RESUMEN
BACKGROUND: Foods contain lipids that are easily susceptible to oxidation, which can modify their sensory properties. Although these compounds provide characteristic flavours and odours, there are also unwanted compounds, such as volatile secondary oxidation products, representing a recurring problem for both the industry and consumers. Synthetic antioxidants are often employed to prevent this but their chronic consumption can be detrimental to human health. The present study evaluates the antioxidant potential of ethanolic extracts from Ganoderma resinaceum and Phlebopus bruchii using an accelerated oxidation test. RESULTS: The composition profile of the extracts was investigated, identifying the presence of tryptophan, quinic acid, caffeic acid and 3,4-dihydroxyphenylglycol-phenolic acid. The antioxidant capacity of the extracts was compared with that of butylated hydroxytoluene (BHT) in sunflower oil that was oven-heated at 60 °C. Chemical (peroxide value, p-anisidine value and conjugated dienes) and volatile (2-octenal, 2-heptenal and 2,4-decadienal) indicators were measured over 28 days. The peroxide value decreased for both extracts at a similar level to that of BHT 0.02% w/w, and conjugate dienes decreased in the presence of G. resinaceum 0.1% w/w. Meanwhile, p-anisidine exhibited a slightly greater decrease for P. bruchii 0.1% w/w than for BHT. The sample with 0.1% w/w of extracts showed a reduction in volatile secondary oxidation compounds, indicating significant antioxidant activity. CONCLUSION: Based on these results, both extracts could be proposed as potential antioxidants in foods with a high lipid content. © 2024 Society of Chemical Industry.
Asunto(s)
Antioxidantes , Ganoderma , Oxidación-Reducción , Antioxidantes/química , Ganoderma/química , Peroxidación de Lípido/efectos de los fármacos , Lípidos/químicaRESUMEN
Secreted phospholipases (sPLA2s) in plants are a growing group of enzymes that catalyze the hydrolysis of sn-2 glycerophospholipids to lysophospholipids and free fatty acids. Until today, around only 20 sPLA2s were reported from plants. This review discusses the newly acquired information on plant sPLA2s including molecular, biochemical, catalytic, and functional aspects. The comparative analysis also includes phylogenetic, evolutionary, and tridimensional structure. The observations with emphasis in Glycine max sPLA2 are compared with the available data reported for all plants sPLA2s and with those described for animals (mainly from pancreatic juice and venoms sources).
RESUMEN
The cyclic ketones, thujone and dihydrocarvone, are lipophilic components of essential oils extracted from different plants, which have proven insecticidal activity. The GABAA receptor is activated by the neurotransmitter GABA and is the action site of widely used neurotoxic pesticides. Many compounds that regulate GABAA receptor function interact with membrane lipids, causing changes in their physical properties and consequently, in the membrane dynamic characteristics that modulate receptor macromolecules. In the present study, the biophysical effects of thujone (a gabaergic reference compound) and dihydrocarvone (structurally very similar) were explored by using monomolecular films of DPPC as a model membrane system, to gain insight into membrane-drug interaction. The compression isotherms showed that both ketones expand the DPPC isotherms and increase membrane elasticity. They penetrate the monolayer but their permanence depends on the possibility of establishing molecular interactions with the film component, favored by defects present in the membrane at the phase transition. Finally, by using Brewster angle microscopy (BAM) as a complementary technique for direct visualization of the study films, we found that incorporating ketone seems to reduce molecular repulsion among phospholipid headgroups. Our results reinforce the notion that changes in membrane mechanics may be occurring in the presence of the assayed ketones, suggesting that their interaction with the receptor's surrounding membrane may modulate or affect its functionality, possibly as part of the mechanism of the bioactivity described for thujone and DHC.
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Membrana Celular/química , Membrana Celular/efectos de los fármacos , Monoterpenos/farmacología , 1,2-Dipalmitoilfosfatidilcolina/química , 1,2-Dipalmitoilfosfatidilcolina/metabolismo , Adsorción , Monoterpenos Bicíclicos , Membrana Celular/metabolismo , Monoterpenos Ciclohexánicos , Monoterpenos/química , ReologíaRESUMEN
Secretory phospholipase A2 (sPLA2) are soluble enzymes that catalyze the conversion of phospholipids to lysophospholipids and free fatty acids at membrane interfaces. The effect of IAA and IPA auxins over the activity of recombinant sPLA2 isoforms from Glycine max was studied using membrane model systems including mixed micelles and Langmuir lipid monolayers. Both phytohormones stimulate the activity of both plant sPLA2 using DLPC/Triton mixed micelles as substrate. To elucidate the mechanism of action of the phytohormones, we showed that both auxins are able to self-penetrate lipid monolayers and cause an increment in surface pressure and an expansion of lipid/phytohormone mixed interfaces. The stimulating effect of auxins over phospholipase A2 activity was still present when using Langmuir mixed monolayers as organized substrate regardless of sPLA2 source (plant or animal). All the data suggest that the stimulating effect of auxins over sPLA2 is due to a more favorable interfacial environment rather to a direct effect over the enzyme.
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Glycine max/efectos de los fármacos , Ácidos Indolacéticos/farmacología , Fosfolipasas A2 Secretoras/metabolismo , Reguladores del Crecimiento de las Plantas/farmacología , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Micelas , Fosfolipasas A2 Secretoras/genética , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Glycine max/enzimología , Especificidad por SustratoRESUMEN
Two secretory phospholipase A2 (sPLA2s) from Glycine max, GmsPLA2-IXA-1 and GmsPLA2-XIB-2, have been purified as recombinant proteins and the activity was evaluated in order to obtain the optimum conditions for catalysis using mixed micelles and lipid monolayers as substrate. Both sPLA2s showed a maximum enzyme activity at pH 7 and a requirement of Ca(2+) in the micromolar range. These parameters were similar to those found for animal sPLA2s but a surprising optimum temperature for catalysis at 60 °C was observed. The effect of negative interfacial charges on the hydrolysis of organized substrates was evaluated through initial rate measurements using short chain phospholipids with different head groups. The enzymes showed subtle differences in the specificity for phospholipids with different head groups (DLPC, DLPG, DLPE, DLPA) in presence or absence of NaCl. Both recombinant enzymes showed lower activity toward anionic phospholipids and a preference for the zwitterionic ones. The values of the apparent kinetic parameters (Vmax and KM) demonstrated that these enzymes have more affinity for phosphatidylcholine compared with phosphatidylglycerol, in contrast with the results observed for pancreatic sPLA2. A hopping mode of catalysis was proposed for the action of these sPLA2 on mixed phospholipid/triton micelles. On the other hand, Langmuir-monolayers assays indicated an optimum lateral surface pressure for activity in between 13 and 16 mN/m for both recombinant enzymes.
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Biocatálisis , Membrana Celular/enzimología , Glycine max/enzimología , Fosfolipasas A2 Secretoras/metabolismo , Aire , Estabilidad de Enzimas , Cinética , Fosfolipasas A2 Secretoras/química , Especificidad por Sustrato , Temperatura , Agua/químicaRESUMEN
At the present, no secreted phospholipase A2 (sPLA2) from soybean (Glycine max) was investigated in detail. In this work we identified five sequences of putative secreted sPLA2 from soybean after a BLAST search in G. max database. Sequence analysis showed a conserved PA2c domain bearing the Ca²âº binding loop and the active site motif. All the five mature proteins contain 12 cysteine residues, which are commonly conserved in plant sPLA2s. We propose a phylogenetic tree based on sequence alignment of reported plant sPLA2s including the novel enzymes from G. max. According to PLA2 superfamily, two of G. max sPLA2s are grouped as XIA and the rest of sequences as XIB, on the basis of differences found in their molecular weights and deviating sequences especially in the N- and C-terminal regions of the isoenzymes. Furthermore, we report the cloning, expression and purification of one of the putative isoenzyme denoted as GmsPLA2-XIA-1. We demonstrate that this mature sPLA2 of 114 residues had PLA2 activity on Triton:phospholipid mixed micelles and determine the kinetic parameters for this system. We generate a model based on the known crystal structure of sPLA2 from rice (isoform II), giving first insights into the three-dimensional structure of folded GmsPLA2-XIA-1. Besides describing the spatial arrangement of highly conserved pair HIS-49/ASP-50 and the Ca⺲ loop domains, we propose the putative amino acids involved in the interfacial recognition surface. Additionally, molecular dynamics simulations indicate that calcium ion, besides its key function in the catalytic cycle, plays an important role in the overall stability of GmsPLA2-XIA-1 structure.