{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T12:21:39Z","timestamp":1774614099297,"version":"3.50.1"},"reference-count":111,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T00:00:00Z","timestamp":1774483200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"PhotoBioValue\u2014Light effect on photobioreactor design for microalgae cultivation: enhancement of photosynthetic efficiency and biomass value","award":["PTDC\/BTA-BTA\/2902\/2021"],"award-info":[{"award-number":["PTDC\/BTA-BTA\/2902\/2021"]}]},{"name":"SeaProPep\u2014SEAweed as a sustainable source of PROtein and bioactive PEPtides","award":["2022.05853.PTDC"],"award-info":[{"award-number":["2022.05853.PTDC"]}]},{"name":"national funds through FCT\/MCTES","award":["UIDB\/00511\/2020"],"award-info":[{"award-number":["UIDB\/00511\/2020"]}]},{"name":"national funds through FCT\/MCTES","award":["LA\/P\/0045\/2020"],"award-info":[{"award-number":["LA\/P\/0045\/2020"]}]},{"name":"national funds through the FCT\/MECI","award":["UIDB\/50006\/2020"],"award-info":[{"award-number":["UIDB\/50006\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>Interest in microalgae-based technologies has emerged in recent years as a response to environmental challenges and the global food crisis, for providing alternative and sustainable food products. This study used temperature variations between 18 and 32 \u00b0C and nitrogen-to-phosphorus (N:P) ratios between 1.9 and 42.6 to model and optimize growth and composition of Chlorella vulgaris, a nutritionally interesting species. Lower temperatures appear ideal for this strain. An increase in average biomass productivity was observed with decreasing temperature, leading to a maximum of 122.27 mgdw L\u22121 d\u22121 at 18 \u00b0C on the fourth day of cultivation. The maximum productivities for total proteins, fatty acids, carbohydrates, and pigments were, respectively, 26.9 mg L\u22121 d\u22121, 26.4 mg L\u22121 d\u22121, 16.0 mg L\u22121 d\u22121, and 2.41 mg L\u22121 d\u22121, all referring to 18 \u00b0C. The fatty acid, carotenoid, and amino acid profiles were also ascertained; several indicators suggested that cultivation of these microalgae under the aforementioned optimal conditions holds potential for the food industry. The high proportion of polyunsaturated fatty acids\u2014including two essential fatty acids; the high production of lutein, and the presence of several essential amino acids are among the favorable indicators. Overall, the information generated by this study is helpful to support future pilot studies aimed at the commercial production of microalgae-derived products.<\/jats:p>","DOI":"10.3390\/md24040124","type":"journal-article","created":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T11:32:47Z","timestamp":1774611167000},"page":"124","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["On the Design of Chlorella vulgaris Composition for Potential Food Uses via Manipulation of Cultivation Conditions"],"prefix":"10.3390","volume":"24","author":[{"given":"Ana S.","family":"Pinto","sequence":"first","affiliation":[{"name":"LEPABE (Laboratory for Process Engineering, Environment, Biotechnology and Energy), ALiCE (Associate Laboratory in Chemical Engineering), Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias s\/n, 4200-465 Porto, Portugal"}]},{"given":"Joana","family":"Oliveira","sequence":"additional","affiliation":[{"name":"LEPABE (Laboratory for Process Engineering, Environment, Biotechnology and Energy), ALiCE (Associate Laboratory in Chemical Engineering), Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias s\/n, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6192-2760","authenticated-orcid":false,"given":"Ana F.","family":"Esteves","sequence":"additional","affiliation":[{"name":"LEPABE (Laboratory for Process Engineering, Environment, Biotechnology and Energy), ALiCE (Associate Laboratory in Chemical Engineering), Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias s\/n, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1686-3850","authenticated-orcid":false,"given":"Susana","family":"Casal","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE (Associated Laboratory for Green Chemistry-Network of Chemistry and Technology), Faculty of Pharmacy, University of Porto, Rua de Jorge de Viterbo Ferreira 228, 4050-313 Porto, Portugal"}]},{"given":"Gustavo","family":"Mil-Homens","sequence":"additional","affiliation":[{"name":"LEPABE (Laboratory for Process Engineering, Environment, Biotechnology and Energy), ALiCE (Associate Laboratory in Chemical Engineering), Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias s\/n, 4200-465 Porto, Portugal"}]},{"given":"Francisco X.","family":"Malcata","sequence":"additional","affiliation":[{"name":"LEPABE (Laboratory for Process Engineering, Environment, Biotechnology and Energy), ALiCE (Associate Laboratory in Chemical Engineering), Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias s\/n, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2738-5102","authenticated-orcid":false,"given":"Jos\u00e9 C. M.","family":"Pires","sequence":"additional","affiliation":[{"name":"LEPABE (Laboratory for Process Engineering, Environment, Biotechnology and Energy), ALiCE (Associate Laboratory in Chemical Engineering), Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias s\/n, 4200-465 Porto, Portugal"}]},{"given":"T\u00e2nia G.","family":"Tavares","sequence":"additional","affiliation":[{"name":"LEPABE (Laboratory for Process Engineering, Environment, Biotechnology and Energy), ALiCE (Associate Laboratory in Chemical Engineering), Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias s\/n, 4200-465 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"144590","DOI":"10.1016\/j.scitotenv.2020.144590","article-title":"Revisiting carbon, nitrogen, and phosphorus metabolisms in microalgae for wastewater treatment","volume":"762","author":"Su","year":"2021","journal-title":"Sci. Total Environ."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"108493","DOI":"10.1016\/j.biotechadv.2024.108493","article-title":"Is it possible to shape the microalgal biomass composition with operational parameters for target compound accumulation?","volume":"79","author":"Esteves","year":"2025","journal-title":"Biotechnol. Adv."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"103601","DOI":"10.1016\/j.eti.2024.103601","article-title":"Enhancing urban wastewater treatment: Chlorella vulgaris performance in tertiary treatment and the impact of anaerobic digestate addition","volume":"34","author":"Sousa","year":"2024","journal-title":"Environ. Technol. Innov."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"119736","DOI":"10.1016\/j.enconman.2025.119736","article-title":"Exploring microalgae species for integrated bioenergy Production: A Multi-Fuel cascade valorisation approach","volume":"332","author":"Oliveira","year":"2025","journal-title":"Energy Convers. Manag."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Esteves, A.F., Soares, O.S.G.P., Vilar, V.J.P., Pires, J.C.M., and Gon\u00e7alves, A.L. (2020). The Effect of Light Wavelength on CO2 Capture, Biomass Production and Nutrient Uptake by Green Microalgae: A Step Forward on Process Integration and Optimisation. Energies, 13.","DOI":"10.3390\/en13020333"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"158689","DOI":"10.1016\/j.scitotenv.2022.158689","article-title":"Role of microalgae in achieving sustainable development goals and circular economy","volume":"854","author":"Olabi","year":"2023","journal-title":"Sci. Total Environ."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.algal.2018.08.005","article-title":"Microalgae research worldwide","volume":"35","year":"2018","journal-title":"Algal Res."},{"key":"ref_8","first-page":"725","article-title":"Culturversuche mit Zoochlorellen, Lichenengonidien und anderen niederen Algen","volume":"47","author":"Beijerinck","year":"1890","journal-title":"Bot. Ztg."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"105543","DOI":"10.1016\/j.envexpbot.2023.105543","article-title":"Evidence of physiological adaptation of Chlorella vulgaris under extreme salinity\u2014New insights into a potential halotolerance strategy","volume":"216","author":"Mountourakis","year":"2023","journal-title":"Environ. Exp. Bot."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.biortech.2015.06.050","article-title":"Saline wastewater treatment by Chlorella vulgaris with simultaneous algal lipid accumulation triggered by nitrate deficiency","volume":"193","author":"Shen","year":"2015","journal-title":"Bioresour. Technol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"012007","DOI":"10.1088\/1742-6596\/1338\/1\/012007","article-title":"Fatty Acid Analysis of Marine Microalgae Chlorella vulgaris in Modified Medium Used GC-FID","volume":"1338","author":"Kurnia","year":"2019","journal-title":"J. Phys. Conf. Ser."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Mendes, A.R., Sp\u00ednola, M.P., Lordelo, M., and Prates, J.A.M. (2024). Chemical Compounds, Bioactivities, and Applications of Chlorella vulgaris in Food, Feed and Medicine. Appl. Sci., 14.","DOI":"10.3390\/app142310810"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1263\/jbb.101.87","article-title":"Commercial applications of microalgae","volume":"101","author":"Spolaore","year":"2006","journal-title":"J. Biosci. Bioeng."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1007\/s10811-005-1641-4","article-title":"An evaluation of methods for extraction and quantification of protein from marine macro- and microalgae","volume":"17","author":"Barbarino","year":"2005","journal-title":"J. Appl. Phycol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Barkia, I., Saari, N., and Manning, S.R. (2019). Microalgae for High-Value Products Towards Human Health and Nutrition. Mar. Drugs, 17.","DOI":"10.3390\/md17050304"},{"key":"ref_16","unstructured":"Esteves, A.F.C. (2025, May 17). Shaping Microalgal Biomass Composition for Targeted Product Accumulation: Impact of Operational Parameters. Available online: https:\/\/repositorio-aberto.up.pt\/handle\/10216\/162001."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/j.rser.2014.04.007","article-title":"Morphology, composition, production, processing and applications of Chlorella vulgaris: A review","volume":"35","author":"Safi","year":"2014","journal-title":"Renew. Sustain. Energy Rev."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s10811-020-02126-z","article-title":"Lipid accumulation and profiling in microalgae Chlorolobion sp. (BIOTECH 4031) and Chlorella sp. (BIOTECH 4026) during nitrogen starvation for biodiesel production","volume":"33","author":"Arguelles","year":"2021","journal-title":"J. Appl. Phycol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1007\/s00449-020-02284-x","article-title":"Characteristics of lipid biosynthesis of Chlorella pyrenoidosa under stress conditions","volume":"43","author":"Dong","year":"2020","journal-title":"Bioprocess Biosyst. Eng."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"121783","DOI":"10.1016\/j.biortech.2019.121783","article-title":"Effect of light quality on growth rate, carbohydrate accumulation, fatty acid profile and lutein biosynthesis of Chlorella sp. AE10","volume":"291","author":"Li","year":"2019","journal-title":"Bioresour. Technol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e23238","DOI":"10.1016\/j.heliyon.2023.e23238","article-title":"Improvement of N and P ratio for enhanced biomass productivity and sustainable cultivation of Chlorella vulgaris microalgae","volume":"10","author":"Magyar","year":"2024","journal-title":"Heliyon"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"116076","DOI":"10.1016\/j.envres.2023.116076","article-title":"Microalgal cultures for the remediation of wastewaters with different nitrogen to phosphorus ratios: Process modelling using artificial neural networks","volume":"231","author":"Salgado","year":"2023","journal-title":"Environ. Res."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Montoya-Vallejo, C., Guzm\u00e1n Duque, F.L., and Quintero D\u00edaz, J.C. (2023). Biomass and lipid production by the native green microalgae Chlorella sorokiniana in response to nutrients, light intensity, and carbon dioxide: Experimental and modeling approach. Front. Bioeng. Biotechnol., 11.","DOI":"10.3389\/fbioe.2023.1149762"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1082","DOI":"10.1016\/j.renene.2019.09.071","article-title":"Experimental evaluation of the culture parameters for optimum yield of lipids and other nutraceutically valuable compounds in Chloroidium saccharophillum","volume":"147","author":"Santhakumaran","year":"2020","journal-title":"Renew. Energy"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1007\/s10863-019-09813-8","article-title":"Fatty acid synthesis by Chlamydomonas reinhardtii in phosphorus limitation","volume":"52","author":"Qari","year":"2020","journal-title":"J. Bioenerg. Biomembr."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1007\/s13201-019-0969-x","article-title":"Response surface methodology as a statistical tool for optimization of physio-biochemical cellular components of microalgae Chlorella pyrenoidosa for biodiesel production","volume":"9","author":"Bajwa","year":"2019","journal-title":"Appl. Water Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"8255","DOI":"10.1007\/s00253-019-10051-6","article-title":"Effects of multi-temperature regimes on cultivation of microalgae in municipal wastewater to simultaneously remove nutrients and produce biomass","volume":"103","author":"Xu","year":"2019","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1146","DOI":"10.1016\/j.cep.2009.03.006","article-title":"Effect of temperature and nitrogen concentration on the growth and lipid content of Nannochloropsis oculata and Chlorella vulgaris for biodiesel production","volume":"48","author":"Converti","year":"2009","journal-title":"Chem. Eng. Process. Process Intensif."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"33262","DOI":"10.1021\/acsomega.0c04961","article-title":"Effect of Temperature on Biological Macromolecules of Three Microalgae and Application of FT-IR for Evaluating Microalgal Lipid Characterization","volume":"5","author":"Zhao","year":"2020","journal-title":"ACS Omega"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"129386","DOI":"10.1016\/j.biortech.2023.129386","article-title":"Acclimation-driven microalgal cultivation improved temperature and light stress tolerance, CO2 sequestration and metabolite regulation for bioenergy production","volume":"385","author":"Chauhan","year":"2023","journal-title":"Bioresour. Technol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"124850","DOI":"10.1016\/j.biortech.2021.124850","article-title":"Stepwise-incremental physicochemical factors induced acclimation and tolerance in oleaginous microalgae to crucial outdoor stresses and improved properties as biodiesel feedstocks","volume":"328","author":"Maneechote","year":"2021","journal-title":"Bioresour. Technol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"102027","DOI":"10.1016\/j.algal.2020.102027","article-title":"Haematococcus pluvialis bioprocess optimization: Effect of light quality, temperature and irradiance on growth, pigment content and photosynthetic response","volume":"51","author":"Pereira","year":"2020","journal-title":"Algal Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.algal.2016.03.001","article-title":"Effect of temperature and nitrogen concentration on lipid productivity and fatty acid composition in three Chlorella strains","volume":"16","author":"Stirk","year":"2016","journal-title":"Algal Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"20170082","DOI":"10.1515\/cppm-2017-0082","article-title":"Application of central composite design to optimize culture conditions of Chlorella vulgaris in a batch photobioreactor: An efficient modeling approach","volume":"13","author":"Kazeem","year":"2018","journal-title":"Chem. Prod. Process Model."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.fbp.2022.12.004","article-title":"Multivariable optimization process of heterotrophic growth of Chlorella vulgaris","volume":"138","author":"Geada","year":"2023","journal-title":"Food Bioprod. Process."},{"key":"ref_36","unstructured":"Dunn, J., and Grider, M.H. (2023). Physiology, Adenosine Triphosphate, StatPearls. Available online: http:\/\/europepmc.org\/books\/NBK553175."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Pacheco-Torgal, F., Ivanov, V., and Tsang, D. (2020). Building integrated photobioreactor. Bio-Based Materials and Biotechnologies for Eco-Efficient Construction, Woodhead Publishing.","DOI":"10.1016\/B978-0-12-819481-2.00001-5"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"106355","DOI":"10.1016\/j.resconrec.2022.106355","article-title":"Overview on stress-induced strategies for enhanced microalgae lipid production: Application, mechanisms and challenges","volume":"183","author":"Song","year":"2022","journal-title":"Resour. Conserv. Recycl."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"8813","DOI":"10.1007\/s13399-021-02086-5","article-title":"Enhancing the efficiency of thermal conversion of microalgae: A review","volume":"13","author":"Mustapha","year":"2023","journal-title":"Biomass Convers. Biorefin."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.tibtech.2019.07.011","article-title":"Dunaliella Microalgae for Nutritional Protein: An Undervalued Asset","volume":"38","author":"Sui","year":"2020","journal-title":"Trends Biotechnol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"102104","DOI":"10.1016\/j.algal.2020.102104","article-title":"Microalgae under environmental stress as a source of antioxidants","volume":"52","author":"Gauthier","year":"2020","journal-title":"Algal Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1007\/s11157-021-09571-3","article-title":"Fatty acids of microalgae: Diversity and applications","volume":"20","author":"Maltseva","year":"2021","journal-title":"Rev. Environ. Sci. Bio\/Technol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1186\/s13068-019-1458-z","article-title":"Hormesis effects of phosphorus on the viability of Chlorella regularis cells under nitrogen limitation","volume":"12","author":"Fu","year":"2019","journal-title":"Biotechnol. Biofuels"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"121912","DOI":"10.1016\/j.cej.2019.121912","article-title":"Enhancing lipid production in microalgae Chlorella PY-ZU1 with phosphorus excess and nitrogen starvation under 15% CO2 in a continuous two-step cultivation process","volume":"375","author":"Chu","year":"2019","journal-title":"Chem. Eng. J."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Oslan, S.N.H., Shoparwe, N.F., Yusoff, A.H., Rahim, A.A., Chang, C.S., Tan, J.S., Oslan, S.N., Arumugam, K., Bin Ariff, A., and Sulaiman, A.Z. (2021). A Review on Haematococcus pluvialis Bioprocess Optimization of Green and Red Stage Culture Conditions for the Production of Natural Astaxanthin. Biomolecules, 11.","DOI":"10.3390\/biom11020256"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"125523","DOI":"10.1016\/j.biortech.2021.125523","article-title":"Transcriptomic analysis unravels the modulating mechanisms of the biomass and value-added bioproducts accumulation by light spectrum in Eustigmatos cf. polyphem (Eustigmatophyceae)","volume":"338","author":"Zhang","year":"2021","journal-title":"Bioresour. Technol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1186\/s13213-020-01588-3","article-title":"Effects of different abiotic stresses on carotenoid and fatty acid metabolism in the green microalga Dunaliella salina Y6","volume":"70","author":"Wu","year":"2020","journal-title":"Ann. Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"101732","DOI":"10.1016\/j.algal.2019.101732","article-title":"Improved production of lutein and \u03b2-carotene by thermal and light intensity upshifts in the marine microalga Tetraselmis sp. CTP4","volume":"45","author":"Wu","year":"2020","journal-title":"Algal Res."},{"key":"ref_49","unstructured":"Pandey, A., Lee, D.-J., Chisti, Y., and Soccol, C. (2014). An Open Pond System for Microalgal Cultivation. Biofuels from Algae, Elsevier."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1186\/s13068-022-02249-0","article-title":"The growth, lipid accumulation and adaptation mechanism in response to variation of temperature and nitrogen supply in psychrotrophic filamentous microalga Xanthonema hormidioides (Xanthophyceae)","volume":"16","author":"Gao","year":"2023","journal-title":"Biotechnol. Biofuels Bioprod."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"101821","DOI":"10.1016\/j.algal.2020.101821","article-title":"Co-production of lutein and fatty acid in microalga Chlamydomonas sp. JSC4 in response to different temperatures with gene expression profiles","volume":"47","author":"Ma","year":"2020","journal-title":"Algal Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"102127","DOI":"10.1016\/j.algal.2020.102127","article-title":"Interactive effect of light quality and temperature on Chlamydomonas reinhardtii growth kinetics and lipid synthesis","volume":"53","author":"Li","year":"2021","journal-title":"Algal Res."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"129611","DOI":"10.1016\/j.biortech.2023.129611","article-title":"Utilization of lipidic food waste as low-cost nutrients for enhancing the potentiality of biofuel production from engineered diatom under temperature variations","volume":"387","author":"Yang","year":"2023","journal-title":"Bioresour. Technol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1080\/02757540.2019.1688308","article-title":"Antioxidant response mechanism of freshwater microalgae species to reactive oxygen species production: A mini review","volume":"36","author":"Ugya","year":"2020","journal-title":"Chem. Ecol."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Pinto, A.S., Maia, C., Sousa, S.A., Tavares, T., and Pires, J.C.M. (2025). Amino Acid and Carotenoid Profiles of Chlorella vulgaris During Two-Stage Cultivation at Different Salinities. Bioengineering, 12.","DOI":"10.3390\/bioengineering12030284"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"102008","DOI":"10.1016\/j.algal.2020.102008","article-title":"Bioprospecting and characterization of temperature tolerant microalgae from Bonaire","volume":"50","author":"Barten","year":"2020","journal-title":"Algal Res."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"113023","DOI":"10.1016\/j.envres.2022.113023","article-title":"Physiological and transcription level responses of microalgae Auxenochlorella protothecoides to cold and heat induced oxidative stress","volume":"211","author":"Xing","year":"2022","journal-title":"Environ. Res."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"111852","DOI":"10.1016\/j.rser.2021.111852","article-title":"Continuous cultivation of microalgae in photobioreactors as a source of renewable energy: Current status and future challenges","volume":"154","author":"Peter","year":"2022","journal-title":"Renew. Sustain. Energy Rev."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1016\/j.tibtech.2018.09.006","article-title":"Current Bottlenecks and Challenges of the Microalgal Biorefinery","volume":"37","author":"Gifuni","year":"2019","journal-title":"Trends Biotechnol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1016\/j.algal.2016.06.016","article-title":"Influence of temperature on Chlorella vulgaris growth and mortality rates in a photobioreactor","volume":"18","author":"Bernard","year":"2016","journal-title":"Algal Res."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"114","DOI":"10.33714\/masteb.717126","article-title":"Determination of growth conditions for Chlorella vulgaris","volume":"9","year":"2020","journal-title":"Mar. Sci. Technol. Bull."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.enconman.2017.06.077","article-title":"Semicontinuous outdoor cultivation and efficient harvesting of marine Chlorella vulgaris BDUG 91771 with minimum solid co-precipitation and high floc recovery for biodiesel","volume":"149","author":"Mathimani","year":"2017","journal-title":"Energy Convers. Manag."},{"key":"ref_63","unstructured":"Cassidy, K.O. (2011). Evaluating Algal Growth at Different Temperatures. [Master\u2019s Thesis, University of Kentucky]. Available online: https:\/\/uknowledge.uky.edu\/bae_etds\/3."},{"key":"ref_64","first-page":"36","article-title":"Cultivation of microalgae (Chlorella vulgaris) for biodiesel production","volume":"23","year":"2015","journal-title":"Res. Pap. Fac. Mater. Sci. Technol. Slovak Univ. Technol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/0044-8486(89)90218-4","article-title":"Growth and \u03c93 fatty acid and amino acid composition of microalgae under different temperature regimes","volume":"77","author":"James","year":"1989","journal-title":"Aquaculture"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"126227","DOI":"10.1016\/j.biortech.2021.126227","article-title":"Recycling spent water from microalgae harvesting by fungal pellets to re-cultivate Chlorella vulgaris under different nutrient loads for biodiesel production","volume":"344","author":"Chu","year":"2022","journal-title":"Bioresour. Technol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/j.algal.2015.07.014","article-title":"Towards sustainable microalgal biomass production by phycoremediation of a synthetic wastewater: A kinetic study","volume":"11","author":"Silva","year":"2015","journal-title":"Algal Res."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"107818","DOI":"10.1016\/j.bej.2020.107818","article-title":"Nitrogen supplementation for the production of Chlorella vulgaris biomass in secondary effluent from dairy industry","volume":"165","author":"Nunes","year":"2021","journal-title":"Biochem. Eng. J."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1002\/biot.201400594","article-title":"Improving protein production of indigenous microalga Chlorella vulgaris FSP-E by photobioreactor design and cultivation strategies","volume":"10","author":"Chen","year":"2015","journal-title":"Biotechnol. J."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1007\/s10499-011-9477-1","article-title":"High-density growth and crude protein productivity of a thermotolerant Chlorella vulgaris: Production kinetics and thermodynamics","volume":"20","author":"Mahboob","year":"2012","journal-title":"Aquac. Int."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1466","DOI":"10.1002\/jctb.5899","article-title":"Effect of nitrogen concentration, two-stage and prolonged cultivation on growth rate, lipid and protein content of Chlorella vulgaris","volume":"94","author":"Metsoviti","year":"2019","journal-title":"J. Chem. Technol. Biotechnol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.biortech.2018.12.046","article-title":"Light regime and growth phase affect the microalgal production of protein quantity and quality with Dunaliella salina","volume":"275","author":"Sui","year":"2019","journal-title":"Bioresour. Technol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/0044-8486(89)90057-4","article-title":"Biomass production and variation in the biochemical profile (total protein, carbohydrates, RNA, lipids and fatty acids) of seven species of marine microalgae","volume":"83","author":"Ferreiro","year":"1989","journal-title":"Aquaculture"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.rser.2018.07.050","article-title":"Microalgae lipid and biomass for biofuel production: A comprehensive review on lipid enhancement strategies and their effects on fatty acid composition","volume":"97","author":"Sajjadi","year":"2018","journal-title":"Renew. Sustain. Energy Rev."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"122875","DOI":"10.1016\/j.biortech.2020.122875","article-title":"Growth phase-dependent biochemical composition of green microalgae: Theoretical considerations for biogas production","volume":"303","author":"Klin","year":"2020","journal-title":"Bioresour. Technol."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"580","DOI":"10.1016\/j.renene.2020.02.047","article-title":"Regulation of lipid accumulation using nitrogen for microalgae lipid production in Schizochytrium sp. ABC101","volume":"153","author":"Ju","year":"2020","journal-title":"Renew. Energy"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1016\/j.biortech.2011.11.133","article-title":"Effect of light intensity and nitrogen starvation on CO2 fixation and lipid\/carbohydrate production of an indigenous microalga Scenedesmus obliquus CNW-N","volume":"113","author":"Ho","year":"2012","journal-title":"Bioresour. Technol."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.algal.2015.04.023","article-title":"The effect of nitrogen limitation on the physiology and metabolism of Chlorella vulgaris var L3","volume":"10","author":"Ikaran","year":"2015","journal-title":"Algal Res."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/0076-6879(87)48036-1","article-title":"Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes","volume":"148","author":"Lichtenthaler","year":"1987","journal-title":"Methods Enzymol."},{"key":"ref_80","unstructured":"(2025, September 06). 11.1: Fatty Acids\u2014Chemistry LibreTexts. Available online: https:\/\/chem.libretexts.org\/Courses\/American_River_College\/CHEM_309%3A_Applied_Chemistry_for_the_Health_Sciences\/11%3A_Lipids_-_An_Introduction\/11.01%3A_Fatty_Acids."},{"key":"ref_81","doi-asserted-by":"crossref","unstructured":"Patel, A., Rova, U., Christakopoulos, P., and Matsakas, L. (2020). Introduction to essential fatty acids. Nutraceutical Fatty Acids from Oleaginous Microalgae: A Human Health Perspective, Wiley.","DOI":"10.1002\/9781119631729"},{"key":"ref_82","doi-asserted-by":"crossref","unstructured":"Moradi-Kheibari, N., Ahmadzadeh, H., and Lyon, S.R. (2022). Correlation of Total Lipid Content of Chlorella vulgaris With the Dynamics of Individual Fatty Acid Growth Rates. Front. Mar. Sci., 9.","DOI":"10.3389\/fmars.2022.837067"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"e04447","DOI":"10.1016\/j.heliyon.2020.e04447","article-title":"Characterization of Chlorella sorokiniana and Chlorella vulgaris fatty acid components under a wide range of light intensity and growth temperature for their use as biological resources","volume":"6","author":"Yun","year":"2020","journal-title":"Heliyon"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1007\/s13668-021-00363-3","article-title":"Dietary Polyunsaturated Fatty Acids (PUFAs): Uses and Potential Health Benefits","volume":"10","author":"Kapoor","year":"2021","journal-title":"Curr. Nutr. Rep."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"8231","DOI":"10.1111\/1750-3841.17529","article-title":"Chlorella vulgaris as a food substitute: Applications and benefits in the food industry","volume":"89","author":"Wang","year":"2024","journal-title":"J. Food Sci."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"2325","DOI":"10.1007\/s10811-018-1430-5","article-title":"An efficient method for the sequential production of lipid and carotenoids from the Chlorella Growth Factor-extracted biomass of Chlorella vulgaris","volume":"30","author":"Anthony","year":"2018","journal-title":"J. Appl. Phycol."},{"key":"ref_87","doi-asserted-by":"crossref","unstructured":"Wu, K., Lai, J., Zhang, Q., Wang, Y., Cui, X., Liu, Y., Wu, X., Yu, Z., and Ruan, R. (2024). Optimizing Chlorella vulgaris Cultivation to Enhance Biomass and Lutein Production. Foods, 13.","DOI":"10.3390\/foods13162514"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"101707","DOI":"10.1016\/j.algal.2019.101707","article-title":"Pilot-scale production of lutein using Chlorella vulgaris","volume":"44","author":"McClure","year":"2019","journal-title":"Algal Res."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"474","DOI":"10.3945\/an.113.004028","article-title":"Renzi Carotenoids","volume":"4","author":"Hammond","year":"2013","journal-title":"Adv. Nutr."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1016\/S0306-9877(03)00198-1","article-title":"Are lutein and zeaxanthin conditionally essential nutrients for eye health?","volume":"61","author":"Semba","year":"2003","journal-title":"Med. Hypotheses"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"104107","DOI":"10.1016\/j.jff.2020.104107","article-title":"Lutein protected the retina from light induced retinal damage by inhibiting increasing oxidative stress and inflammation","volume":"73","author":"Yang","year":"2020","journal-title":"J. Funct. Foods"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"992","DOI":"10.1093\/jn\/133.4.992","article-title":"Lutein and Zeaxanthin Dietary Supplements Raise Macular Pigment Density and Serum Concentrations of these Carotenoids in Humans","volume":"133","author":"Bone","year":"2003","journal-title":"J. Nutr."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"282","DOI":"10.3390\/colorants3040020","article-title":"Trade-Off Between Growth Regimes in Chlorella vulgaris: Impact on Carotenoid Production","volume":"3","author":"Caetano","year":"2024","journal-title":"Colorants"},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/0960-8524(96)00058-2","article-title":"Evolution of pigment composition in Chlorella vulgaris","volume":"57","author":"Gouveia","year":"1996","journal-title":"Bioresour. Technol."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.jpba.2017.09.018","article-title":"Separation, identification and quantification of carotenoids and chlorophylls in dietary supplements containing Chlorella vulgaris and Spirulina platensis using High Performance Thin Layer Chromatography","volume":"148","author":"Hynstova","year":"2018","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1093\/advances\/nmy054","article-title":"Nutritionally Essential Amino Acids","volume":"9","author":"Hou","year":"2018","journal-title":"Adv. Nutr."},{"key":"ref_97","doi-asserted-by":"crossref","unstructured":"Wu, G. (2021). Amino Acids: Biochemistry and Nutrition, Second Edition. Amino Acids: Biochemistry and Nutrition, CRC Press. [2nd ed.].","DOI":"10.1201\/9781003092742"},{"key":"ref_98","doi-asserted-by":"crossref","unstructured":"Lorenzo, K., Santocildes, G., Torrella, J.R., Magalh\u00e3es, J., Pag\u00e8s, T., Viscor, G., Torres, J.L., and Ramos-Romero, S. (2023). Bioactivity of Macronutrients from Chlorella in Physical Exercise. Nutrients, 15.","DOI":"10.3390\/nu15092168"},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1007\/s10811-016-0974-5","article-title":"Algae as nutritional and functional food sources: Revisiting our understanding","volume":"29","author":"Wells","year":"2017","journal-title":"J. Appl. Phycol."},{"key":"ref_100","doi-asserted-by":"crossref","unstructured":"Wu, G. (2017). Principles of Animal Nutrition. Principles of Animal Nutrition, CRC Press.","DOI":"10.1201\/9781315120065"},{"key":"ref_101","unstructured":"(2025, May 12). FMT DSA SERIES|Bench Tio Fermenter, Pilot Scale Fermenter, Air Lift Bioreactor, Beer Fermenter|FERMENTEC. Available online: http:\/\/fermentec.co.kr\/eng\/product\/bench-top-fermenter\/fmt-dsa-series\/?ckattempt=2."},{"key":"ref_102","doi-asserted-by":"crossref","unstructured":"Oliveira, J., Pardilh\u00f3, S., Dias, J.M., and Pires, J.C.M. (2023). Microalgae to Bioenergy: Optimization of Aurantiochytrium sp. Saccharification. Biology, 12.","DOI":"10.3390\/biology12070935"},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1023\/A:1008046023487","article-title":"An optical method for the rapid measurement of micromolar concentrations of nitrate in marine phytoplankton cultures","volume":"11","author":"Collos","year":"1999","journal-title":"J. Appl. Phycol."},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"2485","DOI":"10.5012\/bkcs.2009.30.10.2485","article-title":"Efficient Colorimetric Assay of RNA Polymerase Activity Using Inorganic Pyrophosphatase and Ammonium Molybdate","volume":"30","author":"Lee","year":"2009","journal-title":"Bull. Korean Chem. Soc."},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/S0021-9258(19)52451-6","article-title":"Protein Measurement with the Folin Phenol Reagent","volume":"193","author":"Lowry","year":"1951","journal-title":"J. Biol. Chem."},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1002\/elsc.201200212","article-title":"Carbon dioxide biofixation by Chlorella vulgaris at different CO2 concentrations and light intensities","volume":"14","author":"Lopes","year":"2014","journal-title":"Eng. Life Sci."},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1021\/ac60111a017","article-title":"Method for Determination of Sugars and Related Substances","volume":"28","author":"Dubois","year":"1956","journal-title":"Anal. Chem."},{"key":"ref_108","unstructured":"Van Wychen, S., and Laurens, L.M.L. (2025, June 09). Determination of Total Carbohydrates in Algal Biomass: Laboratory Analytical Procedure (LAP) (Revised), Available online: www.nrel.gov\/publications."},{"key":"ref_109","doi-asserted-by":"crossref","unstructured":"Pagels, F., Pereira, R.N., Vicente, A.A., and Guedes, A.C. (2021). Extraction of Pigments from Microalgae and Cyanobacteria\u2014A Review on Current Methodologies. Appl. Sci., 11.","DOI":"10.3390\/app11115187"},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/0021-9673(93)E0821-B","article-title":"Applications of amino acid derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate: Analysis of feed grains, intravenous solutions and glycoproteins","volume":"661","author":"Cohen","year":"1994","journal-title":"J. Chromatogr. A"},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"1379","DOI":"10.1007\/s10811-021-02412-4","article-title":"Effects of irradiance of red and blue:red LEDs on Scenedesmus obliquus M2-1 optimization of biomass and high added-value compounds","volume":"33","author":"Pagels","year":"2021","journal-title":"J. Appl. Phycol."}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/24\/4\/124\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T11:43:41Z","timestamp":1774611821000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/24\/4\/124"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,26]]},"references-count":111,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["md24040124"],"URL":"https:\/\/doi.org\/10.3390\/md24040124","relation":{},"ISSN":["1660-3397"],"issn-type":[{"value":"1660-3397","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,26]]}}}