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<p><i>Phaeodactylum tricornutum</i> is a promising candidate for the production of recombinant therapeutic proteins including antibodies. So far, low secretion yields remain a significant bottleneck for pharmaceutical applications. Towards the objective to increase production yield of secreted recombinant protein, this study investigates the optimization of environmental and nutritional parameters, under laboratory conditions, to maximize the production of a model secreted recombinant protein (eGFPsec) under the control of the Nitrate Reductase promoter (P<sub>NR</sub>). We evaluated the interplay between nitrate availability, Photosynthetic Photon Flux Density (PPFD), and pH level within both natural and artificial seawater matrices. Our results demonstrate that while P<sub>NR</sub> activity does not increase with nitrate concentrations beyond non-limiting levels, a PPFD of 150 µmol photons m<sup>− 2</sup> s<sup>− 1</sup> significantly enhances eGFPsec accumulation in the medium. Interestingly, cultures grown in Natural Sea Water under pH-regulated conditions (pH = 8) exhibited markedly higher secreted-eGFP fluorescence compared to those in artificial seawater. These findings highlight the importance of optimizing key environmental and nutritional parameters to improve secretion recombinant protein yields. By identifying key drivers of eGFP fluorescence-based productivity and accumulation in the medium, this work provides actionable insights for process optimization, reinforcing the potential of <i>P. tricornutum</i> as a robust and cost-effective platform for industrial microalgae-based biomanufacturing.</p>

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Enhancing recombinant eGFP secretion in P. tricornutum by optimizing culture conditions

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月16日

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  • Nesrine Gargouch,
  • Aurélie Charrier,
  • Charlotte Toustou,
  • Gaël Bougaran,
  • Bruno Saint-Jean,
  • Muriel Bardor

摘要

Phaeodactylum tricornutum is a promising candidate for the production of recombinant therapeutic proteins including antibodies. So far, low secretion yields remain a significant bottleneck for pharmaceutical applications. Towards the objective to increase production yield of secreted recombinant protein, this study investigates the optimization of environmental and nutritional parameters, under laboratory conditions, to maximize the production of a model secreted recombinant protein (eGFPsec) under the control of the Nitrate Reductase promoter (PNR). We evaluated the interplay between nitrate availability, Photosynthetic Photon Flux Density (PPFD), and pH level within both natural and artificial seawater matrices. Our results demonstrate that while PNR activity does not increase with nitrate concentrations beyond non-limiting levels, a PPFD of 150 µmol photons m− 2 s− 1 significantly enhances eGFPsec accumulation in the medium. Interestingly, cultures grown in Natural Sea Water under pH-regulated conditions (pH = 8) exhibited markedly higher secreted-eGFP fluorescence compared to those in artificial seawater. These findings highlight the importance of optimizing key environmental and nutritional parameters to improve secretion recombinant protein yields. By identifying key drivers of eGFP fluorescence-based productivity and accumulation in the medium, this work provides actionable insights for process optimization, reinforcing the potential of P. tricornutum as a robust and cost-effective platform for industrial microalgae-based biomanufacturing.