• 제목/요약/키워드: MALDI TOF

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Identification of Differentially Expressed Proteins at Four Growing Stages in Chicken Liver

  • Lee, K.Y.;Jung, K.C.;Jang, B.G.;Choi, K.D.;Jeon, J.T.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권10호
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    • pp.1383-1388
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    • 2008
  • Because of high growth rate and large deposition of fat in the abdomen, the chicken has been used as a model organism for understanding lipid metabolism, fattening and growing. In this study, differentially expression of proteins in chicken liver, one of the important organs for lipid metabolism, has been investigated at four different growing stages. After separation of proteins using two-dimensional electrophoresis (2-DE), more than 700 protein spots were detected. Among them, 13 growing stage specific proteins in chicken liver were selected and further investigated by matrix-assisted laser adsorptions ionization-time of flight mass spectrometry (MALDI-TOF MS). Of these, 12 proteins were matched to existing proteins based on a database search. The identified fat-related proteins in this study were fatty acid synthase (FASN) and malic enzyme (ME1). These proteins were more highly expressed at week 32 than at other weeks. In order to confirm the differential expression, one of the proteins, FASN, was confirmed by western blotting. The identified proteins will give valuable information on biochemical roles in chicken liver, especially for lipid metabolism.

Production of Biosurfactant Lipopeptides Iturin A, Fengycin, and Surfactin A from Bacillus subtilis CMB32 for Control of Colletotrichum gloeosporioides

  • Kim, Pyoung-Il;Ryu, Jae-Won;Kim, Young-Hwan;Chi, Youn-Tae
    • Journal of Microbiology and Biotechnology
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    • 제20권1호
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    • pp.138-145
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    • 2010
  • A bacterial strain isolated from soil for its potential to control the anthracnose disease caused by Colletotrichum gloeosporioides was identified as a Bacillus subtilis. Bacillus subtilis CMB32 produced antifungal agents on M9 broth at $30^{\circ}C$. Biosurfactant lipopeptides produced by Bacillus subtilis CMB32 were precipitated by adjusting to pH 2 and extracting using chloroform/methanol, and then were purified using column chromatography and reverse-phase HPLC. The molecular masses of the lipopeptides were estimated by MALDI-TOF mass spectrometry as (a) 1,080, (b) 1,486, and (c) 1,044 Da, respectively. They had cyclic structures and amino acid compositions of (a) Pro, Asx, Ser, Tyr, Glx, (b) Glx, Tyr, Thr, Ala, Pro, lie, and (c) Glx, Leu, Val, Asx, respectively. Further analysis revealed that Bacillus subtilis CMB32 produced three antifungal lipopeptides: (a) iturin A, (b) fengycin, and (c) surfactin A.

붕장어(Conger myriaster)의 피부로부터 평활근 수축작용을 지닌 신경성 펩타이드의 정제 (Purification of Neuropeptide with the Contractile Activity on the Smooth Muscle from the Skin of Conger Eel Conger myriaster)

  • 고혜진;박남규
    • 한국수산과학회지
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    • 제45권4호
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    • pp.358-366
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    • 2012
  • A novel neuropeptide was isolated from the skin of the conger eel Conger myriaster using hagfish Eptatretus burgeri intestine as a bioassay system. The sequence of the purified peptide was analyzed using automated amino acid sequencing and MALDI-TOF mass spectrophotometry. The molecular ion peak in the MALDI-TOF mass spectrum of the peptide was at m/z 962.89 $(M+H)^+$. The sequence of the peptide was determined to be L-P-M-L-E-T-Q-M, and was tentatively named comyrin. To investigate the complete primary structure of comyrin, comyrin-OH and comyrin-$NH_2$ were synthesized and the chemical and pharmacological properties of the synthetic peptides were compared with those of the native peptide. However, the elution time of synthetic peptides did not match that of the native peptide on the reverse-phase HPLC chromatogram. In addition, the synthetic peptides did not cause contractile activity in the intestinal smooth muscle of the hagfish. Based on these results, one possible reason for this disagreement may be the presence of a D-amino acid in comyrin.

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.

Vibrio anguillarum O1이 생산하는 Outer Membrane Vesicle (OMV)의 분리 및 OMV 내의 단백질 특성 (Isolation and characterization of the outer membrane vesicle (OMV) protein from Vibrio anguillarum O1)

  • 홍경은;김동균;민문경;공인수
    • 한국해양바이오학회지
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    • 제2권2호
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    • pp.123-125
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    • 2007
  • Vibrio anguillarum is a gram-negative bacterium that causes vibriosis in approximately 80 different fish species. V. anguillarum produces several exotoxins are correlated with the pathogenesis of vibriosis. This study is focused on the composition of the outer membrane vesicle. Most of gram-negative bacteria produce outer membrane vesicle (OMV) during cell growth. OMV was formed from the outer membrane surface of cell and than released to extracellular environment. OMV consists of outer membrane lipids, outer membrane protein (OMP), LPS, and soluble periplasmic components. Also, they contain toxins, adhesions, and immunomodulatory. Many gram-negative bacteria were studied out forming OMV. In Vibrio sp., formation of OMV by electron microscopy has been reported from V. cholerae and V. parahaemolyticus. In present study, we isolated OMV from V. anguillarum and OMV protein was separated by SDS-PAGE. Magor band was sliced and analyzed by MALDI-TOF. The major protein band of 38kDa was identified as OmpU by MALDI-TOF MS analysis.

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A Polymeric Micellar Carrier for the Solubilization of Biphenyl Dimethyl Dicarboxylate

  • Chi, Sang-Cheol;Yeom, Dae-Il;Kim, Sung-Chul;Park, Eun-Seok
    • Archives of Pharmacal Research
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    • 제26권2호
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    • pp.173-181
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    • 2003
  • A polymeric micelle drug delivery system was developed to enhance the solubility of poorly-water soluble drug, biphenyl dimethyl dicarboxylate, DDB. The block copolymers consisting of poly(D,L-lactide) (PLA) as the hydrophobic segment and methoxy poly(ethylene glycol) (mPEG) as the hydrophilic segment were synthesized and characterized by NMR, DSC and MALDI-TOF mass spectroscopy. The size of the polymeric micelles measured by dynamic light scattering showed a narrow monodisperse size distribution with the average diameter less than 50 nm. The MW of mPEG-PLA, 3000 (MW of mPEG, 2 K; MW of PLA, 1K), and the presence of hydrophilic and hydrophobic segments on the polymeric micelles were confirmed by MALDI-TOF mass spectroscopy and NMR, respectively. Polymeric micelle solutions of DDB were prepared by three different methods, i.e. the matrix method, emulsion method and dialysis method. In the matrix method, DDB solubility was reached to 13.29 mg/mL. The mPEG-PLA 2K-1K micelle system was compared with the poloxamer 407 micelle system for their critical micelle concentration, micelle size, solubilizing capacity, stability in dilution and physical state. DDB loaded-polymeric micelles prepared by the matrix method showed a significantly increased aqueous solubility (>5000 fold over intrinsic solubility) and were found to be superior to the poloxamer 407 micelles as a drug carrier.

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.

쌀겨 추출물로부터 스핑고당지질의 분리와 구조결정 (Isolation and Characterization of Major Glycosphingolipid from Rice Bran Extract)

  • ;;강병원
    • Applied Biological Chemistry
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    • 제50권1호
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    • pp.72-76
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    • 2007
  • 대표적인 스핑고지질인 글리코실세라미드의 생리적 기능을 조사하기 위하여 쌀겨 추출물로부터 cerebroside를 분리하였다. 정제하지 않은 글리코실세라미드를 flash system 칼럼으로 분리한 후 ELSD를 검출기로 사용하여 순상 HPLC로 정제하였다. 클로로포름 : 메탄올 : 증류수(99:11:1, v/v/v)을 용출용매로 사용하여 주요 cerebroside를 얻을 수 있었고, MALDI/TOF-MS, FAB-MS, GC, 600MHz $^1$H-NMR로 구조를 분석하였다. 구성당은 글루코오스였고 cerebroside의 구성 지방산은 2-hydroxy-arachidic산이었다. 장쇄 염기는 sphinga-4,8-dienine이었다.

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.

경북지역 개와 고양이에서 methicillin-resistant Staphylococcus. pseudintermedius (MRSP) 검출률 조사 (Detection of methicillin-resistant S. pseudintermedius (MRSP) isolated from dogs and cats in Kyungpook Area)

  • 변준호;손준형;이영미;정효훈;오태호
    • 한국동물위생학회지
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    • 제41권4호
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    • pp.229-237
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    • 2018
  • Identification of antibiotic resistant bacteria isolated from animals is necessary for seeking a proper treatment and for preventing the spread of the bacteria among animals. Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is of worldwide concern in veterinary medicine. This study was conducted to investigate the antimicrobial susceptibility of Staphylococcus pseudintermedius (S. pseudintermedius) isolated from dogs and cats. Out of the total number of 150 specimens, 35 isolates (23.3%) were identified as S. pseudintermedius when tested by MALDI-TOF MS (Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectroscopy). Among them, 5 isolates (14.3%) were the case of MRSP. In the results of the antimicrobial susceptibility test, the isolates of S. pseudintermedius were susceptible to amikacin (100%), cephalothin (100%), vancomycin (100%), amoxicillin (85.7%), oxacillin (85.7%), enrofloxacin (82.8%), chloramphenicol (80%). On the other hand, they were resistant to penicillin (71.4%), tetracycline (48.6%), ampicillin (37.1%), kanamycin (31.5%), erythromycin (31.4%), respectively.