분자표지 이용 Polyphenol Oxidase 저활성 국산밀 유전자원 선발

Song Joong Yun,Kyeong-Hoon Kim,Chon-Sik Kang,Young-Keun Cheong,Chul Soo Park,Kyung-Ho Kim,Hak-Shin Kim,Young-Jin Kim 한국육종학회 2012 한국육종학회지 Vol.44 No.1

The color of wheat flour and dough affect significantly on the quality of wheat. The major factor causing darkening of dough color is activity of polyphenol oxidase (PPO). Therefore, we examined the relationship between the PPO activities and the genotypes of the PPO-specific molecular markers in Korean wheat cultivars, and tested the usefulness of the selected markers for the selection of Korean wheat germplasm lines low in PPO activity. Five sequence-tagged site (STS) markers specific to wheat PPO genes, PPO-05, PPO-16, PPO-18, PPO-30 and PPO-43, were examined for their levels of polymorphism in Korean wheat cultivars. The genotypes of the marker PPO-18, which is linked to Ppo-A1 locus, were correlated significantly highly with the levels of grain PPO activity. Furthermore, Korean wheat cultivars could be classified into the high or low PPO activity group by the allele type of Ppo-A1 which was differentiated by the PPO-18. Grain PPO activity was lower about twice in the cultivars with Ppo-A1b allele than in those with Ppo-A1a allele. The cultivars with Ppo-A1b allele had lower grain weight and wide grain width, also they had less ash, protein, Fe and total polyphenol than those with Ppo-A1a allele. Thus, the lightness of flour and dough sheet was brighter in the cultivars with Ppo-A1b allele. The four wheat cultivars including Suwon252, which had lowest PPO activity and Ppo-A1b allele, were selected from 200 genetic lines as valuable genetic resources for the breeding of wheat variety with bright white-colored flour.

분자표지 이용 Polyphenol Oxidase 저활성 국산밀 유전자원 선발

김경훈,강천식,정영근,박철수,김경호,김학신,김영진,윤성중 한국육종학회 2012 한국육종학회지 Vol.44 No.1

밀가루와 면대 색이 밝은 품종의 육종효율을 증진하기 위하여 PPO 활성과 연관된 sequence-tagged site(STS) 분자표지인 PPO-05, PPO-16, PPO-18, PPO-30과 PPO-43을 이용하여 국내의 밀 품종의 PPO활성 수준과 유전자형 간의 관계를 분석하였다. Ppo-A1에 연관된 PPO-18이 국내 밀 품종의 PPO 활성 판별과 선발에 유용한 분자표지로 확인되었다. PPO 활성은 Ppo-A1b 유전자형 품종(0.446)이 P The color of wheat flour and dough affect significantly on the quality of wheat. The major factor causing darkening of dough color is activity of polyphenol oxidase (PPO). Therefore, we examined the relationship between the PPO activities and the genotypes of the PPO-specific molecular markers in Korean wheat cultivars, and tested the usefulness of the selected markers for the selection of Korean wheat germplasm lines low in PPO activity. Five sequence-tagged site (STS) markers specific to wheat PPO genes, PPO-05, PPO-16, PPO-18, PPO-30 and PPO-43, were examined for their levels of polymorphism in Korean wheat cultivars. The genotypes of the marker PPO-18, which is linked to Ppo-A1 locus, were correlated significantly highly with the levels of grain PPO activity. Furthermore, Korean wheat cultivars could be classified into the high or low PPO activity group by the allele type of Ppo-A1 which was differentiated by the PPO-18. Grain PPO activity was lower about twice in the cultivars with Ppo-A1b allele than in those with Ppo-A1a allele. The cultivars with Ppo-A1b allele had lower grain weight and wide grain width, also they had less ash, protein, Fe and total polyphenol than those with Ppo-A1a allele. Thus, the lightness of flour and dough sheet was brighter in the cultivars with Ppo-A1b allele. The four wheat cultivars including Suwon252, which had lowest PPO activity and Ppo-A1b allele, were selected from 200 genetic lines as valuable genetic resources for the breeding of wheat variety with bright white-colored flour.

Characterization of PPO(Polyphenol Oxidase) cDNA in Sweet Persimmon(Diospyros kaki)

Bahn, Sung Chul,Chung, Young Soo,Shin, Jeong Sheop,Lee, Chang Hoo,Nam, In Hyung 한국식물학회 1999 Journal of Plant Biology Vol.42 No.1

Polyphenol oxidases (PPOs) are copper metalloproteins which play a critical role in producing various derivatives of polyphenolic compounds. These enzymes are encoded in the nuclear genome and subsequently transported into the plastid. A partial cDNA clone encoding polyphenol oxidase was isolated from immature sweet persimmon leaves. Using two degenerate primers, a single band of PCR product was produced by the touchdown PCR method. In sequence analysis, this clone (define SPO; 1381bp) revealed high similarities to the previously reported plant PPO genes. Moreover, this SPO clone was comprised of copper A and copper B-binding sites and a histidine-rich region, indicating that this clone contained the key conserved elements of PPO. To characterize the spatial and temporal expression of PPO, total RNAs were extracted from leaf and fruit peel tissues at different developmental stages. In northern analysis, transcripts of PPO were detected in all the samples tested, but the strongest expression levels were detected in mature fruit peels and leaves. Expression of PPO was inhibited in the transitional stage from chloroplast to chromoplast. In addition, the expression of PPO was highly induced in wounded immature fruit peel tissues.

Enzymatic browning and genome-wide polyphenol oxidase gene identification in three contrasting avocado accessions

Higuera-Rubio Jesús M.,Ibarra-Laclette Enrique,Reyes-López Miguel A.,Sandoval-Castro Eduardo,Cruz-Mendívil Abraham,Vega-García Misael O.,Calderón-Vázquez Carlos L. 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.4

This study aims to disentangle avocado enzymatic browning by identifying and analyzing the PPO coding genes. Two avocado accessions (AVO48 and San Miguel) and the Hass cultivar with contrasting browning kinetics and enzyme activity levels were selected for gene characterization. Upon 90 min of light exposure, Hass and San Miguel showed a greater decrease in luminosity retention (closer to 40% of initial luminosity) compared to AVO48 (85% of luminosity). PPO activity in crude extracts was significantly higher (P < 0.05) in San Miguel (696 U μg-1 protein) than Hass (174 U μg-1 protein) and AVO48 (46–56 U μg-1 protein). San Miguel showed a higher Vmax Km-1 ratio (20.88 min-1), followed by Hass (14.29 min-1) and AVO48 (1.64 min-1), suggesting that San Miguel and Hass have higher substrate affinity. Four PPO coding genes: PamPPO1, PamPPO2, PamPPO3 and PamPPO4 were identified in the Hass genome, all of them containing the main features of plant PPOs, but with specific amino acid combinations in the catalytic pocket of the tyrosinase domain; suggesting that PPO1, PPO2 and PPO4 have monophenolase activity, whereas PPO3, has o-diphenolase activity. The evidence of transcription of PPO3 in fruit of the three genotypes suggests an important role for this gene in avocado pulp browning. PPO2 expression was only found in AVO48. This research provides gene candidates for selective silencing to reduce enzymatic browning.

열수 침지 처리가 치콘 절단면의 적변 억제에 미치는 영향

정현진(Hyun Jin Jung),강호민(Ho-Min Kang) 한국원예학회 2014 원예과학기술지 Vol.32 No.3

치콘의 판매 중 상품성 저하의 원인이 되는 치콘 절단면 적변 현상을 억제하기 위해 열수 침지 실험을 실시하였다. 열수침지처리로는 대조구(20℃ 1분)과 38℃에서의 4분, 8분, 42℃에서의 2분, 4분, 그리고 45℃에서의 1분, 2분을 두었다. 색차계로 측정한 △a<SUP>*</SUP>과 △h값은 42℃ 2분 처리에서는 가장 낮게 유지되었으며, 42℃의 2분과 45℃의 1분 처리구에서는 대조구보다 유의 있는 낮은 값을 나타내었다. 10℃에 저장 6일째 관능 평가한 절단면의 적변지수는 42℃의 2분과 4분 처리 그리고 45℃의 1분 처리에서 낮았다. 적변의 원인 물질이 되는 총페놀함량을 저장 6일째 조사하였는데, 이 또한 절단면의 색변화가 적었던 42℃의 2분과 4분 처리 그리고 45℃의 1분 처리에서 낮았다. 페놀물질 합성에 관여하는 PAL 활성은 42℃ 2분 처리에서는 가장 낮은 값을 보였으며, 42℃의 2분와 45℃의 1분 처리구에서는 대조구보다 유의 있는 낮은 수치를 보였다. 그러나 가장 강한 열처리였던 45℃의 2분 처리에서는 절단면의 적변이 대조구와 유사한 수준으로 진행되었으며, 페놀함량과 PAL 활성도 대조구와 차이가 없었다. 공기에 노출된 절단조직에서 페놀물질을 산화시켜 적변을 일으키는 PPO의 활성은 42℃와 45℃의 모든 처리구에서 낮았는데, 특히 다른 조사항목에서는 모두 대조구와 유사한 수준을 보였던 45℃의 2분 처리구가 가장 낮은 수치를 보였다. 또한 각 조사항목간 상관관계에서, PPO를 제외한 모든 조사항목간에서는 대부분 고도의 상관관계를 보였으며, 특히 관능평가한 적변지수와 △h값은 r = 0.927<SUP>***</SUP>를, PAL 활성과 총 페놀물질 함량과는 r = 0.942<SUP>***</SUP>의 고도의 상관관계를 보였다. Hot water dipping test was conducted for chicon to restrict red discoloration of its basal part which impairs the product value during sales. Hot water dipping treatment was given to chicon for 4 min and for 8 min at 38℃ and for 2 min and 4 min at 42℃, and for 1 min and 2 min at 45℃, along with control (for one min at 20℃). The red discoloration indices of basal part of chicon during sensory evaluation on the sixth day of storage under the storage temperature at 10℃ was lower at 42℃ for 2 min, 42℃ for 4 min and 45℃ for 1 min treatments. The color change value of the basal part in chicon measured by colorimeter showed that the lowest △a<SUP>*</SUP> and △h were maintained in the basal part of chicon treated at 42℃ for 2 min. Whereas, color changes in 42℃ for 2 min and 45℃ for 1 min treatments were significantly low as compared with that of control. The contents of total phenolic compounds which are the substances that cause red discoloration of basal part in chicon were lowest at 42℃ for 2 min, 42℃ for 4 min and 45℃ for 1 min treatments. The activity of phenylalanine ammonia lyase (PAL) resposible for in the synthesis of phenolic substances was the least in 42℃ for 2 min treatment. Whereas, PAL activity of the chicons treated a t 42℃ for 2 min and at 4 5℃ for 1 min were significantly lower than t hat of c ontrol. However, r ed d iscoloration was progressed as similar level with that of control in the basal part of chicon at 45℃ for 2 min. The contents of total phenolic compounds and PAL activity in this treatment were not significantly different from those in control. The polyphenol oxidase (PPO) activity which causes red discoloration of cut tissues was low in all the treatments including 42℃ and 45℃ treatment at which no inhibition effects of the red discoloration of basal part of chicon were observed. When the correlation coefficient between each investigated index was tested, most of them showed high correlation except the PPO activity and particularly and the red discoloration index and sensory evaluation △h values, and PAL activity and total phenolic compounds content were r = 0.927<SUP>**</SUP>, and r = 0.942<SUP>**</SUP>, respectively.

식품첨가제를 첨가한 전해산화수의 세정효과

정승원,정진웅 한국식품저장유통학회(구 한국농산물저장유통학회) 2002 한국식품저장유통학회지 Vol.9 No.2

세정수로서의 전해산화수 효능을 증대시키기 위하여 다양한 식품첨가제를 첨가하여 제조한 0℃ 이하의 세정수에 대한 냉각특성, 살균 및 갈변억제 효과를 조사하였다. 식품첨가제의 첨가비율은 NaCl 0.85%(w/v), ethanol 0.5%(v/v), 레몬과즙 0.5%(v/v), 유자과즙 0.5%(v/v), polysorbate 80은 1 ppm으로 결정하였다. 미생물 사멸효과는 초기 5.63×10exp(8) CFU/mL인 Escherichia coli KCTC 1039가 모든 첨가구에서 15∼30초 이내에 전부 사멸하였으며, Bacillus cereus KCTC 1012는 polysorbate 80 및 ethanol 첨가구에서 2분 후, Lactobacillus plantarum KCTC 3108은 polysorbate 80, 유자 및 레몬과즙 첨가구에서 30초만에, 그리고 Erwinia carotovora subsp. carotovora KCTC 2776은 polysorbate 80 및 레몬과즙 첨가구에서 30초만에 사멸하는 효과를 나타내었다. 갈변억제효과는 polyphenol oxidase의 활성을 측정하여 비교한 결과, ascorbic acid 0.5% 첨가구에 5분간 침지하였을 때 57%의 활성저해를 제외한 모든 처리구에서 62∼84%의 높은 활성저해 효과를 보였으며, 그 중에서도 NaCl 및 유자과즙 첨가구에서 30분 침지시에 각각 64, 91 units로 가장 낮은 활성을 보여주었다. 절단 감자를 30분간 침지처리한 후의 색도에서도 대부분의 처리구에서 색차(ΔE)가 3 이하를 나타내었다. This study, to enhance the sterilization, browning inhibition and precooling effect of electrolyzed oxidizing water(EOW) as cleaning water on food industry, was carried out to investigate the efficacy of electrolyzed oxidizing water(EOW) with 0.85% NaCl, 0.5% ethanol, polysorbate 80 of 1 ppm, 0.5% lemon juice and 0.5% citron juice. Escherichia coli KCTC 1039 with initial count of 5.63×10exp(8) CFU/mL were reduced to <10^1 CFU/mL after 15∼30 sec when it was treated by electrolyzed oxidizing water added with various food additives. Bacillus cereus KCTC 1012 were reduced to <10^1 CFU/mL after 2 minutes treatment with electrolyzed oxidizing water containing polysorbate 80 and ethanol. Lactobacillus plantarum KCTC 3108 were reduced to <10^1 CFU/mL after 30 sec treatment with electrolyzed oxidizing water containing polysorbate 80, citron juice and lemon juice, respectively. Erwinia carotovora subsp. carotovora KCTC 2776 were reduced to <10^1 CFU/mL after 30 sec treatment with electrolyzed oxidizing water containing polysorbate 80 and lemon juice. Browning inhibition effect was determined by comparison of polyphenol oxidase activity. Inhibition ratio of polyphenol oxidase was approximately 62∼84% in most treatments with the exception of 57% and 25% inhibition by 0.5% ascorbic acid and polysorbate 80, respectively. Sliced potato dipped in electrolyzed oxidizing water containing NaCl and citron juice for 30 minutes showed significantly low PPO activity, 64 units in treatment with NaCl and 91 units in treatment with citron juice. At the same time, changes in color value(ΔE) of sliced potato was below 3 in most treatments.

Effect of Polyphenol Oxidase Activity on Discoloration of Noodle Dough Sheet Prepared from Korean Wheats

Chon-Sik Kang,Young-Keun Cheong,Sun-Lim Kim,Dae-Ki Kim,Jung-Gon Kim,Chul Soo Park 韓國作物學會 2008 Korean journal of crop science Vol.53 No.2

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.

Effect of Polyphenol Oxidase Activity on Discoloration of Noodle Dough Sheet Prepared from Korean Wheats

Kang, Chon-Sik,Cheong, Young-Keun,Kim, Sun-Lim,Kim, Dae-Ki,Kim, Jung-Gon,Park, Chul-Soo The Korean Society of Crop Science 2008 한국작물학회지 Vol.53 No.2

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.

국산밀의 폴리페놀 산화 효소 활성과 국수 색과의 관계

강천식,김경훈,박종철,김경호,박광근,정영근,윤성중,박철수 한국육종학회 2011 한국육종학회지 Vol.43 No.5

밀가루 및 국수 면대의 색깔 변화에 영향을 미치는 요인을 분석하여 소비자가 선호하는 밝은 색택의 밀가루 및 국수 제조에 적합한 품종의 육성에 유용한 간편 분석법을 확립하고자 본 연구를 수행하였다. 국산밀 25 품종을 이용하여 종실 및 밀가루 특성과 저장기간 동안 국수 면대의 밝기 변화를 조사하였다. 종실의 PPO 활성은 천립중이 낮을수록, 종실 회분과 단백질 함량이 낮을수록 낮은 경향을 나타내었다. 종실의 PPO 활성은 총폴리페놀 함량(r=0.609** This study was carried out to find major factors affecting the color of wheat flour and noodles and develop simple analysis method for the breeding of wheat cultivars suitable for producing flour and noodles with bright color which are preferred by consumers. Customers who buy white noodle or flour prefer bright-colored food to dark-colored products. We evaluated grains of 25 Korean wheat cultivars for chemical composition, grain characteristics, and color change of noodle dough sheets during storage. Polyphenol oxidase (PPO) has been connected to discoloration of white salted noodles and other wheat end products. The grain PPO activity was reduced as the 1,000 grain weight, grain ash content, and protein content decreased. The grain PPO activity was positively correlated with the total polyphenol content (r=0.609**) and iron content (r=0.655**). Lightness, protein, polyphenol and Fe content of flour were positively correlated with PPO activity of grain. Cultivars with higher grain PPO activity showed darker noodles and were more easily discolored during the storage (from 2 hr to 48 hr). Thus, PPO activity can be used as a selection index in a breeding program for wheat cultivars of bright-colored flour.

콩나물의 저장 중 온도변이에 따른 품질변화

전승호,이세훈,김영주,오세윤,김경문,정종일,심상인 한국작물학회 2010 한국작물학회지 Vol.55 No.3

본 연구는 콩나물용 플라스틱 필름봉지(OPP+PE)에 포장되어 유통되는 콩나물을 냉장고 온도인 3℃ 와 대형 마트의 진열대 온도인 13℃ 로 저장한 콩나물의 품질변화에 미치는 요인을 분석하여, 유통 기간 중 콩나물의 품질변화에 대한 기초 자료를 알아보고자 하였으며, 그 결과는 다음과 같다. 1. 플라스틱 필름 봉지에 포장된 콩나물은 냉장 온도인 3℃ 에 비해 마트 진열대 온도인 13℃ 에서 세근의 발생수가 많았으며, 하배축과 뿌리의 길이도 더 길어졌으며, 수분함량과 생체중에서도 13℃ 에서 더 늘어났다. 경도 조사에서는 13℃ 에서 경도 값의 증가로 하배축이 질겨졌으며, 색도 조사에서도 3℃ 의 자엽과 하배축에서는 거의 변화를 보이지 않았으나, 13℃ 에서는 하배축은 갈색으로 변하고, 자엽은 푸르게 변하였다. 2. 자엽과 하배축의 환원당 함량변화에서는 저장 기간이 경과함에 따라 감소하였는데, 저장 기간 3일차부터 13℃ 저장조건에서 하배축에서 변화 폭이 크게 나타났다. 3. 유리당의 성분 중에서는 galactose와 fructose의 함량이 높았고, 유리당 함량의 변화는 3℃ 에 비해서 13℃ 에서 더욱 민감하게 나타났으며, galactose와 fructose의 함량은 2~3배정도 증가하였다. 4. 유리 아미노산의 함량 변화도 13℃ 에서 크게 증가하였으며, 저장 3일차부터 변화의 폭이 크게 조사되었다. 5. 조단백질의 함량 변화에서는 30.9~35.4%로 증가하였으며, 13℃ 에서 변화의 폭이 크게 나타났다. 6. PPO 활성에서는 3℃ 에서는 4일차부터 크게 증가하여 저장 9일차에서 계속 증가되었으나, 13℃ 에서는 저장 1일차부터 PPO 활성이 크게 증가하여 저장 7일 차부터는 더 이상의 증가가 없었다. Soybean sprouts produced at optimal temperature are placed or displayed for several days in market shelf of relatively cool temperature (ca. 13~circC ). During this period a number of changes occur including changes in color, smell, taste, nutritional quality, etc. In order to investigate the changes of these factors, soybean sprouts packed in plastic film bag (OPP+PE) were stored at the two different temperature (3~circC and 13~circC ). Morphological characters, physicochemical changes and enzymes activity related to visible quality (color) of soybean sprouts were examined. The numbers of fine roots were greater and hypocotyls were longer in soybean sprouts stored at 13~circC , although there was no significant difference in diameter, fresh weight and dry weight of hypocotyls between the two storage temperatures. Browning of hypocotyl, as an indicator of a typical deterioration in sprout quality, was highly dependent on the activity of polyphenol oxidase (PPO). Considering the low level of soluble protein in hypocotyls, the relatively higher activity of PPO suggested a critical role of PPO in stored soybean sprouts. PPO activity of sprouts stored at 13~circC was 2-fold higher than that of sprouts stored at 3~circC after 4 days. In sprouts stored at 13~circC , the PPO activity was increased from day 0 until 6 days and since then, it was not detected. Crude protein content was increased to 30.9~35.4% based on dry weight with extended storage period. The change in crude protein was greater in sprouts stored at high temperature (13~circC ). Total free amino acid content was increased in both temperatures. However, the changing rate of free amino acid was greater in sprouts stored at 13~circC .