Applied Molecular Biosciences Unit

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Organizational Unit

Publications

Random mutagenesis as a promising tool for microalgal strain improvement towards industrial production

Publication . Trovão Dos Santos, Mafalda; Schüler, Lisa M.; Machado, Adriana; Bombo, Gabriel; Navalho, Sofia; Barros, Ana; Pereira, Hugo; Silva, Joana; Freitas, Filomena; Varela, João

Microalgae have become a promising novel and sustainable feedstock for meeting the rising demand for food and feed. However, microalgae-based products are currently hindered by high production costs. One major reason for this is that commonly cultivated wildtype strains do not possess the robustness and productivity required for successful industrial production. Several strain improvement technologies have been developed towards creating more stress tolerant and productive strains. While classical methods of forward genetics have been extensively used to determine gene function of randomly generated mutants, reverse genetics has been explored to generate specific mutations and target phenotypes. Site-directed mutagenesis can be accomplished by employing different gene editing tools, which enable the generation of tailor-made genotypes. Nevertheless, strategies promoting the selection of randomly generated mutants avoid the introduction of foreign genetic material. In this paper, we review different microalgal strain improvement approaches and their applications, with a primary focus on random mutagenesis. Current challenges hampering strain improvement, selection, and commercialization will be discussed. The combination of these approaches with high-throughput technologies, such as fluorescence-activated cell sorting, as tools to select the most promising mutants, will also be discussed.

2022-06-30Journal article

Open access

Valorization of brewery waste through Polyhydroxyalkanoates production supported by a metabolic specialized microbiome

Publication . Carvalheira, Mónica; Amorim, Catarina L.; Oliveira, Ana Catarina; Guarda, Eliana C.; Costa, Eunice; Ribau Teixeira, Margarida; Castro, Paula M. L.; Duque, Anouk F.; Reis, Maria A. M.

Raw brewers’ spent grain (BSG), a by-product of beer production and produced at a large scale, presents a composition that has been shown to have potential as feedstock for several biological processes, such as polyhydroxyalkanoates (PHAs) production. Although the high interest in the PHA production from waste, the bioconversion of BSG into PHA using microbial mixed cultures (MMC) has not yet been explored. This study explored the feasibility to produce PHA from BSG through the enrichment of a mixed microbial culture in PHA-storing organisms. The increase in organic loading rate (OLR) was shown to have only a slight influence on the process performance, although a high selectivity in PHA-storing microorganisms accumulation was reached. The culture was enriched on various PHA-storing microorganisms, such as bacteria belonging to the Meganema, Carnobacterium, Leucobacter, and Paracocccus genera. The enrichment process led to specialization of the microbiome, but the high diversity in PHA-storing microorganisms could have contributed to the process stability and efficiency, allowing for achieving a maximum PHA content of 35.2 ± 5.5 wt.% (VSS basis) and a yield of 0.61 ± 0.09 CmmolPHA/CmmolVFA in the accumulation assays. Overall, the production of PHA from fermented BSG is a feasible process confirming the valorization potential of the feedstock through the production of added-value products.

2022-08-30Journal article

Open access

Does predation exacerbate the risk of endosymbiont loss in heat stressed hermatypic corals? molecular cues provide insights into species-specific health outcomes in a multi-stressor ocean

Publication . Madeira, Carolina; Dias, Marta; Ferreira, Ana; Gouveia, Raúl; Cabral, Henrique; Diniz, Mário S.; Vinagre, Catarina

Ocean warming has been a major driver of coral reef bleaching and mass mortality. Coupled to other biotic pressures, corals' ability for acclimatization and adaptation may become compromised. Here, we tested the combined effects of warming scenarios (26, 30, and 32 degrees C) and predation (wound vs. no wound) in coral health condition (paleness, bleaching, and mortality), cellular stress responses (heat shock protein 70 kDa Hsp70, total ubiquitin Ub, and total antioxidant capacity TAC), and physiological state (integrated biomarker response index, IBR) of seven Scleractinian coral species, after being exposed for 60 days. Results show that although temperature was the main factor driving coral health condition, thermotolerant species (Galaxea fascicularis, Psammocora contigua, and Turbinaria reniformis) displayed increased paleness, bleaching, and mortality in predation treatments at high temperature, whereas thermosensitive species (Acropora tenuis, Echinopora lamellosa, and Montipora capricornis brown and green morphotypes) all died at 32 degrees C, regardless of predation condition. At the molecular level, results show that there were significant main and interactive effects of species, temperature, and predation in the biomarkers assessed. Temperature affected Hsp70, Ub, and TAC, evidencing the role of protein folding and turnover, as well as reactive oxygen species scavenging in heat stress management. Predation increased Hsp70 and Ub, suggesting the activation of the pro-phenoloxidase system and cytokine activity, whereas the combination of both stressors mainly affected TAC during moderate stress and Ub under severe stress, suggesting that redox balance and defense of homeostasis are crucial in tissue repair at high temperature. IBR levels showed an increasing trend at 32 degrees C in predated coral fragments (although non-significant). We conclude that coral responses to the combination of high temperature and predation pressure display high inter-species variability, but these stressors may pose a higher risk of endosymbiont loss, depending on species physiology and stress intensity.

2022-03Journal article

Open access

Comprehensive metabolomics and lipidomics profiling of prostate cancer tissue reveals metabolic dysregulations associated with disease development

Publication . Lima, Ana Rita; Carvalho, Márcia; Aveiro, Susana; Melo, Tânia; Domingues, M. Rosário; Macedo-Silva, Catarina; Coimbra, Nuno; Jerónimo, Carmen; Henrique, Rui; Bastos, Maria de Lourdes; Guedes de Pinho, Paula; Pinto, Joana

Prostate cancer (PCa) is a global health problem that affects millions of men every year. In the past decade, metabolomics and related subareas, such as lipidomics, have demonstrated an enormous potential to identify novel mechanisms underlying PCa development and progression, providing a good basis for the development of new and more effective therapies and diagnostics. In this study, a multiplatform metabolomics and lipidomics approach, combining untargeted mass spectrometry (MS) and nuclear magnetic resonance (NMR)-based techniques, was applied to PCa tissues to investigate dysregulations associated with PCa development, in a cohort of 40 patients submitted to radical prostatectomy for PCa. Results revealed significant alterations in the levels of 26 metabolites and 21 phospholipid species in PCa tissue compared with adjacent nonmalignant tissue, suggesting dysregulation in 13 metabolic pathways associated with PCa development. The most affected metabolic pathways were amino acid metabolism, nicotinate and nicotinamide metabolism, purine metabolism, and glycerophospholipid metabolism. A clear interconnection between metabolites and phospholipid species participating in these pathways was observed through correlation analysis. Overall, these dysregulations may reflect the reprogramming of metabolic responses to produce high levels of cellular building blocks required for rapid PCa cell proliferation.

2021-03Journal article

Restricted access

Priority effects, environmental filtering and neutral coexistence explain large- to small-scale distribution of invasive sun corals in the SW Atlantic

Publication . Barbosa, A. C.C.; Vinagre, Catarina; Kitahara, MV; Flores, A. A. V.

Two sun coral species, Tubastraea tagusensis and T. coccinea, have successfully colonized reef habitats along the Southwest Atlantic. However, their invasive biology has been largely addressed without considering species-specific distribution patterns. Here, we assessed the distribution and abundance of Tubastraea spp. at vertical rocky reef sites within a number of islands along 120 km of coastline off the northern coast of Sao Paulo State, Brazil, to (1) investigate possible mechanisms underlying the invasion dynamics in the region, (2) test species-specific distributions according to a key environmental filter (depth), and (3) examine within-patch patterns to assess whether competition, niche-based or neutral processes are best candidates to modulate local species coexistence. Sun corals were found in the great majority of the studied locations, and the probability of finding them at any given reef site was estimated to be 0.54. There was substantial species segregation across locations, consistent with primary priority effects. Within locations, results suggest environmental filtering, with T. coccinea apparently advantaged in more hydrodynamic environments just below the surf zone. At sun coral patches with extensive co-occurrence of T. tagusensis and T. coccinea, the presence of each species can be, remarkably, modeled as an independent event, suggesting neutral coexistence. The spread of sun corals is an ongoing and increasingly invasive process that may be explained by the enemy-release hypothesis and the lack of negative interactions between Tubastraea species. The stochastic nature of small-scale distributions sets an additional challenge to predict (and thus control) sun coral invasion.

2023Journal article

Restricted access