TY - JOUR T1 - Determination of changes in biodiesel production by Gloeocystis vesiculosa cultured in wastewater with the artificial sweetener aspartame and its metabolites AU - Onay, Melih PY - 2026 DA - April Y2 - 2026 DO - 10.3153/AR26008 JF - Aquatic Research JO - Aquat Res PB - Nuray ERKAN ÖZDEN WT - DergiPark SN - 2618-6365 SP - 78 EP - 89 VL - 9 IS - 2 LA - en AB - Microalgae can grow and form biomass in wastewater to produce various end products. In this study, the biomass, lipid content, and lipid yield changes of Gloeocystis vesiculosa cultured in wastewater containing different concentrations of the artificial sweetener APM and its metabolites DKP and PHE were evaluated. The highest biomass amount was 1080 ± 20 mg/L at 250 mg/L of DKP. Conversely, the highest lipid percentage was 31 ± 2% at 250 mg/L APM. There was no significant difference in lipid yield due to the lower biomass content relative to the higher lipid percentages. APM acts as a stress trigger. In addition, the antioxidant enzyme activities of Gloeocystis vesiculosa in wastewater with artificial sweeteners were determined. At 250 mg/L APM, the highest levels of SOD, CAT, APX, and MDA activity were 91 ± 4 U/mg protein, 78 ± 4 U/mg protein, 18 ± 2 U/mg protein, and 3.9 ± 0.2 nmol/mg protein. Future studies using a large-scale photobioreactor will investigate the effects of Gloeocystis vesiculosa on biodiesel production and wastewater treatment in wastewater contaminated with APM, DKP, and PHE. KW - Biodiesel KW - Gloeocystis vesiculosa KW - Aspartame KW - Wastewater KW - Antioxidant activity CR - Abiusi, F., Trompetter, E., Hoenink, H., Wijffels, R.H., & Janssen, M. (2021). Autotrophic and mixotrophic biomass production of the acidophilic Galdieria sulphuraria ACUF 64. Algal Research, 60, 102513. https://doi.org/10.1016/j.algal.2021.102513 CR - Al-Badri, S.G. (2021). Bioethanol Production from Gloeocystis vesiculosa in Synthetic Wastewater. Van Yuzuncu Yil University, Institute of Natural and Applied Science, Department of Environmental Engineering, M. Sc Thesis, Van, Türkiye. CR - Ameri, M., Baron-Sola, A., Khavari-Nejad, R. A., Soltani, N., Najafi, F., Bagheri, A., Martínez, F., & Hernández, L. E. (2020). Aluminium triggers oxidative stress and antioxidant response in the microalgae Scenedesmus sp. Journal of Plant Physiology, 246–247, 153114. https://doi.org/10.1016/j.jplph.2020.153114 CR - Capek, L., Uhliariková, I., Košťálová, Z., Hindáková, A., & Capek, P. (2023). Structural properties of the extracellular biopolymer (β-D-xylo-α-D-mannan) produced by the green microalga Gloeocystis vesiculosa Nägeli. Carbohydrate Research, 525, 108766. https://doi.org/10.1016/j.carres.2023.108766 CR - Çeliktaş, M.O. (2020). Düz-Panel Hava Kaldırmalı Fotobiyoreaktörün Tasarımı ve Çeşitli Dalgaboylarında LED Işık ile Mikroalglerden Yüksek Biyodizel Üretimi. Van Yuzuncu Yil University, Institute of Natural and Applied Science, Department of Environmental Engineering, M. Sc Thesis, Van, Türkiye CR - Choudhary, M., Jetley, U. K., Abash Khan, M., Zutshi, S., & Fatma, T. (2007). Effect of heavy metal stress on proline, malondialdehyde, and superoxide dismutase activity in the cyanobacterium Spirulina platensis-S5. Ecotoxicology and Environmental Safety, 66(2), 204–209. https://doi.org/10.1016/j.ecoenv.2006.02.002 CR - Dias, R.R., Deprá, M.C., De Menezes, C.R., Zepka, L.Q., & Jacob-Lopes, E. (2025). Microalgae cultivation in wastewater: How realistic is this approach for value-added product production? Processes, 13(7), 2052. https://doi.org/10.3390/pr13072052 CR - Duan, Y., Guo, X., Yang, J., Zhang, M., & Li, Y. (2020). Nutrients recycle and the growth of Scenedesmus obliquus in synthetic wastewater under different sodium carbonate concentrations. Royal Society Open Science, 7(1), 191214. https://doi.org/10.1098/rsos.191214 CR - Fal, S., Aasfar, A., Rabie, R., Smouni, A., & Arroussi, H. El. (2022). Salt induced oxidative stress alters physiological, biochemical and metabolomic responses of green microalga Chlamydomonas reinhardtii. Heliyon, 8(1), e08811. https://doi.org/10.1016/j.heliyon.2022.e08811 CR - Folch, J., Lees, M., & Stanley, G.H.S. (1957). A simple method for the isolation and purification of total lipides from animal tissues. Journal of Biological Chemistry, 226(1), 497–509. https://doi.org/10.1016/S0021-9258(18)64849-5 CR - Gao, G., Wu, M., Fu, Q., Li, X., & Xu, J. (2019). A two-stage model with nitrogen and silicon limitation enhances lipid productivity and biodiesel features of the marine bloom-forming diatom Skeletonema costatum. Bioresource Technology, 289, 121717. https://doi.org/10.1016/j.biortech.2019.121717 CR - Geng, Y., Shaukat, A., Azhar, W., Raza, Q.-U.-A., Tahir, A., Abideen, M. Z. U., Zia, M. A. B., Bashir, M. A., & Rehim, A. (2025). Microalgal biorefineries: A systematic review of technological trade-offs and innovation pathways. Biotechnology for Biofuels and Bioproducts, 18(1), 93. https://doi.org/10.1186/s13068-025-02694-7 CR - Hamed, S.M., Zinta, G., Klöck, G., Asard, H., Selim, S., & AbdElgawad, H. (2017). Zinc-induced differential oxidative stress and antioxidant responses in Chlorella sorokiniana and Scenedesmus acuminatus. Ecotoxicology and Environmental Safety, 140, 256–263. https://doi.org/10.1016/j.ecoenv.2017.02.055 CR - Ismaiel, M.M.S., & Piercey-Normore, M.D. (2023). Antioxidant enzymes of Pseudochlorella pringsheimii under two stressors: Variation of SOD isoforms activity. Journal of Plant Research, 136(5), 755–767. https://doi.org/10.1007/s10265-023-01473-5 CR - Jeffers, T.L., McCombs, R., Schmollinger, S., Tirumani, S., Upadhyaya, S., Merchant, S.S., Niyogi, K.K., & Roth, M.S. (2025). An algal nutrient‐replete, optimized medium for fast growth and high triacylglycerol accumulation. Plant Direct, 9(9), e70106. https://doi.org/10.1002/pld3.70106 CR - Kuo, E.Y., Cai, M.-S., & Lee, T.-M. (2020). Ascorbate peroxidase 4 plays a role in the tolerance of Chlamydomonas reinhardtii to photo-oxidative stress. Scientific Reports, 10(1), 13287. https://doi.org/10.1038/s41598-020-70247-z CR - Li, X., Yang, C., Zeng, G., Wu, S., Lin, Y., Zhou, Q., Lou, W., Du, C., Nie, L., & Zhong, Y. (2020). Nutrient removal from swine wastewater with growing microalgae at various zinc concentrations. Algal Research, 46, 101804. https://doi.org/10.1016/j.algal.2020.101804 CR - Maltsev, Y., Kulikovskiy, M., & Maltseva, S. (2023). Nitrogen and phosphorus stress as a tool to induce lipid production in microalgae. Microbial Cell Factories, 22(1), 239. https://doi.org/10.1186/s12934-023-02244-6 CR - Mihiraj, R.A.I.S., Abinayan, I., Madhusanka, H.G.D., Ketheesan, B., & Sivakanthan, S. (2022). Enhancement of lipid yield in Chlorella sp. under different stress conditions for the production of biodiesel. Journal of Dry Zone Agriculture, 8(2), 48–67. https://doi.org/10.4038/jdza.v8i2.62 CR - Morales-Sánchez, D., Tinoco-Valencia, R., Kyndt, J., & Martínez, A. (2013). Heterotrophic growth of Neochloris oleoabundans using glucose as a carbon source. Biotechnology for Biofuels, 6(1), 100. https://doi.org/10.1186/1754-6834-6-100 CR - Muhetaer, G., Jayasanka, S., & Fujino, T. (2020). Oxidative stress and antioxidant responses of Phormidium ambiguum and Microcystis aeruginosa under diurnally varying light conditions. Microorganisms, 8(6), 890. https://doi.org/10.3390/microorganisms8060890 CR - Nur, M.M.A., & Buma, A.G.J. (2019). Opportunities and challenges of microalgal cultivation on wastewater, with special focus on palm oil mill effluent and the production of high value compounds. Waste and Biomass Valorization, 10(8), 2079–2097. https://doi.org/10.1007/s12649-018-0256-3 CR - Oğuz, A., Köker, L., Özbayram, E.G., Akcaalan, R., & Albay, M. (2024). Biodiesel production from Botryococcus sudeticus and Chlorella vulgaris: Assessment of nitrogen deficiency on lipid, fame yield and biodiesel properties. Waste and Biomass Valorization, 15(5), 2757–2768. https://doi.org/10.1007/s12649-023-02359-2 CR - Onay, M. (2020). Biomass and bio-butanol production from Borodinellopsis texensis CCALA 892 in synthetic wastewater: determination of biochemical composition. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 24(2), 306–316. https://doi.org/10.19113/sdufenbed.573432 CR - Onay, M., & Ayas, Z.S. (2024). Coproduction of biofuel and pigments from Micractinium sp. using UV-induced mutagenesis and adding abscisic acid and salicylic acid for biorefinery concepts. Arabian Journal for Science and Engineering, 49(6), 7929–7944. https://doi.org/10.1007/s13369-023-08531-z CR - Onay, M., Sonmez, C., Oktem, H.A., & Yucel, A.M. (2014). Thermo-resistant green microalgae for effective biodiesel production: Isolation and characterization of unialgal species from geothermal flora of central Anatolia. Bioresource Technology, 169, 62–71. https://doi.org/10.1016/j.biortech.2014.06.078 CR - Qiu, C., Wang, W., Zhang, Y., Zhou, G.-J., & Bi, Y. (2022). Response of antioxidant enzyme activities of the green microalga Chlorococcum sp. AZHB to Cu2+ and Cd2+ Stress. Sustainability, 14(16), 10320. https://doi.org/10.3390/su141610320 CR - Sabatini, S.E., Juárez, Á.B., Eppis, M.R., Bianchi, L., Luquet, C.M., & Ríos De Molina, M.D.C. (2009). Oxidative stress and antioxidant defenses in two green microalgae exposed to copper. Ecotoxicology and Environmental Safety, 72(4), 1200–1206. https://doi.org/10.1016/j.ecoenv.2009.01.003 CR - Salgado, E.M., Esteves, A.F., Gonçalves, A.L., & Pires, J.C.M. (2023). Microalgal cultures for the remediation of wastewaters with different nitrogen to phosphorus ratios: Process modelling using artificial neural networks. Environmental Research, 231, 116076. https://doi.org/10.1016/j.envres.2023.116076 CR - Satpati, G.G., Chandra Gorain, P., Paul, I., & Pal, R. (2016). An integrated salinity-driven workflow for rapid lipid enhancement in green microalgae for biodiesel application. RSC Advances, 6(113), 112340–112355. https://doi.org/10.1039/C6RA23933A CR - Serrano, G., Miranda-Ostojic, C., Ferrada, P., Wulff-Zotelle, C., Maureira, A., Fuentealba, E., Gallardo, K., Zapata, M., & Rivas, M. (2021). Response to static magnetic field-induced stress in Scenedesmus obliquus and Nannochloropsis gaditana. Marine Drugs, 19(9), 527. https://doi.org/10.3390/md19090527 CR - Sharma, A.K., Jaryal, S., Sharma, S., Dhyani, A., Tewari, B.S., & Mahato, N. (2025). Biofuels from microalgae: A review on microalgae cultivation, biodiesel production techniques and storage stability. Processes, 13(2), 488. https://doi.org/10.3390/pr13020488 Shen, G., Lei, S., Li, H., Yu, Q., Wu, G., Shi, Y., Xu, K., Ren, H., & Geng, J. (2023). Occurrence and removal of four artificial sweeteners in wastewater treatment plants of China. Environmental Science: Processes & Impacts, 25(1), 75–84. https://doi.org/10.1039/D2EM00351A CR - Vishwakarma, R., Dhar, D.W., & Saxena, S. (2019). Influence of nutrient formulations on growth, lipid yield, carbon partitioning and biodiesel quality potential of Botryococcus sp. and Chlorella sp. Environmental Science and Pollution Research, 26(8), 7589–7600. https://doi.org/10.1007/s11356-019-04213-2 CR - Wawryk, N.J.P., Huang, G., Craven, C., Qiu, J., Jmaiff Blackstock, L.K., & Li, X.-F. (2023). Aspartame-sweetened tap water: transformation products and 2,6-dichloro-1,4-benzoquinone formation. Environmental Science & Technology, 57(3), 1332–1341. https://doi.org/10.1021/acs.est.2c07156 CR - Xu, L., Cheng, X., & Wang, Q. (2018). Enhanced lipid production in Chlamydomonas reinhardtii by co-culturing with Azotobacter chroococcum. Frontiers in Plant Science, 9, 741. https://doi.org/10.3389/fpls.2018.00741 CR - Yang, L., Chen, J., Qin, S., Zeng, M., Jiang, Y., Hu, L., Xiao, P., Hao, W., Hu, Z., Lei, A., & Wang, J. (2018). Growth and lipid accumulation by different nutrients in the microalga Chlamydomonas reinhardtii. Biotechnology for Biofuels, 11(1), 40. https://doi.org/10.1186/s13068-018-1041-z CR - Yusuf, N., Athirah, N.M., & A.S. (2022). Antioxidative responses of Chlorella vulgaris under different growth phases. Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology, 17(3), 111–120. https://doi.org/10.15578/squalen.692 UR - https://doi.org/10.3153/AR26008 L1 - https://dergipark.org.tr/en/download/article-file/5378485 ER -