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bioRxiv · 10.1101/2025.10.28.685034

Generation of Chlorella vulgaris starch mutants and their biomass and lipid productivities under different culture media

Ramos, A. C. E.·Hamilton, A.·Molina, I.·Mcginn, P.·Regan, S.
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Abstract

BackgroundMicroalgae are an important feedstock for the production of a wide variety of products, including biodiesel. Biodiesel, composed of fatty acid alkyl esters, is produced through the transesterification reaction of triacylglycerol (TAG). Microalgae store their energy reserves primarily as starch and TAGs. Therefore, several studies have focused on understanding the partitioning of carbon precursors between starch and TAG biosynthetic pathways. In this study, 5 starch mutants of Chlorella vulgaris were developed and cultured on different culture media. ResultsChlorella vulgaris starch mutants were generated through UV-random mutagenesis. Five starch mutants were selected for this study: four low-starch producing mutants (st27, st29, st43 and st54) and one high-starch producing mutant (st80). The starch mutants were cultured on media with different organic carbon sources, and lipid and biomass productivity were measured. Mixotrophic growth on glucose resulted in the highest lipid productivity in all the mutants, including st80, without compromising growth, whereas photoautotrophic growth generally did not result in changes in lipid productivity of the starch mutants. The highest increase in lipid productivity was observed for st27, with a 3.8-fold higher lipid productivity than wildtype. ConclusionsAll starch mutants increased their lipid productivities when grown mixotrophically on glucose, suggesting the overflow hypothesis could explain the partitioning of carbon between starch and TAGs. Out of the mutants generated in this work, st27 resulted in the highest increases in lipid productivities, reaching an increase of 380% when grown mixotrophically on glucose, without compromising growth. The high-starch producing mutant st80 provides insight into a possibility to develop starch- and TAG-rich microalgal biomass.

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Ramos, A. C. E., Hamilton, A., Molina, I., Mcginn, P., Regan, S.. 2025-10-29. Generation of Chlorella vulgaris starch mutants and their biomass and lipid productivities under different culture media. https://doi.org/10.1101/2025.10.28.685034

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