. Appl. Phycol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.algal.2016.11.005","article-title":"Wet Lipid Extraction from the Microalga Nannochloropsis sp.: Disruption, Physiological Effects and Solvent Screening","volume":"21","author":"Angles","year":"2017","journal-title":"Algal Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1258","DOI":"10.3390\/md12031258","article-title":"A Novel Lipid Extraction Method from Wet Microalga Picochlorum sp. at Room Temperature","volume":"12","author":"Yang","year":"2014","journal-title":"Mar. Drugs"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"102586","DOI":"10.1016\/j.algal.2021.102586","article-title":"Supercritical Fluid Extraction and Pressurized Liquid Extraction Processes Applied to Eicosapentaenoic Acid-Rich Polar Lipid Recovery from the Microalga Nannochloropsis sp.","volume":"61","year":"2022","journal-title":"Algal Res."},{"key":"ref_44","first-page":"15519","article-title":"Obtaining EPA-Rich Polar Lipids from Microalga Nannochloropsis sp. by Silica-Gel Chromatography Using Non-Toxic Solvents","volume":"13","author":"Medina","year":"2022","journal-title":"Biomass Convers. Biorefin."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/j.fbp.2024.09.002","article-title":"Large-Scale Production of Nannochloropsis-Derived EPA: Current Status and Perspectives via a Biorefinery Context","volume":"148","author":"Piyatilleke","year":"2024","journal-title":"Food Bioprod. Process."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"103073","DOI":"10.1016\/j.algal.2023.103073","article-title":"Obtaining Eicosapentaenoic Acid-Enriched Polar Lipids from Microalga Nannochloropsis sp. by Lipase-Catalysed Hydrolysis","volume":"71","year":"2023","journal-title":"Algal Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/j.algal.2018.10.023","article-title":"Extraction and Fractionation of Microalgae-Based Protein Products","volume":"36","author":"Stoykova","year":"2018","journal-title":"Algal Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1186\/1476-511X-12-102","article-title":"Acute Appearance of Fatty Acids in Human Plasma\u2014A Comparative Study between Polar-Lipid Rich Oil from the Microalgae Nannochloropsis oculata and Krill Oil in Healthy Young Males","volume":"12","author":"Kagan","year":"2013","journal-title":"Lipids Health Dis."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1089\/ind.2017.29102.wmo","article-title":"Cutting Out the Middle Fish: Marine Microalgae as the Next Sustainable Omega-3 Fatty Acids and Protein Source","volume":"13","author":"Moomaw","year":"2017","journal-title":"Ind. Biotechnol."}],"container-title":["Processes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2227-9717\/14\/3\/420\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T13:08:32Z","timestamp":1769432912000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2227-9717\/14\/3\/420"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,25]]},"references-count":49,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["pr14030420"],"URL":"https:\/\/doi.org\/10.3390\/pr14030420","relation":{},"ISSN":["2227-9717"],"issn-type":[{"value":"2227-9717","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,25]]}}}