Random UV Mutagenesis for the production of Chlorella vulgaris mutants with low chlorophyll content for the food industry
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Abstract
Microalgae is currently gaining attention as an alternative source for food production. The market is currently demanding colorless algae with high protein and lutein content as an alternative to currently available commodities. This research was aimed at performing random UV mutagenesis on Chlorella vulgaris to obtain mutants with enhanced growth rates and increased growth characteristics. A total of seven mutants were selected to be analyzed after the random mutagenesis. Small (40 mL) and larger-scale (1,000 mL) reactors were used to analyze the production of C. vulgaris mutant biomass, focusing on the dry matter, starch, chlorophyll, protein, fatty acids, and lutein contents. Results implied that mutants showed a higher specific growth rate (2.1–2.5-fold higher) as compared to the wild type. The three mutants (MT 1, 2, and 3) that exhibited a yellow color were subsequently chosen for further scalability. In larger-scale reactors, all mutants exhibited higher protein contents while displaying lower carbohydrate and chlorophyll contents in comparison to the wild type. Moreover, MT 1 exhibited the highest concentration of lutein (0.37%–0.38%) and the lowest concentration of chlorophyll (0.1–0.14%), both of which are of significance for potential applications in the food industry.
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Conclusion
Congratulations on this exciting result of isolating multiple mutants with enhanced heterotrophic growth through random UV mutagenesis. I'm particularly curious about how these mutants would perform under mixotrophic conditions compared to the wild type. While your approach of developing chlorophyll-deficient Chlorella strains certainly streamlines the culture-to-food pipeline, it would be valuable to understand whether the growth advantages persist across different cultivation modes. If there's substantial growth reduction compared to wild type performance in mixotrophic or photoautotrophic settings, perhaps post-harvest photobleaching methods of a faster growing WT strain could offer complementary efficiency benefits. I'd be especially interested in future genetic characterization to identify which specific genes have …
Conclusion
Congratulations on this exciting result of isolating multiple mutants with enhanced heterotrophic growth through random UV mutagenesis. I'm particularly curious about how these mutants would perform under mixotrophic conditions compared to the wild type. While your approach of developing chlorophyll-deficient Chlorella strains certainly streamlines the culture-to-food pipeline, it would be valuable to understand whether the growth advantages persist across different cultivation modes. If there's substantial growth reduction compared to wild type performance in mixotrophic or photoautotrophic settings, perhaps post-harvest photobleaching methods of a faster growing WT strain could offer complementary efficiency benefits. I'd be especially interested in future genetic characterization to identify which specific genes have been disrupted in these promising mutants.
Very exciting and thanks for sharing the work openly!
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One of the major benefits of UV mutagenesis, in76comparison to chemical methods, is that UV mutagenesis is random (Trovão et al., 2022).77.CC-BY 4.0 International licenseavailable under awas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is madeThe copyright holder for this preprint (whichthis version posted April 30, 2025.;https://doi.org/10.1101/2025.04.28.651037doi:bioRxiv preprint
While it is true that UV mutagenesis is random, there are many methods of chemical mutagenesis that are also random. Chemical mutagens like EMS, MMS, and sodium azide all produce stochastic genetic alterations throughout the genome. The statement suggesting randomness as a distinctive benefit of UV over chemical methods is not entirely accurate, as both approaches …
One of the major benefits of UV mutagenesis, in76comparison to chemical methods, is that UV mutagenesis is random (Trovão et al., 2022).77.CC-BY 4.0 International licenseavailable under awas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is madeThe copyright holder for this preprint (whichthis version posted April 30, 2025.;https://doi.org/10.1101/2025.04.28.651037doi:bioRxiv preprint
While it is true that UV mutagenesis is random, there are many methods of chemical mutagenesis that are also random. Chemical mutagens like EMS, MMS, and sodium azide all produce stochastic genetic alterations throughout the genome. The statement suggesting randomness as a distinctive benefit of UV over chemical methods is not entirely accurate, as both approaches fundamentally create unpredictable mutations.
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Unlike other methods that introduce68external genetic material into the target cell, it does not produce genetically modified69organisms (Bleisch et al., 2022; Kanakdande et al., 2021; Trovão et al., 2022).
The statement that random mutagenesis 'does not produce genetically modified organisms' seems misleading. While certain regulatory frameworks may classify these differently than transgenic methods, random UV mutagenesis fundamentally alters genetic material. This regulatory distinction appears based more on historical precedent than biological reality. For scientific accuracy, perhaps acknowledging these as genetically modified organisms that fall under different regulatory classifications would be more appropriate.
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The Petri dishes with the cultures were then irradiated with UV light for 5 minutes in119a sterile hood.
It would be helpful to know what the power output of the light is, what wavelength it emits, and how far the light source is from the petri dish.
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