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- Isolation and selection of protein-rich mutants of chlorella vulgaris by fluorescence-activated cell sorting with enhanced biostimulant activity to germinate garden cress seedsPublication . Trovão dos Santos, Mafalda; Schüler, Lisa; Pedroso, Humberto; Reis, Ana; Santo, Gonçalo Espírito; Barros, Ana; Correia, Nádia; Ribeiro, Joana; Bombo, Gabriel; Gama, Florinda; Viana, Catarina; Costa, Monya; Ferreira, Sara; Cardoso, Helena; Varela, João; Silva, Joana; Freitas, Filomena; Pereira, HugoMicroalgae are a promising feedstock with proven biostimulant activity that is enhanced by their biochemical components (e.g., amino acids and phytohormones), which turns them into an appealing feedstock to reduce the use of fertilisers in agriculture and improve crop productivity and resilience. Thus, this work aimed to isolate protein-rich microalgal mutants with increased biostimulant activity. Random mutagenesis was performed with Chlorella vulgaris, and a selection of protein-rich mutants were sorted through fluorescence-activated cell sorting (FACS), resulting in the isolation of 17 protein-rich mutant strains with protein contents 19-34% higher than that of the wildtype (WT). Furthermore, mutant F4 displayed a 38%, 22% and 62% higher biomass productivity, growth rate and chlorophyll content, respectively. This mutant was then scaled up to a 7 L benchtop reactor to produce biomass and evaluate the biostimulant potential of this novel strain towards garden cress seeds. Compared to water (control), the germination index and the relative total growth increased by 7% and 19%, respectively, after the application of 0.1 g L-1 of this bioproduct, which highlights its biostimulant potential.
- Improving the heterotrophic media of three chlorella vulgaris mutants toward optimal color, biomass and protein productivityPublication . Trovão dos Santos, Mafalda; Cunha, Miguel; Santo, Gonçalo Espírito; Pedroso, Humberto; Reis, Ana; Barros, Ana; Correia, Nádia; Schüler, Lisa; Costa, Monya; Ferreira, Sara; Cardoso, Helena; Ventura, Márcia; Varela, João; Silva, Joana; Freitas, Filomena; Pereira, HugoThe high production costs and unappealing sensory properties still limit the widespread commercialization of microalgae feedstocks. Therefore, this work focused on fine-tuning the heterotrophic medium composition to cultivate novel green, yellow, and white Chlorella vulgaris mutant strains. Screening assays were carried out to select the most significant factors, and different nutrient concentrations were optimized by modelling biomass and protein productivity, specific growth rate, and color, via response surface methodology. The biomass and protein productivities achieved by these strains were improved by up to 70% and 94%, respectively. Additionally, biomass color was correlated with medium composition for the first time, allowing the improvement of the yellow and white mutant colorations by 20%. Overall, the findings of this study are vital to overcoming the challenges of the biobased industry, allowing the enhancement of the cost-effectiveness, attractiveness, and nutritional profiles of microalgae-based products in different markets and applications.