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Pyee, Jae ho 한국식물학회 1996 Journal of Plant Biology Vol.39 No.3
The major cuticular components have been shown to be synthesized in the epidermis. Therefore, cloning of epidermis-specific genes could yield information to be used to isolate and characterize the enzymes involved in the cuticle biosynthesis. A subtractive cDNA library was prepared from Senecio odorus in which epidermis-specific cDNAs were enriched. Differential screening of the library using epidermal and non-epidermal probes revealed two cDNAs. One of them designated epi425 was identified, based on the sequence homology, as a member of a new class in the LTP gene family and the other clone designated epi23 as a gene encoding an aldehyde decarbonylase. Northern blot analyses showed that epi425 and epi23 cDNAs hybridized with a transcript of about 600 and 2,100 nucleotides, respectively, from the epidermis but not from the non-epidermal tissues. Further characterization of these clones will provide more information on the mechanism of the cuticle biosynthesis.
A Molecular Switch for Induction of Resveratrol Biosynthesis in Grapes
Jae ho Pyee,Mi Sook Lee 한국생약학회 2004 Natural Product Sciences Vol.10 No.5
Resveratrol has been reported to possess a variety of biological and pharmaceutical activities. Regardless of its beneficial effects on health, the amount of resveratrol in grapes is very low. In order to induce the resveratrol biosynthesis, the promoter region of a genomic fragment encoding the resveratrol synthase was isolated and a molecular switch was identified which provides us with defining biotic or abiotic inducers that transcriptionally up-regulate the gene expression involved in the resveratrol biosynthesis. We could successfully increase the amount of resveratrol in grapes up to 3-fold by using these environmental factors.
포도과피에 특이적으로 발현하는 stilbene synthase cDNA의 분리 및 과실발달 단계별 발현양상 분석
피재호,이호림 단국대학교 1999 論文集 Vol.34 No.-
Resveratrol (stilbene), a phytoalexin, is produced by stilbene synthase from malonyl-CoA and p-coumaroyl-CoA as substrates and its biosynthesis is induced by environmental stresses such as fungal elicitor and UV. In order to elucidate its role as an anti-stress compound, stilbene synthase (stsy) cDNAs were isolated from Jyoho grape skin, which has been known to contain high quantity of resveratrol, and their expressions were analyzed during the berry development. RT-PCR analysis showed that stilbene synthase gene was expressed in higher level in leaves and flowers than in berries, and its expression in berries was specific to skin. The expression was first detected 10 weeks postflowering in berry skin and increased as berries grew and ripened. Interestingly, another homolog was amplified from the RT-PCR analysis. Its expression was observed at the early developmental stage and reached at the maximum level 8 weeks postflowering, but was not detected thereafter. Both of the PCR produts were subcloned and they shared 70% homology in the nucleotide sequence. These results suggest that there are two or more stilbene synthase genes differentially regulated during the berry develepment.
Park, Jinsun,Pyee, Jaeho,Park, Heonyong Canadian Science Publishing 2014 Canadian journal of physiology and pharmacology Vol.92 No.12
<P> Pinosylvin is a known functional compound of the Pinus species. Pinosylvin at low concentrations (∼pmol/L) was reported to promote cell proliferation in endothelial cells. However, this study found that pinosylvin at a high concentration (100 μmol/L) induces cell death in bovine aortic endothelial cells. Therefore, we examined how pinosylvin was associated with apoptosis, autophagy, and necrosis. Pinosylvin at a high concentration appeared to promote caspase-3 activation, nuclear condensation, and the “flip-flop” of phosphatidylserine, indicating that pinosylvin induces apoptosis. However, based on flow cytometry data obtained from double-staining with annexin V and propidium iodide, pinosylvin was shown to inhibit necrosis, a postapoptotic process. Pinosylvin induced LC3 conversion from LC3-I to LC3-II and p62 degradation, which are important indicators of autophagy. In addition, AMP-activated protein kinase (AMPK) appeared to be activated by pinosylvin, and an AMPK inhibitor was markedly shown to reduce the LC3 conversion. The inhibitory effect of an AMPK inhibitor was reversed by pinosylvin. These results suggest that pinosylvin induces autophagy via AMPK activation. Further, necrosis was found to be promoted by an autophagy inhibitor and then restored by pinosylvin, while the caspase-3 inhibitor had no effect on necrosis. These findings indicate that pinosylvin-induced autophagy blocks necrotic progress in endothelial cells. </P>
Streptomyces coelicolor A3(2)의 actinorhodin생성에 미치는 아미노산의 영향
김재헌,피재호,오충훈,최정은 단국대학교 1996 論文集 Vol.30 No.-
We investigated the fatty acid compositions before and after the actinorhodin producrion in Streptomyces coelicolor A3(2). The relatice proportion of 15 and 17 carbon branched-chain fatty acids decreased signifi-cantly during the actinorhodin production. Fatty acid compositions in the mycelia grown in the presence of L-arginine as a sole nitrogen source, which supported actinorhodin production, showed higher relative pro-portion of the i15:0, i17:0 and ai17:0 branched-chain fatty acids. Therefore, it was concluded that common precursors derived from the amino acid were used for actinorhodin and fatty acid biosynthesis.
Kisuk Bae,Jaeho Pyee 한국산업식품공학회 2004 산업 식품공학 Vol.8 No.4
The roots of Polygonum cuspidatum have been used as herbal medicines in Asia and resveratrol is one of the main active chemicals of root extract. Although a great number of studies have reported its biological and pharmacological activities and purification, resveratrol biosynthesis has never been investigated at the biochemical or genetic level in P. cuspidatum. Hence, we tested resveratrol content in various tissues. Roots accumulated approximately 20 to 50-fold higher levels of resveratrol than other parts of P. cuspidatum plant. Resveratrol content in roots was increased by about 1.84-fold by irradiation with ultraviolet as compared to untreated roots. RT-PCR analysis also showed that PcSTSY, a gene encoding resveratrol synthase (STSY) was expressed at the highest level in roots compared to other tissues in P. cuspidatum plant. These results suggest that resveratrol synthesis is spatially regulated in this plant and that the biosynthesis is induced by irradiation with ultraviolet. In conclusion, this biochemical and genetic results will be applicable to development of foods and beverages containing a high level of resveratrol with beneficial effects on health. The roots of Polygonum cuspidatum have been used as herbal medicines in Asia and resveratrol is one of the main active chemicals of root extract. Although a great number of studies have reported its biological and pharmacological activities and purification, resveratrol biosynthesis has never been investigated at the biochemical or genetic level in P. cuspidatum. Hence, we tested resveratrol content in various tissues. Roots accumulated approximately 20 to 50-fold higher levels of resveratrol than other parts of P. cuspidatum plant. Resveratrol content in roots was increased by about 1.84-fold by irradiation with ultraviolet as compared to untreated roots. RT-PCR analysis also showed that PcSTSY, a gene encoding resveratrol synthase (STSY) was expressed at the highest level in roots compared to other tissues in P. cuspidatum plant. These results suggest that resveratrol synthesis is spatially regulated in this plant and that the biosynthesis is induced by irradiation with ultraviolet. In conclusion, this biochemical and genetic results will be applicable to development of foods and beverages containing a high level of resveratrol with beneficial effects on health.
Lee, Hyang-Hwa,Pyee, Jae-Ho The Korean Society of Medicinal Crop Science 2004 韓國藥用作物學會誌 Vol.12 No.5
Microsomal ${\omega}-3$ fatty acid desaturase (FAD3) is an essential enzyme in the production of the n-3 polyunsaturated fatty acid ${\alpha}-linolenic$ acid during the seed developing stage. To understand the regulatory mechanism of the gene encoding the ${\omega}-3$ fatty acid desaturase, a genomic fragment corresponding to the previously isolated perilla seed PfFAD3 cDNA was amplified from perilla (Perilla frutescens Britt) by GenomeWalker PCR. Sequence analysis of the fragment provided with identification of a 1485-bp 5'-upstream region and a 241-bp intron in the open reading frame. To determine the tissue-specificity of the PfFAD3 gene expression, the 5'-upstream region was fused to the ${\beta}-glucuronidase$ (GUS) gene and incorporated into Arabidopsis thaliana. Histochemical assay of the transgenic plants showed that GUS expression was restricted to seed and pollen, showing that PfFAD3 gene was exclusively expressed in those tissues.