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- Response surface methodology in the optimization of extraction conditions for Gracilaria gracilis extracts for use in thermoplastic food coatingsPublication . Reboleira, João; Andrade, Mariana A.; Sanches-Silva, Ana; Ganhão, Rui; Mendes, Susana; Vilarinho, Fernanda; Félix, Rafael; Pinheiro, Joaquina; Bernardino, Susana M.Incorporation of antioxidant agents in edible films and packages often relies in the usage of essential oils and other concentrated hydrophobic liquids, with reliable increases in antimicrobial and antioxidant activities of the overall composite, and consequent improvements in product shelf life. These oils are often obtained from plant sources and the usage of organic solvents in their extraction has been criticized as to whether it constitutes a health hazard for consumers [1, 2, 3]. The use of water-soluble antioxidant agents is less common, due to lower activities and extraction yields. As such, using commonly available seaweeds as the source for both the main polymers and the supplementing antioxidant/antimicrobial agents in bioactive films can potentially reduce production costs and create a safer, more sustainable product [1]. Hydroethanolic extracts of commercially available red macroalgae Gracilaria gracilis were evaluated for their antioxidant potential and phenolic content, as part of the preliminary assays for the selection of algal biomass for the enrichment of thermoplastic films. The extracts were obtained through use of solid-liquid extractions, over which yield, DPPH radical reduction capacity, total phenolic content, and FRAP activity assays were measured [4, 5]. Solid to liquid ratio (SLR), extraction time, and ethanol to water ratio were selected as independent variables with experimental ranges and configurations obtained using a Box-Behnken design with three factors, resulting in 15 experimental conditions. Extraction duplicates were used [4]. Response surface methodology was then used to estimate the effect of each extraction condition on the tested bioactivities. Bioactivities were significantly (p<0.05) and positively affected by the presence of ethanol in the extraction solvent, while extraction yield was reduced. Lower solid to liquid ratios (higher solvent proportions) resulted in higher yields, but had no significant (p<0.05) impact on bioactivities. Time was not a relevant factor in any of the measured variables, and as such will be minimized in any further work. 100% ethanol with minimum extraction time (10 minutes) and 1g to 5mL SLR maximize antioxidant activities per gram of dry extract. Maximum ethanolic yields were obtained with a theoretical extraction of 100 minutes and a 1g to 25mL SLR. Aqueous extractions displayed similar results in terms of optimum conditions, but with overall lower bioactivities and higher yields, very likely due to high amounts of soluble polysaccharides. While the high antioxidant activities from the ethanolic extracts encourage the selection of this solvent for extraction of bioactives, low yields may make them later unfeasible. Future studies over the compatibility of these extracts in the polymeric matrix of the films, as well as further optimization of the extraction process will be necessary before the definitive choice for bioactive origin and processing is made.
- Morphological and mechanical characterization of films incorporating Porphyran extracted from Porphyra dioicaPublication . Baptista, Ricardo S.; Teles, Marco; Adão, Pedro; Afonso, Clélia; Bernardino, Raul; Bernardino, Susana; Ferro, Alberto C.; Elias, Sara; Guedes, MafaldaThis paper studies the effects of glycerol plasticizers and/or alginate, pectin, and carboxymethylcellulose polysaccharides on the mechanical and physical properties of porphyran-based films to evaluate the films’ ability to be used as food packaging. Films were characterized in terms of their composition, microstructural and morphological features, thermal properties, water interaction, and mechanical performance. All films are homogeneous, transparent, and slightly brownish in color. The structures are amorphous and crosslinked, showing the films’ thermoset nature. Moisture content and water solubility depend on the second polysaccharide added to the porphyran, but they both increase with the addition of glycerol to the formulations; water vapor permeability is strongly affected by the second polysaccharide in the formulation. The films display stiff and brittle mechanical behavior, but ductility increases significantly in formulations containing glycerol plasticizers. The barrier and mechanical performance values of the materials produced were found to be lower than those reported for commercial food packaging. The formulations containing glycerol displayed lower water vapor permeability values, ranging from 2.98 for porphyran/carboxymethylcellulose/glycerol to 6.65 mm.g.d-1.m-2.kPa-1 for porphyran/alginate/glycerol films. All films, except porphyran/glycerol and porphyran/alginate/glycerol, had ultimate tensile strengths above 10 MPa—the threshold value that ensures that a package is ductile enough to withstand handling and forming operations. Furthermore, the porphyran/ pectin/glycerol and porphyran/carboxymethylcellulose/glycerol films displayed sufficiently high ductility values of 2.94 and 3.10%, respectively. These results indicate that the studied porphyran/pectin/glycerol and porphyran/carboxymethylcellulose/glycerol formulations have a combination of physical and mechanical properties that ensure adequate film integrity and function through the complete food packaging supply chain. The results here reported represent an opportunity to extend the scope of porphyran films to applications in the dry food packaging industry.
- Desenvolvimento de um sumo de fruta com adição de macroalga Gelidium corneum, recorrendo à tecnologia HPP: caracterização físico-química, microbiológica e sensorialPublication . Marques, A. T.; Bernardino, SusanaNo setor alimentar, o controlo da qualidade dos produtos e dos processos é uma etapa essencial, uma vez que é por este meio que se avaliam os padrões exigidos, quer a nível de legislação, quer a nível de mercado. O presente trabalho teve, como objetivos principais, o desenvolvimento de um novo produto alimentar com adição de algas, bem como o controlo de qualidade a nível de uma unidade fabril de produção de sumos e polpas naturais, à base de hortofrutícolas. Considerando a tendência e potencial de crescimento apresentados pelo setor das bebidas, nomeadamente dos sumos de fruta naturais, juntamente com a incessante procura por ingredientes naturais ricos em vários nutrientes e com propriedades bioativas, tem-se vindo a registar o lançamento de um número considerável de bebidas inovadoras, onde as macroalgas têm merecido um papel de destaque. Desta forma, procedeu-se à elaboração de dois protótipos, nomeadamente, sumo de framboesa e sumo de espinafres, com a adição do extrato da alga Gelidium corneum, estabilizados através do processo de Hiperpressão a frio, com o intuito de se avaliar o impacto desse extrato nos diferentes parâmetros de qualidade dos sumos. No que respeita aos parâmetros físico-químicos, o pH e teor de sólidos solúveis (TSS) foram avaliados após produção (t0), no tempo intermédio (t15) e no tempo final (t30). A cor foi determinada no t0 e no t30 e os restantes parâmetros, como acidez titulável, quantificação total de polifenóis, capacidade de redução do radical DPPH, teor de proteína bruta, teor de cinzas, teor de minerais e oligoelementos e teor de vitaminas (A, B1, B2 e C) foram avaliados no tempo final (t30). Foram, também, avaliados os parâmetros microbiológicos mais significativos, nomeadamente, microrganismos totais a 30°C, Escherichia coli, Salmonella spp, bolores e leveduras (ao t0, t15 e t30) e realizou-se uma avaliação sensorial ao 29.º dia após produção. Estes sumos apresentaram-se microbiologicamente estáveis, não se tendo verificado crescimento microbiológico durante o período de armazenamento de 30 dias. A análise dos resultados físico-químicos permitiu constatar que a adição do extrato da alga Gelidium corneum aos sumos influenciou as suas características, tendo havido um aumento significativo do pH com a consequente diminuição da acidez titulável e um aumento do teor de sólidos solúveis, em comparação com os sumos controlo. Em relação à avaliação da cor, no caso das amostras de sumos de framboesa, verificou-se que a adição do extrato de algas influenciou o parâmetro a*. No entanto, ambas as amostras permitiram a manutenção deste parâmetro, ao longo do tempo. Quanto ao parâmetro b*, as duas amostras de sumo com algas demonstraram uma evolução mais estável, em relação aos respetivos controlos. No que respeita ao teor em minerais, a adição do extrato da alga demonstrou aumentar os teores de magnésio, sódio, potássio e iodo. Em termos sensoriais, o sumo de framboesa com algas obteve maior aceitação por parte do painel de provadores, podendo-se constatar que o atributo da cor foi o que mostrou maiores variações, entre as duas amostras.
- Seasonal nutritional profile of Gelidium corneum (Rhodophyta, Gelidiaceae) from the center of PortugalPublication . Cavaco, Mário; Duarte, Adriana; Freitas, Marta; Afonso, Clélia; Bernardino, Susana M.; Pereira, Leonel; Martins, Mendelson; Mouga, TeresaGelidium corneum is a well-known agarophyte, harvested worldwide for its high agar quality. However, the species also exhibits an interesting nutritional profile, but with seasonal variations. Therefore, to evaluate the nutritional value of G. corneum, ash, crude protein, total lipids, and carbohydrates were analyzed at different times of the year. The heavy metals mercury, arsenic, lead, cadmium, and tin, as well as iodine were also measured. Finally, the seasonal antioxidant capacity of G. corneum extracts was evaluated. Our results indicate that the biomass is rich in protein (up to 16.25 ± 0.33%) and carbohydrates (up to 39.5 ± 3.29%), and low in lipids (up to 2.75 ± 0.28%), and especially in the summer, the AI, TI indexes, n-6/n-3 and h/H ratios (0.93, 0.6, 0.88 and 1.08, respectively) are very interesting. None of the contaminants exceeded the legally established limits, and the iodine values were adequate for a healthy diet. Finally, the antioxidant capacity is fair, with the DPPH ≤ 10.89 ± 1.46%, and ABTS ≤ 13.90 ± 1.54% inhibition, FRAP ≤ 0.91 ± 0.22 AAE.g−1, and TPC ≤ 6.82 ± 0.26 GAE.g−1. The results show that G. corneum is an attractive resource, with potential use as food or as a food supplement.
- A new biocatalyst: Penicillin G acylase immobilized in sol‐gel micro‐particles with magnetic propertiesPublication . Bernardino, Susana M. S. A.; Fernandes, Pedro; P. Fonseca, Luís; Bernardino, SusanaThe present work focuses on the development and basic characterization of a new magnetic biocatalyst, namely penicillin G acylase (PGA), immobilized in sol-gel matrices with magnetic properties, ultimately aimed for application in cephalexin (CEX) synthesis. A mechanically stable carrier, based on porous xerogels silica matrixes starting from tetramethoxysilane (TMOS), was prepared leading to micro-carriers with medium sized particles of 30 μm, as determined by scanning electron microscopy. An immobilization yield of 95-100% and a recovered activity of 50-65% at 37°C, as determined by penicillin G (PG) hydrolysis (pH STAT method), were observed. These results clearly exceed those reported in a previous work on PGA immobilization in sol-gel, where only 10% of activity was recovered. The values of activity were kept constant for 6 months. Immobilized PGA (682 U/gdry weight) retained high specific activity throughout ten consecutive runs for PG hydrolysis, suggesting adequate biocatalyst stability. The CEX synthesis was performed at 14°C, using the free and immobilized PGA in aqueous medium. Phenylglycine methyl ester was used as acyl donor at 90 mM and 7-aminodeacetoxycephalosporanic acid was the limiting substrate at 30 mM. The CEX stoichiometric yield after 1-h reaction was close to 68% (23 mM CEX/h) and 65% (19 mM CEX/h), respectively.
- Novel active food packaging films based on whey protein incorporated with seaweed extract: development, characterization, and application in fresh poultry meatPublication . Andrade, Mariana A.; Barbosa, Cássia H.; Souza, Victor G. L.; Coelhoso, Isabel M.; Reboleira, João; Bernardino, Susana; Ganhão, Rui; Mendes, Susana; Fernando, Ana Luisa; Vilarinho, Fernanda; Silva, Ana Sanches; Ramos, FernandoAlgae and seaweeds are used in cookery since the beginnings of human civilization, particularly in several Asian cultures. Phenolic compounds are secondary metabolites produced by aquatic and terrestrial plants for their natural defense against external stimuli, which possess powerful antimicrobial and antioxidant properties that can be very important for the food industry. The main objective of this study was to develop a whey protein concentrate active coating, incorporated with a Fucus vesiculosus extract in order to delay the lipid oxidation of chicken breasts. Ten hydroethanolic extracts from F. vesiculosus were obtained and their antioxidant capacity was evaluated through two antioxidant activity assays: the DPPH radical scavenging activity and -carotene leaching assay. The total content in phenolics compounds was also determined by Folin-Ciocalteu method. The chosen extract was the one obtained from the freeze-dried F. vesiculosus using 75% (v/v) ethanol as extraction solvent. The extract was successfully incorporated into a whey protein film and successfully strengthened the thickness, tensile strength, and elastic modulus. The active film also was able to inhibit the chicken breasts lipid oxidation for 25 days of storage.
- New applications for Gelidium corneum: Antimicrobial and antioxidant activity within a biorefinery conceptPublication . Mouga, Teresa; Cavaco, Mário; Duarte, Adriana; Correia, Ana Patrícia; Freitas, Marta V.; Bernardino, Susana; Afonso, CléliaThere are more than four hundred endogenous seaweeds that occur in Portugal, and red seaweeds (Rhodophyta) are the most abundant. Among these, Gelidium corneum, common in the Atlantic coast of Portugal, is currently being collected in the wild, in S. Martinho do Porto, exclusively for the extraction of agar. During the summer season this species is harvested from the bottom of the ocean, dried and sold, representing an important economic local activity. Yet, besides agar, this species is important for its bioactivities, namely for its antioxidant and antimicrobial capacities, thus most of the biomass is currently being disregarded as an important marine resource. Therefore, to further research the potential of G. corneum as a source of bioactive compounds, the biomass was seasonally harvested and characterized (protein, ash, moisture), and different extract were made (aqueous and ethanolic). These extracts were analyzed for the antioxidant potential (DDPH, ABTS, TPC and FRAP), and for antimicrobial capacity (against Escherichia coli, Bacillus subtilis, Candida albicans, Saccharomyces cerevisiae and Staphylococcus epidermidis). Preliminary results show that there are seasonal variations in the chemical composition of the biomass. Extracts tested also show interesting free radical scavenging capacity and antimicrobial activity against some of the strains tested. The attained results demonstrate that the biorefinery concept should be implemented to the agar industry, valuing the seaweeds’ biomass so to allow its use in other industries, such as textiles or cosmetics.
- Enhancement of the antioxidant and antimicrobial activities of porphyran through chemical modification with tyrosine derivativesPublication . Adão, Pedro; Reboleira, João; Teles, Marco; Santos, Beatriz; Ribeiro, Nádia; Teixeira, Carlos M.; Guedes, Mafalda; Pessoa, João Costa; Bernardino, Susana M.The chemical modification of porphyran hydrocolloid is attempted, with the objective of enhancing its antioxidant and antimicrobial activities. Sulfated galactan porphyran is obtained from commercial samples of the red algae Porphyra dioica using Soxhlet extraction with water at 100ºC and precipitation with isopropyl alcohol. The extracted porphyran is then treated with modified L-tyrosines in aqueous medium in the presence of NaOH, at ca. 70ºC. The modified tyrosines L1 and L2 are prepared through a Mannich reaction with either thymol or 2,4-di-tert-butylphenol, respectively. While the reaction with 2,4-di-tert-butylphenol yields the expected tyrosine derivative, a mixture of products is obtained with thymol. The resulting polysaccharides are structurally characterized and the respective antioxidant and antimicrobial activities are determined. Porphyran treated with the N-(2-hydroxy-3,5-di-tert-butyl-benzyl)-L-tyrosine derivative, POR-L2, presents a noticeable superior radical scavenging and antioxidant activity compared to native porphyran, POR. Furthermore, it exhibited some antimicrobial activity against S. aureus. The surface morphology of films prepared by casting with native and modified porphyrans is studied by SEM/EDS. Both POR and POR-L2 present potential applicability in the production of films and washable coatings for food packaging with improved protecting characteristics.
- Marine-Derived Polysaccharides and Their Potential Health Benefits in Nutraceutical ApplicationsPublication . Carrasqueira, Joana; Bernardino, Susana; Bernardino, Raul; Afonso, CléliaMarine-derived polysaccharides have sparked immense interest in the nutraceutical industry as they possess a wide range of bioactivities which are highlighted in this review. These include antioxidants, anti-inflammatory, anti-cancer, gut microbiota regulator, anti-diabetic, and anti-obesity. Algae, marine invertebrates, vertebrates, and microorganisms are the main sources of marine polysaccharides, such as alginate, fucoidan, laminarin, carrageenan, chitosan, glycosaminoglycans, and exopolysaccharides. The structure and functional groups of these compounds influence their bioactive properties. Moreover, the functional properties of polysaccharides, such as gelling, thickening, and stabilising capabilities, are also crucial in product development, where they can serve as gluten substitutes in bakery goods and stabilisers in icings, sauces, and yoghurts. The potential of commercial products under development, such as marine polysaccharide supplements, is discussed, along with already commercialised products in the nutraceutical market. This review emphasises the enormous potential of marine-derived polysaccharides as bioactive compounds with health benefits and commercial value.
- Promoção do Sucesso e Redução do Abandono Académico: Plano de AçãoPublication . Lázaro, Alexandra; Pereira, Ana Patrícia; Henriques, Carolina; Toneca, Cláudia; Amaro, Dina; Tavares, Dina; Rodrigues, Filipa; Seco, Graça; Marques, Gorete; Silva, Joana Carrilho da; Varela, Jorge; Brites, Leonel; Rosálio, Lina; Magueta, Lúcia; Filipe, Luís Pereira; Caetano, Marta; Frade, Patrícia; Vaz, Paula; Gonçalves, Pedro; Alves, Sandra; Mina, Sónia; Reis, Susana; Almeida, Susana; Bernardino, Susana; Fonseca, Susana; Ferreira, Tiago; Tavora, VítorDe forma de criar e potenciar, no Instituto Politécnico de Leiria, um trabalho colaborativo a partir da identificação e utilização de uma metodologia integrada de promoção do sucesso escolar, que permita identificar, no contexto académico, todas as dimensões do problema e um roteiro de intervenção futuro que inclua a identificação de estratégias para promoção do sucesso académico em Instituições de Ensino Superior nacionais e internacionais, foi desenvolvido o “Laboratório da Mudança”. Através de um método sistémico, participativo e criativo para enfrentar, gerir e mitigar problemas sociais complexos, como são o insucesso e abandono escolar, foi desenvolvido um trabalho em conjunto com os diferentes intervenientes da comunidade académica do Instituto Politécnico de Leiria, no qual foi utilizada a ferramenta Teoria da Mudança para a criação de um plano de ação face a problemas sociais complexos. No Laboratório da Mudança foram convidados a participar vários atores da comunidade académica do IP Leiria, desde direções e escolas, coordenações pedagógicas e demais serviços, bem como docentes e estudantes. A equipa multidisciplinar constituída para o desenvolvimento do Laboratório trouxe contributos variados relevantes e inovadores no desenho de uma estratégia colaborativa e na construção do presente plano de ação. Aquando da identificação do problema, e após o término do projeto, estes atores contribuirão na promoção de outras estratégias, além das propostas em desenvolvimento neste projeto, que possam ser concebidas no futuro visando a resolução dos problemas identificados do insucesso e abandono escolar.
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