• Title/Summary/Keyword: MALDI-TOF Mass

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

  • Byun, Jun Ho;Sohn, Jun Hyung;Lee, Young Mi;Jeong, Hyo Hoon;Oh, Tae Ho
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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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.

Bacterial Identification and Detection of Equol in Korean Soybean Paste (한국 된장에서 Equol의 검출 및 미생물 동정)

  • Woo, Seung-Gyun;Lee, So-Yeon;Choi, Go-Woon;Hong, You-Jin;Lee, So-Min;Park, Kang Gyun;Eom, Yong-Bin
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.286-291
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    • 2015
  • Equol has beneficial effects on human health. Fermented soy products contain equol, and many microbes participate in the equol production process. This study investigated fermented Korean soybean paste, doenjang. Thirty seven doenjang samples collected from different manufacturers were examined. Equol was detected in 3 samples (D2, D13, and D19) at the maximum content of 507 ng/100 g by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fifteen microbial species were isolated and identified by 16S rRNA gene sequence analysis and by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Bacillus spp, Paenibacillus spp, Tetragenococcus spp, Stapylococcus spp, and Clostridium species were the predominant bacteria in equol containing doenjang samples.

Proteomic Analysis of Cytokinin Induced Proteins in Arabidopsis (단백체를 이용한 애기장대 Cytokinin 유도 단백질의 분석)

  • Liang Ying-Shi;Cha Joon-Yung;Ermawati Netty;Jung Min-Hee;Bae Dong-Won;Lee Chang-Won;Son Dae-Young
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.251-256
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    • 2005
  • Cytokinins are essential plant hormones that play crucial roles in various aspects of plant growth and development. To better understand the molecular mechanisms of cytokinin action, we identified cytokinin related proteins by a proteomic approach. Proteins extracted from control and trans-zeatin treated Arabidopsis seedlings were separated and analyzed by two dimensional gel analysis. Differentially expressed protein spots were identified with peptide mass fingerprinting based on matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and database searching, We obtained ten up-regulated and one down-regulated proteins upon t-zeatin treatment. The expression of the following proteins was induced; pollen allergen like protein, L-ascorbate peroxidase, tetrapyrrole methylase family protein, SGT1 protein homolog, disease resistance related protein, maternal embryogenesis control protein, paxneb related protein, gluthathione S-transferase and IAA amino acid hydrolase homolog.

Purifications and Characterizations of a Ferredoxin and Its Related 2-Oxoacid:Ferredoxin Oxidoreductase from the Hyperthermophilic Archaeon, Sulfolobus solfataricus P1

  • Park, Young-Jun;Yoo, Chul-Bae;Choi, Soo-Young;Lee, Hee-Bong
    • BMB Reports
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    • v.39 no.1
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    • pp.46-54
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    • 2006
  • The coenzyme A-acylating 2-oxoacid:ferredoxin oxidoreductase and ferredoxin (an effective electron acceptor) were purified from the hyperthermophilic archaeon, Sulfolobus solfataricus P1 (DSM1616). The purified ferredoxin is a monomeric protein with an apparent molecular mass of approximately 11 kDa by SDS-PAGE and of $11,180{\pm}50$ Da by MALDI-TOF mass spectrometry. Ferredoxin was identified to be a dicluster, [3Fe-4S][4Fe-4S], type ferredoxin by spectrophotometric and EPR studies, and appeared to be zinc-containing based on the shared homology of its N-terminal sequence with those of known zinc-containing ferredoxins. On the other hand, the purified 2-oxoacid: ferredoxin oxidoreductase was found to be a heterodimeric enzyme consisting of 69 kDa $\alpha$ and 34 kDa $\beta$ subunits by SDS-PAGE and MALDI-TOF mass spectrometry. The purified enzyme showed a specific activity of 52.6 units/mg for the reduction of cytochrome c with 2-oxoglutarate as substrate at $55^{\circ}C$, pH 7.0. Maximum activity was observed at $70^{\circ}C$ and the optimum pH for enzymatic activity was 7.0 -8.0. The enzyme displays broad substrate specificity toward 2-oxoacids, such as pyruvate, 2-oxobutyrate, and 2-oxoglutarate. Among the 2-oxoacids tested (pyruvate, 2-oxobutyrate, and 2-oxoglutarate), 2-oxoglutarate was found to be the best substrate with $K_m$ and $k_{cat}$ values of $163\;{\mu}M$ and $452\;min^{-1}$, respectively. These results provide useful information for structural studies on these two proteins and for studies on the mechanism of electron transfer between the two.

Purification and Characterization of Manganese Peroxidase of the White-Rot Fungus Irpex lacteus

  • Shin Kwang-Soo;Kim Young Hwan;Lim Jong-Soon
    • Journal of Microbiology
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    • v.43 no.6
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    • pp.503-509
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    • 2005
  • The production of manganese peroxidase (MnP) by Irpex lacteus, purified to electrophoretic homogeneity by acetone precipitation, HiPrep Q and HiPrep Sephacryl S-200 chromatography, was shown to correlate with the decolorization of textile industry wastewater. The MnP was purified 11.0-fold, with an overall yield of $24.3\%$. The molecular mass of the native enzyme, as determined by gel filtration chromatography, was about 53 kDa. The enzyme was shown to have a molecular mass of 53.2 and 38.3 kDa on SDS-PAGE and MALDI-TOF mass spectrometry, respectively, and an isoelectric point of about 3.7. The enzyme was optimally active at pH 6.0 and between 30 and $40^{\circ}C$. The enzyme efficiently catalyzed the decolorization of various artificial dyes and oxidized Mn (II) to Mn (III) in the presence of $H_2O_2$. The absorption spectrum of the enzyme exhibited maxima at 407, 500, and 640 nm. The amino acid sequence of the three tryptic peptides was analyzed by ESI Q- TOF MS/MS spectrometry, and showed low similarity to those of the extracellular peroxidases of other white-rot basidiomycetes.

Multiple Intramuscular Abscesses Caused by Nocardia abscessus in a Patient with Chronic Obstructive Lung Disease: Clinical Microbiology Considerations (만성 폐쇄성 폐질환으로 저용량 스테로이드 유지 중인 환자에게 발생한 Nocardia abscessus에 의한 다발성 근육 농양 1예)

  • Jung-Ah Kim;Hyunjoo Dong;Eunjung Lee;Jongtak Jung;Yae Jee Baek;Tae Hyong Kim;Tae Youn Choi
    • The Korean Journal of Medicine
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    • v.99 no.1
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    • pp.50-56
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    • 2024
  • Nocardiosis is uncommon. Immunocompromising conditions predispose individuals to pulmonary and disseminated nocardiosis of the brain, skin, and subcutaneous tissues. The most common pathogens are Nocardia cyriacigeorgica, Nocardia nova, and Nocardia farcinica. The speciation of Nocardia to determine antimicrobial susceptibility is difficult using traditional biochemical methods. Here, we report the case of a 73-year-old man with chronic obstructive lung disease who developed a rapidly progressing intramuscular abscess around the left hip and thigh. Within 3 days, the lesions progressed to an epidural abscess at the L4 to S1 level. Although he was treated with broad-spectrum antibiotics and extensive incision and drainage, he died of rapidly progressive respiratory failure. Nocardia abscessus (N. abscessus) was identified in pus samples using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). This case shows that the diagnosis of an intramuscular abscess caused by N. abscessus is challenging and that using MALDI-TOF MS may facilitate the diagnosis and ensure appropriate treatment.

Identification of Lactobacillus spp. associated with nematodes in peach farm soil (복숭아 농장 토양에서 Nematodes와 연관된 Lactobacillus spp.의 분리 및 동정)

  • Lee, Woo-Hyun;Choi, Jae Im;Lee, Jin Il;Lee, Won-Pyo;Yoon, Sung-Sik
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.163-169
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    • 2017
  • Strains D4 and D5 were isolated from peach-rotten soil during the peach harvest season. The isolates were identified based on morphological and biochemical characterization, and identification was determined by 16S rRNA gene sequencing. Results showed that D4 has high similarity to Lactobacillus plantarum ATCC $14917^T$ and Lactobacillus pentosus ATCC $8041^T$ at 99.05% and 98.98%, respectively. D5 was also similar to Lactobacillus pentosus ATCC $8041^T$ and Lactobacillus plantarum ATCC $14917^T$ at 98.71% and 98.64%, respectively. In contrast, isolates showed differences in carbohydrate utilization in comparison to Lactobacillus plantarum ATCC $14917^T$ and Lactobacillus pentosus ATCC $8041^T$. In view of this we performed VITEK MS matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis, multiplex PCR fingerprinting, and random amplified polymorphic DNA (RAPD)-PCR to further confirm the identification of D4 and D5. The results of these analyses showed that both strains were most similar to Lactobacillus plantarum.

Identification of Cold Stress-related Proteins in Rice Leaf Tissue (벼의 잎 조직에서 발현되는 저온 스트레스 관련 단백질의 분리 동정)

  • Lee Dong-Gi;Lee Sang-Hoon;Lee Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.4
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    • pp.287-296
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    • 2005
  • To investigation protein expression pattern in rice leaves exposed to cold stress, the soluble proteins extracted from leaf tissue were fractionated with $15\%$ PEG and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Differentially expressed proteins were identified by peptide mass fingerprinting using matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Eight proteins up-regulated and 10 down-regulated were found in $15\%$ PEG supernatant fraction. In addition, 13 proteins up-regulated and 14 down-regulated were found in $15\%$ PEG pellet fraction. It was identified the differentially expressed proteins in $15\%$ PEG supernatant fraction as pimerase/dehydratase fructokinase, ribose-5-phosphate isomerase (Rpi), chaperonin 21 precursor, probable photosystem II oxygen-envolving complex (PS II OEC) protein 2 precursor and thioredoxin h-type (Trx-h) and those in $15\%$ PEG pellet fraction as OSINBb0059K02.15, hypothetical protein, putative mitogen-activated protein kinase kinase (MAPKK), beta 7 subunit of 205 proteasome, ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit. These proteins are involved in metabolism, energy, protein synthesis, disease/defense and signal transduction-related proteins.