Browsing by Author "Pedroso, Humberto"
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- Heterotrophic cultivation of Chlorella vulgaris yellow mutant on sidestreams: Medium formulation and process scale-upPublication . Trovão, Mafalda; Barros, Ana; Machado, Adriana; Reis, Ana; Pedroso, Humberto; Espírito Santo, Gonçalo; Correia, Nádia; Costa, Monya; Ferreira, Sara; Varela, João; Cardoso, Helena; Silva, Joana; Pereira, Hugo; Freitas, FilomenaMicroalgal protein is a promising feedstock to complement and/or replace other protein sources. Besides requiring less land and water usage, microalgae production is a more sustainable process, especially if industrial sidestreams are used as nutrient sources. Additionally, the heterotrophic cultivation of microalgae, such as Chlorella vulgaris, enables the achievement of much higher biomass productivity and lower areal footprint than autotrophic cultivation. Chlorophyll-deficient strains of C. vulgaris, as the yellow strain 7Y, provide microalgal biomass with improved sensory properties. In line with this, a waste-based medium was formulated to cultivate this strain, aiming at maximum biomass productivity. In this context, several industrial sidestreams were screened, and two food wastes and corn molasses were selected for their high nitrogen and glucose concentrations, respectively. The waste-based medium formulated was compared to the inorganic optimised medium at laboratory scale in Erlenmeyer flasks and 7-L reactors. The results obtained in the 7-L fermenters revealed that both conditions achieved similar biomass productivities and growth rates of approximately 14 g L-1 d-1 and 0.8 d-1, respectively. The biomass and protein productivities were further enhanced by supplying a higher nitrogen concentration in the feeding solution when the process was scaled-up to 200-L reactors, reaching 22 and 6.3 g L-1 d-1, respectively, thus validating the developed industrial waste-based medium for the efficient cultivation of C. vulgaris under heterotrophic conditions.
- 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.
- Oxyfluorfen: a novel metabolic inhibitor to select microalgal chlorophyll-deficient mutant strains for nutritional applicationsPublication . Trovão dos Santos, Mafalda; Cardoso, Lucas; Schüler, Lisa; Machado, Adriana; Santo, Gonçalo Espírito; Pedroso, Humberto; Reis, Ana; Barros, Ana; Correia, Nádia; Costa, Monya; Ferreira, Sara; Cardoso, Helena; Mateus, Marília; Silva, Joana; Pereira, Hugo; Freitas, Filomena; Varela, JoãoNowadays, there is an increasing demand for novel feedstocks and alternative protein sources to meet global needs. Because of their rich nutritional profiles and high protein contents, microalgae-based food products and supplements are being developed. Nonetheless, these products present organoleptic characteristics such as taste, smell and colour that are often considered unpleasant by human and animal consumers. To address this constraint, strain improvement approaches such as random mutagenesis have been used, which combined with the right selection strategy, lead to more appealing microalgal biomass. In this work, a novel selection strategy using oxyfluorfen, an inhibitor of the chlorophyll synthesis pathway, was applied for the first time to isolate chlorophyll-deficient strains of Scenedesmus rubescens and Chlorella vulgaris upon treatment with ethyl methanesulfonate (EMS). With this approach, one S. rubescens brownish (37Y01) mutant strain, as well as two C. vulgaris mutant strains, one yellow (31Y15) and one white (31W62), were obtained. S. rubescens 37Y01 displayed a reduced protein content of 19.1% dry weight (DW) compared to that of the wildtype, which presented a protein content of 25.0% DW. C. vulgaris wildtype and mutants exhibited higher protein contents, in the 42.844.3% DW range, compared to Scenedesmus rubescens (p < 0.05). The selective pressure of this inhibitor allowed the selection of S. rubescens and C. vulgaris mutants displaying 55% and 95% decrease in chlorophyll content, respectively. The reduced chlorophyll content greatly improves the sensory properties and consumer acceptance of established mutants, increasing the potential of both strains as feedstocks to develop novel food products.