• Title/Summary/Keyword: MALDI MS

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Identification of LAB and Fungi in Laru, a Fermentation Starter, by PCR-DGGE, SDS-PAGE, and MALDI-TOF MS

  • Ahmadsah, Lenny S.F.;Kim, Eiseul;Jung, Youn-Sik;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.32-39
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    • 2018
  • Samples of Laru (a fermentation starter) obtained from the upper part of Borneo Island were analyzed for their lactic acid bacteria (LAB) and fungal diversity using both a culture-independent method (PCR-DGGE) and culture-dependent methods (SDS-PAGE and MALDI-TOF MS). Pediococcus pentosaceus, Lactobacillus brevis, Saccharomycopsis fibuligera, Hyphopichia burtonii, and Kodamaea ohmeri were detected by all three methods. In addition, Weissella cibaria, Weissella paramesenteroides, Leuconostoc citreum, Leuconostoc mesenteroides, Lactococcus lactis, Rhizopus oryzae/Amylomyces rouxii, Mucor indicus, and Candida intermedia were detected by PCR-DGGE. In contrast, Lactobacillus fermentum, Lactobacillus plantarum, Pichia anomala, Candida parapsilosis, and Candida orthopsilosis were detected only by the culture-dependent methods. Our results indicate that the culture-independent method can be used to determine whether multiple laru samples originated from the same manufacturing region; however, using the culture-independent and the two culture-dependent approaches in combination provides a more comprehensive overview of the laru microbiota.

Purification and Characterization of PC-Like Cadmium-Binding Peptide from Root of Rumex crispus

  • Chang, Ju-Youn;Lee, In-Sook;Park, Jin-Sung;Chang, Yoon-Young;Bae, Bum-Han
    • The Korean Journal of Ecology
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    • v.26 no.5
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    • pp.263-266
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    • 2003
  • This research investigated the process of removing cadmium and tested the detoxification mechanism of the cadmium-binding peptide (Cd-BP) from Rumex crispus. Phytochelatin-like cadmium-binding peptide (PC-Cd-BP) of Rumex crispus was purified and identified. Rumex crispus was exposed to 4.3 mg Cd/L for seven days. Heat-treated supernatant fraction taken by root tissues showed traces of PC-Cd-BP An analysis of the material through Gel-filteration chromatography on the Sephadex G-75 column showed two symmetrical Cd-BP peaks. The major peak with the smaller molecular weight was further purified by $C_{18}$ reverse-phase HPLC to produce apparent homogeneity. The amino acid composition of Cd-BP from Rumex crispus included cysteine (22.6%), glutamate and glutamate acid (20%), and glycine (12%). It was similar the amino acid composition of most PC. The molecular weight of the purified peptide was determined at 568-706 Da by MALDI-TOF MS. Therefore, the Cd-BP of Rumex crispus was PC-Cd-BP consisting of isopeptides.

Proteome analysis of chloroplast proteins in stage albinism line of winter wheat (triticum aestivum) FA85

  • Hou, Dian-Yun;Xu, Hong;Du, Guang-Yuan;Lin, Jun-Tang;Duan, Min;Guo, Ai-Guang
    • BMB Reports
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    • v.42 no.7
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    • pp.450-455
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    • 2009
  • The "stage albinism line of winter wheat" FA85 was a specific natural mutant strain on leaf color. This physiological mutation was controlled by cytogene. In order to reveal the genetic and biochemical mechanism of albinism, 2-DE was used to investigate the difference of chloroplast protein expression pattern between FA85 and its parent wheat Aibian 1. From the results of 2-DE gels analysis, approximately 683 spots were detected on each gel, and 57 spots were expressed differently at least two-fold. Using MALDI-TOF/TOF MS, 14 of 57 spots were identified, which could be categorized into four classes: carbon metabolism, energy metabolism, defense/stress response and signal transduction. Compared with the parent wheat, the expression of ATPase-$\gamma$ and GP1-$\alpha$ was up-regulated in FA85, and of other proteins was down-regulated. Together, we concluded that the expression of chloroplast proteins had changed obviously in FA85, which might be related to the leaf color mutant.

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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    • v.21 no.10
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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.

SFRP Synthesis of Acenaphthylene Oligomers and Block Copolymers. Potential Light Harvesting Structures

  • Ali, Dildar;Ahmed, Zaheer;Dust, Julian M.;Kazmaier, Peter M.;Buncel, Erwin
    • Bulletin of the Korean Chemical Society
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    • v.32 no.7
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    • pp.2377-2384
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    • 2011
  • Azo-acenaphthylene oligomers with repeating acenaphthylene units "n" up to 4, 5, 7, 17 and 19 have been prepared successfully using nitroxide mediated Stable Free Radical Polymerization (SFRP). Azo-acenaphthylene oligomers, reversibly end-capped by the stable nitroxide 2,2,6,6-tetramethyl-1-piperidinoxyl (TEMPO), were further reacted via radical addition to 4-(naphthalenemethoxy)styrene monomer for diblock co-polymer formation. Characterization of the oligomers and diblock co-polymers was accomplished using MALDI-MS supported by GPC (Gel Permeation Chromatography) and $^1H$ NMR spectrometry. MALDI-MS afforded definitive results by providing an inter-peak interval of 152 (m/z), corresponding to acenaphthylene monomer, and inter-peak interval of 260 (m/z) for the naphthalenemethoxystyrene monomer unit in block copolymers. Our study opens the way to control the number of repeat units in the oligomers. Further these oligomers can be tailored with various monomers for the formation of block copolymers.

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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    • v.14 no.2
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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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    • v.45 no.3
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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.

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

  • Ko, Weon-Bae;Park, Byoung-Eun;Lee, Young-Min
    • Elastomers and Composites
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    • v.43 no.1
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    • pp.31-38
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    • 2008
  • Sonochemical reaction of fullerene oxides, $[C_{70}(O)_n](n\geq1)$ with several aromatic amines such as 4-nitroaniline, 3-nitroaniline, and 4-isopropylaniline, in the presence of $FeCl_3$ were investigated under ultrasonic irradiation. This method is applicable to a wide variety of aromatic amines especially ring deactivated, to afford the corresponding cleavage products under mild conditions. The aminated fullerenes were confirmed by MALDI-TOF-MS and UV-vis spectra.

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

  • Jin Young Yun
    • Biomedical Science Letters
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    • v.29 no.4
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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.

Alteration of Phospholipids during the Mitophagic Process in Lung Cancer CellsS

  • Lee, Jae Won;Cho, Kyung Mi;Jung, Jae Hun;Tran, Quangdon;Jung, Woong;Park, Jongsun;Kim, Kwang Pyo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1790-1799
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    • 2016
  • Matrix assisted laser desorption ionization (MALDI)-time of flight/mass spectrometry (TOF/MS) was applied to investigate alterations in phospholipids in mitophagic cancer cells. Several phospholipids, including phosphatidylcholines (PCs), sphingomyelins (SMs), and phosphatidylinositols (PIs), were successfully analyzed in control and mitophagy-induced H460 cells in the positive and negative ion modes. Principal component analysis was applied to differentiate the two groups. The upregulated and downregulated phospholipid species in the mitophagic cells were also represented in a heatmap. In the volcano plot (fold change > 1.3 and p value < 0.01), individual species of seven PCs, two SMs, and three PIs were selected as differentially regulated phospholipids. In particular, almost all the molecular species of PC, SM, and PI were downregulated in the mitophagic cells. Quantification of these lipids indicated that mitophagy induces altered metabolism of phospholipids. Therefore, phospholipid alterations during the mitophagic process of lung cancer cells were well characterized by MALDI-TOF/MS.