• Title/Summary/Keyword: metabolites analysis

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Simple and Rapid Evaluation System for Endosulfan Toxicity and Selection of Endosulfan Detoxifying Microorganism Based on Lumbricus rubellus (Lumbricus rubellus를 이용한 endosulfan의 간편, 신속 독성 평가 및 endosulfan 분해 미생물의 선별)

  • Sohn Ho-Yong;Kim Hong-Ju;Kum Eun-Joo;Lee Jung-Bok;Kwon Gi-Seok
    • Journal of Life Science
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    • v.16 no.1
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    • pp.108-113
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    • 2006
  • To compensate the problems of chemical assay in detoxification of recalcitrant and a practical approach in selection of bioremediation bacteria, a simple and rapid toxicity evaluation system was constructed based on Lumbricus rubellus. Long term-culture and specific equipment are not necessary, and semi-quantitative analysis of toxicity at sub-lethal concentration is possible by measuring of dose-dependent increased yellowish secreted compounds. When the toxicity of endosulfan, its metabolites and structurally related chemicals were measured for 24 h, the results were coincided with previous reports. Toxicity was found in endosulfan, endosulfan sulfate, aldrin, and dieldrin, respectively. Rapid and economic selection of endosulfan-detoxifying bacteria was possible using our system. Klebsiella pneumoniae KE-1, K. oxytoca KE-8 and Pseudomonas sp. KS-2P, reported endosulfan degrading bacteria, ameliorated the endosulfan toxicity, whereas E. coli, B. subtilis and other bacteria failed to protect the toxicity of endosulfan in L. rubellus. Our results suggest that the constructed system is useful to selection of microorganism as well as toxicity evaluation against toxic recalcitrants.

A1E Induces Apoptosis via Targeting HPV E6/E7 Oncogenes and Intrinsic Pathways in Cervical Cancer Cells

  • Ham, Sun Young;Bak, Ye Sol;Kwon, Tae Ho;Kang, Jeong Woo;Choi, Kang Duk;Han, Tae Young;Han, Il Young;Yang, Young;Jung, Seung Hyun;Yoon, Do Young
    • Journal of Applied Biological Chemistry
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    • v.57 no.2
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    • pp.103-111
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    • 2014
  • A1E is an extract from traditional Asian medicinal plants that has therapeutic activities against cancers, metabolic disease, and other intractable conditions. However, its mechanism of action on cervical cancer has not been studied. In order to ascertain if A1E would have pronounced anti-cervical cancer effect, cervical cancer cells were incubated with A1E and apoptosis was detected by nuclear morphological changes, annexin V-FITC/PI staining, cell cycle analysis, western blotting, Reverse-transcription polymerase chain reaction, and measurement of mitochondrial membrane potential. Expression of human papiloma virus E6 and E7 oncogenes was down-regulated in A1E-treated cervical cancer cells, while p53 and retinoblastoma protein levels were enhanced. A1E also perturbed cell cycle progression at sub-G1 and altered cell cycle regulatory factors in SiHa cervical cancer cells. A1E activated apoptotic intrinsic pathway markers such as caspase-9, caspase-3 and poly ADP-ribose polymerase, and down-regulated expression of Bcl-2 and Bcl-xl. A1E induced mitochondrial membrane potential collapse and cytochrome c release, and inhibited phosphatidylinositol 3-kinase (PI3K)/Akt, key factors involved in cell survival signaling. Taken all these results, A1E induced apoptosis via activation of the intrinsic pathway and inhibition of the PI3K/Akt survival-signaling pathway in SiHa cervical cancer cells. In conclusion, A1E exerts anti-proliferative action growth inhibition on cervical cancer cells through apoptosis which demonstrates its anti-cervical cancer properties.

Screening of Edible Mushrooms for the Production of Lovastatin and its HMG-CoA Reductase Inhibitory Activity (Lovastatin을 생산하는 식용버섯 선발과 HMG-CoA reductase 저해 효과)

  • Lee Jae-Won;Lee Soo-Min;Gwak Ki-Seob;Lee Ji-Yoon;Choi In-Gyu
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.83-88
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    • 2006
  • This research was performed to determine the production of lovastatin and its HMG-CoA reductase activity produced by fruit bodies and mycelial liquid cultures of domestic edible mushrooms (8 fungal strains). By deter-mining TLC analysis for the confirmation of the presence of lovastatin, all the extracts from fruit bodies and mycelial liquid culture showed same Rf value (0.46), whick was identical to that of the standard lovastatin. In order to extract lovastatin from fruit body, the mixture of water/acetonitrile/methanol was chosen as the most effective solvent. Extracts from fruit body and mycelial liquid culture of pleurotus ostreatus produced the high-est lovastatin 0.98 mg/g based on dry biomass, and 21.90 mg/L, respectively. In the inhibition rate of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, the highest was obtained in P. ostreatus as 67.8% among fruit bodies, and the rates of mycelial liquid culture extracts from P. ostreatus and Laetiporus sulphureus were 37.2% and 29.1%, respectively. Unusually L. sulphureus showed high inhibition rate with low content of lovastatin due to the contribution of campesterol and gamma-sitosterol with hypocholesterolemic activity as metabolites.

Quantitative analysis of water-soluble vitamins and polyphenolic compounds in tomato varieties (Solanum lycopersicum L.) (토마토(Solanum lycopersicum L.) 품종 간 수용성 비타민과 폴리페놀계 성분 함량 변이 분석)

  • Kim, Daen;Son, Beunggu;Choi, Youngwhan;Kang, Jumsoon;Lee, Yongjae;Je, Beungil;Park, Younghoon
    • Journal of Plant Biotechnology
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    • v.47 no.1
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    • pp.78-89
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    • 2020
  • Tomato fruit quality is determined by the contents of various functional metabolites in addition to fruit appearance. To develop tomato cultivars with higher amounts of functional compounds, an efficient quantification method is required to identify the natural variations in the compounds in the tomato germplasm. In this study, we investigated tomato varieties, which included 23 inbred lines and 12 commercial F1 cultivars, for their contents of seven watersoluble vitamins (vitamin C, vitamins B1, B2, B3, B5, B6, and B9) and five polyphenolic compounds (quercetin, rutin, kaempferol, myricetin, and naringenin chalcone). The results of high performance liquid chromatography and liquid chromatography-mass spectrometry showed that vitamin C and naringenin chalcone were the major water-soluble vitamins and polyphenolic compounds, respectively, and their abundance was highly variable depending on the cultivar. By contrast, the contents of vitamin B1, quercetin, and kaempferol were lowest among the cultivars. With regard to the relationship between metabolic compounds and fruit characteristics, a significant association was found in fruit size, indicating that cherry tomato varieties contain higher amounts of the compounds compared to large fresh-type varieties. However, no direct association was detected in fruit color, except for naringenin chalcone. The results of this study provide new insights on the quantification of metabolic compounds and the selection of breeding materials, which are prerequisites for the development of functional tomato varieties.

Isolation of Stenotrophomonas rhizopilae Strain GFC09 with Ginsenoside Converting Activity and Anti-wrinkle Effects of Converted Ginsenosides (사포닌 전환 활성 Stenotrophomonas rhizopilae Strain GFC09 균주의 분리 동정 및 전환 사포닌의 주름 개선 효과)

  • Min, Jin Woo;Kim, Hye-Jin;Joo, Kwang-Sik;Kang, Hee-Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.375-382
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    • 2015
  • Ginsenosides (ginseng saponin) as the one of important pharmaceutical compounds of ginseng and is responsible for the pharmacological and biological activities. These ginsenoside produces diverse small molecules ginsenoside which have more pharmacological activities including anti-wrinkle, anti-cancer and anti-oxidant effects. In the present study, we isolated bacteria using esculin agar, to produce ${\beta}$-glucosidase, and we focused on the bio-transformation of ginsenoside. Phylogenetic tree analysis was performed by comparing the 16S rRNA sequences; we identified the strain as Stenotrophomonas rhizopilae strain GFC09. In order to determine the optimal conditions for enzyme activity, the crude enzyme was incubated with 1 mM ginsenoside $Rb_1$. Bioconversion of ginsenoside $Rb_1$ were analyzed using TLC and HPLC. The crude enzyme hydrolyzed the ginsenoside $Rb_1$ along the following pathway: LB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$ into compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. The structure of the hydrolyzed metabolites were identified by NMR. The activity screening tests showed that the conversion product induced the production of type I procollagen in a dose-dependent manner. These results suggested that hydrolyzed ginseng product containing the ginsenoside $F_2$ and compound K could be useful as an active ingredient for wrinkle-care cosmetics.

Analysis of Tubulysin Biosynthetic Genes in Archangium gephyra (Archangium gephyra의 tubulysin 생합성 유전자 분석)

  • Choi, Juo;Park, Taejoon;Kang, Daun;Lee, Jeongju;Kim, Yungpil;Lee, Pilgoo;Chung, Gregory J.Y.;Cho, Kyungyun
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.458-465
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    • 2021
  • Tubulysins are a group of bioactive secondary metabolites from myxobacteria exhibiting strong anticancer activity against various cancer cell lines. In this study, we describe the identification of putative tubulysin biosynthetic gene clusters (tubA~tubF) in the genome sequences of two tubulysin-producing myxobacterial strains, Archangium gephyra MEHO_002 and MEHO_004. The inactivation of the putative tubulysin biosynthetic genes resulted in a tubulysin-production defect. The DNA sequences of the A. gephyra MEHO_002 and MEHO_004 tubulysin biosynthetic genes were 97% identical, and the amino acid sequences of the encoded proteins shared a similarity of 97-100%. The nucleotide sequences of the tubulysin biosynthetic gene clusters in MEHO_002 and MEHO_004 were 86% identical to that in Cystobacter sp. SBCb004 known as a tubulysin-producing myxobacterium, and the organization of the clusters was identical except for the lack of a tubZ gene in the clusters in MEHO_002 and MEHO_004. The amino acid sequences of the proteins encoded by each gene were 88-97% similar to those encoded by SBCb004, and the domain compositions of the proteins were also identical.

Analysis of the seed metabolite profiles and antioxidant activity of perilla varieties (대사체분석 기술을 이용한 들깨 종자의 품질 특성 및 항산화 효과)

  • Gu, Suyeon;Son, Yejin;Park, Ji Yeong;Choi, Sung-Gil;Lee, Myoung Hee;Kim, Hyun-Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.3
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    • pp.193-199
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    • 2019
  • Perilla seed oil is considered to be a rich source of omega-3 fatty acids and phenolic compounds, which have various health benefits. To improve its oil content and nutritional quality, 29 varieties of perilla have been bred, but their nutritional qualities, including the fatty acid and phenolic compound contents, have not been investigated. Therefore, in this study, the seed metabolite profiles, including the fatty acids, of these 29 perilla varieties were analyzed. Palmitic acid, stearic acid, linoleic acid, linolenic acid, rosmarinyl glucoside, and rosmarinic acid were identified as the major metabolites of perilla. The fatty acid contents were higher in the Okdong, Deulsaem, Daesil, Joim, Yeupsil, Danjo, and Joongmo5103 varieties (in order). However, the total phenolic compounds and antioxidative activities of Okdong and Deulsaem were relatively low, whereas the fatty acid and total phenol contents of Joim, Yeupsil, and Danjo were relatively high. These results may be useful for comparing the nutritional quality of the 29 perilla varieties.

Differential Expression of Isoflavone Biosynthetic Genes in Soybean During Germination (콩 발아기간 중 isoflavone 생합성 유전자 발현 변이)

  • Lim, Jin-Su;Kim, Seo-Young;Kim, Yong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.4
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    • pp.365-374
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    • 2021
  • Soybean isoflavones are essential secondary metabolites synthesized through the phenylpropanoid pathway, and they play vital roles in human health. Isoflavone content is a complex quantitative trait controlled by multiple genes, and the genetic mechanisms underlying isoflavone biosynthesis remain largely unknown. Therefore, the present study analyzed the content of isoflavone and expression of six key genes involved in its biosynthesis (i.e., CHS6, HID, IF7GT, IF7MaT, GmIMaT1, and GmIMaT3) during soybean seed germination. Isoflavone content was quantified using high-performance liquid chromatography, and isoflavone biosynthetic gene expression was analyzed using quantitative real-time PCR. Two cultivars, namely 'Daepung2ho' and 'Pungsannamulkong', which are high- and low-isoflavone cultivars, respectively, were used. Isoflavone accumulation gradually increased with the progression of the germination period. As such, malonyl glucosides accounted for over 80% of the total content, whereas acetyl glucosides were present at trace amounts. Transcriptional analysis of isoflavone biosynthetic genes demonstrated expression patterns parallel to isoflavone content; however, there was no clear correlation between isoflavone content and gene expression. Moreover, most isoflavone biosynthetic genes showed different expression patterns depending on the individual gene or genotypes. Among the tested genes, HID showed consistently higher expression, except at 3 days after germination, and its expression was upregulated in 'Daepung2ho' but downregulated in 'Pungsannamulkong'. In addition, all tested genes exhibited different expression patterns between cotyledons and hypocotyls and responded differently to the germination period. These findings suggest that the expression levels of isoflavone biosynthetic genes are not consistent with the germination period and appear to be genotype-dependent.

Analysis of Chemical Components for Aerial and Underground Parts of Wild Ginseng and Evaluation of Skin Anti-aging Efficacy (야생 산삼 지상부 및 지하부의 화학성분 분석과 피부 항노화 효능 평가)

  • Seok-Seon Roh;Gwang Jin Lee;Byunghyun Kim;Bo Kyoung Hwang;Hyojin Kim;Yun Hee Chang;Jae-kun Yoou;Young-Sung Ju
    • The Korea Journal of Herbology
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    • v.38 no.5
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    • pp.85-95
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    • 2023
  • Objectives : This study was intended to reveal the chemical profiles of aerial(leaf, stem) and underground(rhizome, radix) parts of wild ginseng, and to investigate their anti-aging effects on human skin cells. Methods : Wild ginseng, estimated for over 20 years, was divided into the aerial and underground parts. Total phenolic contents of each extracts were measured using a Folin-ciocalteu method. The contents of 18 amino acids, 8 minerals and 27 ginsenosides were determined by GC-FID, ICP-MS and LC-MS, respectively. The anti-aging effects, including the radical scavenging activity, the activation of mitochondrial function on human fibroblasts, and the proliferation activity on human keratinocyte progenitor cells, for the whole plant and underground part of wild ginseng were evaluated. Results : The total phenolic acids, amino acids, and minerals in the aerial part were more than twice as high as in the underground part. Compared to the cultivated ginseng root, there were various types of ginsenosides in both parts of wild ginseng, and the total amount was more than twice as high. In particular, the aerial part significantly contained ginsenoside F1, F2, C-Mc1, and C-O, and the distinctive patterns that distinguish each parts of wild ginseng from the cultivated ginseng root were derived. The whole plant and underground part of wild ginseng exhibited significant antioxidant effect(14.3-45.6%), activation of mitochondrial membrane potential(105.5-120.1%), and cell proliferation(112.1-125.4%). Conclusions : The entire plant and underground part of wild ginseng are high value-added plants and have beneficial effects on skin anti-aging properties through its abundant metabolites.

Effects of Short Microwave Irradiation Time at the Seedlings Stage on the Growth and Secondary Metabolite Contents of Lettuce (Lactuca sativa L.) (유묘단계에서 단시간 마이크로웨이브 처리가 상추의 생육 및 이차대사산물 함량에 미치는 영향)

  • Yong Jae Lee;Su Yong Park;Ju Hyung Shin;Seung Yong Hahm;Gwang Ya Lee;Jong Seok Park
    • Journal of Bio-Environment Control
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    • v.32 no.3
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    • pp.217-225
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    • 2023
  • This experiment was conducted to investigate the effects of microwave irradiation on the growth and secondary metabolite contents of lettuce seedlings. Seedlings at three weeks after sowing were treated by a microwave oven for 0, 4, 8, and 12 seconds with 200 W. After cultivation in a close plant production system for 4 weeks, plant growth measurements and secondary metabolite analysis were performed. The results showed that the fresh and dry weights of the shoot and root, leaf area, leaf length, and the number of leaves were decreased as increasing the microwave treatment times. Chlorophyll a and b, total carotenoids were increased and total phenolics were decreased at the 12-second treatment compared to the 4-second treatment. Total flavonoid contents were decreased at the 8-second treatment compared to the control. These results suggest that the changes in the levels of secondary metabolites were caused by oxidative stress. Although there was no significant difference in secondary metabolite contents excluding total flavonoid contents on the microwave treatments compared to the control, the significant difference suggests that the microwave treatment of 200 W and 2.45 GHz may alter secondary metabolite contents of lettuce after 4 weeks.