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PPO Activity (ppo + activity)
Selected AbstractsEffect of Nonthermal Treatment on the Molecular Properties of Mushroom PolyphenoloxidaseJOURNAL OF FOOD SCIENCE, Issue 5 2003N.K. Sun ABSTRACT To elucidate the mechanism of inactivation of mushroom polyphenoloxidase (PPO) by nonthermal treatment, PPO solutions were irradiated up to 10 kGy or pressurized at 600 MPa and 800 MPa, respectively. PPO activities were significantly affected by , irradiation, and treatment at 5 kGy decreased the activity by 93%. Treatment of PPO at 600 MPa decreased the activity slightly, yet 10 and 20 min treatments at 800 MPa decreased the activities by 28% and 43%, respectively. Circular dichroism study showed that nonthermal treatment altered the ellipticity values in the range of 210 and 225 nm, resulting in decrease of the ordered structure. Fluorescence spectroscopy indicated that nonthermal treatment quenched the emission intensity. [source] Polyphenol oxidase activity in grass and its effect on plant-mediated lipolysis and proteolysis of Dactylis glomerata (cocksfoot) in a simulated rumen environmentJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2006Michael RF Lee Abstract Little is known about the level or activity of polyphenol oxidase (PPO) in grasses and its potential impact on proteolysis and lipolysis. Six grass species were initially screened for PPO activity (740.6, 291.9, 213.6, 119.0, 16.3 and 6.5 U g,1 fresh weight (FW) for cocksfoot, hybrid ryegrass, Italian ryegrass, perennial ryegrass, timothy and tall fescue respectively). Cocksfoot, which expressed the highest activity, was then used to determine the effect of PPO on plant-mediated proteolysis and lipolysis in a simulated rumen environment. Sourced cocksfoot was macerated and incubated in an antibiotic-containing anaerobic medium with or without ascorbate to deactivate PPO in the dark at 39 °C over five time points. At each time point (0, 1, 2, 6 and 24 h), six replicate samples were destructively harvested; three of the replicates were used for lipid analysis and the other three for protein, free amino acid and bound phenol determination. Characterisation of the herbage showed PPO activities of 649.6 and 0 U g,1 FW, which were reflected in the extent of phenol (derived from quinones) binding to protein after 24 h of incubation, namely 65.1 and 29.6 mg bound phenol g,1 protein (P < 0.001) for cocksfoot and cocksfoot + ascorbate respectively. Proteolysis, measured as free amino acids released into the incubation buffer, was significantly reduced (P < 0.001) with increasing PPO activity, with values after the 24 h incubation of 0.03 and 0.07 mmol L,1 g,1 FW for cocksfoot and cocksfoot + ascorbate respectively. Lipolysis, measured as the proportional decline in the membrane lipid polar fraction, was likewise reduced (P < 0.001) with increasing PPO activity, with values after the 24 h incubation of 0.43 and 0.65 for cocksfoot and cocksfoot + ascorbate respectively. Changes that occurred in protein and the lipid fractions (polar fraction, monoacylglycerol + diacylglycerol, triacylglycerol and free fatty acids) during the incubations are also reported and discussed. These results support the selection of forages high in PPO activity to reduce protein and lipid losses in silo and potentially in the rumen. Copyright © 2006 Society of Chemical Industry [source] Degradative enzymatic activities in fresh-cut blood-orange slices during chilled-storageINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 5 2009Anna Eghle Catalano Summary Blood-orange fruits are suitable to fresh-cut fruit production because of their chemical compositions. Nevertheless, the main limitation of using freshly cut oranges is their susceptibility to juiciness loss and ascorbic acid degradation because of enzymatic alterations. The aim of this work is: to identify some of the enzymes causing the qualitative decay in blood-orange slices during 15 days of chilled storage (at 4 ± 0.5 °C and 85% RH); to investigate the susceptibility to the previous alterations of five blood-orange clones (Moro nucellare, Sanguinello nucellare, Tarocco arcimusa, Tarocco gallo and Tarocco meli) to select the most suitable one for fresh-cut production. The enzymes studied were: pectinmethylesterase (PME) as index of juiciness loss, ascorbate oxidase (AAO) as index of ascorbic acid's degradation and polyphenol oxidase (PPO) as browning index. As far as we know, the changes of AAO activity during chilled storage of blood-orange fresh-cut slices has not previously reported and studied. Different clones showed different enzymatic activities and quality changes during chilled-storage. In particular a low juiciness loss in orange slices was correlated with a lower PME activity, as described in T. meli clone, while a high degradation of ascorbic acid was correlated with an higher AAO activity, as described in T. gallo clone; PPO activity seemed to have no significant action in quality degradation. Tarocco meli was the most suitable clone to the fresh-cut blood-orange production because it has the lowest enzymatic activity (PME, PPO and AAO) and the highest sensorial quality. [source] POLYPHENOLOXIDASE ACTIVITY OF MINIMALLY PROCESSED ,JONAGORED' APPLES (MALUS DOMESTICA)JOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 1 2005A.M.C.N. ROCHA ABSTRACT The influence of three chemical dips using ascorbic acid (AA), citric acid (CA) and calcium chloride (CC) on the polyphenoloxidase (PPO) activity and on the total phenolic content of minimally processed (MP) apple (Malus domestica, cv. Jonagored) during cold storage was evaluated and a potential relationship with enzymatic browning was investigated. An ascorbic acid dip (42.6 mM) of 5 min duration was the most efficient chemical treatment in reducing the PPO activity of apple cubes. A 92% inhibition was achieved after 7 days of storage at 4C. All treatments were advantageous in comparison to the control in reducing color changes. Color changes, determined by absorbance at 420 nm (soluble pigments) and lightness (L) (insoluble pigments) of apple cubes treated with ascorbic acid were correlated with total phenolic content. No correlation was observed between PPO activity and tristimulus color parameters, browning index or total phenolic content of AA-treated apple cubes. [source] PHYSICAL, PHYSIOLOGICAL AND CHEMICAL CHANGES IN POTATO AS INFLUENCED BY ERWINIA CAROTOVORA INFECTIONJOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 5 2002F. NOURIAN Bacterial soft rot, caused by Erwinia carotovora ssp. carotovora (Ecc), is a major disease in stored potatoes. The pathogen causes different physical, physiological and chemical changes in potatoes, which may affect the acceptability of raw and processed products. This study was carried out to evaluate the effect of disease severity on different physico-chemical and physiological properties of raw and cooked potatoes and to select the parameters most responsive to disease severity. Potatoes were inoculated with bacteria and incubated at 20C for different lengths of time to produce different levels of disease. As incubation time increased the volume of disease (VDS) increased, which in turn influenced the respiration rate (RR). In both raw and cooked potatoes, the physical changes (texture and color) associated with the progress of disease were reduced hardness, firmness and L value, and increased a and b values and total color difference (,E). The chemical changes were reduced ascorbic acid and pH, and increased reducing sugars, total sugars and titrable acidity along with the activities of peroxidase and polyphenol oxidase. The changes in physical and chemical parameters of raw and cooked potatoes during storage were described by fractional conversion equation models. All parameters were quite sensitive to disease except reducing sugars, peroxidase and PPO activity. The correlation matrix indicated that several of the quality parameters were related and thus most of them could be successfully used to predict tuber quality from disease. [source] Browning Prevention by Ascorbic Acid and 4-Hexylresorcinol: Different Mechanisms of Action on Polyphenol Oxidase in the Presence and in the Absence of SubstratesJOURNAL OF FOOD SCIENCE, Issue 9 2007E. Arias ABSTRACT:, We have investigated the mechanism of action of 4-hexylresorcinol (4-HR) and ascorbic acid (AA) on the polyphenol oxidase (PPO) catalyzed oxidation of phenolic substrates. Incubation of PPO with 4-HR diminishes strongly PPO activity. This effect can be erroneously interpreted, due to the high affinity of 4-HR for PPO, as irreversible inactivation of PPO. However, PPO activity can be recovered by dialysis after incubation with 4-HR. 4-hexylresorcinol is a canonical enzyme inhibitor that binds preferentially to the oxy form of PPO. It is a mixed-type inhibitor, because it influences both apparent Vmax (1.26 compared with 0.4 units in the absence and presence of 4-HR, respectively) and Km values (0.28 mM compared with 0.97 mM in the absence and in the presence of 4-HR, respectively) of PPO. AA can prevent browning by 2 different mechanisms: In the absence of PPO substrates it inactivates PPO irreversibly, probably through binding to its active site, preferentially in its oxy form. In the presence of PPO substrates, AA reduces PPO oxidized reaction products, which results in a lag phase when measuring PPO activity by monitoring dark product formation but not when monitoring O2 consumption. The simultaneous use of both 4-HR and AA on PPO results in additive prevention of browning. [source] Stability of Copigmented Anthocyanins and Ascorbic Acid in Muscadine Grape Juice Processed by High Hydrostatic PressureJOURNAL OF FOOD SCIENCE, Issue 4 2007D. Del Pozo-Insfran ABSTRACT:, Intermolecular copigmentation is one of the mechanisms of stabilization of anthocyanins in nature and is also responsible for the characteristic color and stability of aged red wines. In the present study, the effect of polyphenol oxidase (PPO) activity on phytochemical stability of an ascorbic acid-fortified muscadine grape juice following high hydrostatic pressure (HHP) processing (400 and 550 MPa for 15 min) and after 21 d of storage at 25 °C was investigated. Addition of rosemary and thyme polyphenolic extracts (copigmentation) was evaluated as a means to stabilize anthocyanins and ascorbic acid during pressurization and subsequent storage. Polyphenolic extracts were partially purified in order to reduce their content of PPO substrates, and improve their stabilization properties within juice matrix. Overall PPO activity increased (3- and 2.5-fold) following HHP at 400 and 550 MPa, respectively, although it was significantly lower in copigmented treatments. Higher anthocyanin losses occurred at 400 (,70%) than at 550 MPa (,46%), which were correlated to antioxidant losses (r= 0.89). Similarly, greater ascorbic acid losses were observed at 400 (84%) than at 550 MPa (18%). Copigmentation increased anthocyanin retention in reference to pressurized controls (3- and 3.2-fold for rosemary and thyme treatments, respectively) and decreased ascorbic degradation (20 to 32%). In stored samples, higher anthocyanin content (>2-fold) and antioxidant capacity (>1.5-fold) was observed for copigmented treatments when compared to control juices. Addition of partially purified copigments increased muscadine grape juice color, antioxidant activity and also reduced phytochemical losses during HHP processing and storage. [source] Polyphenol Oxidase from Apple (Malus domestica Borkh. cv Bramley's Seedling): Purification Strategies and CharacterizationJOURNAL OF FOOD SCIENCE, Issue 1 2006Deirdre M. Ni Eidhin ABSTRACT Polyphenol oxidase (PPO) was isolated from Bramley's Seedling apples with 75.7-fold purification and 26.5% recovery by ammonium sulfate precipitation, phenyl sepharose chromatography, ion exchange chromatography, and hydroxyapatite chromatography. Molecular weight was estimated to be about 45 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE). Optimum PPO activity was at pH 6.5 and greater than 50% activity was retained during storage for 72 h at pH 5.5 to 6.5. Optimum temperature for activity was 30 °C and the enzyme had residual activity of greater than 50% during storage for 72 h at 20 °C to 30 °C and for 24 h at 40 °C to 50 °C. Of the substrates tested, activity was greatest with 4-methylcatechol followed by catechol, pyrogallol, and (,)epicatechin. The most effective inhibitors tested were sodium metabisulfite and ascorbic acid. [source] Morphological and Physiological Differentiation among Vegetative Compatibility Groups of Verticillium dahliae in Relation to V. longisporumJOURNAL OF PHYTOPATHOLOGY, Issue 7-8 2001K. Zeise Abstract Twenty-four isolates of Verticillium dahliae from various geographic regions and host origins were assigned to vegetative compatibility groups (VCG) based on complementation between nitrate-non-utilizing (nit) mutants. The VCG assignment was associated with clearly distinguishable morphological and physiological characteristics. Most isolates of VCG 2B produced deep black colonies, small conidia (3.62 × 0.02 , m), and spherical microsclerotia. They exhibited polyphenol oxidase (PPO) activity, high sporulation rate in shake cultures, fluorescence on sanguinarine-amended PDA (snPDA) and excretion of dark pigment on Czapek Dox agar. The isolates assigned to VCG 4B had significantly longer conidia (4.73 × 0.04 , m), spherical microsclerotia and white, fluffy colonies due to enhanced production of aerial mycelium. They excreted only traces of dark pigment, exhibited PPO activity, did not fluoresce on snPDA and had limited ability to sporulate in shake cultures. Except for lacking PPO activity the only heterokaryon self-incompatible isolate (HSI) was similar to VCG 4B. Ten isolates of Verticillium longisporum from cruciferous hosts did not produce nit mutants. They had clearly longer conidia (7.41 × 0.05 , m), formed elongate microsclerotia, and only poorly sporulated in shake cultures. They did not excrete dark pigment, lacked PPO, and failed to fluoresce on snPDA. The results indicate a clear morphological and physiological differentiation not only between the two species V. longisporum and V. dahliae but also among the VCGs of V. dahliae. [source] Inhibitory effect of phenolics extracted from sorghum genotypes on Aspergillus parasiticus (NRRL 2999) growth and aflatoxin productionJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 6 2007CV Ratnavathi Abstract The inhibitory activity of bioactive polyphenols present in six sorghum genotypes,two red (AON 486 and IS 620), two yellow (LPJ and IS 17779) and two white (SPV 86 and SPV 462) varieties,on Aspergillus parasiticus (NRRL 2999) growth and aflatoxin production was evaluated. In the first experiment the production of aflatoxins in the six sorghum genotypes after removal of surface phenolics by acidic methanol treatment was studied and compared with that in untreated grains. Aflatoxin production was found to be fourfold higher in treated grains. The total phenols and bioactive polyphenols extracted by acidic methanol were quantified using the Folin,Denis method and the bovine serum albumin,benzidine conjugate procedure respectively. In the second experiment the effect of extracted sorghum phenolics under in vitro conditions on fungal growth and aflatoxin production was studied at two concentrations (0.01% and 0.1%) of phenolics. Extracted phenolics added to yeast extract sucrose (YES) medium at 0.1% concentration showed an inhibitory effect on aflatoxin production. At 0.01% phenolic concentration, aflatoxin production was minimal on day 3 after infection. At other time points the aflatoxin content was similar to that in the control. At 9 days after infection the fungal biomass in IS 620 was significantly lower than that in the control. At 0.1% phenolic concentration, aflatoxin production was minimal and the red genotype IS 620 showed maximum resistance. Fungal biomass was lowest at all growth stages in IS 620 as compared with the control. Polyphenol oxidase (PPO) activity was not detected in A. parasiticus grown on YES medium (control). PPO activity was not induced in A. parasiticus by the addition of phenolics to the liquid culture medium (no PPO activity was detected in the culture medium). The inhibitory activity of bioactive polyphenols could be attributed to the lack of PPO enzyme in this fungus. Copyright © 2007 Society of Chemical Industry [source] Modelling the effect of superatmospheric oxygen concentrations on in vitro mushroom PPO activity,JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 14 2006Perla A Gómez Abstract The kinetics of polyphenol oxidase (PPO, EC 1.14.18.1) with respect to oxygen concentrations from 5 to 100% using chlorogenic acid (CGA) as substrate was examined. In vitro mushroom PPO activity was determined by measuring the consumption of oxygen during the oxidation reaction. A differential Michaelis,Menten model was fitted to the obtained total depletion curves. The product concentration as well as the concentration of oxygen had a clear inhibitory effect on the reaction rate. However, the inhibitory effect of oxygen was more evident at low product concentration. A linear mixed inhibition model that considered both the product (oxidised CGA) and oxygen as inhibitors was developed. A model with the product as a competitive inhibitor and oxygen as an uncompetitive inhibitor was the most appropriate to explain the reaction kinetics. The values of the inhibition constants calculated from the model were 0.0032 mmol L,1 for Km (Michaelis,Menten constant related to oxygen), 0.023 mmol L,1 for Kmc (constant for competitive inhibition due to the product), 1.630 mmol L,1 for Kmu (constant for uncompetitive inhibition due to oxygen) and 1.77 × 10,4 mmol L,1 s,1 for Vmax (maximum reaction rate). The results indicate that superatmospheric oxygen concentrations could be effective in preventing enzymatic browning by PPO. Copyright © 2006 Society of Chemical Industry [source] Polyphenol oxidase activity in grass and its effect on plant-mediated lipolysis and proteolysis of Dactylis glomerata (cocksfoot) in a simulated rumen environmentJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2006Michael RF Lee Abstract Little is known about the level or activity of polyphenol oxidase (PPO) in grasses and its potential impact on proteolysis and lipolysis. Six grass species were initially screened for PPO activity (740.6, 291.9, 213.6, 119.0, 16.3 and 6.5 U g,1 fresh weight (FW) for cocksfoot, hybrid ryegrass, Italian ryegrass, perennial ryegrass, timothy and tall fescue respectively). Cocksfoot, which expressed the highest activity, was then used to determine the effect of PPO on plant-mediated proteolysis and lipolysis in a simulated rumen environment. Sourced cocksfoot was macerated and incubated in an antibiotic-containing anaerobic medium with or without ascorbate to deactivate PPO in the dark at 39 °C over five time points. At each time point (0, 1, 2, 6 and 24 h), six replicate samples were destructively harvested; three of the replicates were used for lipid analysis and the other three for protein, free amino acid and bound phenol determination. Characterisation of the herbage showed PPO activities of 649.6 and 0 U g,1 FW, which were reflected in the extent of phenol (derived from quinones) binding to protein after 24 h of incubation, namely 65.1 and 29.6 mg bound phenol g,1 protein (P < 0.001) for cocksfoot and cocksfoot + ascorbate respectively. Proteolysis, measured as free amino acids released into the incubation buffer, was significantly reduced (P < 0.001) with increasing PPO activity, with values after the 24 h incubation of 0.03 and 0.07 mmol L,1 g,1 FW for cocksfoot and cocksfoot + ascorbate respectively. Lipolysis, measured as the proportional decline in the membrane lipid polar fraction, was likewise reduced (P < 0.001) with increasing PPO activity, with values after the 24 h incubation of 0.43 and 0.65 for cocksfoot and cocksfoot + ascorbate respectively. Changes that occurred in protein and the lipid fractions (polar fraction, monoacylglycerol + diacylglycerol, triacylglycerol and free fatty acids) during the incubations are also reported and discussed. These results support the selection of forages high in PPO activity to reduce protein and lipid losses in silo and potentially in the rumen. Copyright © 2006 Society of Chemical Industry [source] Cloning and some properties of Japanese pear (Pyrus pyrifolia) polyphenol oxidase, and changes in browning potential during fruit maturation,JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 11 2003Makiyo Nishimura Abstract A PCR-amplified genomic DNA fragment encoding Japanese pear (Pyrus pyrifolia) polyphenol oxidase (PPO) was cloned and sequenced. The DNA appears to encode a 66 kDa precursor protein consisting of a 56 kDa mature protein and a 9.5 kDa N-terminal transit peptide. The amino acid sequence showed high homology with apple PPO. The PPO mainly existed as a soluble fraction in cells and was limitedly proteolysed, while the mature form (56 kDa) was detected in plastids. Immature fruits showing high browning potential had high PPO activity and a high level of phenolics, while mature fruits showing little browning had high PPO activity but a low level of phenolics. Copyright © 2003 Society of Chemical Industry [source] | ||||||||||