Influence of cultivation conditions and genetic engineering methods on Phaeodactylum tricornutum biomass growth and high-value metabolites content

Nikita  Nechyporuk; Tetiana  Butkevych; Zhanna  Polova; Natalia  Koziko; Tetiana  Nehoda; Mykola  Shumeiko; Olena  Hlushchenko

doi:10.32345/USMYJ.2(163).2026.54-61

Nikita Nechyporuk Third-year Student, Faculty of Pharmacy, Bogomolets National Medical University, Kyiv, Ukraine https://orcid.org/0009-0007-7370-565X
Tetiana Butkevych Department of Pharmacy and Industrial Technology of Medicines, Bogomolets National Medical University, Kyiv, Ukraine https://orcid.org/0000-0002-7570-6150
Zhanna Polova Department of Pharmacy and Industrial Technology of Medicines, Bogomolets National Medical University, Kyiv, Ukraine https://orcid.org/0000-0002-1874-2841
Natalia Koziko Department of Pharmacy and Industrial Technology of Medicines, Bogomolets National Medical University, Kyiv, Ukraine https://orcid.org/0000-0003-0035-491X
Tetiana Nehoda Department of Pharmacy and Industrial Technology of Medicines, Bogomolets National Medical University, Kyiv, Ukraine https://orcid.org/0000-0001-8254-0737
Mykola Shumeiko Department of Pharmacy and Industrial Technology of Medicines, Bogomolets National Medical University, Kyiv, Ukraine https://orcid.org/0000-0002-1069-7652
Olena Hlushchenko Department of Pharmacy and Industrial Technology of Medicines, Bogomolets National Medical University, Kyiv, Ukraine https://orcid.org/0000-0002-5423-3737

DOI: https://doi.org/10.32345/USMYJ.2(163).2026.54-61

Keywords: biomass, cannabinoids, chlorophyll binding proteins, culture media, eicosapentaenoic acid, metabolic engineering, microalgae

Abstract

Introduction. The modern pharmaceutical industry is increasingly shifting from chemical synthesis to bio-oriented technologies, creating demand for new sustainable sources of biologically active substances. The diatom microalga Phaeodactylum tricornutum has emerged as a promising cellular biofactory due to its ease of cultivation, high growth rates, metabolic versatility, and the availability of reproducible protocols for genetic transformation and cryopreservation.

Aim. This narrative review aims to summarise the influence of cultivation conditions and genetic engineering methods on biomass yield and the production of both native high-value metabolites, including eicosapentaenoic acid, fucoxanthin, and chrysolaminarin, and non-native compounds, such as recombinant proteins and cannabinoid precursors, in Phaeodactylum tricornutum for pharmaceutical biotechnology.

Materials and Methods. This narrative review analysed data on cultivation conditions, culture media composition, temperature and light regimes, genetic engineering approaches, and preclinical and clinical evidence related to Phaeodactylum tricornutum and its biotechnological applications.

Results. The key findings indicate that biomass growth and metabolite accumulation are strongly dependent on the composition of culture media, including nitrogen, phosphorus, and silicates, as well as temperature and light regimes. For example, nitrogen starvation increases lipid content but reduces phenolic compounds and carotenoids, whereas higher phosphate levels enhance both biomass production and fucoxanthin accumulation. Two-stage cultivation strategies help mitigate the trade-off between biomass productivity and stress-induced metabolite yield. Genetic engineering approaches, including the overexpression of endogenous genes, the introduction of plant transcription factors, clustered regularly interspaced short palindromic repeats interference, and epigenetic editing using human fat mass and obesity-associated protein demethylase, have successfully increased the yields of native lipids, eicosapentaenoic acid, and fucoxanthin without compromising biomass growth. Furthermore, Phaeodactylum tricornutum has been engineered to produce non-native compounds of pharmaceutical interest, including recombinant vaccine antigens, such as hepatitis B surface antigen and salmon alphavirus antigen, as well as cannabinoid precursors, such as olivetolic acid and cannabigerolic acid. However, cannabinoid production remains at an early experimental stage. Preclinical and clinical studies confirm the safety, bioavailability, and anti-inflammatory effects of Phaeodactylum tricornutum biomass, while fucoxanthin shows promise against inflammatory and neurodegenerative diseases.

Conclusions. In conclusion, Phaeodactylum tricornutum represents a controllable and versatile platform for pharmaceutical biotechnology. However, challenges related to strain stability, scalability, downstream processing costs, and regulatory frameworks must be addressed before its widespread industrial application.

References

Toustou C, Boulogne I, Gonzalez AA, Bardor M. Comparative RNA-Seq of ten Phaeodactylum tricornutum accessions: unravelling criteria for robust strain selection from a bioproduction point of view. Mar Drugs. 2024;22(8):353. https://doi.org/10.3390/md22080353

Celi C, Fino D, Savorani F. Phaeodactylum tricornutum as a source of value-added products: a review on recent developments in cultivation and extraction technologies. Bioresour Technol Rep. 2022;19:101122. https://doi.org/10.1016/j.biteb.2022.101122

Russo MT, Rogato A, Jaubert M, Karas BJ, Falciatore A. Phaeodactylum tricornutum: an established model species for diatom molecular research and an emerging chassis for algal synthetic biology. J Phycol. 2023;59(6):1114-1122. https://doi.org/10.1111/jpy.13400

Castaldi A, Triba MN, Le Moyec L, Hubas C, Le Pennec G, Bourguet-Kondracki ML. Multiblock metabolomics responses of the diatom Phaeodactylum tricornutum under benthic and planktonic culture conditions. Mar Drugs. 2025;23(8):314. https://doi.org/10.3390/md23080314

Elshobary ME, Abo-Shanab WA, Ende SSW, Alquraishi M, El-Shenody RA. Optimizing Phaeodactylum tricornutum cultivation: integrated strategies for enhancing biomass, lipid, and fucoxanthin production. Biotechnol Biofuels Bioprod. 2025;18(1):7. https://doi.org/10.1186/s13068-024-02602-5

Curcuraci E, Manuguerra S, Messina CM, Arena R, Renda G, Ioannou T, et al. Culture conditions affect antioxidant production, metabolism and related biomarkers of the microalgae Phaeodactylum tricornutum. Antioxidants (Basel). 2022;11(2):411. https://doi.org/10.3390/antiox11020411

Song Z, Lye GJ, Parker BM. Morphological and biochemical changes in Phaeodactylum tricornutum triggered by culture media: implications for industrial exploitation. Algal Res. 2020;47:101822. https://doi.org/10.1016/j.algal.2020.101822

Wang S, Hu Z. The marine diatom Phaeodactylum tricornutum as a versatile bioproduction chassis: current progress, challenges, and perspectives. Plant Commun. 2025;6(11):101519. https://doi.org/10.1016/j.xplc.2025.101519

Stiefvatter L, Neumann U, Rings A, Frick K, Schmid-Staiger U, Bischoff SC. The microalgae Phaeodactylum tricornutum is well suited as a food with positive effects on the intestinal microbiota and the generation of SCFA: results from a pre-clinical study. Nutrients. 2022;14(12):2504. https://doi.org/10.3390/nu14122504

Nieri P, Carpi S, Esposito R, Costantini M, Zupo V. Bioactive molecules from marine diatoms and their value for the nutraceutical industry. Nutrients. 2023;15(2):464. https://doi.org/10.3390/nu15020464

Lee AH, Shin HY, Park JH, Koo SY, Kim SM, Yang SH. Fucoxanthin from microalgae Phaeodactylum tricornutum inhibits pro-inflammatory cytokines by regulating both NF-κB and NLRP3 inflammasome activation. Sci Rep. 2021;11(1):543. https://doi.org/10.1038/s41598-020-80748-6

Stiefvatter L, Frick K, Lehnert K, Vetter W, Montoya-Arroyo A, Frank J, et al. Potentially beneficial effects on healthy aging by supplementation of the EPA-rich microalgae Phaeodactylum tricornutum or its supernatant: a randomized controlled pilot trial in elderly individuals. Mar Drugs. 2022;20(11):716. https://doi.org/10.3390/md20110716

Yang Y, Yang M, Zhou Y, Chen X, Huang B. Effect of RNA demethylase FTO overexpression on biomass and bioactive substances in diatom Phaeodactylum tricornutum. Biology (Basel). 2025;14(4):414. https://doi.org/10.3390/biology14040414

Yang R, Wei D. Improving fucoxanthin production in mixotrophic culture of marine diatom Phaeodactylum tricornutum by LED light shift and nitrogen supplementation. Front Bioeng Biotechnol. 2020;8:820. https://doi.org/10.3389/fbioe.2020.00820

Lovio-Fragoso JP, de Jesús-Campos D, López-Elías JA, Medina-Juárez LÁ, Fimbres-Olivarría D, Hayano-Kanashiro C. Biochemical and molecular aspects of phosphorus limitation in diatoms and their relationship with biomolecule accumulation. Biology (Basel). 2021;10(7):565. https://doi.org/10.3390/biology10070565

Ozcan DO, Ovez B. Evaluation of the interaction of temperature and light intensity on the growth of Phaeodactylum tricornutum: kinetic modeling and optimization. Biochem Eng J. 2020;154:107456. https://doi.org/10.1016/j.bej.2019.107456

Ding W, Ye Y, Yu L, Liu M, Liu J. Physiochemical and molecular responses of the diatom Phaeodactylum tricornutum to illumination transitions. Biotechnol Biofuels Bioprod. 2023;16(1):103. https://doi.org/10.1186/s13068-023-02352-w

Butler T, Kapoore RV, Vaidyanathan S. Phaeodactylum tricornutum: a diatom cell factory. Trends Biotechnol. 2020;38(6):606-622. https://doi.org/10.1016/j.tibtech.2019.12.023

Frick K, Yeh YC, Schmid-Staiger U, et al. Comparing three different Phaeodactylum tricornutum strains for the production of chrysolaminarin in flat panel airlift photobioreactors. J Appl Phycol. 2023;35:11-24. https://doi.org/10.1007/s10811-022-02893-x

Ebbing T, Kopp L, Frick K, Simon T, Würtz B, Pfannstiel J, et al. Exploring Phaeodactylum tricornutum for nutraceuticals: cultivation techniques and neurotoxin risk assessment. Mar Drugs. 2025;23(2):58. https://doi.org/10.3390/md23020058

Gao S, Zhou L, Yang W, Wang L, Liu X, Gong Y, et al. Overexpression of a novel gene (Pt2015) endows the commercial diatom Phaeodactylum tricornutum high lipid content and grazing resistance. Biotechnol Biofuels Bioprod. 2022;15(1):131. https://doi.org/10.1186/s13068-022-02221-y

Bao M, Yang W, Li X, Yu G, Gu W, Gao S, et al. A genetically modified triradiate strain of Phaeodactylum tricornutum demonstrates considerable advantages in a 60-L photobioreactor. J Appl Phycol. 2025;37(2):757-764. https://doi.org/10.1007/s10811-025-03473-5

Chen Y, Geng L, Hao Z, Ding N, Di J, Hou H, et al. Heterologous expression of AtLEC1 and AtLEC1-LIKE transcription factors redirects carbon flux toward lipid accumulation in diatom. Microb Cell Fact. 2026;25(1):12. https://doi.org/10.1186/s12934-025-02893-9

Guo W, Weng Y, Ma W, Chang C, Gao Y, Huang X, et al. Improving lipid content in the diatom Phaeodactylum tricornutum by the knockdown of the enoyl-CoA hydratase using CRISPR interference. Curr Issues Mol Biol. 2024;46(10):10923-10933. https://doi.org/10.3390/cimb46100649

Seo S, Chang KS, Choi MS, Jin E. Overexpression of PtVDL1 in Phaeodactylum tricornutum increases fucoxanthin content under red light. J Microbiol Biotechnol. 2024;34(1):198-206. https://doi.org/10.4014/jmb.2309.09018

Csalane Besenyei G. Engineering Phaeodactylum for vaccine production [dissertation]. London: University College London; 2024. Available from: https://discovery.ucl.ac.uk/id/eprint/10195454/

Fantino E, Awwad F, Merindol N, Diaz-Garza AM, Gelinas SE, Robles GCG, et al. Bioengineering Phaeodactylum tricornutum, a marine diatom, for cannabinoid biosynthesis. Algal Res. 2024;77:103379. https://doi.org/10.1016/j.algal.2023.103379

Sene N, Gonçalves Dos Santos KC, Merindol N, Gélinas SE, Custeau A, Awwad F, et al. Impact of heterologous expression of Cannabis sativa tetraketide synthase on Phaeodactylum tricornutum metabolic profile. Biotechnol Biofuels Bioprod. 2025;18(1):42. https://doi.org/10.1186/s13068-025-02638-1

Awwad F, Fantino EI, Héneault M, Diaz-Garza AM, Merindol N, Custeau A, et al. Bioengineering of the marine diatom Phaeodactylum tricornutum with Cannabis genes enables the production of the cannabinoid precursor, olivetolic acid. Int J Mol Sci. 2023;24(23):16624. https://doi.org/10.3390/ijms242316624

Yan Q, Chen YG, Yang XW, Wang A, He XP, Tang X, et al. Engineering a promiscuous prenyltransferase for selective biosynthesis of an undescribed bioactive cannabinoid analog. Commun Biol. 2025;8(1):173. https://doi.org/10.1038/s42003-025-07509-x

Ding YK, Ning Y, Xin D, Fu YJ. Dual cytoplasmic-peroxisomal compartmentalization engineering and multiple metabolic engineering strategies for high yield non-psychoactive cannabinoid in Saccharomyces cerevisiae. Biotechnol J. 2024;19(2):e2300590. https://doi.org/10.1002/biot.202300590

Stiefvatter L, Lehnert K, Frick K, Montoya-Arroyo A, Frank J, Vetter W, et al. Oral bioavailability of omega-3 fatty acids and carotenoids from the microalgae Phaeodactylum tricornutum in healthy young adults. Mar Drugs. 2021;19(12):700. https://doi.org/10.3390/md19120700

Méresse S, Fodil M, Fleury F, Chénais B. Fucoxanthin, a marine-derived carotenoid from brown seaweeds and microalgae: a promising bioactive compound for cancer therapy. Int J Mol Sci. 2020;21(23):9273. https://doi.org/10.3390/ijms21239273