• Journal of Microbiology and Biotechnology
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• 제24권5호
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• pp.614-618
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• 2014

Resveratrol, which is a polyphenolic antioxidant, is dose-dependent when used to provide health benefits, to enhance stress resistance, and to extend lifespans. However, even though resveratrol has therapeutic benefits, its clinical therapeutic effect is limited owing to its low oral bioavailability. An Escherichia coli system was developed that contains an artificial biosynthetic pathway that produces resveratrol glucoside derivatives, such as resveratrol-3-Oglucoside (piceid) and resveratrol-4'-O-glucoside (resveratroloside), from simple carbon sources. This artificial biosynthetic pathway contains a glycosyltransferase addition (YjiC from Bacillus) with resveratrol biosynthetic genes. The produced glucoside compounds were verified through the presence of a product peak(s) and also through LC/MS analyses. The strategy used in this research demonstrates the first harnessing of E. coli for de novo synthesis of resveratrol glucoside derivatives from a simple sugar medium.

• Journal of Microbiology and Biotechnology
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• 제28권7호
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• pp.1105-1111
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• 2018

The flavin-dependent monooxygenase Sam5 was previously reported to be a bifunctional hydroxylase with coumarate 3-hydroxylase and resveratrol 3'-hydroxylase activities. In this article, we showed the Sam5 enzyme has 3'-hydroxylation activities for methylated resveratrols (pinostilbene and pterostilbene), hydroxylated resveratrol (oxyresveratrol), and glycosylated resveratrol (piceid) as substrates. However, piceid, a glycone-type stilbene used as a substrate for bioconversion experiments with the Sam5 enzyme expressed in Escherichia coli, did not convert to the hydroxylated compound astringin, but it was converted by in vitro enzyme reactions. Finally, we report a novel catalytic activity of Sam5 monooxygenase for the synthesis of piceatannol derivatives, 3'-hydroxylated stilbene compounds. Development of this bioproduction method for the hydroxylation of stilbenes is challenging because of the difficulty in expressing P450-type hydroxylase in E. coli and regiospecific chemical synthesis.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.167-167
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• 2017

Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_ Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.105-105
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• 2017

Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

• Plant Biotechnology Reports
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• 제3권2호
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• pp.135-138
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• 2009

Cell cultures of Cayratia trifolia (Vitaceae), a tropical lianas, were maintained in Murashige and Skoog's medium containing $0.25mg\;1^{-1}$ NAA, $0.2mg\;1^{-1}$ kinetin and casein hydrolysate $250mg\;1^{-1}$. Cell suspension cultures of C. trifolia accumulate stilbenes (piceid, resveratrol, viniferin, ampelopsin), which on elicitation by any of $500{\mu}M$ salicylic acid, $100{\mu}M$ methyl jasmonate, $500{\mu}M$ ethrel and $500mg\;1^{-1}$ yeast extract, added on the 7th day, were enhanced by 3- to 6-fold ($5-11mg\;1^{-1}$) by the 15th day.

• 한국식품영양과학회지
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• 제39권10호
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• pp.1467-1475
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• 2010

현재 국내에서 재배되는 대표적인 뽕나무(Morus alba L.) 품종별 오디의 영양 및 기능성성분의 함량과 이화학적 품질 특성을 측정하여 비교하였다. 오디 일반성분의 함량은 수분 83.93~86.27%, 가용성 무질소물 8.63~11.76%, 조단백질 1.03~2.54%, 조지방 0.33~0.92%, 조섬유 0.89~2.87% 및 조회분 0.76~0.98% 범위로 익수뽕, 과상2호 및 청일뽕은 다른 품종(대성뽕, 용뽕, 대자뽕, 과상1호)보다 가용성 무질소물 함량이 높은 반면, 단백질, 지방 및 섬유소 함량이 낮았다. 총 당 및 환원당 함량은 과상2호, 청일뽕 및 익수뽕이 높은 반면, 과상1호 및 용뽕은 가장 낮게 나타났다. 총 폴리페놀 및 총 플라보노이드 함량은 과상1호, 대성뽕 및 용뽕이 다른 품종보다 높았으며, 총 안토시아닌 함량은 과상1호 및 2호, 용뽕, 대성뽕이 높았다. 또한, 오디의 anthocyanin 구성성분 즉, C3G 및 C3R 함량은 대성뽕, 과상1호 및 과상2호가 모두 높았다. 한편, 오디의 4가지 플라보노이드 즉, rutin, isoquercitrin, quercitrin, quercetin 함량은 품종에 따라 다소 차이가 있었으며, 과상2호가 2가지 주된 플라보노이드인 rutin 및 quercetin 함량이 높은 반면, 익수뽕 및 청일뽕은 낮았다. 오디 품종별 piceid 및 4-PM 함량을 측정한 결과, piceid 함량은 대성뽕 및 용뽕이 높았으며, 대자뽕, 익수뽕 및 청일뽕은 검출되지 않았다. 한편, 항당뇨 성분인 4-PM은 청일뽕 및 대성뽕이 높은 반면, 그 외 오디 품종들은 함량이 비슷하였다. 항고혈압성 GABA 함량은 청일뽕, 대성뽕 및 과상2호가 높은 반면, 익수뽕 및 용뽕은 낮았다. 오디 품종별 이화학적 품질 특성 중 과실의 중량 2.11~6.38 g, pH 4.08~4.80, 산도 0.39~0.73%, 당도 5.83~15.34oBrix, 색도 L값 26.31~31.49, a값 1.04~3.85, b값 -0.02~4.09 범위로 품종간에 유의적인 차이를 보였다. 특히 오디 품종 중 익수뽕 및 과상2호는 열매의 크기가 적당히 크고 다른 품종보다 산도는 다소 낮은 반면, 당도는 13~15%로 높았다.

• 식물병연구
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• 제20권3호
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• pp.189-195
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• 2014

Genetic engineering is being used to enhance disease resistance and nutritional value of crops including rice plant. Considering the fast-growing agricultural biotechnology and rapidly increasing global area of transgenic crops, the risk evaluation on environment is necessary. In this study, we surveyed the difference of disease occurrence between transgenic rice variety, Iksan526 transformed with peanut stilbene synthase gene and non-transgenic rice varieties, Dongjin and Nampyeong in the field. Moreover, the possibility of gene transfer from transgenic rice to bacterial and fungal pathogens was investigated. The results of this study indicated that there was no significant difference in the occurrence and severity of the diseases between Iksan526 and Dongjin or Nampyeong. In addition, the results suggested that rice pathogen, such as Xanthomonas oryzae pv. oryzae, Rhizoctonia solani and Magnaporthe grisea did not take up stilbene synthase and bar genes under natural conditions. Moreover the transformed DNA was not transferred to the pathogens even in repetitive contacts.

• Food Science and Biotechnology
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• 제18권5호
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• pp.1284-1287
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• 2009

Tumor necrosis factor (TNF)-${\alpha}$ is chronically elevated in adipose tissues of obese rodents and humans. Increased levels of TNF-${\alpha}$ have been implicated in both the induction of atherogenic adipokines, such as plasminogen activator inhibitor (PAI)-1, and the inhibition of the anti-atherogenic adipokine, adiponectin. In this study, we investigated the effects of trans-stilbene, piceatannol, rhaponticin, and piceid on the TNF-${\alpha}$-induced atherogenic changes of adipokines in 3T3-L1 cells. Exposure to TNF-${\alpha}$ for 24 hr increased PAI-1 secretion and decreased adiponectin secretion. Among stilbenoids, piceatannol significantly inhibited the increased secretion of PAI-1 induced by TNF-${\alpha}$. Adiponectin secretion decreased by TNF-${\alpha}$ was recovered after trans-stilbene and rhaponticin treatments. Our results showed that stilbenoids exerted different effects on TNF-${\alpha}$-induced changes in adipokines secretion in 3T3-L1 adipocytes according to their structural characteristics.

• 원예과학기술지
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• 제29권4호
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• pp.374-381
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• 2011

Stilbene 화합물은 항산화 및 항균 활성을 가지는 폴리페놀계의 천연물이다. 포도를 포함하는 몇몇 종류의 식물에서 stilbene 화합물은 resveratrol의 유도체로서 매우 다양한 형태로 존재한다. 자외선 조사 포도 잎에서 stibene 화합물을 정량 분석하기에 앞서, 먼저 각 stilbene 화합물을 동정하기 위한 실험을 수행하였다. 이를 위하여, 자외선을 심하게 조사한 잎에서 stilbene 배당체를 추출하였다. 추출한 배당체 시료는 ${\beta}$-glucosidase를 이용하여 가수분해한 후, 특정 stilbene 화합물의 질량에 해당하는 m/z에서 HPLC-mass spectrometer를 이용하여 분석하였다. 효소적 가수분해에 의해 chromatogram상에는 glycoside에 해당할 것으로 예상되는 peak의 감소와 aglycone에 해당할 것으로 예상되는 peak 증가가 나타났다. 또한 stilbene 화합물의 광 이성질화를 유도하기 위하여 시료를 일광에 노출하였으며, 광 노출에 의해 trans-isomer에 해당할 것으로 예상되는 peak의 소멸과 cis-isomer에 해당할 것으로 예상되는 peak의 생성이 나타났다. Chromatogram상의 peak의 이러한 증감으로부터 각 peak의 성분을 유추하였다. 이러한 방법으로 포도 잎에서 16종의 stilbene 화합물을 동정할 수 있었으며 자외선을 조사한 포도 잎에서 동정된 화합물에 대한 정량적 분석을 수행하였다. 자외선 조사는 포도 잎에서 총 stilbene 함량의 상당한 증가를 가져왔는데 특히 trans-resveratrol은 수백 배 증가하였다. 또한, resveratrol보다 더 강한 radical 소거 활성을 가지지만 무처리 잎에서는 단지 미량으로만 존재하는 piceatannol의 함량 역시 수십배 증가하였다. 자외선 조사에 의한 이러한 stilbene 함량의 증가는 hormesis 현상으로서 포도의 스트레스 대응 반응의 하나로 생각된다.