2. Academic Validation
  3. Multifactorial interaction and influence of culture conditions on yellow fluorescent protein production in Phaeodactylum tricornutum

Multifactorial interaction and influence of culture conditions on yellow fluorescent protein production in Phaeodactylum tricornutum

  • Bioresour Technol. 2025 Jun:425:132336. doi: 10.1016/j.biortech.2025.132336.
Arun Augustine 1,  Anis Messaabi 1,  Elisa Fantino 2,  Natacha Merindol 1,  Fatma Meddeb-Mouelhi 2,  Isabel Desgagné-Penix 3
Affiliations

Affiliations

  • 1 Department of Chemistry, Biochemistry and Physics, Université du Québec à Trois-Rivières, 3351 boulevard des Forges, Trois-Rivières, QC G9A 5H7, Canada.
  • 2 Department of Chemistry, Biochemistry and Physics, Université du Québec à Trois-Rivières, 3351 boulevard des Forges, Trois-Rivières, QC G9A 5H7, Canada; Plant Biology Research Group, Université du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada.
  • 3 Department of Chemistry, Biochemistry and Physics, Université du Québec à Trois-Rivières, 3351 boulevard des Forges, Trois-Rivières, QC G9A 5H7, Canada; Plant Biology Research Group, Université du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada. Electronic address: Isabel.Desgagne-Penix@uqtr.ca.
PMID: 40044057 DOI: 10.1016/j.biortech.2025.132336
Abstract

Phaeodactylum tricornutum is a promising host for light-driven synthesis of heterologous proteins. However, the marine cold-water environment and alkaline-acidic pH shifts in the culture, necessitated by the diatom's growth requirements. In this study, we analyzed the influence of growth condition on maturation and dynamics of the yellow fluorescent protein (YFP) in episomal-transformant P. tricornutum. A mathematical model was developed to detect the parameters that affect biomass and YFP production. Optimized conditions increased YFP mean fluorescence intensity (MFI) per cell by 4.2-fold (3.6 ± 0.6 to 15.4 ± 1.1) and total protein levels in the culture by 1.8-fold (123 ± 4 to 219 ± 9 µg L-1), without affecting biomass. YFP stability studies in P. tricornutum showed that the ubiquitin-proteasome system contributes the degradation of the recombinant protein, whereas newly synthesized YFP remains stable for up to 12 h. This optimization provides insights into the fluorescent protein-based heterologous production in diatoms.

Keywords

Diatoms; Genetic engineering; Microalgae; Protein inhibitors; Response surface methodology.

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