Francisca Acevedo

Extracts of Maqui (Aristotelia chilensis) and Murta (Ugni molinae Turcz.): Sources of Antioxidant Compounds and α-Glucosidase/α-Amylase Inhibitors

The objective of this work was to evaluate the antioxidant and antihemolytic activities of crude, aqueous, and organic-aqueous extracts of maqui ( Aristotelia chilensis ) and murta ( Ugni molinae Turcz.), together with their inhibiting effect on enzymes involved in the metabolism of carbohydrates. Radical scavenging activity, inhibition of linoleic acid oxidation in a micellar system, antihemolytic activity, and inhibition of α-amylases and α-glucosidases were analyzed. Crude extracts of maqui leaves and fruits were found to be important sources of polyphenolic compounds, showing 69.0 ± 0.9 and 45.7 ± 1.1 mg GAE/g dm, respectively. Polyphenols from maqui leaves were active as antioxidants and antihemolytic compounds (p < 0.05), showing a noncompetitive inhibiting effect on α-glucosidase. Flavan-3-ol polymers and glycosylated flavonols, such as quercetin glucoside and kaempferol glucoside, were tentatively identified in extracts. This preliminary observation provides the basis for further examination of the suitability of polyphenol-enriched extracts from maqui and murta as nutritional or medicinal supplements with potential human health benefits.

Enzymes as useful tools for environmental purposes.

In the environment enzymes may play important and different roles at least in three cases: as main agents (as isolated, cell-bound or immobilized enzymes) in charge of either the transformation and/or degradation of compounds polluting the environment and the restoration of the polluted environment; as reliable and sensitive tools to detect and measure the amount and concentration of pollutants before, during and after the restoration process; as reliable, easy and sensitive indicators of quality and health status of the environment subjected to the restoration process. To our knowledge papers or reviews integrating findings on these three functions of enzymes are missing in literature. Therefore the main scope of the present paper is to briefly encompass general and specific concepts about roles of enzymes as decontaminating agents, pollutant assaying agents and indicators of environment safety. Examples chosen among those published very recently, supporting and confirming peculiarities, features, and performance of enzymatic agents will be illustrated.

Degradation of polycyclic aromatic hydrocarbons by free and nanoclay-immobilized manganese peroxidase from Anthracophyllum discolor

Manganese peroxidase (MnP) produced by Anthracophyllum discolor, a Chilean white rot fungus, was immobilized on nanoclay obtained from volcanic soil and its ability to degrade polycyclic aromatic hydrocarbons (PAHs) compared with the free enzyme was evaluated. At the same time, nanoclay characterization was performed. Nanoclay characterization by transmission electronic microscopy showed a particle average size smaller than 100 nm. The isoelectric points (IEP) of nanoclay and MnP from A. discolor were 7.0 and 3.7, respectively, as determined by micro electrophoresis migration and preparative isoelectric focusing. Results indicated that 75% of the enzyme was immobilized on the nanoclay through physical adsorption. As compared to the free enzyme, immobilized MnP from A. discolor achieved an improved stability to temperature and pH. The activation energy (Ea) value for immobilized MnP (51.9 kJ mol(-1)) was higher than that of the free MnP (34.4 kJ mol(-1)). The immobilized enzyme was able to degrade pyrene (>86%), anthracene (>65%), alone or in mixture, and to a less extent fluoranthene (<15.2%) and phenanthrene (<8.6%). Compared to free MnP from A. discolor, the enzyme immobilized on nanoclay enhanced the enzymatic transformation of anthracene in soil. Overall results indicate that nanoclay, a carrier of natural origin, is a suitable support material for MnP immobilization. In addition, immobilized MnP shows an increased stability to high temperature, pH and time storage, as well as an enhanced PAHs degradation efficiency in soil. All these characteristics may suggest the possible use of nanoclay-immobilized MnP from A. discolor as a valuable option for in situ bioremediation purposes.

Crop seed oil bodies: from challenges in protein identification to an emerging picture of the oil body proteome

Oleaginous seeds store lipids in specialized structures called oil bodies (OBs). These organelles consist of a core of neutral lipids bound by proteins embedded in a phospholipid monolayer. OB proteins are well conserved in plants and have long been grouped into only two categories: structural proteins or enzymes. Recent work, however, which identified other classes of proteins associated with OBs, clearly shows that this classification is obsolete. Proteomics‐mediated OB protein identification is facilitated in plants for which the genome is sequenced and annotated. However, it is not clear whether this knowledge can be dependably transposed to less well‐characterized plants, including the well‐established commercial sources of seed oil as well as the many others being proposed as novel sources for biodiesel, especially in Africa and Asia. Toward an update of the current data available on OB proteins this review discusses (i) the specific difficulties for proteomic studies of organelles; (ii) a 2012 census of the proteins found in seed OBs from various crops; (iii) the oleosin composition of OBs and their role in organelle stability; (iv) PTM of OB proteins as an emerging field of investigation; and finally we describe the emerging model of the OB proteome from oilseed crops.

High carotenoid bioaccessibility through linseed oil nanoemulsions with enhanced physical and oxidative stability

Carotenoid (astaxanthin or lycopene) emulsions obtained by high pressure homogenization were investigated for their physical, oxidative and storage stability and biological fate on an in vitro digestion model of bioaccessibility. Emulsion stability evaluated at various processing environments (20-50°C, 2-10 pH, 0-500 mM NaCl, and 0-35 days storage at 25°C) depended on carotenoid and homogenization pressures (5, 10, 100 MPa). Trolox increased the oxidative stability of nanoemulsions (100 MPa) and acted synergistically with BHT in increasing the stability of lycopene nanoemulsion. Intestinal digestibility depended on homogenization pressures with the fastest release and lower amount of free fatty acids observed at 100 MPa. Carotenoid nanoemulsions (100 MPa) were partially (66%) digested and highly bioaccessible (>70%). Therefore, nanoemulsions provide an effective and stable system for efficient astaxanthin or lycopene delivery and bioavailability in foods, beverages, nutraceuticals and/or other agriproducts.

Oil bodies as a potential microencapsulation carrier for astaxanthin stabilisation and safe delivery

Abstract Astaxanthin (AST) is a valued molecule because of its high antioxidant properties. However, AST is extremely sensitive to oxidation, causing the loss of its bioactive properties. The purposes of this study were to define conditions for microencapsulating AST in oil bodies (OB) from Brassica napus to enhance its oxidative stability, and to test the bioactivity of the microencapsulated AST (AST-M) in cells. Conditions for maximising microencapsulation efficiency (ME) were determined using the Response Surface Methodology, obtaining a high ME (>99%). OB loaded with AST showed a strong electrostatic repulsion in a wide range of pH and ionic strengths. It was found that AST-M exposed to air and light was more stable than free AST. In addition, the protective effect of AST against intracellular ROS production was positively influenced by microencapsulation in OB. These results suggest that OB offer a novel option for stabilising and delivering AST.

Bacterial polyhydroxybutyrate for electrospun fiber production

Nano- and microfibers obtained by electrospinning have attracted great attention due to its versatility and potential for applications in diverse technological fields. Polyhydroxyalkanoates (PHAs) are biopolymers synthesized by microorganisms such as the bacterium Burkholderia xenovorans LB400. In particular, LB400 cells are capable to synthesize poly(3-hydroxybutyrate) (PHB) from glucose. The aim of this study was to produce and characterize electrospun fibers obtained from bacterial PHBs. Bacterial strain LB400 was grown in M9 minimal medium using xylose and mannitol (10gL-1) as the sole carbon sources and NH4Cl (1gL-1) as the sole nitrogen source. Biopolymer-based films obtained were used to produce fibers by electrospinning. Diameter and morphology of the microfibers were analyzed by scanning electron microscopy (SEM) and their thermogravimetric properties were investigated. Bead-free fibers using both PHBs were obtained with diameters of less than 3μm. The surface morphology of the microfibers based on PHBs obtained from both carbon sources was different, even though their thermogravimetric properties are similar. The results indicate that the carbon source may determine the fiber structure and properties. Further studies should be performed to analyze the physicochemical and mechanical properties of these PHB-based microfibers, which may open up novel applications.

Seed Oil Bodies from Gevuina avellana and Madia sativa

In this study, oil bodies (OBs) from Gevuina avellana (OBs-G) and Madia sativa (OBs-M) were isolated and characterized. Microscopic inspection revealed that the monolayer on OB-G was thinner compared to that on OB-M. Cytometric profiles regarding size, complexity, and staining for the two OB sources were similar. Fatty acid to protein mass ratio in both OBs was near 29, indicating high lipid enrichment. OBs-G and OBs-M showed a strong electrostatic repulsion over wide ranges of pH (5.5-9.5) and NaCl concentration (0-150 mM). Proteins displaying highly conserved sequences (steroleosins and aquaporins) in the plant kingdom were identified. The presence of oleosins was immunologically revealed using antibodies raised against Arabidopsis thaliana oleosins. OBs-G and OBs-M exhibited no significant cytotoxicity against the cells. This is the first report about the isolation and characterization of OBs-G and OBs-M, and this knowledge could be used for novel applications of these raw materials.

In vitro activity on human gut bacteria of murta leaf extracts (Ugni molinae turcz.), a native plant from Southern Chile.

Despite the fact that murta infusions have been used to treat gut/urinary infections by native Chileans for centuries, the mechanisms promoting such effects still remain unclear. As a first attempt to unravel these mechanisms, human fecal samples were incubated in a medium containing water extract of murta leaves (ML) and the growth of different bacterial groups was evaluated. Control incubations were made in media containing fructooligosaccharides (FOS) and glucose as a carbon source. Phenolic compounds in the ML extract, likely promoters of bioactivity, were identified by HPLC-DAD-MS(n) . Concentrations (log₁₀ CFU/mL) of bifidobacteria and lactobacilli in media containing the extract and FOS were 7.33 ± 0.05/4.95 ± 0.20 and 6.44 ± 0.22/6.05 ± 0.06, respectively. Clostridia, anaerobes and Enterobacteriaceae grew to a similar extent in media containing murta extract and FOS. In vitro tests (disk diffusion) showed that Gram-positive (Bacillus and Paenibacillaceae) and Gram-negative (Enterobacteriaceae) bacteria isolated from fecal samples were sensitive to both water and 50/50 ethanol/water extracts of ML (28.4 μg gallic acid equivalents). At this concentration, the antimicrobial activity of ML extracts was significantly (P < 0.05) lower than that of penicillin (10 U), whereas the difference between activity of ML extracts and gentamicine (10 μg) was no significant (P > 0.05). No evidence of dependency between the antimicrobial activity of ML extracts and the enzymatic capability of the sensitive strains was found.

A practical culture technique for enhanced production of manganese peroxidase by Anthracophyllum discolor Sp4

In this study, different growth conditions of Anthracophyllum discolor Sp4 including the effect of agitation, additions of lignocellulosic support, inducer and surfactant were evaluated on the MnP production in Kirk medium using a culture system made up of the tubes containing the glass bead . The highest MnP production (1,354 U/L on day 13) was obtained when the medium was supplemented with wheat grain and 0.25 mM MnSO4 as inducer, under static conditions at 30°C. Two isoenzymes were purified (35 and 38 kDa respectively). MnP presented a maximal activity in the pH range between 4.5 and 5.5, a relatively high temperature tolerance (50oC) and a high catalytic activity for 2,6-dimethoxyphenol and hydrogen peroxide.