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Cultivation of heterotrophic Euglena gracilis: The effects of recycled media on culture growth and associations with growth regulating phytohormone profiles

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Abstract

In response to the rapid expansion of the worldwide population, there is an urgent need for the development of new, more environmentally-conscious, food sources. In this context, microalgae, such as Euglena, are of interest due to their capacity for large-scale cultivation and flexible bioproduct output. Media recycling techniques have been proposed as waste- and cost-reducing measures for large-scale algal cultivation. Culture dynamics and phytohormone production during novel fermentation conditions can enhance growth and biomass production; yet, little is known about the impacts on growth regulating phytohormones such as cytokinins (CKs) and abscisic acid (ABA) under these conditions. Using Euglena gracilis, an organic culture medium was recycled at 25, 50 and 75% inclusion rates through multiple growth cycles to evaluate impacts on biomass production and phytohormone profiles. A 25% recycled inclusion rate resulted in biomass accumulation similar to the control over one growth cycle. Subsequent recycled rates and growth cycling reduced E. gracilis biomass accumulation relative to the control, even with cell counts continuing to increase throughout, highlighting that the recycled medium inclusion and the number of repeated culture cycles impacts the biomass output of E. gracilis. Phytohormone analysis indicated that E. gracilis synthesized CKs via the tRNA-degradation pathway with increased hormone levels corresponding to higher recycle rates; ABA increased in the supernatant at higher recycle rates but was not detected in any pellet fractions. This research expands upon the roles of CKs and ABA as signaling molecules and adds guidance for building a suite of conditions specific for industrial product development.

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The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors wish to acknowledge the financial support from the Natural Sciences and Engineering Council of Canada Discovery Grant (NSERC RGPIN-05436) and NSERC College-University Idea to Innovation Grant Program (CUI2I 470778-14). The authors thank Canadian Foundation for Innovation: Leaders Opportunity Fund and Ontario Research Fund: Research Infrastructure for funding instrumentation, including the Thermo Fisher QExactive Orbitrap, in the Water Quality Centre at Trent University.

Funding

Financial support from the Natural Sciences and Engineering Council of Canada Discovery Grant (NSERC RGPIN-05436) to RJNE, ENM and TS and NSERC College-University Idea to Innovation Grant Program (CUI2I 470778–14) to RJNE, ABK and AMK is gratefully acknowledged. Partial funding to AMK was provided by Noblegen Inc. The authors thank Canadian Foundation for Innovation: Leaders Opportunity Fund and Ontario Research Fund: Research Infrastructure for funding instrumentation, including the Thermo Fisher QExactive Orbitrap, in the Water Quality Centre at Trent University.

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Authors and Affiliations

  1. Environmental and Life Sciences Graduate Program, Trent University, Peterborough, ON, K9J 0G2, Canada

    Alexandra M. Kuhne, Erin N. Morrison, Tamanna Sultana, Anna B. Kisiala & R. J. Neil Emery

  2. Biology Department, Trent University, Peterborough, ON, K9J 0G2, Canada

    Erin N. Morrison, Anna B. Kisiala & R. J. Neil Emery

  3. Noblegen Inc., Peterborough, ON, K9L 1Z8, Canada

    Kathleen Horlock-Roberts & Adam Noble

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Contributions

AMK: Methodology, Formal analysis, Investigation, Writing Original Draft; ENM: Data analysis, Figure and Table creation, Writing-review and Editing; TS: Writing Draft; ABK: Formal analysis; KHR: Investigation; AN: Funding acquisition; RJN: Conceptualization, Project administration, Supervision, Funding acquisition. All authors read and approved the final manuscript.

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Correspondence to Erin N. Morrison.

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Competing interests

Noblegen Inc. is a commercial, for-profit organization that is using E. gracilis in future food products and biosolutions. Partial funding for this research was provided by Noblegen Inc. KHR is an employee of Noblegen Inc. AN is the CEO and founder of Noblegen Inc. and is a shareholder.

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Kuhne, A.M., Morrison, E.N., Sultana, T. et al. Cultivation of heterotrophic Euglena gracilis: The effects of recycled media on culture growth and associations with growth regulating phytohormone profiles. J Appl Phycol 35, 2161–2175 (2023). https://doi.org/10.1007/s10811-023-03062-4

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