• Title/Summary/Keyword: LC-ESI MS

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Identification and Quantification of Glucosinolates in Rapeseed (Brassica napus L.) Sprouts Cultivated under Dark and Light Conditions

  • Lee, Min-Ki;Arasu, Mariadhas Valan;Chun, Jin-Hyuk;Seo, Jeong Min;Lee, Ki-Teak;Hong, Soon-Taek;Kim, In Ho;Lee, Yong-Hwa;Jang, Young-Seok;Kim, Sun-Ju
    • Korean Journal of Environmental Agriculture
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    • v.32 no.4
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    • pp.315-322
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    • 2013
  • BACKGROUND: This study was performed for the identification and quantification of glucosinolate (GSL) contents in seven varieties of rapeseed (Brassica napus L.) sprouts cultivated under dark and light conditions. METHODS AND RESULTS: Crude glucosinolates (GSLs) were desulfated by treating with aryl sulfatase and purified using diethylaminoethyl sepharose (DEAE) anion exchange column. Individual GSLs were quantified using high-performance liquid chromatography (HPLC) with electrospray ionization-tandem mass spectrometry (ESI-MS/MS). Eleven GSLs including six aliphatic (progoitrin, sinigrin, glucoalyssin, gluconapoleiferin, gluconapin, and glucobrassicanapin), four indolyl (4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin) and one aromatic (gluconasturtiin) were identified based on the fragmentation patterns of MS spectrum. Aliphatic GSLs were noted as the predominant group with average 85.2% of the total contents. The most abundant GSLs were progoitrin which was ranged at $8.14-118.68{\mu}mol/g$ dry weight (DW). The highest total GSL amounts were documented in 'Hanra' ($146.02{\mu}mol/g$ DW) under light condition and 'Mokpo No. 68' ($86.67{\mu}mol/g$ DW) in dark condition, whereas the lowest was in 'Tamra' (30.13 and $14.50{\mu}mol/g$ DW) in both conditions. The sum of aliphatic GSLs attributed > 80% in all varieties, except 'Tamra' (67.7% and 64.9% in dark and light conditions, respectively) in the total GSL accumulation. Indolyl GSLs were ranged $2.41-15.73{\mu}mol/g$ DW, accounted 2.78-33.6% of the total GSLs in rapeseed varieties. CONCLUSION(S): These results provide valuable information regarding potential beneficial GSL contents individually. This study attempts to contribute to knowledge of the nutritional properties of the different varieties of rapeseed plants. These results may be useful for the evaluation of dietary information.

Proteomics of plant-fungal pathogen interaction: an overview (식물과 곰팡이 병원균과의 상호작용에 대한 프로테오믹스 최근 연구 동향)

  • Kim, Jin Yeong;Lee, So Eui;Oh, Ha Ram;Choi, In Soo;Kim, Yong Chul;Kim, Sun Tae
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.1-9
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    • 2014
  • So far it has been generally considered that proteomic approaches are very useful for studying plant-microbes interaction. In this review, recent studies based on papers published from 2010 to 2013 have investigated proteomics analysis in various interaction during plant-fungal pathogen infection by means of gel-based proteomics coupled with mass spectrometry (MS)-based analysis. In rice, three papers focused on rice-Magnaporthe oryzae interaction were mainly reviewed in this study. Interestingly, another study showed proteomic changes in rice inoculated with Puccinia triticina, which is not only an fungal pathogen in wheat and but also results to the disease resistance with non-host defense manner in rice. Additionally, proteomics analysis has been widely subjected to understand defense mechanism during other crops (wheat, tomato, strawberry and mint) and their fungal pathogen interaction. Crops inoculated are analyzed to identify differentially regulated proteins at various tissues such as leaf and apoplast using 2-DE analysis coupled with various MS approaches such as MALDI-TOF MS, nESI-LC-MS/MS and MudPIT, respectively. Taken together, this review article shows that proteomics is applicable to various organisms to understand plant-fungal pathogen interaction and will contribute to provide important information for crop disease diagnosis and crop protection.

Antioxidative Activity and Component Analysis of Phellinus linteus Extracts (상황버섯 추출물의 항산화 및 성분분석)

  • Kim, A-Reum;Kim, Jung-Eun;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.4
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    • pp.309-318
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    • 2011
  • In this study, the antioxidative effect, antibacterial, inhibitory effects on tyrosinase, inhibitory effects on elastase and component analysis of Phellinus linteus (P. linteus) extracts were investigated. The ethyl acetate fraction of P. linteus extracts ($2.94\;{\mu}g/mL$) showed the highest free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. linteus extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction ($0.0072\;{\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of P. linteus extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The P. linteus extracts showed cellular membrane protective effects in a concentration dependent manner ($5{\sim}50\;{\mu}g/mL$). The inhibitory effect ($IC_{50}$) on tyrosinase of P. linteus extract was the highest at 50 % ethanol extract ($6.34\;{\mu}g/mL$), and the inhibitory effect ($IC_{50}$) on elastase of P. linteus was the highest at ethyl acetate fraction ($14.08\;{\mu}g/mL$). TLC, HPLC chromatogram and LC/ESI-MS of the ethyl acetate fraction obtained from P. linteus extracts were identified interfungin A (PL RPT-1a). These results indicate that extract/fractions of P. linteus can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging ROS, and protect cellular membranes against ROS. Extract/fractions of P. linteus can be applicable to new cosmeceuticals for antioxidant, antiaging, antiwrinkle and whitening.

Protective Effects of Flavonoids from the Boehmeria quelpaertense against H2O2-Induced Cytotoxicity in H9c2 Cardiomyoblast Cells (H9c2 심근세포에서 제주모시풀(Boehmeria quelpaertense)로부터 분리된 flavonoids의 H2O2로 유도된 독성 보호 효과)

  • Woo, Kyeong-Wan;Sim, Mi-Ok;Bak, Ho;Jung, Ho Kyung;An, Byeongkwan;Ham, Seong-Ho;Park, Jong Hyuk;Cho, Hyun-Woo
    • Korean Journal of Plant Resources
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    • v.31 no.1
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    • pp.1-9
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    • 2018
  • As a part of an infrastructure project on medicinal herb-based remedies, we conducted a phytohemical investigation of the 100% MeOH extract from the aerial part of Boehmeria quelpaertense; our findings resulted in the isolation of flavonoids (1-2), isoquercitrin (1) and hyperoside (2). The identification and structural elucidation of these compounds were based on $^1H$-, $^{13}C-NMR$, and LC ESI IT-TOF MS data. All the compounds isolated from this plant were reported for the first time. In this study, we examined the antioxidant activity of the 1 and 2 on the hydrogen peroxide ($H_2O_2$)-induced oxidative stress in a Rat Cardiomyoblast cell line (H9c2). The pretreatment of the flavonoids showed that it protects against $H_2O_2$-mediated cell death in the H9c2 cell line. Also, it decreases the intracellular reactive oxygen species (ROS) levels by the flavonoids in the $H_2O_2$-treated H9c2 cell line. These results showed that the 1 and 2 are a source of antioxidants. As a result, they might be helpful in preventing the progress of various oxidative stress mediated diseases, including myocardial infarction.

The Compositions and Contents of Flavonol Glycosides in Onion (Allium cepa L.) Germplasm (양파(Allium cepa L.) 유전자원의 Flavonol 배당체 조성 및 함량)

  • Lee, Jung-Ro;Lee, Min-Ki;Kim, Heon-Woong;Lee, Sung-Hyeon;Lee, Young-Min;Jang, Hwan-Hee;Hwang, Kyung-A;Gwag, Jae-Gyun;Ko, Ho-Cheol;Kim, Jung-Bong;Cho, Jin-Woong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.3
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    • pp.385-389
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    • 2014
  • Flavonols as a major kind of plant secondary metabolites are known for health-promoting compounds in onions (Allium cepa L.). The objectives of this study are to determine profiles of flavonol glycosides in different 75 onion accessions. A total of five flavonols (quercetin 3,4'-diglucoside, Q34'diG; quercetin 3-glucoside, Q3G; quercetin 4'-glucoside, Q4'G; isorhamnetin 4'-glucoside, I4'G; quercetin, Q) were identified from onion accessions. In positive ion mode using LC-ESI-MS, individual flavonols were confirmed from one and two glycosylation binding with aglycone such as quercetin and isorhamnetin. Total flavonol contents were distributed in white onion (range of 0.18-6.47 mg/g DW) and purple onion accessions (range of 2.39-6.47 mg/g), respectively. The mean of flavonol contents in purple onion (4.41 mg/g) showed 1.4-fold higher than white onion (3.23 mg/g). The Q34'diG and Q4'G were considered as the major compounds of flavonol glycosides in onion accessions.

Polyphenol Profiling and Comparison of Extraction Solvents on Useful Component Content and Antioxidant Activity of Rhododendron micranthum Shoot (꼬리진달래 신초의 폴리페놀 프로파일링과 추출용매에 따른 유용성분 함량 및 항산화 활성 비교)

  • Jungmok Kang;Yong-Woo Park;Eun-Suk Jung;Daeho Choi
    • Korean Journal of Plant Resources
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    • v.37 no.2
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    • pp.110-119
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    • 2024
  • This study was conducted to provide information on the possibility of using Rhododendron micranthum as resource plant. Through polyphenol profiling of R. micranthum shoot extract, useful components were selected and the content of useful components and antioxidant activity by extraction solvent were compared. As a result, 37 polyphenol compounds were identified in R. micranthum shoot extract through LC-ESI-MS analysis. Contents by extraction solvent were compared of chlorogenic acid, astragalin, myricetin, and afzelin. Chlorogenic acid was highest at 6.57±0.12 mg/g in the ethanol extract, astragalin was 2.29±0.02 mg/g in the ethanol extract, myricetin was 4.77±0.06 and 4.52±0.27 mg/g in the methanol, ethanol extracts, and afzelin was highest at 0.10±0.01 mg/g in the methanol extract. Total polyphenol content of the R. micranthum shoot extract according to the extraction solvent was highest in the ethanol extract at 80.01±2.36 mg/g, and total flavonoid content was also highest in the ethanol extract at 78.08±3.44 mg/g. The IC50 value of DPPH radical scavenging activity according to the extraction solvent of R. micranthum shoot was lowest in methanol extract at 943.57±10.68 mg/L, and the IC50 value of ABTS radical scavenging activity was also lowest in methanol extract at 641.60±7.58 mg/L.

Comparative Interactomes of VRK1 and VRK3 with Their Distinct Roles in the Cell Cycle of Liver Cancer

  • Lee, Namgyu;Kim, Dae-Kyum;Han, Seung Hyun;Ryu, Hye Guk;Park, Sung Jin;Kim, Kyong-Tai;Choi, Kwan Yong
    • Molecules and Cells
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    • v.40 no.9
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    • pp.621-631
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    • 2017
  • Vaccinia-related kinase 1 (VRK1) and VRK3 are members of the VRK family of serine/threonine kinases and are principally localized in the nucleus. Despite the crucial roles of VRK1/VRK3 in physiology and disease, the molecular and functional interactions of VRK1/VRK3 are poorly understood. Here, we identified over 200 unreported VRK1/VRK3-interacting candidate proteins by affinity purification and LC-MS/MS. The networks of VRK1 and VRK3 interactomes were found to be associated with important biological processes such as the cell cycle, DNA repair, chromatin assembly, and RNA processing. Interactions of interacting proteins with VRK1/VRK3 were confirmed by biochemical assays. We also found that phosphorylations of XRCC5 were regulated by both VRK1/VRK3, and that of CCNB1 was regulated by VRK3. In liver cancer cells and tissues, VRK1/VRK3 were highly upregulated and its depletion affected cell cycle progression in the different phases. VRK3 seemed to affect S phase progression and G2 or M phase entry and exit, whereas VRK1 affects G1/S transition in the liver cancer, which could be explained by different interacting candidate proteins. Thus, this study not only provides a resource for investigating the unidentified functions of VRK1/VRK3, but also an insight into the regulatory roles of VRK1/VRK3 in biological processes.

Genetically Engineered Biosynthesis of Macrolide Derivatives Including 4-Amino-4,6-Dideoxy-L-Glucose from Streptomyces venezuelae YJ003-OTBP3

  • Pageni, Binod Babu;Oh, Tae-Jin;Liou, Kwang-Kyoung;Yoon, Yeo-Joon;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.88-94
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    • 2008
  • Two sugar biosynthetic cassette plasm ids were used to direct the biosynthesis of a deoxyaminosugar. The pOTBP1 plasmid containing TDP-glucose synthase (desIII), TDP-glucose-4,6-dehydratase (desIV), and glycosyltransferase (desVII/desVIII) was constructed and transformed into S. venezuelae YJ003, a strain in which the entire gene cluster of desosamine biosynthesis is deleted. The expression plasmid pOTBP3 containing 4-aminotransferase (gerB) and 3,5-epimerase (orf9) was transformed again into S. venezuelae YJ003-OTBP1 to obtain S. venezuelae YJ003-OTBP3 for the production of 4-amino-4,6-dideoxy-L-glucose derivatives. The crude extracts obtained from S. venezuelae ATCC 15439, S. venezuelae YJ003, and S. venezuelae YJ003-OTBP3 were further analyzed by TLC, bioassay, HPLC, ESI/MS, LC/MS, and MS/MS. The results of our study clearly shows that S. venezuelae YJ003-OTBP3 constructs other new hybrid macrolide derivatives including 4-amino-4,6-dideoxy-L-glycosylated YC-17 (3, [M+ $Na^+$] m/z=464.5), methymycin (4, m/z=480.5), novamethymycin (6, m/z=496.5), and pikromycin (5, m/z=536.5) from a 12-membered ring aglycon (10-deoxymethynolide, 1) and a 14-membered ring aglycon (narbonolide, 2). These results suggest a successful engineering of a deoxysugar pathway to generate novel hybrid macrolide derivatives, including deoxyaminosugar.

Comparison of Anthocyanin Content in Seed Coats of Black Soybean [Glycine max(L.) Merr.] Cultivars Using Liquid Chromatography Coupled to Tandem Mass Spectrometry

  • Shin, Sung-Chul;Lee, Soo-Jung;Lee, Sung-Joong;Chung, Jong-Il;Bae, Dong-Won;Kim, Soo-Taek;Sung, Nak-Ju
    • Food Science and Biotechnology
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    • v.18 no.6
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    • pp.1470-1475
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    • 2009
  • The seed coat of the black soybean contains 3 main anthocyanins such as delphinidin-3-O-$\beta$-glucoside, cyanidin-3-O-$\beta$-glucoside, and petunidin-3-O-$\beta$-glucoside. As a part of our effort on discovering and breeding new black soybean cultivars which possesses specific anthocyanin component rich, we determined the anthocyanin profiles of the 2 cultivars recently developed soybean cv. Gaechuck #1 and cv. Gyeongsang #1, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and compared their content and identity with those of previously known 10 cultivar controls. The Cosmosil-$5C_{18}$-AR-II column were selected for the analysis because of the best peak separation. The column temperature was set up at $35^{\circ}C$. The mobile phase consisting of water containing 0.5%(v/v) formic acid and methanol gave good separation between the 3 anthocyanin analytes and internal standard (quercetin 3-O-$\beta$-rutinoside) and peaks with suppressed tail. The MS/MS spectra of each individual anthocyanin standard were detected in positive electron spray ionization (ESI) modes. It was disclosed that the anthocyanin contents of the soybean cv. Gaechuck#1 and cv. Gyeongsang#1 are roughly higher than those of the 10 controls.

Purification and Characterization of Antibacterial Compound Produced by Bacillus subtilis MJP1 (Bacillus subtilis MJP1이 생산하는 항세균 물질의 분리.정제 및 특성규명)

  • Yim, Eun-Jung;Yang, Eun-Ju;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.38 no.1
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    • pp.84-92
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    • 2010
  • Antibacterial compound from Bacillus subtilis MJP1 was purified using C18 Sep-Pak cartridge, ion exchange chromatography, and gel filtration chromatography. The purified antibacterial compound showed antibacterial activity against Listeria monocytogenes, Bacillus subtilis, Staphylococcus aureus subsp. aureus, and Enterococcus faecalis. The purified antibacterial compound was found to be stable at $100^{\circ}C$ for 5 min and in the pH range of 3.0~9.0, but it was unstable at pH 10.0. It was inactivated by proteinase K and pronase E, and heat treatment at $121^{\circ}C$ for 15 min, but it was stable with lipase and $\alpha$-amylase treatment, which indicated its proteineous nature. Ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compound and confirmed the existence of two peptides (3356.54 Da, 3400.5244 Da).