Fucus vesiculosus-Rich Extracts as Potential Functional Food Ingredients: A Holistic Extraction Approach.

Brown macroalgae are rich sources of nutrients and health-promoting compounds. Nevertheless, their consumption is still limited by their strong organoleptic characteristics, thus requiring the development of extraction strategies to profit from their nutritional value. To fulfil this, two sequential extraction approaches were developed, differing in the solvent used in the first extraction step, water in approach 1 or food-grade ethanol in approach 2, to obtain economic and affordable extracts rich in specific compounds from Fucus vesiculosus. The use of water in the first step of extraction allowed us to recover water-soluble phlorotannins, laminarans and mannuronic-rich alginates, making the subsequent 70% ethanol extract richest in fucoxanthin (0.07% algae DW), and the hot water fractions purest in fucoidans and alginates with a lower mannuronic-to-guluronic (M/G) ratio (2.91). Conversely, when beginning extraction procedures with 96% ethanol, the recovered yields of phlorotannins increased (0.43 g PGE/100 g algae DW), but there was a concomitant seven-fold decrease in the recovery of fucoxanthin in the subsequent 70% ethanol extract. This approach also led to less pure hot water fractions containing fucoidans, laminarans and alginates with a higher M/G ratio (5.50). Overall, this work unveiled the potential of the first extraction steps in sustainable and holistic cascade strategies to modulate the composition of food-grade extracts, creating prospects of their application as tailored functional ingredients in food products.

Structure-function relationships of pectic polysaccharides from broccoli by-products with in vitro B lymphocyte stimulatory activity.

To study structure-function relationships of pectic polysaccharides with their immunostimulatory activity, broccoli by-products were used. Pectic polysaccharides composed by 64 mol% uronic acids, 18 mol% Ara, and 10 mol% Gal, obtained by hot water extraction, activated B lymphocytes in vitro (25-250 µg/mL). To disclose active structural features, combinations of ethanol and chromatographic fractionation and modification of the polysaccharides were performed. Polysaccharides insoluble in 80 % ethanol (Et80) showed higher immunostimulatory activity than the pristine mixture, which was independent of molecular weight range (12-400 kDa) and removal of terminal or short Ara side chains. Chemical sulfation did not promote B lymphocyte activation. However, the action of pectin methylesterase and endo-polygalacturonase on hot water extracted polysaccharides produced an acidic fraction with a high immunostimulatory activity. The de-esterified homogalacturonan region seem to be an important core to confer pectic polysaccharides immunostimulatory activity. Therefore, agri-food by-products are a source of pectic polysaccharide functional food ingredients.

Blanching impact on pigments, glucosinolates, and phenolics of dehydrated broccoli by-products.

Because of high water content, the valorisation of broccoli by-products requires dehydration that can preserve bioactive compounds. Blanching pre-treatment has been reported to improve the drying rate of broccoli. As a thermal treatment, it promotes also enzyme inactivation. Therefore, in this study, the impact of pre-dehydration blanching step, freeze-drying, air-drying at 40 °C, and microwave hydrodiffusion and gravity (MHG) dehydration on the levels of pigments, glucosinolates, and phenolics, was evaluated by UHPLC-DAD-ESI/MSn. When compared to freeze-drying, a technique known to preserve compounds, a pre-blanching step increased the extractability of both pigments and phenolics, while air-drying only retained 49% of the pigments and 70% of phenolics, both without affecting glucosinolates. However, when air-drying was preceded by blanching, less than 50% of compounds were retained. On the other hand, MHG dehydration increased the phenolics extractability by 26%, particularly that of kaempferol derivatives while also retaining the amount of the glucosinolate glucoraphanin, when compared to freeze-drying. Nevertheless, only 23% of indole glucosinolates were recovered and pigments were severely reduced, with lutein accounting only for 32% and only chlorophyll b was observed in trace amounts after MHG dehydration. Therefore, to valorise broccoli by-products as ingredients, different drying technologies may be used when targeting different composition richness: freeze-drying is suitable for pigments and glucosinolates, air-drying is suitable for glucosinolates, while MHG promotes the extractability of phenolic compounds.

Salt pan brine water as a sustainable source of sulphated polysaccharides with immunostimulatory activity.

Marine environments are an enormous source of materials with biological interest, such as sulphated polysaccharides, which have relevant biological activities. In this study the potential of salt pan brine water as an easily accessible source of sulphated polysaccharides was evaluated. This water revealed to have a high quantity of polymeric material, five times more than sea water, mainly composed by highly sulphated polysaccharides. Structural analysis identified a diversity of polysaccharides, namely rhamnans, fucans, mannans, xylomannans, glucuronomannans, galactans, and glucans. All these structures seem to form complexes that are resistant to the salt pan conditions along salt production. These polysaccharides showed in vitro stimulatory activity for B cells, suggesting their potential application in nutraceutical and biomedical fields. Salt pan brine water is a valuable source of environmentally friendly and low-cost available bioactive compounds prone to be exploited.

Structural polymeric features that contribute to in vitro immunostimulatory activity of instant coffee.

An instant coffee fraction, rich in arabinogalactans, obtained by ultrafiltration, using 1 and 5kDa membranes, has previously shown in vitro stimulatory activity on BALB/c mice spleen B lymphocytes. The compounds inducing lymphocytic activation were shown to elute, mainly near the void volume by size-exclusion chromatography, using Bio-Gel P6 (1-6kDa). Treatment of the compounds with chymotrypsin, a digestive protease, did not affect the induced B lymphocyte activation. On the contrary, hydrolysis with an α-l-arabinofuranosidase, removing single terminally-linked arabinose residues, decreased the extent of B cell activation. The immunostimulatory activity of these compounds was also observed by in vitro experiments, using bone marrow-derived macrophages and dendritic cells as responders. Altogether, these results show the relevance of single arabinose residues, present at the non-reducing end of polymeric compounds, to the coffee stimulatory activity in cells mediating innate and acquired immunity.

Structure-function relationships of immunostimulatory polysaccharides: A review.

Immunostimulatory polysaccharides are compounds capable of interacting with the immune system and enhance specific mechanisms of the host response. Glucans, mannans, pectic polysaccharides, arabinogalactans, fucoidans, galactans, hyaluronans, fructans, and xylans are polysaccharides with reported immunostimulatory activity. The structural features that have been related with such activity are the monosaccharide and glycosidic-linkage composition, conformation, molecular weight, functional groups, and branching characteristics. However, the establishment of structure-function relationships is possible only if purified and characterized polysaccharides are used and selective structural modifications performed. Aiming at contributing to the definition of the structure-function relationships necessary to design immunostimulatory polysaccharides with potential for preventive or therapeutical purposes or to be recognized as health-improving ingredients in functional foods, this review introduces basic immunological concepts required to understand the mechanisms that rule the potential claimed immunostimulatory activity of polysaccharides and critically presents a literature survey on the structural features of the polysaccharides and reported immunostimulatory activity.

Influence of molecular weight on in vitro immunostimulatory properties of instant coffee.

Instant coffee was prepared and fractionated into higher (>100kDa), medium (5-10, 10-30, 30-100kDa) and lower (1-5, <1kDa) molecular weight fractions. Sugars and linkage composition characteristics of arabinogalactans and galactomannans were recovered in all fractions. Also, amino acid analysis performed after hydrolysis showed similar compositions in all fractions. On the contrary, free chlorogenic acids and caffeine were only detected in the lowest molecular weight fraction (<1kDa). A direct relationship between the melanoidins browning index and the molecular weight was observed. The fractions obtained were incubated in vitro with murine spleen lymphocytes in order to evaluate their possible immunostimulatory abilities. The surface expression of CD69 (early activation marker) on different lymphocyte sub-populations showed that the fraction with 1-5kDa was able to induce activation of B-lymphocytes. This was the only fraction to induce B-lymphocyte activation, since all the other fractions failed, even when higher concentrations were used.