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Medicinas Complementárias
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1.
Food Chem ; 446: 138866, 2024 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-38430769

RESUMEN

Fresh-cut potatoes are prone to surface browning and physiological degradation. Chlorogenic acid (CGA), a natural phenolic antioxidant, has demonstrated preservative properties in various postharvest products. However, the underlying mechanisms of its application on maintaining quality remain unclear. Therefore, the effect of exogenous CGA treatment on quality deterioration of potato slices and the mechanisms involved were investigated. Results revealed CGA treatment retarded the browning coloration, suppressed microbial growth and inhibited the declines in starch, and ascorbic acid contents in potato slices. Meanwhile, the treatment activated the phenylpropanoid pathway but decreased the activities of phenolic decomposition-related enzymes such as polyphenol oxidase (PPO) and tyrosinase and downregulated StPPO expression. Moreover, the treated slices exhibited reduced accumulation of reactive oxygen species and increased activity of antioxidant enzymes. Additionally, they displayed enhanced 2,2-diphenyl-1-picrylhydrazyl radicals scavenging capacity and higher ATP levels. Therefore, these findings indicated that CGA treatment was effective for quality maintenance and antioxidant capacity enhancement in fresh-cut potatoes, thereby providing potential strategies for the preservation and processing of fresh-cut produce.


Asunto(s)
Antioxidantes , Solanum tuberosum , Antioxidantes/metabolismo , Ácido Clorogénico/farmacología , Ácido Clorogénico/metabolismo , Solanum tuberosum/metabolismo , Fenoles/metabolismo , Ácido Ascórbico/metabolismo , Catecol Oxidasa/metabolismo
2.
Compr Rev Food Sci Food Saf ; 22(3): 2267-2291, 2023 05.
Artículo en Inglés | MEDLINE | ID: mdl-37043598

RESUMEN

Polyphenol oxidase (PPO) is a metalloenzyme with a type III copper core that is abundant in nature. As one of the most essential enzymes in the tea plant (Camellia sinensis), the further regulation of PPO is critical for enhancing defensive responses, cultivating high-quality germplasm resources of tea plants, and producing tea products that are both functional and sensory qualities. Due to their physiological and pharmacological values, the constituents from the oxidative polymerization of PPO in tea manufacturing may serve as functional foods to prevent and treat chronic non-communicable diseases. However, current knowledge of the utilization of PPO in the tea industry is only available from scattered sources, and a more comprehensive study is required to reveal the relationship between PPO and tea obviously. A more comprehensive review of the role of PPO in tea was reported for the first time, as its classification, catalytic mechanism, and utilization in modulating tea flavors, compositions, and nutrition, along with the relationships between PPO-mediated enzymatic reactions and the formation of functional constituents in tea, and the techniques for the modification and application of PPO based on modern enzymology and synthetic biology are summarized and suggested in this article.


Asunto(s)
Camellia sinensis , Catecol Oxidasa/metabolismo , Oxidación-Reducción ,
3.
Plant Physiol Biochem ; 198: 107702, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-37099880

RESUMEN

Pollen development is critical in plant reproduction. Polyphenol oxidases (PPOs) genes encode defense-related enzymes, but the role of PPOs in pollen development remains largely unexplored. Here, we characterized NtPPO genes, and then investigated their function in pollen via creating NtPPO9/10 double knockout mutant (cas-1), overexpression 35S::NtPPO10 (cosp) line and RNAi lines against all NtPPOs in Nicotiana tabacum. NtPPOs were abundantly expressed in the anther and pollen (especially NtPPO9/10). The pollen germination, polarity ratio and fruit weights were significantly reduced in the NtPPO-RNAi and cosp lines, while they were normal in cas-1 likely due to compensation by other NtPPO isoforms. Comparisons of metabolites and transcripts between the pollen of WT and NtPPO-RNAi, or cosp showed that decreased enzymatic activity of NtPPOs led to hyper-accumulation of flavonoids. This accumulation might reduce the content of ROS. Ca2+ and actin levels also decreased in pollen of the transgenic lines.Thus, the NtPPOs regulate pollen germination through the flavonoid homeostasis and ROS signal pathway. This finding provides novel insights into the native physiological functions of PPOs in pollen during reproduction.


Asunto(s)
Flavonoides , Nicotiana , Nicotiana/fisiología , Flavonoides/metabolismo , Catecol Oxidasa/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Polen/metabolismo , Homeostasis , Polifenoles/metabolismo
4.
Int J Biol Macromol ; 240: 124353, 2023 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-37059281

RESUMEN

Theaflavins (TFs) are important quality compounds in black tea with a variety of biological activities. However, direct extraction of TFs from black tea is inefficient and costly. Therefore, we cloned two PPO isozymes from Huangjinya tea, termed HjyPPO1 and HjyPPO3. Both isozymes oxidized corresponding catechin substrates for the formation of four TFs (TF1, TF2A, TF2B, TF3), and the optimal catechol-type catechin to pyrogallol-type catechin oxidation rate of both isozymes was 1:2. In particular, the oxidation efficiency of HjyPPO3 was higher than that of HjyPPO1. The optimum pH and temperature of HjyPPO1 were 6.0 and 35 °C, respectively, while those of HjyPPO3 were 5.5 and 30 °C, respectively. Molecular docking simulation indicated that the unique residue of HjyPPO3 at Phe260 was more positive and formed a π-π stacked structure with His108 to stabilize the active region. In addition, the active catalytic cavity of HjyPPO3 was more conducive for substrate binding by extensive hydrogen bonding.


Asunto(s)
Camellia sinensis , Catequina , Camellia sinensis/química , Catequina/química , Catecol Oxidasa/metabolismo , Isoenzimas , Simulación del Acoplamiento Molecular , Antioxidantes , Té/genética , Té/química , Clonación Molecular
5.
Molecules ; 28(4)2023 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-36838710

RESUMEN

Tea polyphenol (TPs) oxidation caused by polyphenol oxidase (PPO) in manufacturing is responsible for the sensory characteristics and health function of fermented tea, therefore, this subject is rich in scientific and commercial interests. In this work, an in vitro catalysis of TPs in liquid nitrogen grinding of sun-dried green tea leaves by PPO was developed, and the changes in metabolites were analyzed by metabolomics. A total of 441 metabolites were identified in the catalyzed tea powder and control check samples, which were classified into 11 classes, including flavonoids (125 metabolites), phenolic acids (67 metabolites), and lipids (55 metabolites). The relative levels of 28 metabolites after catalysis were decreased significantly (variable importance in projection (VIP) > 1.0, p < 0.05, and fold change (FC) < 0.5)), while the relative levels of 45 metabolites, including theaflavin, theaflavin-3'-gallate, theaflavin-3-gallate, and theaflavin 3,3'-digallate were increased significantly (VIP > 1.0, p < 0.05, and FC > 2). The increase in theaflavins was associated with the polymerization of catechins catalyzed by PPO. This work provided an in vitro method for the study of the catalysis of enzymes in tea leaves.


Asunto(s)
Biflavonoides , Catequina , Polifenoles/análisis , Catecol Oxidasa/metabolismo , Catequina/metabolismo , Biflavonoides/metabolismo , Flavonoides , Té/metabolismo , Antioxidantes
6.
Food Res Int ; 164: 112449, 2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-36738009

RESUMEN

In this study, the browning degrees of fresh-cut potatoes of different cultivars were investigated. Fresh-cut potatoes of the 'Huangjin' cultivar exhibited a higher browning index and sensory quality deterioration over time compared with 'Minshu' potatoes. 'Huangjin' exhibited a higher activity of browning-related enzymes such as polyphenol oxidase, tyrosinase, peroxidase, phenylalanine ammonia-lyase, phospholipase D (PLD), and lipoxygenase (LOX) than 'Minshu'. Furthermore, 'Minshu' exhibited lower H2O2 and malonaldehyde (MDA) contents, lower membrane lipid degradation and peroxidation, and delayed browning, attributable to its low PLD and LOX activities. The ultrastructure of 'Minshu' cells remained intact 7 h after cutting, while that of 'Huangjin' cells was severely damaged, and 'Minshu' cells exhibited more Golgi complexes and black particles than 'Huangjin' cells. Moreover, 'Huangjin' cells exhibited numerous multivesicular bodies, which were nonexistent in 'Minshu' cells. The results show that 'Minshu' potatoes feature a lower browning-related enzyme activity than 'Huangjin', and a tough cell structure to resist post-cut browning.


Asunto(s)
Antioxidantes , Solanum tuberosum , Solanum tuberosum/química , Peróxido de Hidrógeno , Peroxidasa/metabolismo , Fenilanina Amoníaco-Liasa/metabolismo , Catecol Oxidasa/metabolismo
7.
Planta ; 257(1): 13, 2022 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-36522558

RESUMEN

MAIN CONCLUSION: Screening for resistance in 40 potato genotypes to Rhizoctonia solani AG-3PT-stem-canker, antioxidant enzymes activity as well as total phenol compounds were documented. Rhizoctonia solani AG-3PT-stem-canker is one of the most devastating diseases that leads to severe economic losses in potatoes, Solanum tuberosum globally. Crop management and eugenic practices, especially the use of resistance can be effective in reducing the disease incidence. However, the information about potato-R. Solani interaction is still limited. This study explored screening for resistance in forty potato genotypes to R. solani, analyzing biomass growth parameters (BGPs), as well as antioxidant enzymes activity of which peroxidase/peroxide-reductases (POXs), superoxide dismutase (SOD), polyphenol oxidase (PPO), catalase (CAT), phenylalanine ammonia-lyase (PAL), ß-1,3-glucanase (GLU) and total phenol compounds (TPCs) were taken into account. In addition, we analyzed up-regulation of two gene markers (PR-1 and Osmotin), using reverse transcription quantitative PCR (RT-qPCR). For which, the resistant 'Savalan', partially resistant 'Agria', partially susceptible 'Sagita' and susceptible 'Pashandi' were selected to explore the trails in their roots and leaves over the time courses of 1, 2 and 3-weeks post inoculation (wpi) following inoculation. Cluster analysis divided potatoes into four distinct groups, based on disease severity scales (0-100%) significance. The BGPs, shoot and root length, fresh and dry weight, and root volume were also significantly higher in infected potatoes compared to non-inoculated controls. Antioxidant enzymes activity also indicated the highest increased levels for POX (fourfold at 3wpi), CAT (1.5-fold at 3wpi), SOD (6.8-fold at 1wpi), and PAL (2.7-fold at 3wpi) in the resistant genotype, 'Savalan', whereas the highest activity was recorded in TPC (twofold at 1 wpi), PPO (threefold at 3wpi), and GLU (2.3-fold at 1wpi) in partially resistant genotypes. Although the defense-related enzymatic activities were sharply elevated in the resistant and partially resistant genotypes following inoculation, no significant correlations were between the activity trends of the related enzymes. The two related gene markers also showed comprehensive transcriptional responses up to 3.4-fold, predominantly in resistant genotypes. Surprisingly, the PR-1 gene marker, basically resistant to Wilting agent Verticillium dahlia was overexpressed in resistant 'Savalan' and 'Agria' against R. solani AG3-PT. Similar results were obtained on Osmotin gene marker resistant to late-blight P. infestans, and early-blight Alternaria solani that similarly modulates immunity against R. solani. Furthermore, there was a significant correlation between resistance, enzyme activity, and gene expression in the aforesaid cultivars. Studying the physiological metabolic pathways of antioxidant enzymes activity appears to be an important direction in research to elucidate resistance to R. solani in potatoes.


Asunto(s)
Solanum tuberosum , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Resistencia a la Enfermedad/genética , Antioxidantes , Enfermedades de las Plantas , Rhizoctonia/fisiología , Fenilanina Amoníaco-Liasa/genética , Fenilanina Amoníaco-Liasa/metabolismo , Catecol Oxidasa/metabolismo , Superóxido Dismutasa , Fenoles , Mecanismos de Defensa
8.
Ultrason Sonochem ; 90: 106205, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36274416

RESUMEN

The shelf life of fresh-cut fruits and vegetables is affected by microbial growth, enzymatic browning, and loss of flavor. Although ultrasound (US) treatment is often used in the preservation of fresh-cut fruits and vegetables, it has limited antibacterial and preservative effects. Here, we used cactus polysaccharides (CP) to enhance the preservative effect of ultrasound treatment and extended the shelf life of fresh-cut potatoes. The results showed that combined treatment (CP + US) exerted better antimicrobial and anti-browning effects than individual treatments (either US or CP alone). In addition, CP + US has no adverse effect on texture and quality properties, as well as reduced the mobility of internal water. Combination treatment not only significantly decreased the activities of polyphenol oxidase and peroxidase (P < 0.05), but also maintained a high level of phenylalanine ammonia lyase activity and total phenol content during storage. It also maintained the integrity of cell membrane and reduced its permeability by inhibiting the peroxidation of membrane lipids. In addition, CP + US treatment significantly inhibited the activity of antioxidant enzymes and maintained a high DPPH scavenging ability. GC-IMS technology was used to evaluate the flavor of fresh-cut potatoes. The results showed that CP + US treatment reduced the production of a peculiar smell during storage and maintained a good flavor by inhibiting the production of aldehydes. Taken together, these results indicate that the effective preservation method of CP + US treatment can be utilized to increase the shelf life of fresh-cut potatoes.


Asunto(s)
Cactaceae , Solanum tuberosum , Solanum tuberosum/química , Catecol Oxidasa/metabolismo , Frutas/química , Verduras , Polisacáridos/farmacología , Polisacáridos/análisis
9.
Food Res Int ; 161: 111884, 2022 11.
Artículo en Inglés | MEDLINE | ID: mdl-36192997

RESUMEN

This work focuses on understanding the action of a novel seaweed extract with anti-browning functionality in fresh-cut apples. Organic fresh-cut apples were coated by immersion in an aqueous Codium tomentosum seaweed extract (0.5 % w/v), packaged under ambient atmospheric conditions in plastic bags, and stored at 4 ˚C for 15 days. Browning-related enzymatic activities, as well as targeted gene expression related to superficial browning, were monitored immediately after coating and followed at five-day intervals, until a final storage period of 15 days. Gene expression was particularly affected one hour after coating application (day 0), with no expression registered for peroxidase (mdPOD) and phenylalanine ammonia-lyase (mdPAL) genes in the coated samples. A reduction in polyphenol oxidase expression levels was also observed. After 15 days of storage, the coated samples developed lower browning levels and presented distinctly lower activities of polyphenol oxidase and peroxidase - the oxidative enzymes predominantly involved in enzymatic browning. The observed post-coating suppression of mdPAL and mdPOD expression, and reduction in mdPPO expression, suggest that the seaweed C. tomentosum extract delays the activation of these genes, and decreases enzymatic activity, which in turn accounts for the coating's anti-browning effect.


Asunto(s)
Malus , Catecol Oxidasa/genética , Catecol Oxidasa/metabolismo , Malus/metabolismo , Peroxidasa/metabolismo , Fenilanina Amoníaco-Liasa/genética , Fenilanina Amoníaco-Liasa/metabolismo , Extractos Vegetales/farmacología , Plásticos
10.
DNA Res ; 29(5)2022 Aug 23.
Artículo en Inglés | MEDLINE | ID: mdl-35980175

RESUMEN

Mucuna pruriens, commonly called velvet bean, is the main natural source of levodopa (L-DOPA), which has been marketed as a psychoactive drug for the clinical management of Parkinson's disease and dopamine-responsive dystonia. Although velvet bean is a very important plant species for food and pharmaceutical manufacturing, the lack of genetic and genomic information about this species severely hinders further molecular research thereon and biotechnological development. Here, we reported the first velvet bean genome, with a size of 500.49 Mb and 11 chromosomes encoding 28,010 proteins. Genomic comparison among legume species indicated that velvet bean speciated ∼29 Ma from soybean clade, without specific genome duplication. Importantly, we identified 21 polyphenol oxidase coding genes that catalyse l-tyrosine to L-DOPA in velvet bean, and two subfamilies showing tandem expansion on Chr3 and Chr7 after speciation. Interestingly, disease-resistant and anti-pathogen gene families were found contracted in velvet bean, which might be related to the expansion of polyphenol oxidase. Our study generated a high-quality genomic reference for velvet bean, an economically important agricultural and medicinal plant, and the newly reported L-DOPA biosynthetic genes could provide indispensable information for the biotechnological and sustainable development of an environment-friendly L-DOPA biosynthesis processing method.


Asunto(s)
Mucuna , Catecol Oxidasa/genética , Catecol Oxidasa/metabolismo , Cromosomas/metabolismo , Dopamina/metabolismo , Levodopa/genética , Levodopa/metabolismo , Mucuna/genética , Mucuna/metabolismo , Preparaciones Farmacéuticas/metabolismo , Investigación , Tirosina/genética , Tirosina/metabolismo
11.
Ultrason Sonochem ; 89: 106111, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35998484

RESUMEN

Lyophyllum decastes is a common mushroom that is prone to browning during prolonged storage. In this study, the effects of ultrasonic treatment on metabolic gene expression, enzyme activity, and metabolic compounds related to L. decastes browning were investigated. Treatment of the fruiting body at 35 kHz and 300 W for 10 min reduced the browning index of L. decastes by 21.0 % and increased the L* value by 11.1 %. Ultrasonic treatment of the fruiting body resulted in higher levels of total phenols, flavonoids, and 9 kinds of amino acid with catalase (CAT) and peroxidase (POD) activities maintained at high levels. Higher cytochrome c oxidase (CCO), succinate dehydrogenase (SDH), phosphofructokinase (PFK), and pyruvate kinase (PK) activities may be ascribed to increased antioxidant capacity. Moreover, ultrasonication retained higher adenosine triphosphate (ATP) concentrations with an increased energy charge, while there were lower levels of adenosine diphosphate (ADP) and reduced and oxidized nicotinamide adenine dinucleotide (NADH and NAD+), respectively. Meanwhile, lower lignin contents were observed, along with retarded polyphenol oxidase (PPO) and lipoxygenase (LOX) activities. Lower PPO activity reduced the fruiting body enzymatic browning rate through decreased expression of LdPpo1, LdPpo2, and LdPpo3 during storage at 4 °C for 16 days. This activity may be used to determine the effectiveness of ultrasonication.


Asunto(s)
NAD , Succinato Deshidrogenasa , Adenosina Difosfato/metabolismo , Adenosina Trifosfato/metabolismo , Agaricales , Aminoácidos/metabolismo , Antioxidantes/metabolismo , Catalasa/metabolismo , Catecol Oxidasa/metabolismo , Complejo IV de Transporte de Electrones/metabolismo , Metabolismo Energético , Flavonoides , Lignina/metabolismo , Lipooxigenasas/metabolismo , NAD/metabolismo , Fenoles/química , Fosfofructoquinasas/metabolismo , Piruvato Quinasa/metabolismo , Succinato Deshidrogenasa/metabolismo , Ultrasonido
12.
Sci Rep ; 12(1): 12870, 2022 07 27.
Artículo en Inglés | MEDLINE | ID: mdl-35896690

RESUMEN

Tea is a widely consumed beverage prepared using the fresh leaves of Camellia sinensis L. Tea plants are classified into small- and large-leaf varieties. Polyphenol oxidase (PPO), a crucial enzyme in tea manufacturing, catalyzes essential phenolic metabolites into different derivatives. To compare the molecular characteristics of CsPPO between cultivars, we cloned the full-length sequence of CsPPO cDNA from four representative tea cultivars in Taiwan. Amino acid sequence alignment analyses indicated that CsPPO is highly conserved. PPO exhibited similar enzymatic activity in different tea cultivars. Quantitative real-time polymerase chain reaction revealed no significant differences in the CsPPO transcript level between the small- and large-leaf varieties. However, tea harvested in summer and from low-altitude areas had a higher CsPPO transcript level than that harvested in winter and from high-altitude areas. Regulation of CsPPO by temperature was more significant in the small-leaf variety than in the large-leaf variety. The content of flavonoids and the expression of chalcone synthase, anthocyanidin synthase, and anthocyanidin reductase genes involved in the tea flavonoid biosynthesis pathway were higher in the large-leaf than in the small-leaf varieties, suggesting that the large-leaf variety had a higher antioxidative capacity than did the small-leaf variety. Our study compared the molecular properties of CsPPO between two tea varieties and clarified the physiological role of PPO in tea.


Asunto(s)
Camellia sinensis , Catecol Oxidasa , Camellia sinensis/metabolismo , Catecol Oxidasa/genética , Catecol Oxidasa/metabolismo , Regulación de la Expresión Génica de las Plantas , Hojas de la Planta/metabolismo , Proteínas de Plantas/metabolismo , Té/genética , Té/metabolismo
13.
Food Chem ; 389: 133076, 2022 Sep 30.
Artículo en Inglés | MEDLINE | ID: mdl-35489264

RESUMEN

Seabuckthorn extract is rich in bioactive compounds and well known for its health benefits. The study investigated the effect of seabuckthorn leaf extract on browning of fresh-cut potatoes. The results showed that seabuckthorn leaf extract significantly inhibited the browning of fresh-cut potatoes compared with seabuckthorn fruit extract. Catechin, hypericin, gallic acid, casuarinin and isorhamnetin were main components in seabuckthorn leaf extract. Further research revealed that seabuckthorn leaf extract competitively inhibited polyphenol oxidase (PPO) with IC50 value of 0.7 mg/mL. Molecular docking indicated that gallic acid stably bound to the active site of PPO, while isorhamnetin had low affinity on PPO. These results also demonstrated that seabuckthorn leaf extract inhibited browning of fresh-cut potatoes by reducing activities of peroxidase and phenylalanine ammonia-lyase, decreasing contents of phenolics and elevating antioxidant capacity. In addition, synergistic anti-browning effect was found with casuarinin, isorhamnetin, gallic acid and pedunculagin.


Asunto(s)
Almacenamiento de Alimentos , Hippophae , Extractos Vegetales , Solanum tuberosum , Catecol Oxidasa/metabolismo , Ácido Gálico/metabolismo , Hippophae/química , Reacción de Maillard , Simulación del Acoplamiento Molecular , Extractos Vegetales/química
14.
Food Chem ; 383: 132277, 2022 Jul 30.
Artículo en Inglés | MEDLINE | ID: mdl-35168045

RESUMEN

Although Galla rhois has been used as a traditional medicine in Asian countries, there was no application of it in anti-browning food additives. Here, we tested whether Galla rhois inhibits apple juice browning. Apple juice browning was blocked at 250-1000 µg/ml of Galla rhois for 16 days but the effect of vitamin C did not last until a day. In vitro assays showed that the antioxidant capacity of Galla rhois was stronger than that of vitamin C. Further analysis by UPLC-MS/MS identified 17 phytochemicals containing gallotannin derivatives. Docking simulation and polyphenol oxidase activity assay indicate that the mechanisms underlying Galla rhois-mediated inhibition of the enzymatic browning include but are not limited to the combined effects of multiple compounds including galloylglucose- and gallate-derivates. Although marketability and long-term toxicity of Galla rhois should be tested, it may be applied as a food additive to elevate food quality.


Asunto(s)
Malus , Ácido Ascórbico , Productos Biológicos , Catecol Oxidasa/metabolismo , Cromatografía Liquida , Aditivos Alimentarios/farmacología , Malus/química , Espectrometría de Masas en Tándem , Agua
15.
Prep Biochem Biotechnol ; 52(9): 1019-1034, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35015975

RESUMEN

Polyphenol oxidase (PPO) was firstly purified from damson plum as a high antioxidant source. PPO was treated by 0-80% ammonium sulfate precipitation and dialysis. Characterization results were determined for catechol, 4-methyl catechol, pyrogallol and caffeic acid as 0.05 M/pH: 7.2/25 °C; 0.2 M/pH: 4.5/10 °C; 0.01 M/pH: 6.8/5 °C, and 0.2 M/pH: 8.5/10 °C, respectively. Vmax and KM values were calculated for same substrates as 17,219.97 U/(mL*min) and 11.67 mM; 7309.72 U/(mL*min) and 5 mM; 12,580.12 U/(mL*min) and 3.74 mM; 12,100.41 U/(mL*min) and 6.25 mM, respectively. Catechol gave the highest Vmax value among substrates. Affinity purification was performed by using Sepharose 4B-L-Tyrosine-p-aminobenzoic acid and Sepharose 6B-L-Tyrosine-p-aminobenzoic acid. Single bands were approximately observed at 50 kDa for each affinity sample in SDS-PAGE and Native-PAGE. 93.88 and 10.46 purification-folds were obtained for PPO by reference Sepharose-4B and original Sepharose-6B gels. Metal effects upon PPO activity were also investigated due to the importance of enzymatic browning in foods. Cu+2 activation and Fe+2 inhibition were observed with a final metal concentration of 1 mM at 219.66 and 43.18%, respectively. PPO purification from damson plum by affinity chromatography, its characterization, stability evaluation by statistically, and effects of metal ions on damson plum PPO have not been investigated in the literature.


Asunto(s)
Catecol Oxidasa , Prunus domestica , Ácido 4-Aminobenzoico , Sulfato de Amonio , Antioxidantes , Catecol Oxidasa/metabolismo , Catecoles , Cromatografía de Afinidad , Geles , Guayacol , Concentración de Iones de Hidrógeno , Cinética , Peso Molecular , Prunus domestica/metabolismo , Pirogalol , Diálisis Renal , Sefarosa , Especificidad por Sustrato , Tirosina
16.
Biotechnol Appl Biochem ; 69(5): 2249-2256, 2022 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-34775655

RESUMEN

Fresh-cut vegetables and fruits have gained attention among consumers because of their fresh appearance, lack of pollution, nutrition, and convenience. However, in fresh-cut foods, enzymatic browning is the main problem. Polyphenol oxidase (PPO) is a vital enzyme involved in the process of enzymatic browning. In this study, PPO was purified from potato using Sepharose 4B-l-tyrosine-p-aminobenzoic acid affinity chromatography and the effect of some indazoles on the enzyme was determined. The enzyme was purified with a specific activity of 52,857.14 EU/mg protein and 21.26-purification fold. Indazoles exhibited inhibitor properties for PPO with IC50 values in the range of 0.11-1.12 mM and Ki values in the range of 0.15 ± 0.04-3.55 ± 0.88 mM. Among these compounds, 7-chloro-1H-indazole was shown as the most potent PPO inhibitor (Ki : 0.15 ± 0.04 mM). Determination of the enzyme's inhibition kinetics will simplify the testing of candidate PPO inhibitors.


Asunto(s)
Catecol Oxidasa , Solanum tuberosum , Catecol Oxidasa/metabolismo , Solanum tuberosum/metabolismo , Indazoles/farmacología , Frutas/metabolismo
17.
Biomolecules ; 11(9)2021 08 24.
Artículo en Inglés | MEDLINE | ID: mdl-34572470

RESUMEN

In this study, the co-application of chitosan and tetramycin against kiwifruit soft rot and its effects on the disease resistance, growth, quality and aroma of kiwifruit were investigated. The results show that chitosan could effectively enhance tetramycin against soft rot of kiwifruit with the field control efficacy of 85.33% for spraying chitosan 100 time + 0.3% tetramycin AS 5000-time dilution liquid, which was higher than 80.99% for 0.3% tetramycin AS 5000-time dilution liquid and significantly (p < 0.01) higher than 40.66% for chitosan 100-time dilution liquid. Chitosan could significantly (p < 0.05) improve the promoting effects of tetramycin on total phenolics, total flavonoids, SOD activity of kiwifruit compared to tetramycin during storage for 0-28 days and enhance the disease resistance of kiwifruit. Moreover, the co-application of chitosan and tetramycin was more effective than tetramycin or chitosan alone in enhancing fruit growth, improving fruit quality and increasing fruit aroma. This study highlights that chitosan can be used as an adjuvant to enhance tetramycin against soft rot of kiwifruit and promote tetramycin's improvement for the single fruit volume and weight, vitamin C, soluble sugar, soluble solid, dry matter, soluble protein, titratable acidity and aroma of kiwifruit.


Asunto(s)
Actinidia/microbiología , Quitosano/farmacología , Frutas/microbiología , Macrólidos/farmacología , Odorantes , Enfermedades de las Plantas/microbiología , Actinidia/efectos de los fármacos , Actinidia/enzimología , Actinidia/crecimiento & desarrollo , Catecol Oxidasa/metabolismo , Quitosano/toxicidad , Flavonoides/análisis , Frutas/efectos de los fármacos , Frutas/enzimología , Macrólidos/toxicidad , Fenoles/análisis , Superóxido Dismutasa/metabolismo
18.
J Food Sci ; 86(8): 3529-3539, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-34272733

RESUMEN

Surface browning is a vital phenomenon that adversely reduces the quality of fresh-cut potatoes. Although many anti-browning methods have been explored, it is unclear whether exogenous catalase (CAT) treatment influences the enzymatic browning. Our results showed that 0.05% CAT immersion for 5 min alleviated browning during cold storage (4°C, 8 days), which was accompanied by a higher lightness and lower redness; additionally, lower H2 O2 and O2 ·- contents were found. The activities of CAT, ascorbate peroxidase, and glutathione peroxidase and the scavenging efficiency of 2,2-diphenyl-1-picrylhydrazyl were also increased. Moreover, CAT treatment inhibited the activities of polyphenol oxidase, peroxidase, and phenylalanine ammonia lyase and reduced phenol accumulation. Treatment with 0.1% hydrogen peroxide (H2 O2 ) achieved the opposite results. This is the first report of CAT application reducing fresh-cut potato browning, providing a safe treatment alternative for enzymatic discoloration and preliminarily revealing the underlying mechanism with insight into antioxidant regulation. PRACTICAL APPLICATION: This research is helpful for fresh-cut potato producers because a novel, safe, easy-to-carry out anti-browning solution was proposed. Dipping in 0.05% catalase solution for 5 min revealed color improvement in the quality of fresh-cut potato slices. The mechanism may rely on enhancing antioxidant ability (ascorbate peroxidase, and glutathione peroxidase, and 2,2-diphenyl-1-picrylhydrazyl scavenging), reducing reactive oxygen species (H2 O2 , O2 ·-, malondialdehyde) and controlling enzymatic browning reaction factors (polyphenol oxidase, peroxidase, and phenylalanine ammonia lyase, and phenol accumulation). This method shows promise for better meeting the requirements and demands of consumers for fresh quality products.


Asunto(s)
Catalasa , Manipulación de Alimentos , Solanum tuberosum , Catalasa/farmacología , Catecol Oxidasa/metabolismo , Activación Enzimática/efectos de los fármacos , Manipulación de Alimentos/métodos , Calidad de los Alimentos , Malondialdehído/metabolismo , Fenilanina Amoníaco-Liasa/metabolismo
19.
Sci Rep ; 11(1): 14166, 2021 07 08.
Artículo en Inglés | MEDLINE | ID: mdl-34238955

RESUMEN

Short vase life, capitulum wilting, neck bending, and postharvest chilling injury (CI) are major disorders have negative impact on quality and marketing of gerbera cut flowers. Low storage temperatures prolonging the vase life, but on the other hand leads serious CI which decreases the quality and consumer preferences. Spermine (SPER) and γ-aminobutyric acid (GABA) were identified as anti-aging factors delay the senescence and elevate the chilling tolerance in many species. Greenhouse-grown gerbera cv. 'Stanza' sprayed with 2 mM SPER and 1 mM GABA twice (2 T) or thrice (3 T). Cut flowers were stored at 1.5 °C and 8 °C postharvest to study the effects of GABA and SPER on senescence and CI. Vase life, CI and quality of cut flowers were improved by GABA and SPER treatments. No CI was observed in GABA-treated flowers at 1.5 °C; while, flowers sprayed with water showed severe CI. GABA treatments efficiently prolonged the vase life for 6-7 days more than the control (15 days). GABA and SPER increased the fresh weight, solution uptake, protein and proline contents, catalase, peroxidase, and superoxide dismutase activities, while decreased the electrolyte leakage, H2O2, and malondialdehyde contents, polyphenol oxidase, lipoxygenase, and phospholipase D activities. GABA and SPER significantly prolonged the vase life and prevented degradation of proteins and chilling damage and increased capacity of detoxifying and scavenging of H2O2 and reactive oxygen species (ROS), led to alleviate the negative consequences of the senescence and CI.


Asunto(s)
Asteraceae/crecimiento & desarrollo , Criopreservación , Flores/fisiología , Espermina/farmacología , Ácido gamma-Aminobutírico/farmacología , Antioxidantes/metabolismo , Asteraceae/efectos de los fármacos , Biomasa , Catalasa/metabolismo , Catecol Oxidasa/metabolismo , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Electrólitos/metabolismo , Flores/efectos de los fármacos , Peróxido de Hidrógeno/metabolismo , Lipooxigenasa/metabolismo , Malondialdehído/metabolismo , Modelos Biológicos , Peroxidasa/metabolismo , Fosfolipasa D/metabolismo , Proteínas de Plantas/metabolismo , Prolina/metabolismo , Superóxido Dismutasa/metabolismo
20.
Food Chem ; 362: 130224, 2021 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-34098439

RESUMEN

This study evaluated the feasibility of curcumin based photodynamic sterilization technology (PDT) applied to fresh-cut potato slices. Potato samples with 30 µmol L-1 curcumin solution were exposed to 420 nm light emitting diodes (LED) at a total dose of 0.7 kJ cm-2. Results showed that PDT inactivated 2.43 log CFU mL-1 of Escherichia coli (BL 21) and 3.18 log CFU mL-1 of Staphylococcus aureus and maintained the color, texture, weight as well as total solid content of treated potatoes. Additionally, loss of phenols and flavonoids was significantly prevented, increasing the total antioxidant capacity. This was attributed to changes in enzyme activity that PDT decreased the activity of polyphenol oxidase (PPO) and peroxidase (POD) by 59.7% and 47.8% and increased the activity of phenylalanine ammonia-lyase (PAL). Therefore, curcumin-based PDT has the potential to maintain the commercial quality of producing and achieving microbiological safety.


Asunto(s)
Antioxidantes/química , Curcumina/farmacología , Conservación de Alimentos/métodos , Solanum tuberosum/química , Solanum tuberosum/microbiología , Antibacterianos/farmacología , Catecol Oxidasa/metabolismo , Color , Escherichia coli , Flavonoides/análisis , Flavonoides/química , Conservación de Alimentos/instrumentación , Calidad de los Alimentos , Peroxidasa/química , Peroxidasa/metabolismo , Fenoles/análisis , Fenoles/química , Fenilanina Amoníaco-Liasa/metabolismo , Solanum tuberosum/efectos de los fármacos , Solanum tuberosum/metabolismo , Staphylococcus aureus
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