Multiple forms of soluble monophenol, dihydroxyphenylalanine: oxygen oxidoreductase (EC 1.14.18.1) from potato tubers (Solanum tuberosum). IV. Association and dissociation phenomena.

The soluble phenol oxidase of various potato juices (adjusted from physiological pH to pH 4.5, 7.0 and 7.8) was separated by gel chromatography into multiple molecular forms. In acid or neutral and alkaline potato juices, low-mol.-wt. (less than 150,000 daltons) or high-mol.-wt. (greater than 150,000 daltons) enzyme forms predominate, respectively. Conversion of the low-mol.-wt. enzyme forms into high-mol.-wt. enzyme forms, and vice versa, was achieved by changing the pH values from acidic to neutral or alkaline pH, and vice versa. This substantiated our previous idea that the enzyme multiplicity arises from association of various subunits. In alkaline potato juice, considerable loss of monophenol oxidase activity (assayed at pH 6.0) occurred. This confirmed our previous findings that o-diphenol oxidase is more alkali-stable than monophenol oxidase.     

Studies on enzymic browning of potatoes (Solanum tuberosum)

Summary Using analytical data from the literature the tyrosine turnover was calculated by a method published previously by us for 72 potato samples with different rates of browning. The samples included 9 varieties grown at three locations in 1969, which were analysed after harvest and after different times of storage at three temperatures. For 58 samples (81%) this calculation led to the same classification of the varieties as did visual observation of the rate of discolouration. It is concluded that enzymic browning of potatoes is correlated rather with tyrosine turnover, which depends on the concentrations of phenol oxidase, tyrosine, chlorogenic acid, and ascorbic acid, than with any single parameter.

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Untersuchungen zur enzymatischen Bräunung bei Kartoffeln (Solanum tuberosum)IV. Beziehung zwischen Tyrosinumsatz und Bräunung

Zusammenfassung Aus Analysendaten der Literatur wurde für 72 unterschiedlich bräunende Kartoffelproben der Tyrosinumsatz nach einer von uns kürzlich veröffentlichten Methode berechnet. Die Proben beinhalten 9 Sorten aus 3 Standorten im Jahr 1969, die nach der Ernte and nach verschiedenen Lagerzeiten bei 3 verschiedenen Temperaturen analysiert wurden. Die Berechnung des Tyrosinumsatzes führte bei 58 Proben (81%) zur gleichen Klassifizierung der Sorten wie die sensorische Beurteilung der Bräunung. Es wird gefolgert, daß die enzymatische Bräunung bei Kartoffeln eher mit dem Tyrosinumsatz, der von den Konzentrationen an Phenoloxydase, Tyrosin, Chlorogensäure und Ascorbinsäure abhängig ist, als mit einem einzelnen Parameter korreliert ist.

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Dedicated to Prof. Dr. L. Acker on his 65 ^th birthday     

Multiple forms of soluble monophenol, dihydroxyphenylalanine: oxygen oxidoreductase (EC 1.14.18.1) from potato tubers (Solanum tuberosum). III. Influence of pH on the molecular weight distribution of enzyme activity in potato juice.

Gel chromatography on Sepharose and on Sephadex was used to separate the soluble phenol oxidase in various potato juices into multiple molecular forms ranging from 36,000 to 800,000 daltons. Adjustment of potato juice from physiological pH (ca. 6) to pH 4.5 or to pH 7.8 resulted in the predominance of low-mol.-wt. (less than 150,000 daltons) or high-mol.-wt. (greater than 150,000 daltons) enzyme forms, respectively. This suggests association phenomena of subunits. In potato juice of physiological pH and in potato juice adjusted to pH 4.5, all enzyme forms exhibited both monophenol and o-diphenol oxidase activities (assayed at pH 6.0). In potato juice adjusted to pH 7.8 considerable loss of monophenol oxidase activity (assayed at pH 6.0) occurred. This suggests that o-diphenol oxidase is more alkali-stable than monophenol oxidase. The significance of these findings for enzyme purifications and for the in vivo action of the enzyme is discussed.     

Multiple forms of soluble monophenol, dihydroxyphenylalanine: oxygen oxidoreductase (EC 1.14.18.1) from potato tubers (Solarium tuberosum)

Summary The soluble phenol oxidase of various potato juices (adjusted from physiological pH to pH 4.5, 7.0 and 7.8) was separated by gel chromatography into mulitple molecular forms. In acid or neutral and alkaline potato juices, low-mol.-wt. (< 150,000 daltons) or high-mol.-wt. (> 150,000 daltons) enzyme forms predominate, respectively. Conversion of the low-mol.-wt. enzyme forms into high-mol.-wt. enzyme forms, and vice versa, was achieved by changing the pH values from acidic to neutral or alkaline pH, and vice versa. This substantiated our previous idea that the enzyme multiplicity arises from association of various subunits. In alkaline potato juice, considerable loss of monophenol oxidase activity (assayed at pH 6.0) occurred. This confirmed our previous findings thato-diphenol oxidase is more alkali-stable than monophenol oxidase.

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Multiple Formen löslicher Monophenol, Dihydroxyphenylalanin: O₂-Oxidoreductase (EC 1.14.18.1) aus Kartoffeln (Solarium tuberosum)IV. Assoziations- und Dissoziations-Phänomene

Zusammenfassung L:osliche Phenoloxydase verschiedener Kartoffelsäfte (von physiologischem pH eingestellt auf pH 4,5, 7,0 und 7,8) wurde durch Gelchromatographie aufgetrennt. In saurem bzw. neutralem bis alkalischem Saft lagen vorwiegend niedermolekulare (< 150 000 Dalton) bzw. hochmolekulare (> 150 000 Dalton) Enzymformen vor. Durch pH-Wechsel vom sauren in den neutralen bis alkalischen Bereich wurden niedermolekulare Formen in hochmolekulare überführt und umgekehrt. Dies bestätigte unsere früheren Vorstellungen, daß die Enzymmultiplizität aus der Assoziation verschiedener Untereinheiten herrührt. Im alkalischen Kartoffelsaft traten beträchtliche Verluste an Monophenoloxydase-Aktivität (gemessen bei pH 6,0) auf. Dies bestätigte unsere früheren Ergebnisse, daß dieo-Diphenoloxydase-Aktivität alkalistabiler ist als die Monophenoloxydase-Aktivität.

     

Multiple forms of soluble monophenol, dihydroxyphenylalamine: oxygen-oxidoreductase (EC 1.14.18.1) from potato tubers (Solanum tuberosum). II. Partial characterization of the enzyme forms with different molecular weights.

Gel chromatography on Sephadex G-200 was used to separate a soluble phenoloxidase from potatoes (var. Maritta) into at least six active fractions with dopa (dihydroxyphenylalanine) as substrate. Only high-molecular-weight-enzyme forms exhibited monophenoloxidase activity. Re-chromatography of the highest-molecular-weight form gave the same molecular weight distribution as with the crude enzyme. The molecular weights indicate association phenomena of subunits with a molecular weight of about 36000 daltons. According to polyacrylamide-gel electrophoresis, several monomeric forms with differnt isoelectric points seem to be present. This suggests that the large number of multiple forms of the enzyme arises from various combinations of identical and/or different subunits. SDS polyacrylamide gel electrophoresis failed to show the monomeric forms; the dimer and higher oligomers were obtained.     

Studies on enzymic browning of potatoes (Solanum tuberosum). II. The quantitative relationship between browning and its causative factors (author's transl)]

Ten potato varieties, with different rates of browning, were analyzed quantitatively for phenoloxidase, tyrosine, chlorogenic acid, caffeic acid, and for reducing substances (ascorbic acid). The rate of tyrosine turnover was calculated from the data. The fact that the further reactions of the primary oxidation products leading to browning only take place after complete oxidation of the reducing substances, was taken into account. This leads to the same classification of the varieties as does visual observation of the rate of discolouration. Thus a clear relationship between browning and potato constituents is demonstrated.     

Studies on enzymic browning of potatoes (Solanum tuberosum). I. Phenoloxidases and phenolic compounds from different varieties (author's transl)]

31 samples of potato varieties with slow, medium and fast rates of browning were studies. Characteristic enzyme patterns were obtained from polyacrylamide gel electrophoresic of the phenoloxidases of varieties with different discolouration rates. The differences lie mainly in the intensities of the enzyme bands. The qualitative determinaiton of the phenols showed no significant differences. Tyrosine, chlorogenic acid and caffeic acid produce coloured oxidation products; the characteristic colour gradations of in vivo browning were only observed in the presence of tyrosine. It is concluded that the same reactions take place during the discolouration of all the varieties.     

Effect of triacontanol on sprout growth in potatoes (Solanum tuberosum)

The effects of a 3-min dip in various concentrations (0–1–100 mg litre^?1) of a triacontanol emulsion on sprout growth from single-eye pieces of seed potato (Solanum tuberosum L) tubers were assessed after 10 days' growth in acid-washed sand. Treatment effects were compared with those resulting from immersion for 3 min in a mixture of gibberellic acid (6 mg litre^?1) and ethrel (4 mg litre^?1). Triacontanol dips were more effective than gibberellic acid and ethrel. Treatments increased the fresh weight of sprouts but had little effect on sprout number per tuber fragment. The optimum concentration of triacontanol was 10 mg litre^?1.     

Determination of anthocyanins,flavonoids and phenolic acids in potatoes. I: Coloured cultivars of Solanum tuberosum L

The major anthocyanins, flavonoids and phenolic acids in the tubers (skin and flesh), flowers and leaves of 26 cultivars of Solanum tuberosum L with coloured skins and/or flesh have been identified and quantified using analytical HPLC. Red tubers contained mostly pelagonidin-3-(p-coumaroyl-rutinoside)-5-glucoside (200–2000 μg g⁻¹ FW) plus lesser amounts of peonidin-3-(p-coumaroyl-rutinoside)-5-glucoside (20–400 μg g⁻¹ FW). Light to medium purple tubers contained petunidin-3-(p-coumaroyl-rutinoside)-5-glucoside (1000–2000 μg g⁻¹ FW) plus small amounts of malvidin-3-(p-coumaroyl-rutinoside)-5-glucoside (20–200 μg g⁻¹ FW) whilst dark purple–black tubers contained similar levels of petunidin-3-(p-coumaroyl-rutinoside)-5-glucoside together with much higher concentrations of malvidin-3-(p-coumaroyl-rutinoside)-5-glucoside (2000–5000 μg g⁻¹ FW). Tuber flesh also contained chlorogenic acid (30–900 μg g⁻¹ FW) and other phenolic acids plus low concentrations of flavonoids (0–30 μg g⁻¹ FW). Tuber skins showed much higher levels (1000–4000 μg g⁻¹ FW) of chlorogenic acid. The major anthocyanins in flowers were present as the rutinosides or other glycosides of pelargonidin, petunidin and malvidin whilst glycosides of cyanidin and delphinidin were found in some flowers, together with many of the same phenolic acids as found in tubers. The commonest flavonoids included rutin, kaempferol-3-rutinoside and two quercetin-rhamnose-glucosides. Flowers and leaves contained higher concentrations of flavonoids which fell into two patterns, with some cultivars containing high concentrations of quercetin-3-glycosides, whilst others had much lower concentrations. © 1998 SCI.     

Determination of copper, zinc, selenium, lead and cadmium in potatoes (Solanum tuberosum L.) using potentiometric stripping methods.

Potentiometric stripping analysis was used to determine simultaneously the content of zinc(II), cadmium(II), lead(II) and copper(II) in potatoes, whereas the concentration of selenium was determined by cathodic stripping potentiometric analysis. Metal cations were extracted from potatoes by hydrogen peroxide/hydrochloric acid treatments. The relative standard deviation of the methods ranged from 2.3 to 4.1% and the detection limits were lower than 2.5 microg kg(-1). The results obtained with the proposed methods were compared with those obtained with graphite furnace atomic absorption spectroscopy, a common method for determining metals. The results of the two methods agreed to within 6.1%. Twelve samples of yellow flesh potatoes from different cultivars were analysed. Of all the metals determined, Cu and Zn were the most abundant with concentrations between 0.5 and 4.6 mg kg(-1). Selenium was only found in three samples in very low amounts (<0.1 mg kg(-1)), whilst Pb and Cd concentrations were in the range 0.01-0.27 mg kg(-1).     

Effect of preservative agents on the respiration rate of minimally processed potato (Solanum tuberosum cv. Monalisa)

ABSTRACT:  The shelf life of minimally processed potatoes (MPP) is limited by enzyme-catalyzed browning reactions, with the increase in respiration being another factor that affects quality retention of this product. Sulfites are commonly used as effective preservative agents in minimally processing potatoes, but ascorbic acid and citric acid are considered natural sulfite substitutes and more accepted by consumers. The aim of this study was to study the effect of combinations of the preservative agents cited above (sodium metabisulfite 0.1% and 0.5%; citric acid 0.1% and 0.5%; ascorbic acid 0.5%) on the respiration rate of MPP (cv. Monalisa) processed at both ambient and refrigerated temperatures. The results have revealed that there is a significant effect of dipping treatment and temperature on respiration rate of MPP. Sodium metabisulfite (SM) reduces respiratory activity up to 0.8 mL/kg/h. The addition of either citric or ascorbic acid enhanced the effect of SM on the reduction of the respiration rate of MPP. The strongest effect (up to 3.3 mL/kg/h) was observed when a combination of all 3 agents at the higher concentrations was employed at a temperature of 18 °C.     

Processing of pigmented‐flesh potatoes (Solanum tuberosum L.) on the retention of bioactive compounds

Retention of bioactive compounds in white‐ (WFP), yellow‐ (YFP), red‐ (RFP) and purple (PFP)‐fleshed potatoes after drum‐drying (DD) and Refractance Window? drying (RW) was compared with freeze‐drying (FD). Processing, generally, did not influence total phenolics. Total carotenoid was high in YFP, but decreased with DD. Both DD and RW decreased lutein concentration in YFP. The RFP and PFP had the highest total anthocyanin, and their concentrations were decreased by DD. RW decreased total anthocyanin in PFP only. Total antioxidant activity in WFP was higher with DD but lower with RW. In contrast, DD and RW drying had no significant effect on total antioxidants in YFP, RFP and PFP. Therefore, drying generally decreased antioxidants, with FD and RW retaining the highest concentration of bioactive compounds.     

Quality of organically and conventionally grown potatoes: Four-year study of micronutrients, metals, secondary metabolites, enzymic browning and organoleptic properties

The quality of potatoes from organic and conventional farming was investigated in this study. Tubers of eight potato varieties, organically and conventionally produced at one or two geographical sites in controlled field trials, were collected in four consecutive harvests from 1996-1999. The parameters analysed included nitrate, trace elements (As, Cd, Co, Cu, Fe, Hg, Mn, Ni, Pb, Se, Zn), vitamin C, potato glycoalkaloids, as well as chlorogenic acid, polyphenol oxidase and rate of tuber enzymic browning. The results indicated lower nitrate content and higher vitamin C and chlorogenic acid content to be the parameters most consistently differentiating organically from conventionally produced potatoes. Elevated concentrations of glycoalkaloids were also observed throughout the experiments in some potato varieties grown in organic farming systems. Principal component analysis (PCA) of the analytical and other data using three PCs confirmed a good separation between the organically and conventionally produced potatoes when studied in single crop years. However, score-plots (objects) and loading-plots (variables) of pooled results from the consecutive harvests showed that between the years' changes and also variety as well as geographical variations are equally or more important factors determining the quality of potatoes than the farming system. Further studies of various marker compounds of potato quality related to the organic or conventional farming systems should be performed before unbiased information can be given to the consumers.     

Effect of cross‐linking on some properties of potato (Solanum tuberosum L.) starches

Starches separated from different potato cultivars were modified using two different cross‐linking agents: epichlorohydrin (EPI) and phosphoryl chloride (POCl₃) at different concentrations (1.0 and 2.0 g kg^?1 POCl₃; 2.5, 5.0 and 10 g kg^?1 EPI). Differential scanning calorimetry, rheological and retrogradation measurements were performed to characterise the influence of cross‐linking on the properties of potato starches. Cross‐linking considerably reduced swelling power, solubility, water‐binding capacity and paste clarity. The decrease became greater as the reagent concentration increased. The starches treated with 1.0 g kg^?1 POCl₃ exhibited exceptionally higher swelling power than their counterpart native starches. Neither cross‐linking agent caused any change in morphology of the starch granules. Studies on the phase transitions associated with the gelatinisation showed significantly higher values for the onset temperature (T_o), peak temperature (T_p), conclusion temperature (T_c) and enthalpy of gelatinisation (ΔH_gel) for the cross‐linked starches than the native starches. Starches treated with both the reagents showed lower peak storage modulus (G′) and loss modulus (G″) than their native counterparts. The tendency of the starch pastes towards retrogradation increased considerably with increases in storage duration. However, the starches treated with 1 g kg^?1 POCl₃ exhibited much lower syneresis than the other cross‐linked starches. Copyright © 2006 Society of Chemical Industry     

A comparative study of polyphenoloxidases from taro (Colocasia esculenta) and potato (Solanum tuberosum var. Romano)

Taro (C. esculenta) is a staple food in many tropical regions. A comparative study of crude polyphenoloxidases from taro (tPPO) and potatoes (pPPO) was carried out to provide information useful for guiding food processing operations. Crude PPO was prepared by cold acetone precipitation using ascorbic acid as antioxidant. The PPO content of taro acetone powder was 770±17 units (mg protein)⁻¹ as compared with 3848±180 units (mg protein)⁻¹ in potato acetone powder. The pH-activity optimum was pH 4.6 for tPPO and pH 6.8 for pPPO. Both enzymes retained >80% activity after incubation at pH 4.5–8 but there was rapid activity loss at pH < 4. The temperature-activity optimum (T_opt) was 30°C for tPPO and 25°C for pPPO with 75 and 27% of their respective maximum activity retained at 60°C. Both tPPO and pPPO were irreversibly inactivated by 10 min heating at 70°C. The activation enthalpy (ΔH^#) and activation entropy (ΔS^#) for tPPO heat-inactivation were 87.4 (±0.1) kJ mol⁻¹ and −56.2 (±4) J mol⁻¹ K⁻¹, respectively. For pPPO, ΔH^# was 59.1 (±0.1) kJ mol⁻¹ whilst ΔS^# was −141 (±4) J mol⁻¹ K⁻¹. The apparent substrate specificity was established from values V_max/K_m as: 4-methylcatechol>chlorogenic acid>dl-dopa>catechol>pyrogallol> dopamine>>caffeic acid for tPPO. There was no detectable activity towards caffeic acid. The substrate specificity for pPPO was: 4-methylcatechol>caffeic acid>pyrogallol>catechol>chlorogenic acid >dl-dopa>dopamine. According to the order of inhibitor effectiveness (sodium metabisulphite>ascorbic acid>NaCl≈ (EDTA), there was a significant lag-phase before increases occurred in the absorbance at 420 nm. Preincubation of PPO with inhibitors increased the extent of inhibition, indicating a direct effect on the structure of the enzyme.     

Effect of environmental stress during tuber development on accumulation of glycoalkaloids in potato (Solanum tuberosum L)

The effects of low temperature and of combined stresses, cool temperature and waterlogging and also warm temperature and drought, on potato tuber glycoalkaloid levels were investigated, with emphasis on comparison of tubers of similar size. The early maturing cultivars Home Guard, Rocket and British Queen were grown under favourable controlled environment conditions (18/14 °C day/night temperature, 14 h photoperiod) until the imposition of treatments after the start of tuber initiation. Transfer to low temperature (12/9 °C) did not increase total glycoalkaloid concentration in any of the three cultivars. However, waterlogging at 12/9 °C increased total glycoalkaloid concentration during the later stages of development of British Queen. Drought stress at 24/18 °C also increased total glycoalkaloid concentration in British Queen, although the response varied between individual tubers of similar size. Total glycoalkaloid concentrations in tubers of Rocket were not affected by the combined stress treatments, and any effects on Home Guard were very much less than on British Queen. © 1999 Society of Chemical Industry