• 생약학회지
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• 제22권2호
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• pp.101-111
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• 1991

Polyphenol oxidase(PPO) was purified from an extract of Puerariae Radix by ammonium sulfate fractionation followed by Sephadex G-150 column chromatography, which resulted in a 56-fold increase in specific activity. The enzyme was optimum of pH 6.5. The optimum temperature of enzymic reaction was about $40^{\circ}$. The enzyme was thermostable with a half-life equal to 32 min at $70^{\circ}$. Km values of the PPO for catechol and pyrogallol from Lineweaver Burk plots were $1.3{\times}10^{-2}M$, $1.16{\times}10^{-2}M$, respectively. The substrate specificity of the Puerariae Radix PPO showed high affinity toward pyrogallol. Reducing reagents such as cysteine, potassium metabisulfite, ascorbic acid, 2-mercaptoethanol completely inhibited the PPO activity at $10^{-2}M$ level. Linewear-Burk analysis of inhibition data revealed that the inhibition by cysteine, 2-mercaptoethanol, 4-nitrocatechol, potassium cyanide was competitive with Ki values of $4.3{\times10^{-2}M,\;0.73{\times}10^{-6}M,\;6.9{\times}10^{-6}M,\;6.4{\times}10^{-7}M$, respectively. The browning reaction by PPO was observed to decrease temporarily with the addition of sodium diethyl dithiocarbamate, a well known copper chelating agent. Among the divalent cations, $Cu^{2+}$ ion was strong activator on PPO and $Mn^{2+},\;Co^{2+}$ ions was effect on PPO activity. $Zn^{2+},\;Mg^{2+}$ ions was inhibitor on PPO.

• 한국식생활문화학회지
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• 제10권2호
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• pp.89-95
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• 1995

Three potato cultivars, Sumi,Daejima and Namjak, were prepared as slices. They were dipped in distilled water for 20 seconds. The potato slices were packed in polyethylene bags and stored at $5^{\circ}C$. Browning degree, total phenol and chlorogenic acid contents and polyphenol oxidase (PPO) activity were measured. And the correlation analysis of browning parameters were conducted. The results were as follows. There were increase in browning degree, phenolic content and PPO activity during cold storage of potato slices with different cultivars. Among three cultivars, Sumi showed the highest browning degree, phenolic content and PPO activity and also showed the highest % increse of browning and PPO activity during cold storage. On the contrary, Daejima was the lowest. But Daejima showed the highest % increase in phenolic contents during cold storage. With Sumi, browning degree was significantly correlated with PPO activity and phenolic contents (p<0.05). With Daejima and Namjak, a significant correlation was found between browning degree and PPO activity (p<0.05). From the above results, enzymatic browning reactions of potato slices and factors affecting them were dependent on cultivar. Among the tested three cultivars, Daejima showed the lowest browning degree during cold storage and thus seems to be desirable for minimal processing.

• 한국환경과학회지
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• 제8권6호
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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.

• 생약학회지
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• 제30권3호
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• pp.306-313
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• 1999

Polyphenol Oxidase(PPO) was purified from an extract of Ginkgo biloba leaves by ammonium sulfate fractionation followed by sephadex G-150 column chromatography, which resulted in a 18-fold increase in specific activity. The enzyme was most active at pH 8.5 and the temperature optimum for the PPO catechol oxidation reaction was $45^{\circ}C$. Heat inactivation studies showed that heating for 7, 9 and 48 min, at 80, 70 and $60^{\circ}C$ respectively caused a 50% loss in enzymatic activity and that the enzyme was completely inactivated after heat treatment at $90^{\circ}C$ for 60 min. Km values of the PPO for catechol, hydroquinone and 4-methylcatechol derived from Lineweaver-Burk plots were $6.06\;{\times}\;10^{-4}M,\;1.02\;{\times}\;10^{-3}M,\;1.41\;{\times}\;10^{-3}M$ respectively. Of the substrates tested, 4-methylcatechol was oxidized most readily and the enzyme did not oxidize monophenols. The enzyme datalyzed browning reaction was completely inhibited in the presence of reducing reagents, namely ascorbic acid, cysteine, glutathione, 2-mercaptoethanol, potassium metabisulfite at 0.5 mM level. Sodium chloride showed very little inhibition effect on Ginkgo biloba leaves PPO. Lineweaver-Burk analysis of inhibition data revealed that the inhibition by cysteine, 2-mercaptoethanol, potassium cyanide was competitive with ki values of $1.1\;{\times}\;10^{-5}M,\;2.4\;{\times}\;10^{-5}M,\;8\;{\times}\;10^{-5}M$, respectively. Among the divalent cations, $Cu^{2+}ion$ was a strong activator on PPO and $Mn^{2+}ion$ was little or no effect on PPO activity $Ni^{2+}ion$ was an inhibitor on PPO.

• 한국식품영양과학회지
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• 제30권6호
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• pp.1301-1304
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• 2001

본 연구는 사과의 구취억제인자를 구명하기 위한 기초자료로써 사과 추출물의 획분에 따른 사과에서 추출한 PPO의 methyl mercaptan에 대한 구취 억제활성을 조사하였다. 사과를 탁즙하여 구취 억 제 활성 을 측정 해 본 결과, 사과 고형 분의 농도가 증가함에 따라 구취억제활성이 증가하는 경향을 나타내었다. 사과의 저분자획분에 PPO를 첨가한 경우, 저분자 획분의 농도증가 및 반응시간에 따른 methyl mercaptan에 대한 구취억제활성이 증가하는 경향을 나타내었다. 이상의 결과는 사과에서 추출한 PP7에 의한 갈변반응의 중간체로 생성되는 o-quinone이 methy mercaptan과 결합하여 meth- yl mercaptan을 비 휘 발성으로 전환시켜 구취억제활성을 나타내는 것으로 생각된다

• 약학회지
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• 제48권6호
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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.

• 한국작물학회지
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• 제53권2호
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• pp.187-195
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• 2008

Polyphenol oxidase (PPO) is implicated in discoloration of white salted noodles and other wheat based foods. PPO activity was evaluated to determine the effect on discoloration of noodle dough sheets prepared from 25 Korean wheat flours during storage and to screen experimental lines with low PPO activity in 52 Korean wheats. PPO activity was assayed with whole-seed and performed with L-dihydroxyphenylalanine (L-DOPA) as substrates. Absorbance by L-DOPA assay of 25 Korean wheats was from 0.285 to 1.368 at 475 nm. PPO activity was significantly related with grain characteristics, including 1000-kernel weight and grain colors. In flour characteristics, PPO activity positively correlated with ash and protein content (r = 0.658, P < 0.001 and r = 0.424, P < 0.05, respectively) and negatively correlated with $L^*$ value of flour (r = 0.412, P < 0.05). In the changes of color of noodle dough sheet, $L^*$ and $b^*$ values consistently decreased and $a^*$ value increased during storage. PPO activity negatively correlated with $L^*$ value of noodle dough sheet during storage (r = 0.566, P < 0.01 at 2 hr, r = 0.547, P < 0.01 at 24 hr, and r = 0.509, P < 0.01 at 48 hr). But, no significant relationship was found in between PPO activity, $a^*$ and $b^*$ values during storage. The 52 Korean wheat lines examined in this study were divided into 3 different groups, low (< 0.500), medium (0.501-0.999) and high level (> 1.000), on the basis of the level of PPO activity. Twenty two Korean wheat lines showed low level of PPO activity and Suwon 252, 277 and 280 showed lower PPO activity (< 0.200) than others.

• 한국식품조리과학회지
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• 제10권4호
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• pp.339-345
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• 1994

Polyphenol oxidase in Yam was partially purified through ammonium sulfate fractionation, DEAE-cellulose column chromatography. The specific activity of purified PPO was 138.22 unit/mg protein. The optimum pH and temperature of purified PPO were respectively 7.0 and 30$^{\circ}C$. The heat treatment at 80$^{\circ}C$ for 6 min, decreased PPO activity to 50%. The enzyme showed high substrate specificity toward catechol. The Km value for catechol was 5 mM. In the Ames test using S. typhimurium TA98 and TA100 catechol-YEBRP, pyrogallol-YEBRP, chlorogenic-YEBRP showed strong antimutagenicity on sodium azide and MECF excepting hydroquinone-YEBRP showed killing effect on both strains.

• Current Research on Agriculture and Life Sciences
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• 제9권
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• pp.51-59
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• 1991

밤과실의 가공 및 저장중 갈변기구를 규명하는 일환으로 밤에 함유된 polyphenol 화합물 및 PPO를 분리, 동정하고 PPO의 생화학적 특성에 대해 연구한 결과는 다음과 같다. 밤의 total phenol 함량은 $6.5{\mu}g/g$이었고 ferulic acid, caffeic acid, synapic acid, p-coumaric acid, gallic acid, salicylic acid 순으로 함량이 높았다. PPO를 분리, 정제하여 전기영동한 결과 단일의 단백질 및 효소활성밴드를 확인하였으며 정제된 효소의 비활성도는 260.9이고 조효소액보다 11.7배 정제되었다. PPO의 최적 pH와 온도는 각각 5.9, $45^{\circ}C$였으며 효소활성은 $80^{\circ}C$에서 15분 처리시 거의 실활되었다. 무기염의 영향을 본 결과 $Mg^{{+}{+}}$, $Ca^{{+}{+}}$, $Zn^{{+}{+}}$들은 효소활성을 증가시켰으나 $Fe^{{+}{+}}$, $K^+$, $Hg^{{+}{+}}$는 활성을 저해시켰다. 저해제로는 L-ascorbic acid, thiourea, sodium chloride, L-cysteine의 저해작용이 강했다. 밤의 PPO는 o-diphenol에 대한 강한 활성을 나타냈으며 특히 catech이에 대해 가장 강했으며 monophenol에는 활성을 나타내지 않았다. PPO의 Km값은 catechol에 대해 5mM이었다.

• 약학회지
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• 제35권3호
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• pp.222-230
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• 1991

Polyphenol oxidase (PPO) was purified from an extract of Perillae Folium by ammonium sulfate fractionation and sephadex G-150 gel filtration, which molecular weight estimated 65,000$\pm$1,000 in SDS-gel electrophoresis, and pI value was 4.8. The pH and temperature optima were 6.0 and $30^{\circ}C$ respectively. $K_{m}$ values of the PPO for various phenolics derived from Lineweaver-Burk plots were 4.0$\times$10$^{-4}$, caffeic acid; 4.2$\times$10$^{-3}$M, 4-methylcatechol. The inhibition by 4-nitrocatechoi, potassium cyanide, cysteine, 2-mercaptoethanol was competitive with $K_{i}$ values of 7.6$\times$10$^{-5}$M, 7.2$\times$10$^{-5}$M, 3.6$\times$10$^{-5}$M, 2.2$\times$10$^{-5}$M, respectively. Among the divalent cations, Cu$^{2+}$ ion was strong activator on PPO and Zn$^{2+}$, Ni$^{2+}$ ions were little effect on PPO activity. In comparing the amino acid composition of Perillae Folium PPO with that of wheat isozyme, grape, spinach showed similarity. But the content of glycine phenylalanine was abundant relatively.