RESUMEN
Anthocyanins are bioactive compounds responsible for various physiological processes in plants and provide characteristic colors to fruits and flowers. Their biosynthetic pathway is well understood; however, the enzymatic degradation mechanism is less explored. Anthocyanase (ß-glucosidase (BGL)), peroxidase (POD), and polyphenol oxidase (PPO) are enzymes involved in degrading anthocyanins in plants such as petunias, eggplants, and Sicilian oranges. The aim of this work was to investigate the physicochemical interactions between these enzymes and the identified anthocyanins (via UPLC-MS/MS) in cranberry (Vaccinium macrocarpon) through molecular docking to identify the residues likely involved in anthocyanin degradation. Three-dimensional models were constructed using the AlphaFold2 server based on consensus sequences specific to each enzyme. The models with the highest confidence scores (pLDDT) were selected, with BGL, POD, and PPO achieving scores of 87.6, 94.8, and 84.1, respectively. These models were then refined using molecular dynamics for 100 ns. Additionally, UPLC-MS/MS analysis identified various flavonoids in cranberries, including cyanidin, delphinidin, procyanidin B2 and B4, petunidin, pelargonidin, peonidin, and malvidin, providing important experimental data to support the study. Molecular docking simulations revealed the most stable interactions between anthocyanase and the anthocyanins cyanidin 3-arabinoside and cyanidin 3-glucoside, with a favorable ΔG of interaction between -9.3 and -9.2 kcal/mol. This study contributes to proposing a degradation mechanism and seeking inhibitors to prevent fruit discoloration.
Asunto(s)
Antocianinas , Catecol Oxidasa , Simulación del Acoplamiento Molecular , Vaccinium macrocarpon , Antocianinas/química , Antocianinas/metabolismo , Catecol Oxidasa/metabolismo , Catecol Oxidasa/química , Vaccinium macrocarpon/química , Peroxidasa/metabolismo , Peroxidasa/química , Espectrometría de Masas en Tándem , Proteínas de Plantas/metabolismo , Proteínas de Plantas/química , Simulación de Dinámica Molecular , Simulación por Computador , Frutas/química , Frutas/metabolismo , Frutas/enzimologíaRESUMEN
Diverse enzymatic reactions taking place after the killing of green vanilla beans are involved in the flavor and color development of the cured beans. The effects of high hydrostatic pressure (HHP) at 50-400 MPa/5 min and blanching as vanilla killing methods were evaluated on the total phenolic content (TPC), polyphenoloxidase (PPO), and peroxidase (POD) activity and the color change at different curing cycles of sweating-drying (C0-C20) of vanilla beans. The rate constants describing the above parameters during the curing cycles were also obtained. The TPC increased from C1 to C6 compared with the untreated green beans after which it started to decrease. The 400 MPa samples showed the highest rate of phenolic increase. Immediately after the killing (C0), the highest increase in PPO activity was observed at 50 MPa (46%), whereas for POD it was at 400 MPa (25%). Both enzymes showed the maximum activity at C1, after which the activity started to decrease. As expected, the L* color parameter decreased during the entire curing for all treatments. An inverse relationship between the rate of TPC decrease and enzymatic activity loss was found, but the relationship with L* was unclear. HHP appears to be an alternative vanilla killing method; nevertheless, more studies are needed to establish its clear advantages over blanching.
Asunto(s)
Vanilla , Presión Hidrostática , Manipulación de Alimentos/métodos , Fenoles , Catecol OxidasaRESUMEN
Enzymatic browning is a major problem in minimally processed banana puree; it reduces consumer's acceptability and affects nutritional quality. The objective of this work was to evaluate the effects ultrasound (40â kHz/10â min), microwave (800â W/ 25â s) and UV-C radiation (1.97â kJ/m2) applied to banana puree. Colour parameters (L*, a*, b*, chroma and hue), browning index (BI), polyphenol oxidase (PPO) and peroxidase (POD) activities, total phenolic compounds (TPC), antioxidant capacity (AOC) and microbiological counts were monitored throughout storage at 4 °C. Ultrasound (US) and microwave (MW) treatments achieved a significant (p < 0.05) reduction in PPO activity and BI; moreover, ultrasound effectively retained phenolic compounds content (75% of initial value). The AOC was in coincidence with TPC values. POD activity was partially inhibited by UV-C while MW and US increase its activity. Although UV-C treatment was not effective to control browning development, it was effective to maintain microbiological stability after 20 days of storage (1.48 ± 0.01 log CFU/g). The evaluated treatments have the advantage of being less aggressive than conventional thermal treatments while maintaining fresh characteristics of the product.
Asunto(s)
Musa , Microondas , Rayos Ultravioleta , Ultrasonido , Catecol Oxidasa , Antioxidantes , Fenoles/análisisRESUMEN
Currently, catalysts with oxidative activity are required to create valuable chemical, agrochemical, and pharmaceutical products. The catechol oxidase activity is a model reaction that can reveal new oxidative catalysts. The use of complexes as catalysts using iron (III) and structurally simple ligands such as pyrazine (pz), quinoxaline (qx), and phenazine (fz) has not been fully explored. To characterize the composition of the solution and identify the abundant species which were used to catalyze the catechol oxidation, the distribution diagrams of these species were obtained by an equilibrium study using a modified Job method in the HypSpec software. This allows to obtain also the UV-vis spectra calculated and the formation constants for the mononuclear and binuclear complexes with Fe3+ including: [Fe(pz)]3+, [Fe2(pz)]6+, [Fe(qx)]3+, [Fe2(qx)]6+, [Fe(fz)]3+, and [Fe2(fz)]6+. The formation constants obtained were log ß110 = 3.2 ± 0.1, log ß210 = 6.9 ± 0.1, log ß110 = 4.4 ± 0.1, log ß210 = 8.3 ± 0.1, log ß110 = 6.4 ± 0.2, and log ß210 = 9.9 ± 0.2, respectively. The determination of the catechol oxidase activity for these complexes did not follow a traditional Michaelis-Menten behavior.
Asunto(s)
Hierro , Metanol , Catecol Oxidasa , Hierro/química , Fenazinas , Pirazinas , QuinoxalinasRESUMEN
The stimulus to the modeling of enzyme functioning sites comes from their potential to give insight into the natural enzyme's mechanistic pathways, ascertain the role of that different metal ion in the active site and construct better catalysts motivated by nature. The presence of metal ion leads to the activation of molecular oxygen in the metalloenzymes. The metalloenzymes such as the catechol oxidase (CO) enzyme that oxidizes the catechol to corresponding quinones which eventually protect damage tissues from plant and pathogen. Thus, the design and characterization of catalysts used as selectively and efficiently oxidation reactions have grown to be unique challenges for modern inorganic chemists. In this work, two novel tetranuclear complexes (1 and 2) have been synthesized in excellent yield. The complexes were characterized using various spectroscopic techniques such as FTIR, UV-Visible and PXRD pattern. The structure of 1 and 2 was elucidated by SC-XRD (single crystal X-ray diffraction) analysis. The magnetic study reveals the presence of the antiferromagnetic nature of 1 and 2. Both 1 and 2 shows a very good catecholase-like activity by oxidizing the catechol to analogous quinone in methanolic solution. Thus, a structure-activity relationship can further help us design other substituted tetranuclear complexes with enhanced catecholase like activity.Communicated by Ramaswamy H. Sarma.
Asunto(s)
Catecol Oxidasa , Cobre , Estructura Molecular , Cobre/química , Catecol Oxidasa/química , Catecol Oxidasa/metabolismo , Cristalografía por Rayos X , CatálisisRESUMEN
BACKGROUND: The present study investigated the sensory acceptance, oxidative damage and protection, and possible anatomical-structural damage of cells from the surface of shapes of minimally processed yam. The tubers were minimally processed into the peeled rondelle, dice and 'chateau cut' (chateau) shapes, the latter of which was obtained after performing the abrasion technique. Control treatment corresponded to the rondelle shape with the periderm. The pieces were kept packed at 5 ± 2 °C for 14 days. RESULTS: Peeled rondelle and chateau were sensorially the most well-accepted yam shapes and achieved the highest purchase intention. The enzymes were partially modulated by the detected H2 O2 levels. Oxidative burst lasted longer in the minimally processed tissues than in the control. Polyphenol oxidase activity showed a clear difference in behavior between the minimally processed pieces and the control. Minimal processing induced transient increases in phenolic compounds, for which the expression was lowest in the abraded pieces. On the other hand, these pieces exhibited greater cell collapse on the surface of the amyliferous parenchyma. CONCLUSION: Based on the results of the trained panel, the abrasion technique is an alternative to provide shapes that are better accepted and marketable, more resistant to browning, and can be stored for up to 12 days. Resistance to browning may be related to a more efficient modulation of enzymatic antioxidant systems and intense deposition of cell debris on the surface of the amyliferous parenchyma. © 2021 Society of Chemical Industry.
Asunto(s)
Comportamiento del Consumidor , Dioscorea/química , Manipulación de Alimentos/métodos , Catecol Oxidasa/análisis , Catecol Oxidasa/metabolismo , Dioscorea/metabolismo , Manipulación de Alimentos/instrumentación , Humanos , Fenoles/análisis , Fenoles/metabolismo , Tubérculos de la Planta/química , Tubérculos de la Planta/metabolismoRESUMEN
Biochemical characterization of polyphenol oxidase (PPO) present in purple sweet potato (PSP) is a key step in developing efficient methodologies to control oxidative damage caused by this enzyme to the valuable components of PSP, such as caffeoylquinic acid derivatives and acylated anthocyanins. Thus, this work focused on the assessment of the effects of pH, temperature, and chemical agents on the PPO activity as well as characterization of the PPO substrate specificity towards major phenolic compounds found in PSP. The optimum conditions of enzyme activity were pH 7 and a temperature range of 20-30 °C at which phenolic substrates were oxidized with 72.5-99.8% yield. Zn2+ ions remarkably reduced PPO activity while Cu2+ ions improved enzyme performance. The highest substrate preference was shown for 3,4,5-tri-caffeoylquinic and 3,5-di-caffeoylquinic acid, followed by 5-caffeoylquinic and caffeic acid, 3,4- and 4,5-di-caffeoylquinic acids, peonidin-3-caffeoyl-p-hydroxybenzoyl-sophoroside-5-glucoside. The highest Km values were found for 4,5-feruloyl-caffeoylquinic acid and catechol.
Asunto(s)
Antocianinas/química , Antocianinas/metabolismo , Catecol Oxidasa/metabolismo , Ipomoea batatas/enzimología , Ácido Quínico/análogos & derivados , Acilación , Unión Proteica , Ácido Quínico/química , Ácido Quínico/metabolismoRESUMEN
Microwave heating has been considered a promising technology for continuous flow thermal processing of fluid foods due to better retention of quality. Considering the importance of açai-berry pulp and its perishability, the inactivation kinetics of peroxidase (POD) and polyphenol oxidase (PPO) were investigated under conventional and microwave heating. First-order two-component model was well fitted to the data, indicating the presence of at least two fractions with different resistances. POD was more thermally resistant (90% inactivation for 40 s at 89 °C) and could be considered as a processing target. Inactivation curves dependency on heating technology suggests specific effects of microwaves on the protein structure. Additionally, the dielectric properties of açai-berry pulp were evaluated at 915 and 2,450 MHz for temperatures up to 120 °C. Power penetration depth dropped with temperature at 915 MHz (from 29 to 11 mm), but was less affected at 2,450 MHz (between 8 and 11 mm).
Asunto(s)
Catecol Oxidasa/metabolismo , Euterpe/metabolismo , Microondas , Peroxidasa/metabolismo , Calor , CinéticaRESUMEN
Emerging technologies, such as focused microwave heating of liquid foods, have been studied to reduce quality losses due to the high temperatures of conventional processing. Besides faster heating, microwaves can also have non-thermal effects on inactivation; however, this is a controversial issue. The objective of this study was to compare conventional and focused microwave heating under similar conditions for the inactivation of two polyphenol oxidases (PPOs): mushroom tyrosinase in buffer and the PPO present in coconut water. Small samples under stirring were treated at temperatures between 50 and 90 °C and three kinetic models were adjusted considering the whole time-temperature history. The Weibull model could best describe inactivation in both heating processes, which was more effective with microwave heating for temperatures over 70 °C. Validation runs show that the model can satisfactorily describe the PPO inactivation. This study contributes for the design of liquid food pasteurization by focused microwave technology.
Asunto(s)
Catecol Oxidasa/química , Pasteurización/métodos , Agaricales/enzimología , Tampones (Química) , Catecol Oxidasa/metabolismo , Cocos/enzimología , Activación Enzimática , Industria de Procesamiento de Alimentos/métodos , Calefacción , Cinética , Microondas , Modelos Teóricos , Monofenol Monooxigenasa/química , Monofenol Monooxigenasa/metabolismo , TemperaturaRESUMEN
Carioca beans contribute to health maintenance around the world, and the evaluation of commercial postharvest storage (CPS) ensures their quality. This study aimed to evaluate the effect of CPS on technological, physicochemical and functional properties of carioca beans. Two genotypes (Pontal-PO and Madreperola-MP beans) were stored under CPS or controlled conditions and were evaluated after harvest and after three- and six-months storage. PO and MP hardened with time, but the cooking time did not differ. PO is darker than MP and both darkened over time. Storage time affected pH and acidity of the beans and MP presented better physicochemical properties than PO, with lower activity of peroxidase (p = 0.004) and polyphenoloxidase (p = 0.001) enzymes. Glycosylated kaempferol was suggested as a possible chemical marker to differentiate the aging of PO and MP beans. In conclusion, besides the technological differences, the storage was able to prevent physicochemical and functional alterations of beans.
Asunto(s)
Almacenamiento de Alimentos/métodos , Phaseolus/química , Catecol Oxidasa/metabolismo , Fibras de la Dieta/análisis , Dureza , Humanos , Humedad , Concentración de Iones de Hidrógeno , Quempferoles/análisis , Quempferoles/química , Nutrientes/análisis , Peroxidasas/metabolismo , Phaseolus/metabolismo , Ácido Fítico/análisis , Espectrofotometría , Temperatura , Factores de TiempoRESUMEN
The research reported herein focuses on the synthesis of two new Cu(II) complexes {[Cu2(2-X-4,6-bis(di-2-picolylamino)-1,3,5-triazine], with X = butane-1,4-diamine (2) or N-methylpyrenylbutane-1,4-diamine (3)}, the latter with a pyrene group as a possible DNA intercalating agent. The structure of complex (3) was determined by X-ray crystallography and shows the dinuclear {CuII(µ-OCH3)2CuII} unit in which the CuII···CuII distance of 3.040 Å is similar to that of 2.97 Å previously found for 1, which contains a {CuII(µ-OH)2CuII} structural unit. Complexes (2) and (3) were also characterized in spectroscopic and electrochemical studies, and catecholase-like activity were performed for both complexes. The kinetic parameters obtained for the oxidation of the model substrate 3,5-di-tert-butylcatechol revealed that the insertion of the spacer butane-1,4-diamine and the pyrene group strongly contributes to increasing the catalytic efficiency of these systems. In fact, Kass becomes significantly higher, indicating that these groups influence the interaction between the complex and the substrate. These complexes also show DNA cleavage under mild conditions with moderate reaction times. The rate of cleavage (kcat) indicated that the presence of butane-1,4-diamine and pyrene increased the activity of both complexes. The reaction mechanism seems to have oxidative and hydrolytic features and the effect of DNA groove binding compounds and circular dichroism indicate that all complexes interact with plasmid DNA through the minor groove. High-resolution DNA cleavage assays provide information on the interaction mechanism and for complex (2) a specificity for the unpaired hairpin region containing thymine bases was observed, in contrast to (3).
Asunto(s)
Biomimética , Catecol Oxidasa/química , Complejos de Coordinación/química , Cobre/química , Endonucleasas/química , Triazinas/química , Cristalografía por Rayos X , Ligandos , Estructura Molecular , Oxidación-Reducción , Potenciometría , Análisis Espectral/métodosRESUMEN
Cold plasma is a potential alternative to traditional thermal conservation methods because of its high efficiency in the preservation and retention of quality parameters. The objective of this study was to evaluate the application of atmospheric cold plasma on some qualitative aspects of apple cubes and apple juice. The research used dielectric barrier discharge plasma and studied different excitation frequencies of plasma: 50, 200, 400, 600, and 900 Hz. The effects of plasma application were evaluated on enzymatic activity (PPO and POD), total phenolic compounds, antioxidant capacity, and colorimetry. Plasma treatment partially inactivated the polyphenol oxidase enzyme in apples cubes and juice. Inactivation of peroxidase occurred only in apple juice. Total phenolic content and antioxidant capacity presented no significant difference between the treated and control samples of apple cubes, while significant changes were observed in apple juice. The changes in color parameters were slight and did not compromise the product quality. Plasma application was able to partially inactivate the enzymes responsible for browning while maintaining the quality and sensory properties of apple cubes and juice.
Asunto(s)
Antioxidantes/análisis , Jugos de Frutas y Vegetales/análisis , Malus/química , Fenoles/análisis , Catecol Oxidasa/metabolismo , Color , Manipulación de Alimentos , Gases em PlasmaRESUMEN
The aim of this paper was to evaluate the effect of cold plasma excitation frequency on camu-camu juice processing. Different levels of frequency (200, 420, 583, 698 and 960 Hz) were applied on camu-camu juice to measure the contents of ascorbic acid and anthocyanins, as well as to evaluate the antioxidant compounds (DPPH, ABTS, FRAP and phenolic compounds), peroxidase and polyphenol oxidase enzymatic activity and color. Furthermore, the juice bioaccessibility was evaluated after simulated digestion. The ascorbic acid concentration was increased when higher excitation frequencies were employed, increasing their bioavailability. Anthocyanins, peroxidase and polyphenol oxidase presented considerable degradation with increasing the plasma excitation frequency. For this reason, the juice processing proposed herein represents an alternative to enhance its nutritional quality. Moreover, the use of cold plasma reduced the activity concentration of endogenous enzymes, presenting considerable degradation for higher excitation frequency.
Asunto(s)
Manipulación de Alimentos , Jugos de Frutas y Vegetales/análisis , Myrtaceae/química , Extractos Vegetales/análisis , Gases em Plasma/química , Antocianinas/análisis , Antioxidantes/análisis , Ácido Ascórbico/análisis , Catecol Oxidasa/metabolismo , Color , Análisis de los Alimentos , Calidad de los Alimentos , Frutas/química , Peroxidasa/metabolismo , Fenoles/análisisRESUMEN
Haemocyanins (Hcs) are copper-containing, respiratory proteins that occur in the haemolymph of many arthropod species. Here, we report the presence of Hcs in the chilopode Myriapoda, demonstrating that these proteins are more widespread among the Arthropoda than previously thought. The analysis of transcriptome of S. subspinipes subpinipes reveals the presence of two distinct subunits of Hc, where the signal peptide is present, and six of prophenoloxidase (PPO), where the signal peptide is absent, in the 75 kDa range. Size exclusion chromatography profiles indicate different quaternary organization for Hc of both species, which was corroborated by TEM analysis: S. viridicornis Hc is a 6 × 6-mer and S. subspinipes Hc is a 3 × 6-mer, which resembles the half-structure of the 6 × 6-mer but also includes the presence of phenoloxidases, since the 1 × 6-mer quaternary organization is commonly associated with hexamers of PPO. Studies with Chelicerata showed that PPO activity are exclusively associated with the Hcs. This study indicates that Scolopendra may have different proteins playing oxygen transport (Hc) and PO function, both following the hexameric oligomerization observed in Hcs.
Asunto(s)
Catecol Oxidasa/metabolismo , Quilópodos/metabolismo , Precursores Enzimáticos/metabolismo , Hemocianinas/química , Hemocianinas/metabolismo , Análisis de Secuencia de ADN/métodos , Animales , Proteínas de Artrópodos/química , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/metabolismo , Catecol Oxidasa/química , Quilópodos/genética , Cromatografía en Gel , Precursores Enzimáticos/química , Regulación de la Expresión Génica , Hemocianinas/genética , Hemolinfa/metabolismo , Modelos Moleculares , Peso Molecular , Filogenia , Conformación Proteica , Multimerización de ProteínaRESUMEN
The super-intensive BioFloc Technology (BFT) system has been highlighted as a promising eco-friendly alternative to the traditional shrimp rearing systems. To gain insight into the impact of environmental rearing conditions on shrimp intestinal immunity, we assessed the expression profile of key immunological genes in the midgut of Litopenaeus vannamei shrimp reared in two contrasting culture systems: the indoor super-intensive BFT and the outdoor intensive Green-Water System (GWS). From the 30 analyzed genes, the expression levels of 25 genes were higher in the midgut of shrimp reared in BFT than in GWS. The main functional categories represented in BFT-shrimp were the prophenoloxidase-activating system, immune signaling, antimicrobial peptides, and RNA interference pathway. Comparatively, only the RNAi pathway gene Dicer-1 (LvDcr1) was more expressed in animals from the GWS group. However, despite the differences in gene expression, the total midgut bacterial abundance was similar between the experimental groups. Altogether, our results suggest that the microbial-rich environment offered by the BFT system can be acting as an immunostimulant by altering the immune expression profile of the midgut. The gene expression level found in GWS animals could be related to the chronic presence of the IMNV in the Brazilian Northeast. Knowing the effects of environmental stress factors on the intestinal immune defenses can provide an in-depth understanding of the relationship between cultivated shrimp and the major pathogens affecting the shrimp industry.
Asunto(s)
Acuicultura/métodos , Tracto Gastrointestinal/fisiología , Penaeidae/inmunología , Animales , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/metabolismo , Brasil , Catecol Oxidasa/genética , Catecol Oxidasa/metabolismo , Ambiente , Precursores Enzimáticos/genética , Precursores Enzimáticos/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/inmunología , Inmunidad Innata , Inmunización , Ribonucleasa III/genética , Ribonucleasa III/metabolismo , Transducción de Señal/inmunologíaRESUMEN
The tri-dentate Schiff base ligand 3-(2-hydroxyethylimino)-1-phenylbut-1-en-1-ol (L) produced the tetra-nuclear Cu(II) distorted cubane complex which contain Cu4O4 core, upon reaction with Cu(II)acetate.H2O. The complex was structurally characterized by X-ray crystallography and found that, in this tetrameric and tetra-nuclear distorted cubane structure, each two-fold deprotonated Schiff base ligand coordinated to a Cu(II) center with their alcoholic oxygens and imine nitrogens and formed six and five-membered chelate rings. At the same time, each ligand bridged to a neighboring Cu(II) atom by its alcoholic oxygen, thus the metal centers became penta-coordinated. The copper(II) complex with µ-ɳ2-hydroxo bridges and Cu .Cu distance about 3â¯Å was structurally similar to the active site of natural catechol oxidase enzyme and exhibited excellent catecholase activity in aerobic oxidation of 3,5-di-tert-butyl catechol to its o-quinone. The kinetics and mechanism of the oxidation of 3, 5-DTBCH2 catalyzed by [CuL]4 complex, were studied at four different temperatures from 283 to 313K by UV-Vis spectroscopy. Interaction of [CuL]4 complex with FS-DNA was investigated by UV-Vis and fluorescence spectroscopy, viscosity measurements, cyclic voltammetry (CV), circular dichroism (CD) and agarose gel electrophoresis. The main mode of binding of the complexes with DNA was intercalation. The interaction between [CuL]4 complex and bovine serum albumin (BSA) was studied by UV-Vis, fluorescence and synchronous fluorescence spectroscopic techniques. The results indicated a high binding affinity of the complex to BSA. In vitro anticancer activity of the complex was evaluated against A549, Jurkat and Ragi cell lines by MTT assay. The complex was remarkably active against the cell lines and can be a good candidate for an anticancer drug. Theoretical docking studies were performed to further investigate the DNA and BSA binding interactions.
Asunto(s)
Complejos de Coordinación/farmacología , Cobre/farmacología , ADN/metabolismo , Sustancias Intercalantes/farmacología , Albúmina Sérica Bovina/metabolismo , Animales , Catálisis , Catecol Oxidasa/química , Catecoles/química , Bovinos , Complejos de Coordinación/química , Cobre/química , Cristalografía por Rayos X , Sustancias Intercalantes/química , Modelos Moleculares , Oxidación-Reducción/efectos de los fármacos , TemperaturaRESUMEN
Before a population becomes extinct, there are hidden costs in the physiology at the individual level that provide valuable insights into their condition. Here, we study two dams with one species in common (Argia anceps Garrison, 1996) to evaluate whether their physiological condition differed (total protein quantity, prophenoloxidase (proPO) and phenoloxidase (PO) activity, and protein carbonylation) during two consecutive years. The first dam, "El Gallinero" (contaminated, C), contains organic input from mines and agricultural activity, whereas the second, "Paso de Vaqueros" (non-contaminated, NC), is part of a biosphere reserve. Although at a phenological level, some physiological differences were observed (2012 vs 2013), individuals from the contaminated population had less total protein (2012, median = 1.815 µg/µL; 2013, 0.081 µg/µL) and more carbonylations in their proteins (2012, median = 19.00 nmol/mg; 2013, median = 121.69 nmol/mg) compared with the non-contaminated population (protein quantity in 2012, median = 3.716 µg/µL; 2013, median = 0.054 µg/µL; protein carbonylations in 2012, median = 0.00 nmol/mg; 2013, median = 99.44 nmol/mg). However, no significant differences were found in prophenoloxidase (C, median = 0.002 Vmax; NC, median = 0.002 Vmax) and phenoloxidase activity (C, median = 0.002 Vmax; NC, median = 0.001 Vmax). In addition, the biological oxygen demand (BOD) and Zn were more elevated in the C than NC population (C, BOD = 11.7, Zn = 0.17; NC, BOD = 8, Zn = 0.14). The results show that the impact of human activity can be observed not only through the extinction of species, but also at the physiological level of the individuals composing the populations through the evaluation of biomolecular damage, which can be observed at a much shorter scale compared with species extinction.
Asunto(s)
Contaminación Ambiental/efectos adversos , Odonata/fisiología , Animales , Organismos Acuáticos , Catecol Oxidasa , Monitoreo del Ambiente , Precursores Enzimáticos , Proteínas de Insectos , México , Monofenol Monooxigenasa , Carbonilación ProteicaRESUMEN
Gladiolus grandiflorus L. is highly susceptible to Fusarium and losses caused by this disease varies from 60% to 100%. Injuries caused during harvest, transport and inadequate storage, facilitate infection. The dynamics of wound healing can reduce infection by Fusarium. The objective was to characterize the wound healing in corms of G. grandiflora stored under refrigeration and how it affects the entry and establishment of F. oxysporum f. sp. gladioli infection. Corms were wounded and stored at 12 ± 4°C and relative humidity of 90 ± 5%. Cell damage, fresh weight loss, respiration, phenolic compounds, tissue darkening, suberization, lignification and resistance to infection were evaluated. Wounds on corms caused transepidermal damage with collapse and cell death. Physiological (increased loss of mass and respiration) and biochemical changes (deposition of lignin and suberin, enzymatic activity) occurred in the cells neighboring those death by the injury. The injury caused gradual darkening of the tissue, injured and neighbor. Fusarium oxysporum infection decreased with wound healing. The healing of injured G. grandiflora corms stored at 12ºC occurs from the 3rd day after injury by the accumulation of suberin, lignin, and melanin, inhibiting F. oxysporum f. sp. gladioli infection.
Asunto(s)
Asparagaceae/microbiología , Fusarium/fisiología , Enfermedades de las Plantas/microbiología , Asparagaceae/enzimología , Biomasa , Catalasa/metabolismo , Catecol Oxidasa/metabolismo , Respiración de la Célula , Humedad , Peroxidasa/metabolismoRESUMEN
The soursop (Annona muricata L.) is a climacteric fruit that may undergo enzymatic browning during ripening, mainly by the activity of polyphenol oxidase (PPO). Soursop PPO was purified 160-fold by hydrophobic interaction and ion-exchange chromatography. The native structure has a molecular weight of 112 kDa corresponding to a dimeric structure. The protein has an optimum pH and temperature of 6.5 and 25°C, respectively; and activation energy of 40.97 kJ·mol-1 . The lowest Km value was observed for caffeic acid (0.47 mM); the best substrate was 4-methyl-catechol (1,067 U·mM-1 min-1 ). Inactivation assays showed that PPO was completely inactivated by tropolone, Na2 S2 O5 and ascorbic acid, and thermally at 55°C for <5 min, microwave exposure reduced activity to 57% at 70 W in 30 s and ultrasound treatment diminished activity to 43% at 120 W in 220 s. This study allows a better understanding of soursop PPO behavior and provides inactivation information. PRACTICAL APPLICATIONS: The conservation of fresh fruits is complicated due to the enzymatic reactions that are present in fruits, such as enzymatic browning. The enzymes responsible for these reactions can be inactivated by, different chemical compounds as well as by the use of emerging technologies, such as microwaves and sonication, which seek to satisfy the consumer needs to obtain fresh products with good nutritional characteristics and adequate safety.
Asunto(s)
Annona/enzimología , Catecol Oxidasa/química , Frutas/efectos de la radiación , Proteínas de Plantas/química , Annona/química , Annona/genética , Annona/efectos de la radiación , Catecol Oxidasa/aislamiento & purificación , Estabilidad de Enzimas , Conservación de Alimentos , Frutas/química , Frutas/enzimología , Frutas/genética , Cinética , Microondas , Peso Molecular , Proteínas de Plantas/aislamiento & purificación , Ondas UltrasónicasRESUMEN
BACKGROUND: Little is known about how human disease vectors will modify their life history patterns and survival capacity as a result of climate change. One case is that of Chagas disease, which has triatomine bugs and Trypanosoma cruzi as vectors and parasite, respectively. This work aimed to determine: (i) the activity of the prophenoloxidase system (prophenoloxidase and phenoloxidase activity, two indicators of immune ability) in three intestine regions (anterior midgut, posterior midgutand rectum) of the triatomine bug Meccus pallidipennis under three temperature conditions (20 °C, 30 °C and 34 °C) against two T. cruzi strains [ITRI/MX/14/CHIL (Chilpancingo) and ITRI/MX/12/MOR (Morelos)], and (ii) whether vector survival varies under these three temperatures after infection by these T. cruzi strains. RESULTS: Our results indicate that prophenoloxidase activity was lower at higher temperatures, that the level of prophenoloxidase activity elicited by each strain was different (higher in Chilpancingo than in Morelos strains), and that prophenoloxidase activity was more intense in the anterior midgut than in the posterior midgut or rectum. Survival rates were lower in insects maintained at higher temperatures and infected by Chilpancingo strains. CONCLUSIONS: These results indicate that climate change could lead to lower prophenoloxidase activity and survival rates in triatomines when infected with different T. cruzi strains, which could reduce the vector capacity of M. pallidipennis.