Light Modulation for Bioactive Pigment Production in Synechocystis salina.

Cyanobacteria are microorganisms that are well-adapted to sudden changes in their environment, namely to light conditions. This has allowed them to develop mechanisms for photoprotection, which encompass alteration in pigment composition. Therefore, light modulation appears to be a suitable strategy to enhance the synthesis of specific pigments (e.g., phycocyanin) with commercial interest, in addition to conveying a more fundamental perspective on the mechanisms of acclimatization of cyanobacterium species. In this study, Synechocystis salina was accordingly cultivated in two light phase stages: (i) white LED, and (ii) shift to distinct light treatments, including white, green, and red LEDs. The type of LED lighting was combined with two intensities (50 and 150 µmolphotons·m-2·s-1). The effects on biomass production, photosynthetic efficiency, chlorophyll a (chl a) content, total carotenoids (and profile thereof), and phycobiliproteins (including phycocyanin, allophycocyanin, and phycoerythrin) were assessed. White light (under high intensity) led to higher biomass production, growth, and productivity; this is consistent with higher photosynthetic efficiency. However, chl a underwent a deeper impact under green light (high intensity); total carotenoids were influenced by white light (high intensity); whilst red treatment had a higher effect upon total and individual phycobiliproteins. Enhanced PC productivities were found under modulation with red light (low intensities), and could be achieved 7 days earlier than in white LED (over 22 days); this finding is quite interesting from a sustainability and economic point of view. Light modulation accordingly appears to be a useful tool for supplementary studies pertaining to optimization of pigment production with biotechnological interest.

Gloeothece sp. as a Nutraceutical Source-An Improved Method of Extraction of Carotenoids and Fatty Acids.

The nutraceutical potential of microalgae boomed with the exploitation of new species and sustainable extraction systems of bioactive compounds. Thus, a laboratory-made continuous pressurized solvent extraction system (CPSE) was built to optimize the extraction of antioxidant compounds, such as carotenoids and PUFA, from a scarcely studied prokaryotic microalga, Gloeothece sp. Following "green chemical principles" and using a GRAS solvent (ethanol), biomass amount, solvent flow-rate/pressure, temperature and solvent volume-including solvent recirculation-were sequentially optimized, with the carotenoids and PUFA content and antioxidant capacity being the objective functions. Gloeothece sp. bioactive compounds were best extracted at 60 °C and 180 bar. Recirculation of solvent in several cycles (C) led to an 11-fold extraction increase of ß-carotene (3C) and 7.4-fold extraction of C18:2 n6 t (5C) when compared to operation in open systems. To fully validate results CPSE, this system was compared to a conventional extraction method, ultrasound assisted extraction (UAE). CPSE proved superior in extraction yield, increasing total carotenoids extraction up 3-fold and total PUFA extraction by ca. 1.5-fold, with particular extraction increase of 18:3 n3 by 9.6-fold. Thus, CPSE proved to be an efficient and greener extraction method to obtain bioactive extract from Gloeothece sp. for nutraceutical purposes-with low levels of resources spent, while lowering costs of production and environmental impacts.

Design of whey protein nanostructures for incorporation and release of nutraceutical compounds in food.

Whey proteins are widely used as nutritional and functional ingredients in formulated foods because they are relatively inexpensive, generally recognized as safe (GRAS) ingredient, and possess important biological, physical, and chemical functionalities. Denaturation and aggregation behavior of these proteins is of particular relevance toward manufacture of novel nanostructures with a number of potential uses. When these processes are properly engineered and controlled, whey proteins may be formed into nanohydrogels, nanofibrils, or nanotubes and be used as carrier of bioactive compounds. This review intends to discuss the latest understandings of nanoscale phenomena of whey protein denaturation and aggregation that may contribute for the design of protein nanostructures. Whey protein aggregation and gelation pathways under different processing and environmental conditions such as microwave heating, high voltage, and moderate electrical fields, high pressure, temperature, pH, and ionic strength were critically assessed. Moreover, several potential applications of nanohydrogels, nanofibrils, and nanotubes for controlled release of nutraceutical compounds (e.g. probiotics, vitamins, antioxidants, and peptides) were also included. Controlling the size of protein networks at nanoscale through application of different processing and environmental conditions can open perspectives for development of nanostructures with new or improved functionalities for incorporation and release of nutraceuticals in food matrices.

Evaluation of the antioxidant activity of cell extracts from microalgae.

A growing market for novel antioxidants obtained from non-expensive sources justifies educated screening of microalgae for their potential antioxidant features. Characterization of the antioxidant profile of 18 species of cyanobacteria (prokaryotic microalgae) and 23 species of (eukaryotic) microalgae is accordingly reported in this paper. The total antioxidant capacity, accounted for by both water- and lipid-soluble antioxidants, was evaluated by the (radical cation) ABTS method. For complementary characterization of cell extracts, a deoxyribose assay was carried out, as well as a bacteriophage P22/Salmonella-mediated approach. The microalga Scenedesmus obliquus strain M2-1 exhibited the highest (p > 0.05) total antioxidant capacity (149 ± 47 AAU) of intracellular extracts. Its scavenger activity correlated well with its protective effects against DNA oxidative damage induced by copper(II)-ascorbic acid; and against decay in bacteriophage infection capacity induced by H2O2. Finally, performance of an Ames test revealed no mutagenic effects of the said extract.

In vitro screening for anti-microbial activity of chitosans and chitooligosaccharides, aiming at potential uses in functional textiles.

Antimicrobial finishing of textiles has been found to be an economical way to prevent (or treat) skin disorders. Hence, this research effort was aimed at elucidating the relationship between molecular weight (MW) of chitosan and its antimicrobial activity upon six dermal reference microorganisms, as well as the influence of the interactions with cotton fabrics on said activity. Using 3 chitosans with different MW, as well as two chitooligosaccharide (COS) mixtures, a relevant antimicrobial effect was observed by 24 h for the six microorganisms tested; it was apparent that the antimicrobial effect is strongly dependent on the type of target microorganism and on the MW of chitosan being higher for lower MW in the case of E. coli, K. pneumoniae and P. aeruginosa, and the reverse in the case of both Gram-positive bacteria. Furthermore, a strong anti-fungal effect was detectable upon C. albicans, resembling the action over Gram-positive bacteria. Interactions with cotton fabric resulted in a loss of COS activity when compared with cultured media, relative to the effect over Gram-negative bacteria. However, no significant differences for the efficacy of all the 5 compounds were observed by 4 h. The three chitosans possessed a higher antimicrobial activity when impregnated onto the fabric, and presented a similar effect on both Gram-positive bacteria and yeast, in either matrix. Pseudomonas aeruginosa showed to be the most resistant microorganism to all five compounds.

Antioxidant activity of sugar molasses, including protective effect against DNA oxidative damage.

Extracts were obtained from molasses, a byproduct of the sugar industry, via a number of chromatographic steps. Their antioxidant capacity was studied, including the inhibitory effect upon DNA oxidative damage; the phenolic compound profile thereof was ascertained as well. Two extracts exhibited significant antioxidant features, expressed by their capacity to decolorize ABTS radical cation and to scavenge hydroxyl free radicals (via deoxyribose assay). Those 2 extracts also brought about protection against induced DNA oxidative damage (via decreasing DNA scission, as assessed by electrophoresis). The phenolic compounds syringic acid, p-hydroxybenzoic acid, vanillic acid, p-hydroxybenzaldehyde, and ferulic acid were positively identified and quantified.

Infusions of Portuguese medicinal plants: Dependence of final antioxidant capacity and phenol content on extraction features.

BACKGROUND: Aqueous extracts of most medicinal plants traditionally employed in Portugal (at the ratio of 1 g plant: 110 mL water) have been assayed for total antioxidant capacity and phenol content, in order to elucidate their claimed medicinal features. RESULTS: The antioxidant activity was assessed by the ABTS(•+) method; the ascorbic acid equivalent values ranged from 1.4280 ± 0.1261 g L(-1) for avocado (Persea americana (Lauraceae)) obtained by infusion of powder, down to 0.0027 ± 0.0012 g L(-1) for olive (Olea europaea (Oleaceae)) obtained by infusion of leaves. Total phenol content was determined by the Folin-Ciocalteu procedure; the gallic acid equivalent values ranged from 0.5541 ± 0.0289 g L(-1) for avocado obtained by infusion of powder, down to 0.0053 ± 0.0014 g L(-1) for olive obtained by boiling leaves. A good correlation between total antioxidant capacity and total phenol content was found. CONCLUSION: The method of powder infusion should be chosen if high concentration of antioxidants are sought. On the other hand, a high antioxidant capacity and a high phenol content correlate well with the empirically established (and widely publicised) capacity to treat respiratory infections. Copyright © 2007 Society of Chemical Industry.

Preparation of fatty acid methyl esters for gas-chromatographic analysis of marine lipids: insight studies.

Assays for fatty acid composition in biological materials are commonly carried out by gas chromatography, after conversion of the lipid material into the corresponding methyl esters (FAME) via suitable derivatization reactions. Quantitative derivatization depends on the type of catalyst and processing conditions employed, as well as the solubility of said sample in the reaction medium. Most literature pertinent to derivatization has focused on differential comparison between alternative methods; although useful to find out the best method for a particular sample, additional studies on factors that may affect each step of FAME preparation are urged. In this work, the influence of various parameters in each step of derivatization reactions was studied, using both cod liver oil and microalgal biomass as model systems. The accuracies of said methodologies were tested via comparison with the AOCS standard method, whereas their reproducibility was assessed by analysis of variance of (replicated) data. Alkaline catalysts generated lower levels of long-chain unsaturated FAME than acidic ones. Among these, acetyl chloride and BF(3) were statistically equivalent to each other. The standard method, which involves alkaline treatment of samples before acidic methylation with BF(3), provided equivalent results when compared with acidic methylation with BF(3) alone. Polarity of the reaction medium was found to be of the utmost importance in the process: intermediate values of polarity [e.g., obtained by a 1:1 (v/v) mixture of methanol with diethyl ether or toluene] provided amounts of extracted polyunsaturated fatty acids statistically higher than those obtained via the standard method.

Modeling the kinetics of whey protein hydrolysis brought about by enzymes from Cynara cardunculus.

The purpose of this research work was to study the proteolytic activity of aqueous crude extracts of flowers of the plant Cynara cardunculus on the major whey proteins, namely, beta-lactoglobulin (beta-Lg) and alpha-lactalbumin (alpha-La). These extracts, containing a mixture of cardosins A and B (i.e., two distinct aspartic proteases), have been employed for many years in traditional cheese-making in Portugal and Spain. Cow's milk sweet whey was incubated for up to 24 h at various ratios of addition of crude enzyme extract, under controlled pH (5.2 and 6.0) and temperature (55 degrees C). The samples collected were assayed by gel permeation chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A mechanistic model was proposed for the kinetics of the hydrolysis process, which is basically a double-substrate, double-enzyme Michaelis-Menten rate expression; the kinetic parameters were estimated by multiresponse, nonlinear regression analysis. The best estimates obtained for the specificity ratio (i.e., k(cat)/K(m)) of each cardosin within the mixture toward each whey protein indicated that said aspartic proteases possess a higher catalytic efficiency for alpha-La (0.42-4.2 mM(-1).s(-1)) than for beta-Lg (0-0.064 mM(-1).s(-1)), at least under the experimental conditions used. These ratios are below those previously reported for caseins and a synthetic hexapeptide. Cardosins are more active at pH 5.2 than at pH 6.0 and (as expected) at higher enzyme-to-substrate ratios.