• Title/Summary/Keyword: PPO

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Isolation, Purification and Some Properties of Polyphenol Oxidase from Pear (배과실(果實)의 Polyphenol Oxidase의 분리(分離) 정제(精製) 및 그 특성(特性))

  • Kang, Yoon Han;Sohn, Tae Hwa;Choi, Jong Uck
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.55-64
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    • 1986
  • Polyphenol oxidase in japanese pear (Pyrus communis var. mansamkil) was isolated, partially purified and its some properties were investigated. Polyacrylamide disc gel electrophoresis indicated two bands with polyphenol oxidase activity in the extract from acetone dry powder of par flesh. These two polyphenol oxidases (PPO A and PPO B) were purified through acetone precipitation and diethylaminoethyl cellulose column chromatography. PPO A and B were purified 7.8 fold and 8.7 fold by the present procedure, respectively. The Rm values of partially purified PPO A and B were estimated to be 0.58 and 0.68, respectively. The optimum temp, and pH of PPO A activity were $33^{\circ}C$ and pH 7.0, while those of PPO B were $30^{\circ}C$ and pH 4.2, respectively. Two PPO were unstable over the temperature of $60^{\circ}C$. The substrate specificity of pear PPO showed high affinity toward o-diphenolic compounds, especially catechol in PPO A and chlorogenic acid in PPO B, but inactive toward m-diphenol, p-diphenol and monophenols. PPO A showed affinity toward the trihydroxyphenolic compound. $Zn^{{+}{+}}$ activated the PPO A activity but $Fe^{{+}{+}}$ inhibited PPO B activity, while $Fe^{{+}{+}}$ and $Zn^{{+}{+}}$ activated the PPO B activity, while $Fe^{{+}{+}}$ and $Zn^{{+}{+}}$ activated the PPO B activity but $K^+$, $Mg^{{+}{+}}$, $Ca^{{+}{+}}$ and $Hg^{{+}{+}}$ inhibited at 10mM concentration. $Cu^{{+}{+}}$ activated the enzyme action at low concentrations but inhibited at high concentration. Inhibition studies indicated that L-ascorbic acid, L-cysteine and thiourea were most potent. The Km values of PPO A and PPO B for catechol were 20mM and 14.3mM, respectively.

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Effects of Wounding and Jasmonic Acid on Polyphenol Oxidase in Tomato Seedlings (토마토 유식물의 Polyphenol Oxidase에 미치는 상해 및 Jasmonic Acid의 영향)

  • Jin Sun-Young;Hong Jung-Hee
    • Journal of Environmental Science International
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    • v.8 no.6
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    • pp.669-676
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    • 1999
  • The effects of wounding and jasmonic acid(JA) on polyphenol oxidase(PPO) in tomato(Lycopersicon esculentum Mill.) seedlings were investigated. PPO was strongly induced by wounding or JA, and the response was also shown to be systemically induced by wounding. Mechanical wounding in cotyledon or leaf produced a signal that caused the concentration of PPO to increase in the unwounded cotyledon, in the first leaves but not in the second leaves. Severity of wounding and light intensity also affected wound induced change in PPO activity, JA showed a stimulatory effect on the loss of chlorophyll and the rapid increase in PPO activity. The PPO was clearly more active in the wounded leaves than in controls. The potency and specificity of the JA indicate a close relationship between JA and wound-induced changes in PPO in tomato species. JA and abscisic acid(ABA) acted similarly on both unwounded and wounded leaves, but the amount of PPO in the wounded leaves was always more than the respective controls. The highest increase in PPO activity occurred in woundand JA-induced leaves of seedlings kept under bright lighting. Benzyladenine(BA) completely abolished JA- and ABA-induced PPO activity. The results suggest that JA-induced PPO activity is due to de novo PPO synthesis. Histochemical tests for PPO in stems of wound- and JA -treated tomato plants indicate that PPO was localized primarily, in the. outer .cortex . and xylem parenchyma. It is concluded that exogenously applied JA acts as stress agents and PPO may be a component of the inducible anti-hervivore defense response.

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Purification and characterization of polyphenol oxidase from fresh ginseng

  • Kim, Jae-Joon;Kim, Woo-Yeon
    • Journal of Ginseng Research
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    • v.37 no.1
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    • pp.117-123
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    • 2013
  • Polyphenol oxidase (PPO) was purified from fresh ginseng roots using acetone precipitation, carboxymethyl (CM)-Sepharose chromatography, and phenyl-Sepharose chromatography. Two isoenzymes (PPO 1 and PPO 2) were separated using an ion-exchange column with CM-Sepharose. PPO 1 was purified up to 13.2-fold with a 22.6% yield. PPO 2 bound to CM-Sepharose, eluted with NaCl, and was purified up to 22.5-fold with a 17.4% yield. PPO 2 was further chromatographed on phenyl-Sepharose. The molecular weight of the purified PPO 2 from fresh ginseng was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was about 40 kDa. The optimum temperature and pH were $20^{\circ}C$ and 7.0, respectively, using catechol as a substrate. Pyrogallol showed the highest substrate specificity. The effect of a PPO inhibitor showed that its activity increased slightly in the presence of a low concentration of citric acid. High concentrations of acidic compounds and sulfite agents significantly inhibited purified ginseng PPO 2.

Changes in Polyphenol Oxidase of Apple during the Storage (저장중(貯藏中) 사과의 Polyphenol Oxidase의 변화(變化)에 대(對)하여)

  • Kim, Ok Yeoun;Sohn, Tae Hwa
    • Current Research on Agriculture and Life Sciences
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    • v.1
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    • pp.141-150
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    • 1983
  • This report decribes the changes of internal browning and PPO in apples, when the method of storage was changed in the condition of low temperature. Internal browning was found more Fuji harvested in November than Fuji in October. The degree of internal browning was the highest during the storage of 0.09mm 2% and 0.06mm 2%, 0.06mm 20% was followed. In the subatmospheric storage with complete remove of $CO_2$ internal browning was not observed. During the storage of P.E. film bag, Fuji havested in November was higher than in October, and Fuji on 0.09mm 2% was higher than in 0.06mm, and Fuji during the storage of P.E. film bag was increased in PPO activity and PPO specific activity. During the subatmospheric storage with complete remove of $CO_2$, PPO activity of Fuji was little changed, but it's PPO specific activity was continuously decreased. PPO activity of Ralls was more higher than PPO activity of Fuji. PPO activity and PPO specific activity during the early stage of storage and the end stage of storage were little changed. Internal browning of Fuji was more effected $CO_2$ injury than chilling injury. There is no relation between internal browning and degree of PPO activity was effected by $CO_2$ and slightly related with internal browning.

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Preparation of Composite Nafion/polyphenylene Oxide(PPO) with Hetropoly Acid(HPA) Membranes for Direct Methanol Fuel Cells (헤테로폴리산을 포함한 직접 메탄올 연료전지용 나피온/폴리페닐렌옥사이드 복합막의 제조)

  • Kim, Donghyun;Sauk, Junho;Kim, Hwayong;Lee, Kab Soo;Sung, Joon Yong
    • Korean Chemical Engineering Research
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    • v.44 no.2
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    • pp.187-192
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    • 2006
  • The preparation and characterization of new polymer composite membranes containing polyphenylene oxide (PPO) thin films with hetropoly acid (HPA) are presented. PPO thin films with phosphotungstic acid (PWA) or phosphomolybdic acid (PMA) have been prepared by using the solvent mixture. The PWA and PPO can be blended using the solvent mixture, because PPO and PWA are not soluble in the same solvent. In this study, methanol was used as a solvent dissolving PWA and chloroform was used as a solvent dissolving PPO. PPO-PWA solutions were cast onto a glass plate with uniform thickness. The composite membranes were prepared by casting Nafion mixture on porous PPO-PWA films. The morphology and structure of these PPO-PWA films were observed with scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS). The composite membranes were characterized by measuring their ion conductivity and methanol permeability. The performance was evaluated with composite membranes as electrolytes in fuel cell conditions. The methanol cross-over of composite membranes containing PPO-PWA barrier films in the DMFC reduced by 66%.

Gas Separation Membranes Prepared from Polystyrene-block-Polybutadiene/Poly(phenylene oxide) Blends for Carbon Dioxide Separation from a Flue Gas (배기가스로부터 이산화탄소 분리를 위한 SB 이종 블록공중합체/즐리페닐렌 옥사이드 블렌드 기체분리막)

  • Jung, You-Sun;Kim, Chang-Keun
    • Polymer(Korea)
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    • v.32 no.6
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    • pp.593-597
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    • 2008
  • To separate carbon dioxide from a flue gas, membranes for gas separation was fabricated from polystyrene-b-polybutadiene (SB) diblock copolymer blends with poly(phenylene oxide), PPO. SB diblock copolymer formed miscible blends with PPO in the experimental range (lower than or equal to 70 wt% PPO). When the blend contained PPO whose composition is in the range of 40-50 wt%, the discontinuous phase of polybutadiene block in SB diblock copolymer, was changed to discrete phase, while polystyrene blocks containing PPO was changed to the continuous phase. A sudden decrease of the gas permeability and a sudden increase of the gas selectivity was observed at these blend compositions. A gas separation membranes having excellent mechanical properties and exhibiting advantages in gas permeability and selectivity could be fabricated from blends containing more than 50 wt% PPO.

Inhibition of Enzymatic Browning of Taro (Colocasia antiquorum var. esculenta) by Maillard Reaction Products from Glycine and Glucose (Glycine과 Glucose의 Maillard Reaction Products에 의한 토란의 효소적 갈변 저해)

  • 이민영;이민경;김춘영;박인식
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.6
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    • pp.1013-1016
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    • 2004
  • The inhibitory effect of MRPs (Maillard reaction products) on enzymatic browning of taro was investigated. The MRPs prepared by heating glycine and glucose at 9$0^{\circ}C$ for 7 hr exhibited a strong inhibitory effect on taro polyphenol oxidase (PPO). The maximum inhibitory activity of MRPs against taro PPO was detected toward (+)-catechin, catechol, 4-methylcatechol followed by L-$\beta$-3,4-dihydroxyphenylalanine (L-DOPA) and pyragallol as a substrate. The MRPs synthesized from fructose and glucose with glycine as a amino acid significantly reduced the taro PPO activity. MRPs prepared by higher glycine or glucose concentration showed stronger inhibition against taro PPO. Increasing reaction time of the glycine and glucose promoted the inhibitory effect of MRPs against the PPO activity of taro, whereas the color formation was gradually increased.

Time-dependent Degradation of Polyphenol Oxidase in Perilla Fructescens Leaves

  • Kim, An-Keun;Kim, Yoo-Kyung
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.142.3-143
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    • 2003
  • Time-dependent PPO activity was determined at $4^\times$C and $30^\times$C. The result of activity determination, PPO extracted by phosphate buffer containing triton x-1l4(tPPO) was more stable than PPO by phosphate buffer(bPPO). The result of electrophoresis, at first a band was appeared at 48kd. After 1-3days a partial degrade band was appeared in bPPO and three partial degrade bands in tPPO. No activity band was appeared in PPOs at $30^\times$C and bPPO at $4^\times$C after 4 days. (omitted)

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Purification and Characterization of Polyphenol Oxidase from Oyster Mushroom (Pleurotus ostreatus) (느타리버섯(Pleurotus ostreatus)의 Polyphenol Oxidase 분리 정제 및 특성 조사)

  • Choi, Ju-Hee;Kim, Hyun-Jin;Park, Sun-Young;Ham, Kyung-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1447-1452
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    • 2011
  • Polyphenol oxidase (PPO) isoforms were partially purified from oyster mushroom (Pleurotus ostreatus) using various chromatography techniques, and their characteristics of heat stability, substrate affinity, optimum pH, and optimum temperature were investigated. Three PPO isoforms named PO-I, PO-II-1, and PO-II-2 were partially purified from oyster mushroom. The molecular weight of PO-II-1 was 70 kDa and PO-I and PO-II-2 were less than 6 kDa each. Characterization was carried out using a PPO isoform partially purified by hydrophobic interaction chromatography. Optimum temperature was $55^{\circ}C$ and optimum pH 5.0. However, the PPO was inactivated at neutral pH or by heating at $80^{\circ}C$ for 30 min, while the 40% PPO still remained active after heating at $60^{\circ}C$ for 45 min. The PPO isoform showed the highest substrate affinity to chlorogenic acid and pyrogallol, in which KM values were 1.01 and 2.06 mM, respectively. Therefore, these results suggested that the mushrooms should be stored at a pH higher than 7.0 and at a low temperature to prevent enzymatic browning.

Influence of Ph and Temperature on Polyphenol Oxidase in the Leaves of Perilla frutescens var. japonica (들깨잎 폴리페놀 산화효소의 pH 및 온도에 의한 영향)

  • Kim, Yoo-Kyung;Kim, An-Keun
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.384-390
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    • 2004
  • Polyphenol oxidase-catalyzed oxidation of substrates (t-butylcatechol, 4-methylcatechol, chlorogenic acid, caffeic acid and pyrocatechol) were performed in the Ph range 4~8. Co ncentrations of substrate's major oxidation products were monitored by high performance liquid chromatograph. The nature and amounts of products formed were highly pH dependent. They also were ifluenced by kinds of substrates. Major oxidation product of 4-methylcatechol appeared the maxium value at pH 5, them of chlorogenic acid, caffeic acid and pyrocatechol at pH 6.0 and that of t-butylcatechol at pH 5~7. Time-dependent PPO activity was determined at $4^{\circ}C\;and\;30^{\circ}C$. PPO extracted by phosphate buffer containing triton X-114 (t-PPO) was more stable than PPO by phosphate buffer (b-PPO). The result of electrophoresis, at first PPO was showed only a band at 48 kd. After 1~3 days a partial degrade band was appeared in b-PPO and three partial degrade bands in t-PPO. No activity band was appeared in PPOs at $30^{\circ}C$ and b-PPO at $4^{\circ}C$ after 4 days. And a band (37 kDa) in t-PPO was remained finally and disappered. PPO from Perillae leaves has two activity bands at 48 and 37 kDa in previous paper. It was supposed that PPO in the leaves of Perilla frutescens was a protein having one molecular weight as 48 kDa. And 37 kDa protein, relatively proteolysis-resistant, was a proteolyzed form of a major form.