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  • Sargassum muticum and Osmundea pinnatifida enzymatic extracts: chemical, structural, and cytotoxic characterization
    Publication . Rodrigues, Dina; Costa-Pinto, Ana R.; Sousa, Sérgio; Vasconcelos, Marta W.; Pintado, Manuela M.; Pereira, Leonel; Rocha-Santos, Teresa A. P.; Costa, João P. da; Silva, Artur M. S.; Duarte, Armando C.; Gomes, Ana M. P.; Freitas, Ana C.
    Seaweeds, which have been widely used for human consumption, are considered a potential source of biological compounds, where enzyme-assisted extraction can be an efficient method to obtain multifunctional extracts. Chemical characterization of Sargassum muticum and Osmundea pinnatifida extracts obtained by Alcalase and Viscozyme assisted extraction, respectively, showed an increment of macro/micro elements in comparison to the corresponding dry seaweeds, while the ratio of Na/K decreased in both extracts. Galactose, mannose, xylose, fucose, and glucuronic acid were the main monosaccharides (3.2–27.3 mg/glyophilized extract) present in variable molar ratios, whereas low free amino acids content and diversity (1.4–2.7 g/100gprotein) characterized both extracts. FTIR-ATR and 1H NMR spectra confirmed the presence of important polysaccharide structures in the extracts, namely fucoidans from S. muticum or agarans as sulfated polysaccharides from O. pinnatifida. No cytotoxicity against normal mammalian cells was observed from 0 to 4 mglyophilized extract/mL for both extracts. The comprehensive characterization of the composition and safety of these two extracts fulfils an important step towards their authorized application for nutritional and/or nutraceutical purposes.
  • Chemical composition of red, brown and green macroalgae from Buarcos bay in Central West Coast of Portugal
    Publication . Rodrigues, Dina; Freitas, Ana C.; Pereira, Leonel; Rocha-Santos, Teresa A. P.; Vasconcelos, Marta W.; Roriz, Mariana; Rodríguez-Alcalá, Luís M.; Gomes, Ana M. P.; Duarte, Armando C.
    Six representative edible seaweeds from the Central West Portuguese Coast, including the less studied Osmundea pinnatifida, were harvested from Buarcos bay, Portugal and their chemical characterization determined. Protein content, total sugar and fat contents ranged between 14.4% and 23.8%, 32.4% and 49.3% and 0.6–3.6%. Highest total phenolic content was observed in Codium tomentosum followed by Sargassum muticum and O. pinnatifida. Fatty acid (FA) composition covered the branched chain C13ai to C22:5 n3 with variable content in n6 and n3 FA; low n6:n3 ratios were observed in O. pinnatifida, Grateloupia turuturu and C. tomentosum. Some seaweed species may be seen as good sources of Ca, K, Mg and Fe, corroborating their good nutritional value. According to FTIR-ATR spectra, G. turuturu was associated with carrageenan seaweed producers whereas Gracilaria gracilis and O. pinnatifida were mostly agar producers. In the brown algae, S. muticum and Saccorhiza polyschides, alginates and fucoidans were the main polysaccharides found.
  • Biodegradation of polyethylene microplastics by the marine fungus zalerion maritimum
    Publication . Paço, Ana; Duarte, Kátia; Costa, João P. da; Santos, Patrícia S.M.; Pereira, R.; Pereira, M. E.; Freitas, Ana C.; Duarte, Armando C.; Rocha-Santos, Teresa A. P.
    Plastic yearly production has surpassed the 300milliontons mark and recycling has all but failed in constituting a viable solution for the disposal of plastic waste. As these materials continue to accumulate in the environment, namely, in rivers and oceans, in the form of macro-, meso-, micro- and nanoplastics, it becomes of the utmost urgency to find new ways to curtail this environmental threat. Multiple efforts have been made to identify and isolate microorganisms capable of utilizing synthetic polymers and recent results point towards the viability of a solution for this problem based on the biodegradation of plastics resorting to selected microbial strains. Herein, the response of the fungus Zalerion maritimum to different times of exposition to polyethylene (PE) pellets, in a minimum growth medium, was evaluated, based on the quantified mass differences in both the fungus and the microplastic pellets used. Additionally, molecular changes were assessed through attenuated total reflectance Fourier transform Infrared Spectroscopy (FTIR-ATR) and Nuclear Magnetic Resonance (NMR). Results showed that, under the tested conditions, Z. maritimum is capable of utilizing PE, resulting in the decrease, in both mass and size, of the pellets. These results indicate that this naturally occurring fungus may actively contribute to the biodegradation of microplastics, requiring minimum nutrients.
  • Impact of enzyme- and ultrasound-assisted extraction methods on biological properties of red, brown, and green seaweeds from the Central West Coast of Portugal
    Publication . Rodrigues, Dina; Sousa, Sérgio; Silva, Aline; Amorim, Manuela; Pereira, Leonel; Rocha-Santos, Teresa A.P.; Gomes, Ana M. P.; Duarte, Armando C.; Freitas, Ana Cristina
    Seaweeds are an excellent source of bioactive compounds, and therefore the use of sustainable and food compatible extraction methods such as enzyme-assisted (EAE) and ultrasound-assisted extraction were applied on Sargassum muticum, Osmundea pinnatifida, and Codium tomentosum. Extracts were evaluated for proximate characterization and biological properties. Higher extraction yields were observed for C. tomentosum EAE (48-62%; p < 0.05 for Cellulase and Viscozyme), followed by O. pinnatifida (49-55%; p < 0.05 except Alcalase) and S. muticum (26-31%; p < 0.05). S. muticum extracts presented the highest nitrogen (25 +/- 2 mg/g(lyoph) (extract)) and total phenolics (261 +/- 37 mu g(catechol) (equiv)/gl(yoph) (extract)) contents, whereas higher sugars (78 +/- 14 mg(glucose) (equiv)/g(lyoph) (extract)) including sulfated polysaccharide (44 +/- 8 mg(Na2SO4) (acid)/g(lyoph) (extract)) contents characterized O. pinnatifida extracts. A higher effect on hydroxyl-radical scavenging activity (35-50%) was observed for all extracts, whereas S. muticum Alcalase and C. tomentosum Cellulase extracts exhibited higher prebiotic activity than fructooligosaccharides. O. pinnatifida and C. tomentosum EAE showed inhibitory potential against alpha-glucosidase (38-49%).
  • Lipolysis in probiotic and synbiotic cheese: The influence of probiotic bacteria, prebiotic compounds and ripening time on free fatty acid profiles
    Publication . Rodrigues, Dina; Rocha-Santos, Teresa A. P.; Gomes, Ana M.; Goodfellow, Brian J.; Freitas, Ana C.
    The influence of probiotic bacteria (Lactobacillus casei-01, Bifidobacterium lactis B94), prebiotic compounds (FOS and inulin) and ripening time (0–60 days) on the free fatty acid (FFA) profile of cheese, with special emphasis on the conjugated linoleic acid (CLA) content, was investigated. After 60 days of ripening, 109–1010 cfu g 1 cheese were recorded in both probiotic and synbiotic cheeses, despite harsh conditions of low pH values (4.1–5.1) and low moisture content (<30%, w/w). Increases in total FFA and CLA were observed throughout the ripening period, especially in synbiotic cheeses containing FOS and inulin (50:50) inoculated with B. lactis B94. The addition of FOS alone or combined with inulin did not significantly affect probiotic strain growth and viability during the ripening period; however, the advantage of the addition of prebiotic compounds in probiotic cheese manufacture is that it may allow the production of cheeses with improved performance as far as functional CLA compounds are concerned, as well as an improved nutritional quality reflected in a lower atherogenicity index.
  • Chemical and structural characterization of Pholiota nameko extracts with biological properties
    Publication . Rodrigues, Dina; Freitas, Ana C.; Sousa, Sérgio; Amorim, Manuela; Vasconcelos, Marta W.; Costa, João P. da; Silva, Artur M. S.; Rocha-Santos, Teresa A. P.; Duarte, Armando C.; Gomes, Ana M. P.
    Edible mushrooms including Pholiota nameko are excellent sources of extractable bioactive compounds with much to explore. Enzymatic extractions with Cellulase and Viscozyme were responsible for highest extraction yields (67-77%). No strong antioxidant activity was observed although extracts were able to scavenge ABTS(+) and OH(+). Potential prebiotic activity was observed in all extracts, some increasing 1.4-2 Log cycles of Lactobacillus acidophilus La-5 and Bifidobacterium animalis BB12. 30-50% α-glucosidase inhibition was observed in ultrasound, Flavourzyme and Cellulase extracts. Flavourzyme and Cellulase extracts are richer in macro (Mg, K and P) and micro elements (Zn, Mn and Fe) in comparison to mushroom. Monosaccharides content and profile varied slightly among both extracts with predominance of glucose, galactose and mannose with no uronic acids detection; Flavourzyme extract reported higher free amino acids content. Presence of α and β-glycosidic structures such as glucans and glucan-protein complexes are among the polysaccharides found in both extracts.
  • Graphene based sensors and biosensors
    Publication . Justino, Celine I. L.; Gomes, Ana R.; Freitas, Ana C.; Duarte, Armando C.; Rocha-Santos, Teresa A. P.
    Graphene has contributed to the fabrication of sensitive sensors and biosensors due to its physical and electrochemical properties. This review discusses the role of graphene and graphene related materials for the improvement of the analytical performance of sensors and biosensors. This paper also provides an overview of recent graphene based sensors and biosensors (2012–2016), comparing their analytical performance for application in clinical, environmental, and food sciences research, and comments on future and interesting research trends in this field.
  • Production of conjugated linoleic acid by food-grade bacteria: A review
    Publication . Andrade, José C.; Ascenção, Kelly; Gullón, Patricia; Henriques, Silvino M. S.; Pinto, Jorge M. S.; Rocha-Santos, Teresa A. P.; Freitas, A. Cristina; Gomes, Ana Maria
    Conjugated linoleic acid (CLA) is a group of isomers of linoleic acid (LA) with several potential health benefits which justifies the efforts to obtain foods with enhanced CLA contents. A possible way to obtain CLA-enriched products is via fermentation as several micro-organisms, many of which are employed in the manufacture of foods, are known to convert free LA into CLA. However, due to several constraints, the use of CLA-producing bacteria still remains challenging and continuous efforts are required. This review summarises the current knowledge on microbial CLA production by food-grade micro-organisms, its technical issues and limitations, potential applications and bioactivities.
  • Development of probiotic tablets using microparticles: viability studies and stability studies
    Publication . Sousa e Silva, J. P.; Sousa, Sérgio C.; Costa, Paulo; Cerdeira, Emília; Amaral, Maria H.; Lobo, José Sousa; Gomes, Ana M. P.; Pintado, Maria M.; Rodrigues, Dina; Rocha-Santos, Teresa; Freitas, A. C.
    Alternative vectors to deliver viable cells of probiotics, to those conferring limited resistance to gastrointestinal conditions, still need to be sought. Therefore the main goal of the study was to develop tablets able to protect entrapped probiotic bacteria from gastric acidity, thus providing an easily manufacturing scale-up dosage form to deliver probiotics to the vicinity of the human colon. Whey protein concentrate microparticles with Lactobacillus paracasei L26 were produced by spray-drying and incorporated in tablets with cellulose acetate phthalate and sodium croscarmellose. The viability of L. paracasei L. 26 throughout tableting as well as its gastric resistance and release from the tablets were evaluated. Storage stability of L. paracasei L26 tablets was also performed by evaluation of viable cells throughout 60 days at 23 degrees C and 33% relative humidity. A decrease of approximately one logarithmic cycle was observed after the acid stage and the release of L. paracasei L26 from the tablets occurred only after 4 h in the conditions tested. Microencapsulated L. paracasei L26 in tablets revealed some susceptibility to the storage conditions tested since the number of viable cells decreased 2 log cycles after 60 days of storage. However, the viability of L. paracasei L26 after 45 days of storage did not reveal significant susceptibility upon exposure to simulated gastrointestinal conditions. The developed probiotic tablets revealed to be potential vectors for delivering viable cells of L. paracasei L26 and probably other probiotics to persons/patients who might benefit from probiotic therapy.
  • Antioxidative peptides: trends and perspectives for future research
    Publication . Freitas, A. C.; Andrade, J. C.; Silva, F. M.; Rocha-Santos, T. A. P.; Duarte, A. C.; Gomes, Ana M. P.
    In recent years, much attention has been given to dietary antioxidants, especially polyphenols. Several peptides derived from protein molecules have also been found to show antioxidant capacity along with other biological properties and thus there is an increasing interest in these compounds as health promoters. This review summarizes and discusses the main sources of antioxidative peptides with focus on food-derived peptides (animal, plant and marine sources), methods of preparation, antioxidant capacity evaluation as well as their proposed mechanisms of action. A discussion of the potential health effects and comments on the different applications for these antioxidants and their potential research interest are also subject of this review.