Scientific Reports of the National University of Life and Environmental Sciences of Ukraine

Chlorella vulgaris microalgae as a potential source of vitamins B₁₂ and D₂

Victor Bortsiukh, Nataliia Golub
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Abstract

The research relevance is determined by the need to summarise current knowledge on the mechanisms of vitamin B₁₂ and D₂ accumulation in Chlorella vulgaris and the factors determining their biochemical variability in microalgae cultures. The study aimed to systematise literature data for 2020-2025 and identify key factors regulating sterol metabolism and vitamin profile in Chlorella vulgaris. The study was based on targeted information search, content analysis, critical comparison, and qualitative synthesis of previous scientific works. Based on the analysis of sources, the study demonstrated that the type of nutrient medium determined the level of micronutrients in the biomass of Chlorella vulgaris. Mineral media provided stable growth but did not contain cobalamin (vitamin B₁₂), while organic substrates – liver extract, whey, permeates, and anaerobic digestates demonstrated a higher potential for cell enrichment due to natural coenzymes, proteins and trace elements. Cobalt ions (Co²⁺) were shown to promote greater accumulation of cobalamin in biomass, while magnesium (Mg²⁺), zinc (Zn²⁺), iron (Fe²⁺/Fe³⁺) and copper (Cu²⁺) were substantial cofactors for enzymes and transport systems that influenced sterogenesis. Regarding vitamin D₂ (ergocalciferol), the literature confirmed the decisive role of ultraviolet radiation in the B range (UV-B), which initiated the photoconversion of ergosterol. The intensity and spectral composition of light determined the rate of sterol accumulation, while salinity, nitrogen or phosphorus deficiency, and other stress factors contributed to increased lipidation and changes in the sterol pool. The study determined that in the stationary phase of the culture, lipid and sterol fractions accumulated more intensively than during the period of active cell division. A generalised analysis of sources showed that the most effective enrichment of Chlorella vulgaris with vitamins B₁₂ and D₂ is achieved by combining organic nutrient media, an optimised lighting spectrum with a UV-B component, and a controlled mineral composition of the medium

Keywords

cobalamin, ergocalciferol, organic extracts, light spectrum, ergosterol, metal ions, stress factors

Suggested citation
Bortsiukh, V., & Golub, N. (2025). Chlorella vulgaris microalgae as a potential source of vitamins B₁₂ and D₂. Scientific Reports of the National University of Life and Environmental Sciences of Ukraine, 21(6),93-113. https://doi.org/10.31548/dopovidi/6.2025.93
References
  1. Abdelkarim, O.H., Wijffels, R.H., & Barbosa, M.J. (2025). Exploiting microalgal diversity for sterol production. Frontiers in Plant Science, 16, article number 1616863. doi: 10.3389/fpls.2025.1616863.
  2. Ahirwar, A., et al. (2024). Fermentation of algal biomass for its nutritional value: Perspectives and revolutions in the food industry. Environmental Technology Reviews, 13(1), 1-28. doi: 10.1080/21622515.2023.2283097.
  3. Ali, H.E.A., El-Fayoumy, E.A., Rasmy, W.E., Soliman, R.M., & Abdullah, M.A. (2021). Two-stage cultivation of Chlorella vulgaris using light and salt stress conditions for simultaneous production of lipid, carotenoids, and antioxidants. Journal of Applied Phycology, 33(1), 227-239. doi: 10.1007/s10811-020-02308-9.
  4. Almutairi, A.W., El-Sayed, A.E.K.B., & Reda, M.M. (2021). Evaluation of high salinity adaptation for lipid bio-accumulation in the green microalga Chlorella vulgaris. Saudi Journal of Biological Sciences, 28(7), 3981-3988. doi: 10.1016/j.sjbs.2021.04.007.
  5. Ampofo, J., & Abbey, L. (2022). Microalgae: Bioactive composition, health benefits, safety and prospects as potential high-value ingredients for the functional food industry. Foods, 11(12), article number 1744. doi: 10.3390/foods11121744.
  6. Ashaari, A., Iehata, S., Kim, H.-J., & Rasdi, N.W. (2024). Recent advancement of zooplankton enriched with nutrients and probiotic isolates from aquaculture systems: A review. Journal of Applied Animal Research, 52(1), article number 2417052. doi: 10.1080/09712119.2024.2417052.
  7. Azhan, A.N.M., Mahari, W.A.W., Razali, W.A.W., Amri, B.K.M., Kamaruzzan, A.S., & Lam, S.S. (2025). Influence of wavelength-specific light on growth, chlorophyll synthesis, fatty acid, and carbohydrate production in Chlorella sorokiniana. Energy & Environment. doi: 10.1177/0958305X251379782.
  8. Barber-Lluch, E., Joglar, V., Moreiras, G., Leão, J.M., Gago-Martínez, A., Fernández, E., & Teira, E. (2021). Variability of vitamin B12 concentrations in waters along the Northwest Iberian shelf. Regional Studies in Marine Science, 42, article number 101608. doi: 10.1016/j.rsma.2020.101608.
  9. Bialevich, V., Zachleder, V., & Bišová, K. (2022). The effect of variable light source and light intensity on the growth of three algal species. Cells, 11(8), article number 1293. doi: 10.3390/cells11081293.
  10. Bunbury, F., Deery, E., Sayer, A.P., Bhardwaj, V., Harrison, E.L., Warren, M.J., & Smith, A.G. (2022). Exploring the onset of B12‐based mutualisms using a recently evolved Chlamydomonas auxotroph and B12‐producing bacteria. Environmental Microbiology, 24(7), 3134-3147. doi: 10.1111/1462-2920.16035.
  11. Carlberg, C. (2022). Vitamin D in the context of evolution. Nutrients, 14(15), article number 3018. doi: 10.3390/nu14153018.
  12. Coronado-Reyes, J.A., & González-Hernández, J.C. (2023). Vitamins from microalgae. In E. Jacob-Lopes, M.I. Queiroz, M.M. Maroneze & L.Q. Zepka (Eds.), Handbook of food and feed from microalgae: Production, application, regulation, and sustainability (pp. 111-115). London: Academic Press. doi: 10.1016/B978-0-323-99196-4.00026-7.
  13. Cunha, A.E.P., Sátiro, J.R., Escobar, B.P., & Simões, R.M.S. (2023). Chlorella vulgaris growth, pigment and lipid accumulation: Effect of progressive light and hydrogen peroxide exposure. Journal of Chemical Technology & Biotechnology, 98(2), 442-450. doi: 10.1002/jctb.7256.
  14. de Brito, B.M., Campos, V.D.M., Neves, F.J., Ramos, L.R., & Tomita, L.Y. (2023). Vitamin B12 sources in non-animal foods: A systematic review. Critical Reviews in Food Science and Nutrition, 63(26), 7853-7867. doi: 10.1080/10408398.2022.2053057.
  15. Der, C., Courty, P.E., Recorbet, G., Wipf, D., Simon-Plas, F., & Gerbeau-Pissot, P. (2024). Sterols, pleiotropic players in plant-microbe interactions. Trends in Plant Science, 29(5), 524-534. doi: 10.1016/j.tplants.2024.03.002.
  16. Durdakova, M., Kolackova, M., Ridoskova, A., Cernei, N., Pavelicova, K., Urbis, P., Richtera, L., Pelcova, P., Adam, V., & Huska, D. (2024). Exploring the potential nutritional benefits of Arthrospira maxima and Chlorella vulgaris: A focus on vitamin B12, amino acids, and micronutrients. Food Chemistry, 452, article number 139434. doi: 10.1016/j.foodchem.2024.139434.
  17. Feng, Y., Xiao, J., Cui, N., Zhao, Y., & Zhao, P. (2021). Enhancement of lipid productivity and self-flocculation by cocultivating Monoraphidium sp. FXY-10 and Heveochlorella sp. Yu under mixotrophic mode. Applied Biochemistry and Biotechnology, 193(10), 3173-3186. doi: 10.1007/s12010-021-03593-x.
  18. Gao, Y., Bernard, O., Fanesi, A., Perré, P., & Lopes, F. (2023). The impact of light/dark regimes on structure and physiology of Chlorella vulgaris biofilms. Frontiers in Microbiology, 14, article number 1250866. doi: 10.3389/fmicb.2023.1250866.
  19. González-González, L.M., & De-Bashan, L.E. (2021). Toward the enhancement of microalgal metabolite production through microalgae-bacteria consortia. Biology, 10(4), article number 282. doi: 10.3390/biology10040282.
  20. Gopalakrishnan, K., Wager, Y.Z., & Roostaei, J. (2024). Co-cultivation of microalgae and bacteria for optimal bioenergy feedstock production in wastewater by using response surface methodology. Scientific Reports, 14, article number 20703. doi: 10.1038/s41598-024-70033-1.
  21. Grishko, V., Zotsenko, V., Bogatko, N., & Ostrovskiy, D. (2024). Biological features, biotechnology, cultivation and areas of stagnation of microscopic single-cell algae of the genus Chlorella: (Review of the literature). Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Agricultural Sciences, 26(101), 329-340. doi: 10.32718/nvlvet-a10150.
  22. Hamouda, R.A., El Latif, A.A., Elkaw, E.M., Alotaibi, A.S., Alenzi, A.M., & Hamza, H.A. (2022). Assessment of antioxidant and anticancer activities of microgreen alga Chlorella vulgaris and its blend with different vitamins. Molecules, 27(5), article number 1602. doi: 10.3390/molecules27051602.
  23. Islam, M. (2024). The consistency of vitamin B12 in marketed microalgae powders. International Journal of Agricultural Research, Innovation and Technology, 14(2), 146-152. doi: 10.3329/ijarit.v14i2.79425.
  24. Ji, X., Luo, X., Zhang, J., & Huang, D. (2021). Effects of exogenous vitamin B12 on nutrient removal and protein expression of algal-bacterial consortium. Environmental Science and Pollution Research, 28(13), 15954-15965. doi: 10.1007/s11356-020-11720-0.
  25. Kamolrat, N., Kamuang, S., Khamket, T., Sangmek, P., & Sitthaphanit, S. (2023). The effect of optimum photoperiod from blue LED light on growth of Chlorella vulgaris in photobioreactor tank. Natural Life Sciences Communications, 22(3), article number e2023038. doi: 10.12982/NLSC.2023.038.
  26. Kashyap, S., Das, N., Kumar, M., Mishra, S., Kumar, S., & Nayak, M. (2025). Poultry litter extract as solid waste supplement for enhanced microalgal biomass production and wastewater treatment. Environmental Science and Pollution Research. doi: 10.1007/s11356-025-35900-y.
  27. Kholif, A.E., & Olafadehan, O.A. (2021). Chlorella vulgaris microalgae in ruminant nutrition: A review of the chemical composition and nutritive value. Annals of Animal Science, 21(3), 789-806. doi: 10.2478/aoas-2020-0117.
  28. Klamann, L., Dutta, R., Ghazaryan, L., Sela-Adler, M., Khozin-Goldberg, I., & Gillor, O. (2023). Cobalamin production in phototrophic and mixotrophic cultures of nine duckweed species (Lemnaceae). bioRxiv. doi: 10.1101/2023.10.31.564895.
  29. Kolesovs, S., & Semjonovs, P. (2023). Microalgal conversion of whey and lactose containing substrates: Current state and challenges. Biodegradation, 34(5), 405-416. doi: 10.1007/s10532-023-10033-6.
  30. Kozhemiaka, O.V., & Peshuk, L.V. (2023). Chlorella as a biologically active component in the technology of health and wellness products. Journal of Chemistry and Technologies, 31(2), 230-239. doi: 10.15421/jchemtech.v31i2.275148.
  31. Kyoto Encyclopedia of Genes and Genomes. (n.d.). Steroid biosynthesis – reference pathway. Retrieved from https://www.kegg.jp/pathway/map00100?utm_source.
  32. Leung, M.F., & Cheung, P.C.K. (2021). Vitamins D and D2 in cultivated mushrooms under ultraviolet irradiation and their bioavailability in humans: A mini-review. International Journal of Medicinal Mushrooms, 23(11), 1-15. doi: 10.1615/IntJMedMushrooms.2021040390.
  33. Liu, C., Wei, H., Ru, S., Wang, W., Li, Z., Liu, L., Song, X., & Wang, J. (2025). Synthetic microalgae community promotes chili growth planted in waste stone powder by enriching beneficial root endophytic bacteria and fungi. Plant and Soil. doi: 10.1007/s11104-025-08011-3.
  34. Maulana, S., et al. (2025). Effect of red and blue light on lipid, protein, carbohydrate, and pigment contents of Navicula sp. Makara Journal of Science, 29(3), 411-421. doi: 10.7454/mss.v29i3.2197.
  35. Mendes, A.R., Spínola, M.P., Lordelo, M., & Prates, J.A.M. (2024). Chemical compounds, bioactivities, and applications of Chlorella vulgaris in food, feed and medicine. Applied Sciences, 14(23), article number 10810. doi: 10.3390/app142310810.
  36. Nef, C., Dittami, S., Kaas, R., Briand, E., Noël, C., Mairet, F., & Garnier, M. (2022). Sharing vitamin B12 between bacteria and microalgae does not systematically occur: Case study of the haptophyte Tisochrysis lutea. Microorganisms, 10(7), article number 1337. doi: 10.3390/microorganisms10071337.
  37. Nowruzi, B., Shishir, A., Porzani, S.J., & Ferdous, U.T. (2022). Exploring the interactions between algae and bacteria. Mini Reviews in Medicinal Chemistry, 22(20), 2596-2607. doi: 10.2174/1389557522666220504141047.
  38. Papadopoulos, K.P., et al. (2023). Vitamin B12 bioaccumulation in Chlorella vulgaris grown on food waste-derived anaerobic digestate. Algal Research, 75, article number 103290. doi: 10.1016/j.algal.2023.103290.
  39. Peng, M., Liu, P., Peng, R., Jiang, X., Li, S., & Jiang, M. (2025). Effects of nitrogen, phosphorus, iron, and silicon on the growth, total lipid content, and fatty acid composition of the diatom Corethron sp. Journal of Applied Phycology. doi: 10.1007/s10811-025-03716-5.
  40. Peshuk, L.V., Prykhodko, D.Y., Kozhemiaka, O.V., & Petrenko, S.A. (2024). Green algae: Focus on innovation, functionality and naturalness. Journal of Chemistry and Technologies, 32(1), 138-152. doi: 10.15421/jchemtech.v32i1.288749.
  41. Pessi, B.A., & Bernard, O. (2024). Dynamical metabolic model for optimizing biotin-regulated lipid production in microalgae-bacteria symbiosis. IFAC-PapersOnLine, 58(14), 151-156. doi: 10.1016/j.ifacol.2024.08.329.
  42. Ribeiro, M., Maciel, C., Cruz, P., Darmancier, H., Nogueira, T., Costa, M., Laranjeira, J., Morais, R.M.S.C., & Teixeira, P. (2023). Exploiting potential probiotic lactic acid bacteria isolated from Chlorella vulgaris photobioreactors as promising vitamin B12 producers. Foods, 12(17), article number 3277. doi: 10.3390/foods12173277.
  43. Ricky, R., Chiampo, F., & Shanthakumar, S. (2022). Efficacy of ciprofloxacin and amoxicillin removal and the effect on the biochemical composition of Chlorella vulgaris. Bioengineering, 9(4), article number 134. doi: 10.3390/bioengineering9040134.
  44. Salbitani, G., & Carfagna, S. (2021). Ammonium utilization in microalgae: A sustainable method for wastewater treatment. Sustainability, 13(2), article number 956. doi: 10.3390/su13020956.
  45. Selvaraj, H. (2024). Unlocking the potential of Chlorella vulgaris: Bioactivity and multifunctional uses. Natural Product Research. doi: 10.1080/14786419.2024.2384087.
  46. Singh, R.P., Yadav, P., Kumar, I., Solanki, M.K., Roychowdhury, R., Kumar, A., & Gupta, R.K. (2023). Advancement of abiotic stresses for microalgal lipid production and its bioprospecting into sustainable biofuels. Sustainability, 15(18), article number 13678. doi: 10.3390/su151813678.
  47. Souza, A.L., & Patti, G.J. (2021). A protocol for untargeted metabolomic analysis: From sample preparation to data processing. In V. Weissing & M. Edeas (Eds.), Mitochondrial medicine: Volume 2: Assessing mitochondria (pp. 357-382). New York: Springer. doi: 10.1007/978-1-0716-1266-8_27.
  48. Sultana, S., Bruns, S., Wilkes, H., Simon, M., & Wienhausen, G. (2023). Vitamin B12 is not shared by all marine prototrophic bacteria with their environment. ISME Journal, 17(6), 836-845. doi: 10.1038/s41396-023-01391-3.
  49. Tong, C.Y., Tomita, H., Miyazaki, K., Chieh, D.C.J., & Honda, K. (2025). Synergistic enhancement of biomass and lipid productivity in Chlorella Sorokiniana co-cultured with vitamins-overproducing Escherichia Coli. Retrieved from https://papers.ssrn.com/sol3/papers.cfm?abstract_id=5351723.
  50. Tzachor, A., van den Oever, S.P., Mayer, H.K., Asfur, M., Smidt-Jensen, A., Geirsdóttir, M., Jensen, S., & Smárason, B.O. (2024). Photonic management of Spirulina (Arthrospira platensis) in scalable photobioreactors to achieve biologically active unopposed vitamin B12. Discover Food, 4(1), article number 69. doi: 10.1007/s44187-024-00152-1.
  51. van den Oever, S.P., & Mayer, H. (2022). Biologically active or just “pseudo”-vitamin B12 as predominant form in algae-based nutritional supplements? Journal of Food Composition and Analysis, 109, article number 104464. doi: 10.1016/j.jfca.2022.104464.
  52. Vitamin B12 in algae: Spirulina, Chlorella and nori. (n.d.). Retrieved from https://www.b12-vitamin.com/algae/.
  53. Vogeler, S., Wikfors, G.H., Li, X., Sauvage, J., & Joyce, A. (2021). Investigation of vitamin B12 concentrations and tissue distributions in larval and adult Pacific oysters and related bivalves. bioRxiv. doi: 10.1101/2021.10.08.463682.
  54. Voshall, A., Christie, N.T., Rose, S.L., Khasin, M., Van Etten, J.L., Markham, J.E., Riekhof, W.R., & Nickerson, K.W. (2021). Sterol biosynthesis in four green algae: A bioinformatic analysis of the ergosterol versus phytosterol decision point. Journal of Phycology, 57(4), 1199-1211. doi: 10.1111/jpy.13164.
  55. Xu, B., Liu, J., Zhao, C., Sun, S., Xu, J., & Zhao, Y. (2021). Induction of vitamin B12 to purify biogas slurry and upgrade biogas using co‐culture of microalgae and fungi. Water Environment Research, 93(8), 1254-1262. doi: 10.1002/wer.1504.
  56. Zhang, T., & Long, J. (2025). Critical assessment of several filamentous algae as bioeconomic energy under different culture conditions. Algal Research, 88, article number 103992. doi: 10.1016/j.algal.2025.103992.
  57. Zhao, D., Poong, S.-W., Ling, T.C., Lai, S.H., Lim, P.-E., & Ngoh, G.C. (2025). Enhanced removal of copper (Cu2+) by microalgae-bacteria consortium: Mechanistic insights, physiological responses, and artificial neural network (ANN)-based prediction. Retrieved from https://papers.ssrn.com/sol3/papers.cfm?abstract_id=5571187.
  58. Zhao, N., Liu, F., Dong, W., Yu, J., Halverson, L.J., & Xie, B. (2024). Quantitative proteomics insights into Chlamydomonas reinhardtii thermal tolerance enhancement by a mutualistic interaction with Sinorhizobium meliloti. Microbiology Spectrum, 12(8), article number e0021924. doi: 10.1128/spectrum.00219-24.