TiO₂ Nano-photocatalyst-driven CO₂ Biofixation and Lipid Accumulation in Brackish Water Cyanobacteria for Sustainable Biodiesel Production
J. Environ. Nanotechnol., Volume 14, No 3 (September 2025) pp. 108-121
Abstract
This study focused on the effect of CO2 concentration on cyanobacterial cultivation using BG-11 medium to enhance biomass and lipid productivity. Different cyanobacterial species isolated from brackish water were screened and identified using molecular characterisation. From the isolated cyanobacterial species, two rapidly growing species, Phormidium uncinatum and Oscillatoria salina were selected. Their biomass and lipid productivity were optimised by varying the CO2 concentration. These strains were subjected to tolerance tests by changing the conditions of the cyanobacterial growth medium, pH, CO2 concentration, and temperature. Biochemical parameters were also estimated. Oscillatoria salina exhibited the highest biomass production, reaching 956 ± 0.04 mg/L at a 15 % CO2 concentration. A high protein content of 25.23 ± 0.32% was found in P. uncinatum compared to O. salina, whereas O. salina showed higher lipid and biomass productivity than P. uncinatum. Furthermore, O. salina was utilised for the production of biodiesel using a nanophotocatalyst (TiO2). It was found to enhance the biodiesel up to 98%. The biodiesel was tested for fuel properties and compared with the ASTM Standards.
Full Text
Reference
Andriopoulos, V., Gkioni, M. D., Koutra, E., Mastropetros, S. G., Lamari, F. N., Hatziantoniou, S. and Kornaros, M., Total Phenolic Content, Biomass Composition, and Antioxidant Activity of Selected Marine Microalgal Species with Potential as Aquaculture Feed, Antioxidants, 11(7), 1320(2022).
https://doi.org/10.3390/antiox11071320
Aoki, J., Sasaki, D. and Asayama, M., Development of a method for phycocyanin recovery from filamentous cyanobacteria and evaluation of its stability and antioxidant capacity, BMC Biotechnol., 21(1), 40(2021).
https://doi.org/10.1186/s12896-021-00692-9
Bhatnagar, M. and Bhatnagar, A., Diversity of Polysaccharides in Cyanobacteria,. In: Satyanarayana T, Johri BN, Das SK (eds.) Microbial Diversity in Ecosystem Sustainability and Biotechnological Applications, Springer, 447–496 (2019).
https://doi.org/10.1007/978-981-13-8315-1_15
Bligh, E.G. and Dyer, W.J., A Rapid Method Of Total Lipid Extraction and Purification, Can. J. Biochem. Physiol., 37(8), 911–917(1959).
https://doi.org/10.1139/o59-099
Brock, T. D., Lower p H Limit for the Existence of Blue-Green Algae: Evolutionary and Ecological Implications, Sci., 179(4072), 480–483(1973).
https://doi.org/10.1126/science.179.4072.480
Cao, Y., Liu, W., Xu, X., Zhang, H., Wang, J. and Xian, M., Production of free monounsaturated fatty acids by metabolically engineered Escherichia coli, Biotechnol Biofuels, 7(1), 59(2014).
https://doi.org/10.1186/1754-6834-7-59
Carpine, R. and Sieber, S., Antibacterial and antiviral metabolites from cyanobacteria: Their application and their impact on human health, Curr. Res. Biotechnol., 365–81(2021).
https://doi.org/10.1016/j.crbiot.2021.03.001
Chatterjee, D., Bhattacharjee, P., Satpati, G.G. and Pal, R., Spray Dried Extract of Phormidium valderianum as a Promising Source of Natural Antioxidant, Int. J. Food Sci., 2014, 1–8(2014).
https://doi.org/10.1155/2014/897497
Chittora, D., Meena, M., Barupal, T., Swapnil, P. and Sharma, K., Cyanobacteria as a source of biofertilisers for sustainable agriculture, Biochem. Biophys. Rep., 22, 100737(2020).
https://doi.org/10.1016/j.bbrep.2020.100737
De-Philippis, R., Sili, C., Paperi, R. and Vincenzini, M., Exopolysaccharide-producing cyanobacteria and their possible exploitation: A review, J. Appl. Phycol., 13(4), 293–299(2001).
https://doi.org/10.1023/A:1017590425924
El-Aty, A. A., Mohamed, A. A. and Samhan, F. A., In vitro antioxidant and antibacterial activities of two freshwater Cyanobacterial species, Oscillatoria agardhii and Anabaena sphaerica, J. Appl. Pharm. Sci., 4, 69-75(2014).
https://doi.org/10.7324/JAPS.2014.40712
Elkomy, R. G. and Ismail, M. M., Crude sulfated polysaccharides extracted from marine cyanobacterium Oscillatoria simplicissima with evaluation antioxidant and cytotoxic activities, Iran. J. Microbiol., 13(4), 553-559 (2021).
https://doi.org/10.18502/ijm.v13i4.6983
Eungrasamee, K., Miao, R., Incharoensakdi, A., Lindblad, P. and Jantaro, S., Improved lipid production via fatty acid biosynthesis and free fatty acid recycling in engineered Synechocystis sp. PCC 6803, Biotechnol. Biofuels, 12(1), 8(2019).
https://doi.org/10.1186/s13068-018-1349-8
Fang, F., Gao, Y., Gan, L., He, X. and Yang, L., Effects of different initial pH and irradiance levels on cyanobacterial colonies from Lake Taihu, China, J. Appl. Phycol., 30(3), 1777–1793(2018).
https://doi.org/10.1007/s10811-018-1394-5
Feng, N., Liu, F., Huang, M., Zheng, A., Wang, Q., Chen, T., Cao, G., Xu, J., Fan, J. and Deng, F., Unravelling the Efficient Photocatalytic Activity of Boron-induced Ti3+ Species in the Surface Layer of TiO2, Sci. Rep., 6(1), 34765(2016).
https://doi.org/10.1038/srep34765
Gadore, V., Mishra, S. R. and Ahmaruzzaman, Md., Advances in photocatalytic biodiesel production: Preparation methods, modifications and mechanisms, Fuel, 362, 130749(2024).
https://doi.org/10.1016/j.fuel.2023.130749
Hamedi, S., Mahdavi, M. A. and Gheshlaghi, R., Improved lipid and biomass productivities in Chlorella vulgaris by differing the inoculation medium from the production medium, Biofuel Res. J., 3(2), 410–416(2016).
https://doi.org/10.18331/BRJ2016.3.2.6
He, X., Wang, A., Wu, P., Tang, S., Zhang, Y., Li, L. and Ding, P., Photocatalytic degradation of microcystin-LR by modified TiO2 photocatalysis: A review, Sci. Tot. Environ., 743, 140694(2020).
https://doi.org/10.1016/j.scitotenv.2020.140694
Hertzberg, S. and Liaaen, J. S., The carotenoids of blue-green algae—I., Phytochem., 5(4), 557–563(1966).
https://doi.org/10.1016/S0031-9422(00)83633-X
Żymańczyk-Duda, E., Samson, S. O., Brzezińska-Rodak, M. and Klimek-Ochab, M., Versatile Applications of Cyanobacteria in Biotechnology, Microorg., 10(12), 2318(2022).
https://doi.org/10.3390/microorganisms10122318
Hotos, G. N. and Antoniadis, T. I., Growth, Phycobiliproteins, Chlorophyll and Carotenoids Content of the Marine Cyanobacteria Phormidium sp. And Cyanothece sp. As Affected by White and Colored Artificial Light in Batch Cultures, xx, x(2022).
https://doi.org/10.20944/preprints202201.0437.v1
Ismaiel, M. M. S., El-Ayouty, Y. M. and Piercey-Normore, M.D., Antioxidants characterisation in selected cyanobacteria, Ann. Microbiol., 64(3), 1223–1230(2014).
https://doi.org/10.1007/s13213-013-0763-1
Jain, S., Kumar, D., Malviya, N., Jain, A., Jain, S. and Jain, V., Estimation of total phenolic, tannins, and flavonoid contents and antioxidant activity of Cedrus deodara heart wood extracts, Egypt. Pharm. J., 14, 10-14(2015).
https://doi.org/10.4103/1687-4315.154690
Janiad, S., Lindblad, P. and Ahmed, M., Lipid Accumulation and Biodiesel Production from Filamentous Cyanobacteria Native to Pakistan, J. Oleo. Sci., 73(10), 1295–1310(2024).
https://doi.org/10.5650/jos.ess23267
Kar, J., Ramrao, D. P., Zomuansangi, R., Lalbiaktluangi, C., Singh, S. M., Joshi, N. C., Kumar, A., Kaushalendra., Mehta, S., Yadav, M. K. and Singh, P. K., Revisiting the role of cyanobacteria-derived metabolites as antimicrobial agent: A 21st century perspective, Front Microbiol, 13, 1034471(2022).
https://doi.org/10.3389/fmicb.2022.1034471
Karageorgou, D., Patel, A., Rova, U., Christakopoulos, P., Katapodis, P. and Matsakas, L., Heterotrophic Cultivation of the Cyanobacterium Pseudanabaena sp. on Forest Biomass Hydrolysates toward Sustainable Biodiesel Production, Microorg., 10(9), 1756(2022).
https://doi.org/10.3390/microorganisms10091756
Karatay, S. E. and Dönmez, G., Microbial oil production from thermophile cyanobacteria for biodiesel production, Appl. Energy, 88(11), 3632–3635(2011).
https://doi.org/10.1016/j.apenergy.2011.04.010
Kebelmann, K., Hornung, A., Karsten, U. and Griffiths, G., Intermediate pyrolysis and product identification by TGA and Py-GC/MS of green microalgae and their extracted protein and lipid components, Biomass Bioenergy, 49, 38–48(2013).
https://doi.org/10.1016/j.biombioe.2012.12.006
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J., Protein measurement with the Folin phenol reagent, J. Biol. Chem., 193(1), 265–275(1951).
https://doi.org/10.1016/S0021-9258(19)52451-6
Ma, J. and Wang, P., Effects of rising atmospheric CO2 levels on physiological response of cyanobacteria and cyanobacterial bloom development: A review, Sci. Total Environ., 754, 141889(2021).
https://doi.org/10.1016/j.scitotenv.2020.141889
Maoka, T., Carotenoids as natural functional pigments, J. Nat. Med., 74(1), 1–16(2020).
https://doi.org/10.1007/s11418-019-01364-x
Moia, I. C., Pereira, S. B., Domizio, P., De-Philippis, R. and Adessi, A., Phormidium ambiguum and Leptolyngbya ohadii Exopolysaccharides under Low Water Availability, Polym., 15(8), 1889(2023).
https://doi.org/10.3390/polym15081889
Molina, A. K., Corrêa, R. C. G., Prieto, M. A., Pereira, C. and Barros, L., Bioactive Natural Pigments’ Extraction, Isolation, and Stability in Food Applications, Mol., 28(3), 1200(2023).
https://doi.org/10.3390/molecules28031200
Mukherjee, G., Nayak, B. K. and Das, D., Cyanobacteria as a valuable source of antiviral, antibacterial and antifungal compounds? an overview, Algol Stud, 143(1), 3–25(2013).
https://doi.org/10.1127/1864-1318/2013/0143
Nainangu, P., Antonyraj, A. P. M., Subramanian, K., Kaliyaperumal, S., Gopal, S., Sampath Renuka, P. and A, W. A., In vitro screening of antimicrobial, antioxidant, cytotoxic activities, and characterisation of bioactive substances from freshwater cyanobacteria Oscillatoria sp. SSCM01 and Phormidium sp. SSCM02, Biocatal. Agric. Biotechnol., 29, 101772(2020).
https://doi.org/10.1016/j.bcab.2020.101772
Nalley, J. O., O’Donnell, D. R. and Litchman, E., Temperature effects on growth rates and fatty acid content in freshwater algae and cyanobacteria, Algal Res., 35, 500–507(2018).
https://doi.org/10.1016/j.algal.2018.09.018
Ni, L., Zhao, F., Li, B., Wei, T., Guan, H. and Ren, S., Antioxidant and Fluorescence Properties of Hydrogenolyzised Polymeric Proanthocyanidins Prepared Using SO42−/ZrO2 Solid Superacids Catalyst, Mol., 23(10), 2445(2018).
https://doi.org/10.3390/molecules23102445
Niangoran, N. U. F., Buso, D., Zissis, G. and Prudhomme, T., Influence of light intensity and photoperiod on energy efficiency of biomass and pigment production of Spirulina (Arthrospira platensis), OCL, 2837(2021).
https://doi.org/10.1051/ocl/2021025
Nicolaus, B., Panico, A., Lama, L., Romano, I., Manca, M.C., De Giulio, A. and Gambacorta, A., Chemical composition and production of exopolysaccharides from representative members of heterocystous and non-heterocystous cyanobacteria, Phytochem., 52(4), 639–647(1999).
https://doi.org/10.1016/S0031-9422(99)00202-2
Olabi, A. G., Shehata, N., Sayed, E. T., Rodriguez, C., Anyanwu, R. C., Russell, C. and Abdelkareem, M. A., Role of microalgae in achieving sustainable development goals and circular economy, Sci. Tot. Environ., 854, 158689(2023).
https://doi.org/10.1016/j.scitotenv.2022.158689
Pade, N. and Hagemann, M., Salt Acclimation of Cyanobacteria and Their Application in Biotechnology, Life, 5(1), 25–49(2014).
https://doi.org/10.3390/life5010025
Pagels, F., Vasconcelos, V. and Guedes, A. C., Carotenoids from Cyanobacteria: Biotechnological Potential and Optimization Strategies, Biomol., 11(5), 735(2021).
https://doi.org/10.3390/biom11050735
Pathak, J., Rajneesh., Maurya, P. K., Singh, S. P., Häder, D. P. and Sinha, R. P., Cyanobacterial Farming for Environment Friendly Sustainable Agriculture Practices: Innovations and Perspectives, Front. Environ. Sci., 67(2018).
https://doi.org/10.3389/fenvs.2018.00007
Ramamurthy, P. C., Singh, S., Kapoor, D., Parihar, P., Samuel, J., Prasad, R., Kumar, A. and Singh, J., Microbial biotechnological approaches: renewable bioprocessing for the future energy systems, Microb. Cell Factories, 20(1), 55(2021).
https://doi.org/10.1186/s12934-021-01547-w
Renugadevi, K., Valli, N. C., Sowmiya, P. and Sunkar, S., Antioxidant activity of phycocyanin pigment extracted from marine filamentous cyanobacteria Geitlerinema sp TRV57, Biocatal. Agric. Biotechnol., 16, 237–242(2018).
https://doi.org/10.1016/j.bcab.2018.08.009
Saini, D.K., Pabbi, S. and Shukla, P., Cyanobacterial pigments: Perspectives and biotechnological approaches, Food Chem. Toxicol., 120, 616–624(2018).
https://doi.org/10.1016/j.fct.2018.08.002
Santhosh, I., Gnanadoss, J. J., Selvarajan, R. and Elumalai, S., Enhanced carotenoid synthesis of Phormidium sp. in stressed conditions, J. Exp. Sci., 2. 38-44(2011).
Scholz, B. and Liebezeit, G., Chemical Screening for Bioactive Substances in Culture Media of Microalgae and Cyanobacteria from Marine and Brackish Water Habitats: First Results, Pharm. Biol., 44(7), 544–549(2006).
https://doi.org/10.1080/13880200600883114
Shahbazi, M., Tohidfar, M. and Azimzadeh Irani, M., Identification of the key functional genes in salt-stress tolerance of Cyanobacterium Phormidium tenue using in silico analysis, 3 Biotech., 11(12), 503(2021).
https://doi.org/10.1007/s13205-021-03050-w
Shaikhoun, S., Zafar, A. M., Wu, Y. H. and Aly Hassan, A., Biodesalination performance of Phormidium keutzingianum concentrated using two methods (immobilisation and centrifugation), Arab. J. Chem., 15(11), 104282(2022).
https://doi.org/10.1016/j.arabjc.2022.104282
Shanmugam, A., Sigamani, S., Venkatachalam, H., Jayaraman, J. and Ramamurthy, D., Antibacterial activity of extracted phycocyanin from Oscillatoria sp, J. App. Pharm. Sci., 7 (03), 062-067(2017).
https://doi.org/10.7324/JAPS.2017.70310
Sharafi, H., Fooladi, J., Tabatabaei, M., Momhed Heravi, M. and Memari, H., Lipid Production Capacity of a Newly Characterised Cyanobacterial Strain Synechocystis sp. MH01: A Comparative Performance Evaluation of Cyanobacterial Lipid-Based Biodiesel, Iran. J. Biotechnol., 19(1), (2021).
https://doi.org/10.30498/IJB.2021.2313
Shawer, E., Elsaied, H., El-Gamal, A. and Sabae, S., Characterisation of cyanobacterial isolates from freshwater and saline subtropical desert lakes, Folia. Microbiol. (Praha), 68(3), 403–414(2023).
https://doi.org/10.1007/s12223-022-01016-w
Silveira, S. T., Burkert, J. F. M., Costa, J. A. V., Burkert, C. A. V. and Kalil, S. J., Optimisation of phycocyanin extraction from Spirulina platensis using factorial design, Bioresour. Technol., 98(8), 1629–1634(2007).
https://doi.org/10.1016/j.biortech.2006.05.050
Singh, D. P., Prabha, R., Verma, S., Meena, K. K. and Yandigeri, M., Antioxidant properties and polyphenolic content in terrestrial cyanobacteria, 3 Biotech., 7(2), 134(2017).
https://doi.org/10.1007/s13205-017-0786-6
Singh, G., Jeyaseelan, C., Bandyopadhyay, K. K. and Paul, D., Comparative analysis of biodiesel produced by acidic transesterification of lipid extracted from oleaginous yeast Rhodosporidium toruloides, 3 Biotech, 8(10), 434(2018).
https://doi.org/10.1007/s13205-018-1467-9
Singh, J. S., Kumar, A., Rai, A. N. and Singh, D. P., Cyanobacteria: A Precious Bio-resource in Agriculture, Ecosystem, and Environmental Sustainability, Front Microbiol., 7(2016).
https://doi.org/10.3389/fmicb.2016.00529
Singh, J. S., Kumar, A. and Singh, M., Cyanobacteria: A sustainable and commercial bio-resource in production of bio-fertiliser and bio-fuel from waste waters, Environ. Sustain. Indic., 3–4, 100008(2019).
https://doi.org/10.1016/j.indic.2019.100008
Sitther, V., Tabatabai, B., Fathabad, S. G., Gichuki, S., Chen, H. and Arumanayagam, A. C. S., Cyanobacteria as a biofuel source: advances and applications, Adv. Cyanobact. Bio., 269–289 (2020).,
https://doi.org/10.1016/B978-0-12-819311-2.00018-8
Srinivasan, M., Sivasubramanian, V., Palanisamy, M. and Chinnu, K., In vitro antioxidant and anti-proliferative activity of Phormidium fragile, J. Algal Biomass Utln., 5(2), 49-57(2014).
Sultana, B., Anwar, F. and Ashraf, M., Effect of Extraction Solvent/Technique on the Antioxidant Activity of Selected Medicinal Plant Extracts, Molecules, 14(6), 2167–2180(2009).
https://doi.org/10.3390/molecules14062167
Truong, D. H., Dinh, H. N., Nhat, T. A. T., Anh, V. B., Tuong, H. D. and Hoang, C. N., Evaluation of the Use of Different Solvents for Phytochemical Constituents, Antioxidants, and In Vitro Anti-Inflammatory Activities of Severinia buxifolia, J. Food Qual., 2019(1), 8178294(2019).
https://doi.org/10.1155/2019/8178294
Thomas, M. K. and Litchman, E., Effects of temperature and nitrogen availability on the growth of invasive and native cyanobacteria, Hydrobiologia, 763(1), 357–369(2016).
https://doi.org/10.1007/s10750-015-2390-2
Tiwari, O. N., Bhunia, B., Chakraborty, S., Goswami, S. and Devi, I., Strategies for improved production of phycobiliproteins (PBPs) by Oscillatoria sp. BTA170 and evaluation of its thermodynamic and kinetic stability, Biochem. Eng. J., 145, 153–161(2019).
https://doi.org/10.1016/j.bej.2019.02.016
Verseux, C., Baqué, M., Lehto, K., De Vera, J. P. P., Rothschild, L.J. and Billi, D., Sustainable life support on Mars – the potential roles of cyanobacteria, Int. J. Astrobiol., 15(1), 65–92(2016).
https://doi.org/10.1017/S147355041500021X
Yadav, G., Sekar, M., Kim, S. H., Geo, V. E., Bhatia, S. K., Sabir, J. S. M., Chi, N. T. L., Brindhadevi, K. and Pugazhendhi, A., Lipid content, biomass density, fatty acid as selection markers for evaluating the suitability of four fast growing cyanobacterial strains for biodiesel production, Bioresour. Technol., 325, 124654(2021).
https://doi.org/10.1016/j.biortech.2020.124654
Yadav, P., Singh, R. P., Rana, S., Joshi, D., Kumar, D., Bhardwaj, N., Gupta, R. K. and Kumar, A., Mechanisms of Stress Tolerance in Cyanobacteria under Extreme Conditions, Stresses, 2(4), 531–549(2022).
https://doi.org/10.3390/stresses2040036
Yang, Z. K., Niu, Y. F., Ma, Y. H., Xue, J., Zhang, M. H., Yang, W. D., Liu, J. S., Lu, S. H., Guan, Y. and Li, H. Y. Molecular and cellular mechanisms of neutral lipid accumulation in diatom following nitrogen deprivation. Biotechnology for Biofuels, 6(1), 67 2013).
https://doi.org/10.1186/1754-6834-6-67
Zampieri, R. M., Adessi, A., Caldara, F., Codato, A., Furlan, M., Rampazzo, C., De-Philippis, R., La Rocca, N. and Dalla Valle, L., Anti-Inflammatory Activity of Exopolysaccharides from Phormidium sp. ETS05, the Most Abundant Cyanobacterium of the Therapeutic Euganean Thermal Muds, Using the Zebrafish Model, Biomol., 10(4), 582(2020).
https://doi.org/10.3390/biom10040582
Yasin, D., Zafaryab, Md., Ansari, S., Ahmad, N., Khan, N.F., Zaki, A., Alam, R. M. M. and Fatma, T., Evaluation of antioxidant and anti-proliferative efficacy of Nostoc muscorum NCCU-442, Biocatal. Agric. Biotechnol., 17, 284–293(2019).
https://doi.org/10.1016/j.bcab.2018.12.001
Young, A. and Lowe, G., Carotenoids—Antioxidant Properties, Antioxidants, 7(2), 28(2018).
https://doi.org/10.3390/antiox7020028
Yu, H. F. and Yang, S. T., Enhancing thermal stability and photocatalytic activity of anatase-TiO2 nanoparticles by co-doping P and Si elements, J. Alloys Compd., 492(1–2), 695–700(2010).
https://doi.org/10.1016/j.jallcom.2009.12.021
Zahra, Z., Choo, D. H., Lee, H. and Parveen, A., Cyanobacteria: Review of Current Potentials and Applications, Environ., 7(2), 13(2020).
