• 제목/요약/키워드: MALDI-TOF MS/MS

검색결과 280건 처리시간 0.031초

Purification and Properties of Chitosanase from Chitinolytic $\beta$-Proteobacterium KNU3

  • Yi, Jae-Hyoung;Jang, Hong-Ki;Lee, Sang-Jae;Lee, Keun-Eok;Choi, Shin-Geon
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.337-343
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    • 2004
  • A bacterial strain concurrently producing extracellular chitosanase and chitinase was isolated from soil and identified as a member of the $\beta$-subgroup of Proteobacteria through its 16S rRNA analysis and some biochemical analyses. The newly discovered strain, named as KNU3, had 99% homology of its 16S rRNA sequence with chitinolytic $\beta$-Proteobacterium CTE108. Strain KNU3 produced 34 kDa of chitosanase in addition to two chitinases of 68 kDa and 30 kDa, respectively. The purified chitosanase protein (ChoK) showed activity toward soluble, colloidal, and glycol chitosan, but did not exhibit any activity toward colloidal chitin. The optimum pH and temperature of ChoK were 6.0 and $70^{\circ}C$, respectively. The chitosanase was stable in the pH 4.0 to 8.0 range at $70^{\circ}C$, while enzyme activity was relatively stable at below $45^{\circ}C$. MALDI-TOF MS and N-terminal amino acid sequence analyses indicated that ChoK protein is related to chitosanases from Matsuebacter sp. and Sphingobacterium multivorum. HPLC analysis of chitosan lysates revealed that glucosamine tetramers and hexamers were the major products of hydrolysis.

Comparative Proteomic Analysis of Virulent Korean Mycobacterium tuberculosis K-strain with Other Mycobacteria Strain Following Infection of U-937 Macrophage

  • Ryoo, Sung-Weon;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Liew, Hyun-Jeong;Kang, Seong-Man;Bai, Gill-Han
    • Journal of Microbiology
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    • 제45권3호
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    • pp.268-271
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    • 2007
  • In Korea, the Mycobacterium tuberculosis K-strain is the most prevalent clinical isolates and belongs to the Beijing family. In this study, we conducted comparative porteomics of expressed proteins of clinical isolates of the K-strain with H37Rv, H37Ra as well as the vaccine strain of Mycobacterium bovis BCG following phagocytosis by the human monocytic cell line U-937. Proteins were analyzed by 2-D PAGE and MALDI-TOF-MS. Two proteins, Mb1363 (probable glycogen phosphorylase GlgP) and MT2656 (Haloalkane dehalogenase LinB) were most abundant after phagocytosis of M. tuberculosis K-strain. This approach provides a method to determine specific proteins that may have critical roles in tuberculosis pathogenesis.

방향족 아민 화합물과 풀러렌 산화물의 [C70(O)n](n≥1)의 초음파 화학 반응 (Sonochemical Reaction of Fullerene Oxides, [C70(O)n](n≥1) with Aromatic Amines)

  • 고원배;박병은;이영민
    • Elastomers and Composites
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    • 제43권1호
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    • pp.31-38
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    • 2008
  • [ $FeCl_3$ ]를 첨가한 초음파 조건에서 4-nitroaniline, 3-nitroaniline, and 4-isopropylaniline등의 방향족 아민 화합물과 풀러렌 산화물$[C_{70}(O)_n](n\geq1)$을 반응시켰다. 이 반응은 방향족 아민 화합물을 사용한 풀러렌 산화물 쪼개짐 반응이다. MALDI-TOF-MS and UV-vis spectra를 사용하여 생성된 화합물이 아민화 풀러렌 유도체임을 확인하였다.

Evaluation of Microbiological Contamination of Water Purifiers at Two Universities in Chungcheong Region

  • Jin Young Yun
    • 대한의생명과학회지
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    • 제29권4호
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    • pp.256-262
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    • 2023
  • The purpose of this study is to investigate microbial contamination in water purifiers from two universities (A and B) in Chungcheong region and to evaluate about the harmfulness of the isolated bacteria to the human. The degree of microbiological contamination of six water purifiers at university A was investigated three times from July 2018 to September 2019, and nine water purifiers at university B were investigated in 2023. The isolated bacteria were biochemically identified using an API kit and Vitek-2 system, and then the bacteria were identified to the species level using MALDI-TOF MS. In addition, the possibility of human infection of the isolated bacteria was evaluated through a literature search. In July 2018 and September 2019, the number of bacteria isolated inside the faucet was below the acceptable standard for hot water, but exceed for cold water in all water purifiers. In January and September 2019, bacteria exceeding the acceptable standards were isolated nine times from the cold water of six water purifies (a total of 12 water purifiers). Bacteria identified by MALDI-TOF MS included anaerobic bacteria (Clostridium novyi, Clostridium themopalmarium etc.), Gram-positive bacilli (Microbacterium testaceum, Arthrobacter woluwensis etc.), and Gramnegative bacilli (Acinetobacter nosocomialis, Comamonas kerstersii etc.), which are difficult identify by biochemical methods. In conclusion, bacteria exceeding the acceptable standard were isolated from the cold water of most of the water purifiers. Most of the isolated bacteria were low-pathogenic bacteria from natural environment, but opportunistic bacteria that can cause infection in humans were also isolated from some water purifiers.

오존산화 콩기름의 구조분석 및 이를 이용한 변성 pMDI 접착제의 중량비에 따른 접착력 변화 (Studies on Chemical Strutures and Adhesion Performance of pMDI Adhesives Modified by Ozonized Soybean Oil with Different Mixing Ratios)

  • 유영삼;이현종;이택준;박헌
    • Journal of the Korean Wood Science and Technology
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    • 제37권1호
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    • pp.56-64
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    • 2009
  • 본 연구의 목적은 콩기름을 오존 산화 처리하여 기능기를 도입하고 이를 바탕으로 친환경 목재 접착제를 개발하는 것이다. 시판용 콩기름을 시간당 약 7.13 g의 오존을 발생시켜 15분, 30분, 60분, 120분간 오존산화 처리한 후 FT-IR, $^1H$-NMR, MALDI-TOF MS, GC/MS 측정하여 구조변화를 분석하였다. 기기분석 결과 오존산화처리에 의하여 콩기름의 불포화 이중결합이 개열되어 카르복실기 또는 알데하이드기가 생성됨을 확인하였다. 오존산화콩기름 변성 pMDI 접착제의 접착력은 오존처리 정도와 pMDI의 혼합비율에 따라 달라지며, KS F 3101 보통합판기준 내수 접착강도를 안정적으로 만족 시키는 조건은 오존산화 처리시간 30분~60분, pMDI와 1:1로 혼합한 경우이며, 준내수 접착강도를 만족시키는 범위는 오존산화 처리시간 30분~60분, pMDI의 혼합비 0.5이상, 상태접착강도를 만족시키는 범위는 오존산화 처리시간 15분~60분, pMDI의 혼합비 0.25 이상임을 알 수 있었다. 위의 실험결과를 종합하여 볼 때, 오존산화 처리한 식용유는 오존처리시간 및 pMDI의 혼합비율을 조절하면 상태접착력 뿐만 아니라 내수접착력이 요구되는 목질복합재료 제조용 접착제 제조가 가능함을 확인할 수 있었다.

Synthesis of Dendrimer Based Polymeric and Macrocyclic Complexes with a Platinum-Acetylide ${\pi}-Conjugated$ Organometallic Core

  • Jang, Woo-Dong
    • Macromolecular Research
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    • 제13권4호
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    • pp.334-338
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    • 2005
  • A three-layered poly(benzyl ether) dendrimer having a bis-ethynylbenzene core was synthesized and characterized with $^{1}H$ NMR and MALDI-TOF-MS spectroscopy. The dendrimer was reacted with platinum complexes to obtain platinum-acetylide based organometallic polymers. When the dendrimer was reacted with trans-[$PtCl_{2}(PEt_{3})_{2}$], a high molecular weight polymeric compound was formed, whereas, with cis-[$PtCl_{2}dppp$], a uniform molecular weight compound was formed, which was found to be a dimeric metallacycle by $^{1}H\;NMR,\;^{31}P\;NMR$ and ESI-TOF-MS spectroscopy. Both these complexes exhibited relatively a strong emission around 440 nm, indicating that they could be potential candidates for blue emitting polymer LEDs.

Highly Dispersed CuO Nanoparticles on SBA-16 Type Mesoporous Silica with Cyclam SBA-16 as a Precursor

  • Prasetyanto, Eko Adi;Sujandi, Sujandi;Lee, Seung-Cheol;Park, Sang-Eon
    • Bulletin of the Korean Chemical Society
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    • 제28권12호
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    • pp.2359-2362
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    • 2007
  • MALDI-TOF-MS technique was applied to obtain structural and compositional information of synthetic polyamides, Nylon6 and Nylon66. Mass spectra of both the original and the hydrolyzed polyamide samples were analyzed using the self calibration method as well as the internal calibration method with the standard materials of known masses. The MALDI-TOF mass spectra of Nylon6 samples showed the presence of protonated, sodiated, and potassiated ions that were assigned to cyclic and NH2/COOH terminated linear oligomers. From the MALDI-TOF mass spectra of Nylon66 samples, the potassiated linear oligomers with three different end groups are identified as well as the cyclic oligomers, i.e., NH2/COOH terminated oligomers, NH2/NH2 terminated oligomers, and COOH/COOH terminated oligomers. Full characterization of the molecular species and end groups present in the polyamide samples has been achieved, and also the changes in mass spectral patterns after the hydrolysis of the samples are presented.

Characterization of Yeast and Bacterial Type Strains with Food and Agricultural Applications by MALDI-TOF Mass Spectrometry Biotyping

  • Harnpicharnchai, Piyanun;Jaresitthikunchai, Janthima;Seesang, Mintra;Jindamorakot, Sasitorn;Tanapongpipat, Sutipa;Ingsriswang, Supawadee
    • 한국미생물·생명공학회지
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    • 제48권2호
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    • pp.138-147
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    • 2020
  • Various microorganisms play important roles in food fermentation, food spoilage, and agriculture. In this study, the biotype of 54 yeast and bacterial strains having high potential for utilization in food and agriculture, including Candida spp., Lactobacillus spp., and Acetobacter spp., were characterized by matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS). This characterization using a fast and robust method provides much-needed information on the selected microorganisms and will facilitate effective usage of these strains in various applications. Importantly, the unique protein profile of each microbial species obtained from this study was used to create a database of fingerprints from these species. The database was validated using microbial strains of the same species by comparing the mass spectra with the created database through pattern matching. The created reference database provides crucial information and is useful for further utilization of a large number of valuable microorganisms relevant to food and agriculture.

Differential Proteomic Analysis of Chinese fir Clone Leaf Response to Salicylic Acid

  • Yang, Mei;Lin, Sizu;Cao, Guangqiu
    • Journal of Forest and Environmental Science
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    • 제26권2호
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    • pp.83-94
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    • 2010
  • Chinese fir (Latin name: Cunninghaimia lanceolata) is one of the major commercial coniferous trees. Most of Chinese fir forests are managed in successive rotation sites, which lead productivity to decline. Autotoxicity is the important reason for soil degradation of Chinese fir plantation, especially, phenolic acids are considered as the major allelopathic toxins which induce autotoxicity in Chinese fir rotation stands. We performed here proteomic approach to investigate the response of proteins in Chinese fir leaves to salicylic acid. The tube plantlets of Chinese fir clone were treated with 120 mg/L salicylic acid for 1, 3 and 5th day. 2-DE, coupled with MALDI-TOF-TOF/MS, was used to separate and identify the responsive proteins. We found 12, 7, and 12 candidate protein spots that were up- or down-regulated by at least 2.5 fold after 1, 3, and 5th day of the stress, respectively. Of these protein spots, 16 spots were identified successfully. According to the putative physiological functions, these proteins were categorized into five classes (1) the proteins involved in protein stability and folding, including 26S proteome, Grp78, Hsp70, Hsp90 and PPIase; (2) the protein involved in photosynthesis and respiration, including OEC 33 kDa subunit, GAPDH; (3) the protein related to cell endurance to acid, F-ATPase; (4) the protein related to cytoskeleton, tubulin; (5) the protein related to protein translation: prolyl-tRNA synthetase. These results give new insights into autotoxic substance stress response in Chinese fir leaves and provide preliminary footprints for further studies on the molecular signal mechanisms induced by the stress.

Protein Expression Profiling of Infected Murine Macrophage Cells (RAW 264.7) by Bacillus anthracis Spores

  • Seo Gwi-Moon;Nam Jeong-Ah;Oh Kwang-Gun;Chai Young-Gyu
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2003년도 International Meeting of the Microbiological Society of Korea
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    • pp.77-79
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    • 2003
  • Current therapeutic strategies far anthrax have had no significant impact on anthrax mortality over the last several decades. This study used a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) discovery platform to generate protein expression profiles in search of overexpressed proteins in murine macrophage cells (RAW264.7) which infected with Bacillus anthracis spores as potentially novel molecular targets. Two differentially expressed proteins were identified in infected murine macrophage cells as Syndapin and CDC46, respectively. Syndapins are potential links between the cortical actin cytoskeleton and endocytosis. Other two proteins were identified from murine macrophage cells infected with avirulent spores as ITBG-2 (CD18) and HSPA5, respectively. These data demonstrate the feasibility of using a MALDI-TOF platform to generate protein expression profiles and identify potential molecular targets for anthrax therapeutics.

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