Semi-continuous cultivation for enhanced protein production using indigenous green microalgae and synthetic municipal wastewater
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Abstract Cultivation of microalgae has gained significant interest as an alternative protein source, potentially becoming a target commodity recovered from microalgae-based wastewater treatment. This study examined a semi-continuous cultivation strategy to optimize protein accumulation of the indigenous freshwater chlorophytes, Lobochlamys segnis and Klebsormidium flaccidum, and simultaneously remove nutrients from wastewater efficiently. A strain-specific regime was made based on a fixed biomass concentration at the start of 24-h cultivation cycle, i.e., a constant initial cell density, which regulated harvesting and fresh medium supply volume according to the dilution rate. Six cultivation cycles were conducted in lab-scale 1L reactors with a synthetic municipal wastewater. Lobochlamys segnis and K. flaccidum grew exponentially in all cycles. The biomass productivity was 573 and 580 mg L–1 day–1, in which the total protein consisted of 62 and 45% of dry cell weight (dw), respectively. When a culture medium deficient in nitrogen and phosphorus was used, protein level was significantly reduced. L. segnis consumed all NH4+ and PO43– supplied by the medium replacement, giving the removal rate of 9.2 and 5.2 mg L–1 day–1. Whereas K. flaccidum removed 13.8 mg L–1 day–1 NH4+ without completing PO43– removal. The amino acid profile of both strains was characterized by glutamic acids content (4–5% dw). We concluded that the designed cultivation regime would support a constant biomass production with stable and high protein content, along with an efficient removal of nutrient from the wastewater.
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Integration of microalgal biotechnology unfolds along two convergent fronts: genetic improvement of strains to enhance the yield of biomolecules of interest, and application of these biological advances in scalable operational regimes with cultivation intensification through higher cell density and integration into existing industrial flows.
“This integration unfolds primarily along two convergent fronts: (i) genetic improvement of strains to enhance the yield of biomolecules of interest—such as proteins, fatty acids, carbohydrates, and pigments—through tools ranging from targeted mutagenesis to synthetic biology and metabolic optimization; and (ii) the application of these biological advances in scalable operational regimes, such as”
From Microalgae biotechnology and its role in sustainable and healthy food design (2025) · cites this paper as Umetani et al., 2024
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Microalgae biotechnology and its role in sustainable and healthy food design
Schneider, A. T., Machado, R. L. S., Dutra, D. A., Machado, E. F., Dias, R. R., Deprá, M. C., Zepka, L. Q., & Jacob-Lopes, E. (2025) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2025.1716473 · Cited by 15 · see citing works
- Authors
- Umetani, I., Sposób, M., & Tiron, O.
- Affiliations
- Norwegian Institute of Bioeconomy Research (NO)
- National Institute for Research and Development for Industrial Ecology (RO)
- Venue
- Journal of Applied Phycology
- Published
- 2024-01-16
- Volume
- 36
- Issue
- 3
- Pages
- 1105-1116
- ISSN
- 0921-8971 (ISSN-L)
- 1573-5176
- OpenAlex
- W4390914192
- Cited by
- 11
- References
- 37
- Citations by year
- 2026: 4 · 2025: 5 · 2024: 2
- FWCI
- 0.74
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Journal of Applied Phycology (publishedVersion, cc-by)
- Funders
- Norwegian Institute of Bioeconomy Research
- Topics
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Physical Sciences › Environmental Science › Environmental Chemistry › Aquatic Ecosystems and Phytoplankton Dynamics
- Life Sciences › Agricultural and Biological Sciences › Aquatic Science › Aquaculture Nutrition and Growth
- Keywords
- semi-continuous cultivation, Klebsormidium flaccidum, municipal wastewater treatment, nutrient removal, nitrogen removal, phosphorus removal, Chlorophyta, biomass productivity, amino acid composition, glutamate, nitrogen and phosphorus deficiency
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