• 원예과학기술지
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• 제29권3호
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• pp.255-259
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• 2011

Polyphenol oxidases (PPO) are copper-containing enzymes responsible for tissue browning in fruits and vegetables including potato, apple and pears. Although these enzymes have been studied for many years, their physiological roles in plants are not yet clear. Therefore, these enzymes need to be purified to characterize further from potato tubers. The classical methods used for the purification of PPO involve several steps. So in this study, we developed a one-step chromatography process for the potato tuber PPO purification. After removal of salts from dissolved ammonium sulfate precipitates of potato tuber extracts using Sephadex-G50 gel filtration, affinity chromatography was carried out on NHS-activated Sepharose 4B using p-aminobenzoic acid as a ligand. The purified enzyme was confirmed by silver staining and a zymogram. The optimum temperature and pH for the purified potato tuber PPO were $15^{\circ}C$ and pH 6.0, respectively. The results obtained in the present study will aid to evaluate PPO from various fruits and vegetables.

• 생약학회지
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• 제20권2호
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• pp.101-109
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• 1989

Purification of polyphenol oxidase(PPO) from Korean native tobacco variety leaves was carried out through the procedure of acetone preciptation, ammonium sulfate fractionation, and Sephadex G-150 gel filtration, resulting in a 84-fold increase in specific activity. The enzyme was stable in a range of pH 7.5 to 8.0 with an optimum of pH 7.5. The optimum temperature for the enzymic reaction was about $60^{\circ}$. It was thermostable with a half-life equal to 20 min at $70^{\circ}$. Km values for (+)-catechin and pyrogallol were $1.6{\times}10^{-3}$ and $0.5{\times}10^{-3}M$, respectively. It possesses high catecholase activity but little or no cresolase activity. Lineweaver-Burk analysis of inhibition data revealed that the inhibition of (+)-catechin oxidation by potassium cyanide, 4-nitrocatechol, cystein and 2-mercaptoethanol was competitive with Ki values of $1.1{\times}10^{-6}$, $1.8{\times}10^{-6}$, $8.9{\times}10^{-6}$ and $1.3{\times}10^{-5}$, respectively.

• 한국초지조사료학회지
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• 제34권1호
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• pp.15-20
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• 2014

To investigate lignification process and its physiological significance under water-deficit condition, the responses of peroxidases, polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) in relation to leaf water status to the short term of water deficit treatment in the leaves with different maturities in forage rape were measured. The significant decrease in relative water content (RWC) and leaf osmotic potential (${\Psi}{\pi}$) were apparent after 5 d of water-deficit treatment. The activity of guaiacol peroxidase (GPOD), ascorbate peroxidase (APOD), coniferyl alcohol peroxidase (CPOD), and syringaldazine peroxidase (SPOD) was depressed especially in middle and old leaves when compared with that of control leaves. On the other hand, in young leaves, a significant increase in CPOD (+34%) and SPOD (+24%) activity as affected by water-deficit treatment was apparent. The activation of PAL and PPO was observed in middle and old leaves for PAL and in young and middle leaves for PPO. These results suggest that peroxidases in middle and old leaves did not involve in lignification under mild water-deficit stress, whereas CPOD and SPOD in young leaves participate in lignification by a coordination with PAL and PPO to incorporate phenol and lignin into the cell walls.

• 약학회지
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• 제45권4호
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• pp.387-396
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• 2001

The effects of ascorbic acid, thiols such as cysteine, n-acetyl-ι-cyteine, glutathione, thiourea, 2-mercaptoethanol and dithiotreithol and organic acids such as magic acid, citric acid, glycolic acid, taurine and kojic acid on polyphenol oxidate (PPO) activity were studied in order to establish if it reacts with oxidized product and/or directly inhibits the enzyme. To investigate the mechanism, the quantification of t-butylcatechol and 4-methylcatechol (phenolic compounds) as substates, their oxidized product and sulphydryl colorless additional compounds were determined by high performance liquid chromatograph (HPLC) method. Chromatographic results indicate that ascorbic acid, organic acids and lower level of cysteine reduced oxidized product of substrates back to their respective positions uf ο-diphenols. On the other hand, other thiols and high level of cysteine reacted with oxidative product of ο-diphenols and then produced sulphydryl colorless compounds. Cysteine apperars to have two types of mechanism of actions in the formation of oxidative products of substrates depending on its concentration; ascorbic acid-type and other thiols-types. The effect of ascorbic acid with thiols on polyphenol oxidase was determined by same method. Chromatographic results indicate that ascorbic acid was more reactive with oxidized product of substrates than thiols.

• 한국식품영양학회지
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• 제33권4호
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• pp.419-427
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• 2020

The purpose of this study was to investigate the qualitative changes of the citron by identifying the type of solution and addition of the solution to prevent the browning reaction of the citron in a way that inhibits the browning of the citron. The browning inhibitor solution was investigated using the individual and mixture, and the results of the degree of browning and chromaticity showed that vitamin C+NaCl+cyclodextrin (CD) had the lowest browning of 0.52. In chromaticity, the ΔE values indicate that the higher the value, the greater the change in color, and the lowest value of the vitamin C+NaCl+CD mixture was 47.0, indicating that there was minimal browning compared to other treatment. The active change of the polyphenol oxidase (PPO) in the citron increased enzyme activity as the browning progressed, and the vitamin C+NaCl+CD solution was the lowest at 68.40 μ/g among the anti-browning solution. Based on these research results, it seems that the CD mixing solution can be used as a citron browning inhibitor.

• 한국식품영양학회지
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• 제30권5호
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• pp.853-859
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• 2017

This study was conducted in an effort to investigate the effect of Maillard reaction products (MRPs) on enzymatic browning of burdock and their anti-oxidant activity. The MRPs were prepared by heating glucose and amino acids at $90^{\circ}C$, which served to produce a strong inhibitory effect on burdock polyphenol oxidase. As the reaction time of the solution containing glucose and amino acid increased at $90^{\circ}C$, the production of MRPs increased and intensity of the brown color deepened. When MRPs were prepared by heating at $90^{\circ}C$ for five hours, the absorbance of MRPs from glucose and lysine was 6.44, while those of glucose and glycine was 1.95. The MRPs synthesized from the glucose and lysine also reduced the pH of MRPs from 5.60 to 4.51, but those from glucose and glycine decreased slightly from 5.57 to 5.33. The Michealis-Menten constant value ($K_m$) of burdock PPO with pyrocatechol as a substrate was 16.0 mM, and MRPs were a non-competitive inhibitor against burdock PPO. The anti-oxidant activity of MRPs was measured by evaluating its radical scavenging activities of DPPH radicals, ABTS radicals and reducing power. The color intensity of MRPs produced by lysine and glucose were deeper than that produced by glucose and glycine. It was also found that MRPs produced from glucose and lysine exhibited stronger anti-oxidant properties than those produced by glucose and glycine.

• 한국식품영양과학회지
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• 제39권7호
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• pp.1018-1023
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• 2010

초임계 이산화탄소를 건조 전처리 방법으로 사용하여 사과의 품질변화를 관찰하고, polyphenol oxidase의 저해효과를 조사하였다. 색도의 경우 L, a, b값은 초임계 이산화탄소의 온도와 압력이 높을 때, 무처리구보다 변화가 적었다. 색차와 갈색도는 초임계 이산화탄소 온도와 압력이 증가할수록 작은 값을 나타내었다. $35^{\circ}C$, 15 MPa와 $45^{\circ}C$, 20 MPa에서 무처리구보다 높은 갈색도를 나타내었다. Polyphenol oxidase(PPO) 활성은 초임계 이산화탄소의 온도와 압력이 증가할수록 활성이 억제되었다. 초임계 이산화탄소 전처리에 따른 springiness는 무처리구보다 증가하였고, hardness는 무처리구보다 감소하였으며, 초임계 이산화탄소의 압력와 온도가 증가할수록 springiness와 hardness는 증가하였다. 이상과 같이 초임계 이산화탄소 전처리가 건조 사과의 색과 조직감에 영향을 미치는 것을 확인할 수 있었다.

• Food Science and Biotechnology
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• 제18권6호
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• pp.1417-1422
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• 2009

This study was conducted to investigate the effect of ultrasound with ascorbic acid on the quality of fresh-cut apples. Prepared apple cubes were dipped in distilled water (US) or in 1% ascorbic acid solution (AS), both were treated with ultrasound at 40 kHz, while the other sample was just dipped in 1% ascorbic acid solution for 1 min (AA). All samples were stored at $4^{\circ}C$ for 12 days. AS treatment had an effect on apple cube color as indicated by its significantly higher L values and lower ${\Delta}E$ value whereas AA- and US-treated samples showed a considerable browning. Higher inhibition rate on browning and polyphenol oxidase (PPO) activity of 46 and 98%, respectively, were observed after AS treatment. The highest total phenolic content of AS-treated samples was shown during whole storage periods. This investigation revealed that the use of ultrasound in combination with ascorbic acid had a positive effect on quality maintaining of fresh-cut apples.

• 한국식품조리과학회지
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• 제12권4호
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• pp.535-540
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• 1996

감자 슬라이스를 CaCl$_2$ 용액과 chitosan 용액에 침지 하거나 혹은 이들 용액의 존재하에서 예열처리 한 후 냉장저장하여 침지 및 예열처리가 감자 슬라이스의 갈변정도, 페놀 함량, PPO 활성의 변화에 대한 영향을 조사한 결과는 다음과 같다. 1. 감자 슬라이스의 L값은 감소하는 경향을 나타내었으나, CaPS는 2주까지 L값이 증가하였고, ChPS는 4주간 L값이 증가하였다(p < 0.05). 또한 예열처리에 의 한 병용 효과에 의해 L값이 가장 증가하였다(p < 0.05). 2. CaPS를 제외한 각 처리군은 total phenol 함량이 감소하였으며(p < 0.05) 예열처 리 한 감자 슬라이스의 total phenol 함량이 더욱 감소하였다(p < 0.05). chloro-genic aicd 함량은 HCaPS를 제외하고 total phenol 함 량의 변화와 같은 경향을 나타내었다. 3. PPO 활성은 침지 및 예열처리를 통하여 감소하였는데 예열처리에 의한 감자 슬라이스의 PPO 활성 이 더욱 감소하였다 이로부터 CaCl$_2$,및 chitosan 용액에 침지 및 예열처리가 갈변억제에 효과적인 것으로 생각되어지며 L값 은 증가하고 페놀함량과 PPO 활성이 감소하였다. 이 러한 효과는 열처리군에서 더 크게 나타났다.

• 한국식품영양과학회지
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• 제15권2호
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• pp.158-164
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• 1986

골덴에서 polyphenol oxidase를 추출(抽出) 및 조정제(粗精製)하여 그 성질(性質)을 조사(調査)한 결과(結果)는 다음과 같다. 최적(最適) pH는 6.5, 최적온도(最適溫度)는 $30^{\circ}C$였으며 효소(酵素)의 기질특이성(基質特異性)은 o-diphenol에 높은 활성(活性)을 나타내었다. 저해제(沮害劑)의 효과(效果)는 Cysteine이 가장 효과(效果)가 좋았고 ascorbate와 sodium metabisulfite는 거의 비슷하였으며 EDTA는 아주 낮은 저해효과(沮害效果)를 보였다. 골덴 PPO를 $20^{\circ}C$ 및 $4^{\circ}C$에서 저장(貯藏)했을때 효소활성(酵素活性)의 변화(變化)는 $20^{\circ}C$에서는 초기(初期) 4일(日)째까지 활성(活性)이 급격히 감소(減少)되었으며 $4^{\circ}C$에서는 초기(初期) 2일(日)까지는 활성(活性)의 변화(變化)가 거의 없었으나 3일(日)째 부터는 급격히 감소(減少)하기 시작(始作)하였다. Polyacrylamide disc gel로 전기영동(電氣泳動)한 결과(結果) 4개(個)의 활성(活性) band가 확인(確認)되었고 열(熱) 처리(處理)한 후(後)에는 각각(各各) 3개(個) 및 1개(個)의 활성(活性) band가 관찰(觀察)되었다. 그리고 효소(酵素)의 열(熱) 안정성(安定性)에 있어서 동일(同一) 처리후(處理後)에 잔존활성(殘存活性)이 40% 및 5% 정도(程度)였으므로 각(各) band의 열(熱) 안정성(安定性)은 효소(酵素)의 열(熱) 안정성(安定性)과 거의 일치(一致)하였다.