Oenococcus oeni allows the increase of antihypertensive and antioxidant activities in apple cider

This study aimed to investigate the impact of the malolactic fermentation (MLF) carried out by Oenococcus oeni on antihypertensive and antioxidant activities in cider. The MLF was induced using three strains of O. oeni. The modification in phenolic compounds (PCs) and nitrogen organic compounds, antioxidant, and antihypertensive activities were determined after MLF. Among the 17 PCs analyzed caffeic acid was the most abundant compound and phloretin, (−)-epicatechin, and myricetin were detected only in malolactic ciders, however, (−)-epigallocatechin was not detected after MLF. The evaluation of nitrogen organic compounds revealed a drop in total protein concentration (from 17.58 to 14.00 mg N/L) concomitantly with a significant release of peptide nitrogen (from 0.31 to a maximum value of 0.80 mg N/L) after MLF. In addition, an extracellular proteolytic activity was evidenced in all MLF supernatants. The FRAP activity increased reaching a maximum of 120.9 μmol FeSO4/mL and the ABTS radical-scavenging activity increased until 6.8 mmol ascorbic acid/L. Moreover, the angiotensin I-converting enzyme inhibitory activity reached a maximum value of 39.8%. The MLF conducted by O. oeni in ciders enables the increase of interesting biological activities and this finding could constitute a valuable tool to add value to final product.Fil: Kristof, Irina. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaFil: Ledesma, Silvana Cecilia. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaFil: Apud, Gisselle Raquel. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaFil: Vera, Nancy Roxana. Universidad Nacional de Tucumán; ArgentinaFil: Aredes Fernández, Pedro Adrián. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentin

Toxigenic potential of Fusarium graminearum isolated from maize of northwest Argentina

Twenty six isolates of Fusarium graminearum from grains of maize hybrids harvested in ±west Argentina were grown on autoclaved rice grain to assess their ability to produce type B trichothecenes. Chemical analysis indicated that 38% of isolates were nivalenol (NIV) producers only, 31% were major NIV producers with high DON(deoxynivalenol)/NIV ratios, 8% were major DON producers with minor NIV production, and 23% were DON producers only. Isolates showed a high variability in their toxigenic potential which was not related to fungal biomass. The distribution of the different chemotypes as well as the high and the low trichothecene-producing Fusarium isolates could not be associated to a geographical origin. Our results confirmed for the first time that isolates of Fusarium graminearum from maize of northwest Argentina are able to produce DON and NIV. A substancial contamination with both NIV and DON is likely in maize from northwest Argentina. Their contents should be quantified in regional surveillances for mycotoxin contamination.Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Apud, Gisselle Raquel. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Belizán, María Melina Eleonora. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vattuone, Marta Amelia. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Catalan, Cesar Atilio Nazareno. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Quimica del Noroeste; Argentina; Argentin

Secondary metabolites form native plants from northwest Argentina: identification and characterization of useful molecules in order to control the mycotoxin from fungi responsible of rot diseases in grapes and crops

Los hongos responsables de prodredumbres reducen el rendimiento de los cultivos y contaminan las cosechas con micotoxinas tóxicas para seres humanos y animales. Los mismos incluyen especies de Fusarium causantes de podredumbres de cereales que son productores de fumonisinas y tricotecenos, y especies de Aspergillus de la sección Nigri responsables de podredumbres de uvas y contaminación con ocratoxina A. El control químico de estos hongos se basa en el uso de fungicidas comerciales los cuales son costosos, contaminan el ambiente y pueden conducir al desarrollo de resistencia fúngica. Los extractos vegetales podrían proveer nuevas moléculas que superen esos problemas. En este trabajo, se investigó el efecto inhibitorio de extractos de partes aéreas (hojas y tallos) de plantas autóctonas del Noroeste Argentino Amphilophium cynanchoides, Tecomastans, Macfadyena cynanchoides y Jacaranda mimosifolia sobre especies de Fusarium (Fusarium graminearum y F. verticillioides) y Aspergillus (A. carbonarius y A. niger). De los 24 extractos obtenidos, 10 alcanzaron a inhibir el 50% del crecimiento fúngico (CI50). Ensayos de microdilución indicaron que los extractos con los menores valores de CI50 fueronlos de acetato de etilo de hojas y tallos de A. cynanchoides contra F.graminearum (CI50=1320 ppm) y F. verticillioides (CI50=1305ppm), respectivamente, y el extracto diclorometano de tallos de M. cynanchoides contra A. niger (CI50=1025 ppm) y A.carbonarius (CI50=1066 ppm). El extracto diclorometano de tallos de M. cynanchoides fue el único que suprimió completamente el crecimiento fúngico (CIM=2500 ppm). El mismo fue antifúngico sobre todas las cepas fúngicas ensayadas en bioautografía por siembra puntual,donde las cepas de Aspergillus fueron más sensibles que las de Fusarium.Los constituyentes antifúngicos del extracto diclorometano se aislaron e identificaron como 2-hidroxi-3-(3-metil-2-butenil)-1,4-naftoquinona (lapachol) y 1-hidroxi-4-metil antraquinona.La antraquinona fue más activa sobre las cepas fúngicas (CIM= 78,1-625 ppm) que el lapachol (CIM=312,5-1250 ppm). El extracto diclorometano y sus constituyentes antifúngicos se ensayaron en mezclas con propiconazol sobre todas las cepas fúngicas, con metabisulfito de sodio contra las cepas de Aspergillus y con sorbato de potasio sobre cepas de Fusarium. El extracto y sus metabolitos aislados tuvieron un efecto sinérgico con el metabisulfito de sodio sobre las cepas de Aspergillus y de propiconazol sobre las cepas de Fusarium.Incorporados en jugo de uva en concentraciones sub-inhibitorias, el extracto diclorometano, el lapachol y la 1-hidroxi-4-metilantraquinona inhibieron la síntesis de OTA mientras que el metabisulfito de sodio estimuló la acumulación de esa micotoxina. Sin embargo, los mismos suprimieron la producción de OTA en mezclas con metabisulfito de sodio. El extracto diclorometano de tallos de M. cynanchoides y sus quinonas identificadas tienen potencial como aditivos de metabisulfito de sodio contra Aspergillus responsables de la podredumbre negra de la uva.Fungi responsible of rot diseases reduce crops yields and contaminate the harvestable food products with mycotoxins noxious for human and animal health. They include the Fusarium species causing cereal rots, which are fumonisin and trichothecene producers, and Aspergillus species of the Nigri section responsible of black grape rots and ochratoxin A (OTA) production. Chemical control of these fungi is based in the use of commercial fungicides which are expensive, generate environmental pollution and led to the development of fungal resistance. Plant extracts might provide new antifungals overcoming these problems. In this work, the inhibitory effect of extracts from aerial parts (leaves and stems) of native plants from northwest Argentina (Amphilophium cynanchoides, Tecoma stans, Macfadyena cynanchoides and Jacaranda mimosifolia) was investigated on Fusarium (F. graminearum and F. verticillioides) and Aspergillus (A. carbonarius and A. niger) species. Ten of a total of 24 extracts inhibited 50% of the fungal growth (IC50). Microdilution assays indicated that the extracts with the lowest IC50 values were the ethyl acetate leaf and stem extracts of A. cynanchoides against F. graminearum (IC50=1320 ppm) and F. verticillioides (IC50=1305 ppm), respectively, and the dichloromethane stem extract of M. cynanchoides against A. niger (IC50=1025 ppm) and A. carbonarius (1066 ppm). The dichloromethane stem extract of M. cynanchoides was the only one that completely suppressed fungal growth (MIC) at 2500 ppm. It was antifungal on all the fungal strains in dot blot assays where Aspergillus strains were more sensitive than the Fusarium strains. The antifungal constituents of the dichloromethane extract were isolated and identified as 2-hydroxy-3- (3-methyl-2-butenyl)-1,4-naphthoquinone (lapachol) and 1-hydroxy-4-methyl anthraquinone. The anthraquinone was more active on the fungal strains (MIC= 78.1-625 ppm) than lapachol (MIC = 312.5-1250 ppm). The dichloromethane extract and the antifungal constituents were tested in mixtures with propiconazole on all the fungal strains, with sodium metabisulfite against the Aspergillus strains and with potassium sorbate on the Fusarium strains. They synergized the effect of sodium metabisulfite on the Aspergillus strains and propiconazole only on Fusarium. Added at sub-MIC concentrations to grape juice, the dichloromethane extract and its antifungal quinines inhibited OTA production while sodium metabisulfite induced OTA accumulation. They suppressed OTA accumulation in mixtures with sodium metabisulfite. The dichloromethane extract of M. cynanchoides and its isolated antifungals have potential as additives of sodium metabisulfite against the Aspergillus responsible of the black grape rotsFil: Apud, Gisselle Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentin

Release of biologically active peptides from grape juice by Oenococcus oeni isolated from argentine wine

The increase in biologically active peptides after proteolytic activity of Oenococcus oeni X2L was studied in grape juice medium (GJM) and in GJM fermented with Saccharomyces cerevisiae mc2. Sequential inoculation of O. oeni X2L with proteolytic activity in GJM before (“I” medium) and after (“F” medium) yeast fermentation allowed peptide release. In the “I” medium, bacterial proteolytic activity (0.083 mM) released 3.88 mg nitrogen (N) of peptides per liter and produced an increase of 203.39 µmol/L in the ferric reducing antioxidant power (FRAP) and 16.49% in the angiotensin I-converting enzyme inhibitory (ACE-I) activity after 48 hrs of incubation. In the “F” medium, a higher proteolytic activity was evidenced after 48 hrs of incubation (0.179 mM), releasing 0.87 mg N of peptides/L. The released peptides in the “F” medium produced an increase in ACE-I activity (22.15%). Calculation of the specific activity (activity expressed per mg N of peptide released) of FRAP, 1, 1-diphenyl-2-picrylhydrazyl scavenging, and ACE-I after 48 hrs of incubation revealed a higher activity in the “F” than in the “I” medium. This finding indicates a higher efficiency of the proteolytic system of O. oeni X2L on grape juice proteins in the release of bioactive peptides after yeast growth.Fil: Stivala, Maria Gilda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Biotecnología; ArgentinaFil: Apud, Gisselle Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Aredes Fernández, Pedro Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Biotecnología; Argentin

Impact of Fungal Diseases in Grapes and Wine: General Aaspects and Recent Advances

Fungi are the most important and prevalent plant pathogens. The infection of plants by several fungi causes destructive and important economical losses of crops in the field and fresh fruits and vegetables during storage and transportation. As a result of fungi metabolism, secondary metabolites, called mycotoxins are produced. The intake of these toxins above certain levels can produce several toxic effects in humans and animals. With respect to wine, fungal diseases reduce yield and quality of grapes and affect chemical and sensory properties of wine. The purpose of this chapter is to describe the main fungal diseases affecting grapes and wine quality, the mycotoxigenic risk associated to wine production and strategies currently performed to prevent and control fungal contamination. Special attention was paid to ochratoxin A, the main mycotoxin present in grapes, must and wine.Fil: Apud, Gisselle Raquel. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaFil: Aredes Fernández, Pedro Adrián. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaFil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentin

Quinones of Macfadyena cynanchoides for Control of Aspergillus carbonarius and Aspergillus niger in Wine

Antiochratoxigenic activity (anti-OTA) of dichloromethane extract of Macfadyena cynanchoides stems, its antifungal metabolites (lapachol and 1-hydroxy-4-methyl anthraquinone), and sodium metabisulfite were evaluated alone and in combination for their effect on Aspergillus species at sub-lethal concentrations. Grape juice inoculated with 5 × 10 3 spores/mL of A. carbonarius or A. niger was supplemented with the extract (2500 to 156.3 µg/mL), the quinones (1250 to 78.1 µg/mL), or sodium metabisulfite (2500 to 156.3 µg/mL). OTA accumulation was measured for the sub-lethal concentrations after incubating in the dark for six days at 15°C. The extract and sodium metabisulfite completely inhibited growth of both fungi at 2500 µg/mL, while lapachol and 1H4MA required 1250 µg/mL. Average OTA biosynthesis declined to 30 to 60% less than the control at extract concentrations of 625 to 1250 μg/mL. Both lapachol and 1H4MA at 625 μg/mL exerted a similar anti-OTA effect, with an average inhibition of 30%. Sodium metabisulfite increased OTA production 60 to 100% at all sub-lethal concentrations. Binary mixtures of extract or lapachol or 1H4MA + sodium metabisulfite, at concentrations of 9.8 + 156.3 µg/mL, 19.5 + 312.5 µg/mL, and 39.1 + 625 µg/mL, completely inhibited OTA production. In addition, the mixture lapachol + sodium metabisulfite (9.8 + 156.3 µg/mL) delayed the time required to complete alcoholic fermentation, but did not reduce Saccharomyces cerevisiae viability or modify physicochemical parameters at the end of fermentation. The mixture sodium metabisulfite + lapachol has potential as an antifungal and anti-OTA agent to reduce the dose of sulfites to control Aspergillus on grapes postharvest.Fil: Apud, Gisselle Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Aredes Fernández, Pedro Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentin

Antifungal activity of Bignoniaceae plants on Aspergillus carbonarius and Aspergillus niger

Twenty four extracts from Bignoniaceae plants of northwest Argentina were tested for antifungal activity against Aspergillus species responsible of the grape black rot. Stems and leaves of Amphilophium cynanchoides, Macfadyena cynanchoides, Tecoma stans and Jacaranda mimosifolia were separately extracted with solvents of increasing polarity to obtain the dichloromethane (fCH2Cl2), ethyl acetate (fEtOAc) and methanol extracts (fMeOH). The fCH2Cl2 from stem of M. cynanchoides had the lowest IC50 (1.0-1.2 mg/mL) and MID values (0.6-1.2 mg) and the highest ID values (5.0-6.8 mm) on A. niger and A. carbonarius. The main contributors of the antifungal activity of fCH2Cl2 were identified as lapachol (MIC = 0.25-1.00 mg/ml) and 1-hydroxy-4-methylanthraquinone (MIC = 0.0625-0.125 mg/mL). These compounds synergized the antifungal activity of sodium metabisulfite and showed an additive effect in mixtures with propiconazol. They might be used as additives of commercial antifungals to protect grapes against A. niger and A. carbonarius.Fil: Apud, Gisselle Raquel. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Universidad San Pablo Tucumán; ArgentinaFil: Aredes Fernández, Pedro Adrián. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Superior de Investigaciones Biologicas. Grupo de Investigacion y Desarrollo del Noroeste Argentino | Universidad Nacional de Tucuman. Instituto Superior de Investigaciones Biologicas. Grupo de Investigacion y Desarrollo del Noroeste Argentino.; ArgentinaFil: Kritsanida, Marina. Universite de Paris, Descartes; FranciaFil: Grougnet, Raphael. Universite de Paris, Descartes; FranciaFil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentin

Proteolytic Activity of Oenococcus oeni Enables the Increase in Antioxidant and Antihypertensive Activities from Wine

Oenococcus oeni is a lactic acid bacterium involved in winemaking where it generally carries out the malolactic fermentation converting the wine's malic acid into lactic acid. In this work were used the strain m of Oenococcus oeni. The culture was inoculated at 108 Log CFU/mL in a synthetic wine medium (SW) supplemented with a fraction of high molecular weight constituted by proteins and polypeptides (FPP) obtained from Cabernet Sauvignon and Syrah wines from Colalao del Valle, Tucumán, Argentine. In presence of FPP, O. oeni maintains viability after 48 h incubation time and release an extracellular proteolytic activity. Therefore, a release peptides of 1.247 and 1.373 mg N/L at 48 h of incubation time was detected in SW supplemented with FPP from Cabernet Sauvignon and Syrah wines respectively. Concomitantly with the maximum peptide release, the “in vitro” biological activities were increased. The released peptides from Cabernet Sauvignon wine enables the increase in the ferric reducing antioxidant power (FRAP) capacity, the scavenging activity of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), and the inhibition of angiotensin I-converting enzyme (ACEI activity) in 392.8 µmol FeSO4/L, 9.7% and 63.9%, respectively. In presence of FPP of Syrah wine, the released peptides increases in 156.5 µmol FeSO4/L, 5.5% and 13.8% the FRAP, DPPH and ACEI activities, respectively. The utilization of Oenococcus oeni m to carry out the malolactic fermentation would contribute to enhance the beneficial biological activities of the final product and provide an additional value to regional wines.Fil: Apud, Gisselle Raquel. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán; ArgentinaFil: Stivala, Maria Gilda. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán; ArgentinaFil: Aredes Fernández, Pedro Adrián. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán; ArgentinaFil: Rodriguez Vaquero, Maria Jose. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán; Argentin

Increase in antioxidant and antihypertensive peptides from Argentinean wines by Oenococcus oeni

Cells from an exponential Oenococcus oeni m1 culture in a grape juice medium were inoculated into a synthetic wine medium (SW) supplemented with a protein and polypeptide fraction (PPF) of high molecular weight (higher than 12,400 Da) obtained from four varietals of Cafayate Argentinean wines. O. oeni maintains viability after 48 h incubation time and enables the increase in extracellular proteolytic activity and the release of low molecular weight peptides by 1.067, 0.397, 0.915 and 0.705 mg N/L in the respective SW supplemented with PPF from Cabernet Sauvignon, Malbec, Tannat and Torrontés wine varietals. After 48 h incubation time, concomitantly with peptide release, an increase in antioxidant and antihypertensive activities was detected in all studied media. The highest increase was detected in the presence of PPF from Cabernet and Tannat wine varietals. Maximum increase in antioxidant activity (366.1 μmol FeSO4/L in the case of ferric reducing antioxidant power and 8.9% in 2,2-diphenyl-1-picrylhydrazyl radical scavenging) was produced by the peptides released from PPF of Cabernet Sauvignon wine. The peptides released from PPF Tannat wine varietal caused the highest increase in antihypertensive activity (56.2% in angiotensin I-converting enzyme inhibitory activity). Oenococcus oeni m1 would provide additional benefits to wine such as an increase in bioactive peptides with multifunctional beneficial activities.Fil: Aredes Fernández, Pedro Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia Para Lactobacilos (i); Argentina; Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina;Fil: Apud, Gisselle Raquel. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina;Fil: Stivala, Maria Gilda. Universidad Nacional de Tucuman. Facultad de Bioquimica, Quimica y Farmacia. Instituto de Quimica Organica. Catedra de Quimica Organica Ii; Argentina;Fil: Rodriguez Vaquero, Maria Jose. Universidad Nacional de Tucuman. Facultad de Bioquimica, Quimica y Farmacia. Instituto de Microbiologia;Fil: Rollan, Graciela Celestina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia Para Lactobacilos (i); Argentina