Dina Rodrigues

Marine biotechnology advances towards applications in new functional foods

The marine ecosystem is still an untapped reservoir of biologically active compounds, which have considerable potential to supply food ingredients towards development of new functional foods. With the goal of increasing the availability and chemical diversity of functional marine ingredients, much research has been developed using biotechnological tools to discover and produce new compounds. This review summarizes the advances in biotechnological tools for production of functional ingredients, including enzymes, for the food industry. Tools involving biotechnological processes (bioreactors, fermentations, bioprocessing) and those involving genetic research designated as molecular biotechnology are discussed highlighting how they can be used in the controlled manipulation and utilization of marine organisms as sources of food ingredients, as well as discussing the most relevant shortcomings towards applications in new functional foods.

Chemical composition of red, brown and green macroalgae from Buarcos bay in Central West Coast of Portugal.

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.

Impact of enzyme- and ultrasound-assisted extraction methods on biological properties of red, brown, and green seaweeds from the central west coast of Portugal.

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/glyoph extract) and total phenolics (261 ± 37 μgcatechol equiv/glyoph extract) contents, whereas higher sugars (78 ± 14 mgglucose equiv/glyoph extract) including sulfated polysaccharide (44 ± 8 mgNa2SO4 acid/glyoph 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 α-glucosidase (38-49%).

Metabolic Profiling of Potential Probiotic or Synbiotic Cheeses by Nuclear Magnetic Resonance (NMR) Spectroscopy

To assess ripening of potential probiotic cheeses (containing either Lactobacillus casei -01 or Bifidobacterium lactis B94) or synbiotic cheeses with fructooligosaccharides (FOS) or a 50:50 mix of FOS/inulin, metabolic profiles have been obtained via classical biochemical analyses and by NMR spectroscopy. The addition of prebiotics to the cheeses resulted in lower proteolysis indices, especially in those synbiotic cheeses inoculated with B. lactis B94. Among synbiotic cheeses the combination of FOS and inulin resulted in an increase in lipolytic activity. The metabolic profiles of the cheeses analyzed by NMR spectroscopy, combined with multivariate statistics, allowed profiles to be distinguished by maturation time, added probiotic bacteria, or, in the case of B. lactis B94 cheese, added prebiotic. The NMR results are in agreement with the biochemical analyses and demonstrate the potential of NMR for the study of metabolic processes in probiotic/synbiotic food matrices.

Toxicity of organic and inorganic nanoparticles to four species of white-rot fungi

The rapid development of nanoparticles (NP) for industrial applications and large-volume manufacturing, with its subsequent release into the environment, raised the need to understand and characterize the potential effects of NP to biota. Accordingly, this work aimed to assess sublethal effects of five NP to the white-rot fungi species Trametes versicolor, Lentinus sajor caju, Pleurotus ostreatus, and Phanerochaete chrysosporium. Each species was exposed to serial dilutions of the following NP: organic-vesicles of SDS/DDAB and of Mo/NaO; gold-NP, quantum dot CdSe/ZnS, and Fe/Co. Fungi growth rate was monitored every day, and at the end of assay the mycelium from each replicate was collected to evaluate possible changes in its chemical composition. For all NP-suspensions the following parameters were characterized: hydrodynamic diameter, surface charge, aggregation index, zeta potential, and conductivity. All tested NP tended to aggregate when suspended in aqueous media. The obtained results showed that gold-NP, CdSe/ZnS, Mo/NaO, and SDS/DDAB significantly inhibited the growth of fungi with effects on the mycelium chemical composition. Among the tested NP, gold-NP and CdSe/ZnS were the ones exerting a higher effect on the four fungi. Finally to our knowledge, this is the first study reporting that different types of NP induce changes in the chemical composition of fungi mycelium.

Encapsulation of probiotic strains in plain or cysteine‐supplemented alginate improves viability at storage below freezing temperatures

Four probiotic bacteria (Lactobacillus paracasei L26, L. casei‐01, L. acidophilus Ki, and Bifidobacterium animalis BB‐12®) were encapsulated in plain alginate or alginate supplemented with L‐cysteine·HCl, and resulting microcapsules were stored at different temperatures, namely 21, 4, −20, or −80°C for a period of up to 6 months. The results showed that the encapsulation in calcium alginate microcapsules was only effective in promoting protection at freezing temperatures, independently of the sensitivity of the strain. Storage of calcium alginate microcapsules at −80°C indicated a protective effect upon viability of all four probiotic strains and the presence of L‐cysteine·HCl in the alginate matrix improved protection upon cell viability of B. animalis BB‐12®. An increase in storage temperature of encapsulated bacteria caused an increase in rate of loss in their viability that was strain dependent. This study suggests that microencapsulation of probiotic cells in calcium alginate can be suitable for sustaining the viability of probiotics in food products that require storage below freezing temperatures, even in the absence of cryoprotectors, contributing to an increased shelf life.

Strategies based on silica monoliths for removing pollutants from wastewater effluents: a review.

Silica monoliths have been used for more than half a century in a wide variety of applications, such as stationary phases for microextraction fibers, capillary columns for chromatography, in the encapsulation of biomolecules for affinity chromatography and for microfluidic or microarray chips and, more recently, and less well known for wastewater treatment. The main objective of this review article is to specifically overview the strategies that use silica monoliths for the removal of chemical pollutants from wastewater effluents or prepared solutions. The discussion of advantages and drawbacks of such strategies will be supported with the main studies carried out so far which have been performed in laboratory environment only. The application and potential research interest in several strategies using composites and biocomposites based silica monoliths as cleaning systems are also discussed.

Chemical and structural characterization of Pholiota nameko extracts with biological properties

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.

Evaluation of tertiary treatment by fungi, enzymatic and photo-Fenton oxidation on the removal of phenols from a kraft pulp mill effluent: a comparative study.

Pulp and paper mills generate pollutants associated to their effluents depending upon the type of process, type of the wood materials, process technology applied, management practices, internal recirculation of the effluent for recovery, the amount of water used in the industrial process and type of secondary treatment. This study is the first that reports a simultaneous evaluation of the effects of tertiary treatments by fungi (Rhizopus oryzae and Pleurotus sajor caju), by enzyme (laccase) and by an oxidation process (photo-Fenton) on individual phenols (vanillin, guaiacol, phloroglucinol, vanillic acid and syringic acid) of a Eucalyptus globulus bleached kraft pulp and paper mill final effluent after secondary treatment (BKPME). The tertiary treatments were applied on BKPME samples and in BKPME samples supplemented with extra concentration of each phenol. Tertiary treatments by Rhizopus oryzae and photo-Fenton oxidation were able of complete removal (100%) of phenols on BKPME samples whereas P. sajor caju and laccase were able of 60–85% removal. On BKPME samples with added concentration of each phenol, photo-Fenton was the only treatment capable of total phenols removal (100%), which suggests a great potential for its application.

Marine Functional Foods

In line with consumer awareness that a relationship between diet, health, and disease prevention exists, the research and development of new functional foods have been the target of many studies over the last years. The increasing ageing of populations, the decrease in quality of life due to stress, the high incidence of so-called modern diseases (cardiovascular disease, obesity, cancer, diabetes, and allergies) represent the driving forces in the quest for different foods and diets to promote healthy active ageing, improve well-being, and counteract the incidence of many diseases. The marine environment provides a huge source of many healthy foods, including seaweeds, which is an example of a marine product that has been part of the diet in several countries around the world. Additionally, marine specimens are also sources of a plethora of chemicals, many of them with biological properties and, therefore, called bioactive compounds. These chemicals can be extracted and incorporated in several food matrices leading to new potential functional foods. This chapter is a review summarizing those marine organisms with healthy properties that are consumed all over the world, as well as the potential of using marine organisms as sources of food ingredients towards applications in new functional foods. Recent findings on functional foods based on marine-derived ingredients are described and discussed.