• Title/Summary/Keyword: MALDI-TOF MS/MS

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Effect of Dihydroxybenzoic Acid Isomers on the Analysis of Polyethylene Glycols in MALDI-MS

  • Lee, Ae-Ra;Yang, Hyo-Jik;Kim, Yang-Sun;Kim, Jeong-Kwon
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1127-1130
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    • 2009
  • The effects of different dihydroxybenzoic acid (DHB) isomers, when used as matrix materials in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), were investigated in analyses of polyethylene glycol (PEG) polymers. PEG polymers ranging from 400 to 8,000 Da were prepared in different DHB isomer matrices using solvent-based and solvent-free methods. PEG samples were detected only in matrices of 2,3-DHB, 2,5-DHB, and 2,6-DHB while the most intense peaks were observed using 2,6-DHB in both solvent-free and solvent-based preparations.

Biochemical mechanisms of fumigant toxicity by ethyl formate towards Myzus persicae nymphs (복숭아혹진딧물(Myzus persicae) 약충에 대한 에틸포메이트 훈증 독성의 생화학적 메커니즘)

  • Kim, Kyeongnam;Lee, Byung-Ho;Park, Jeong Sun;Yang, Jeong Oh;Lee, Sung-Eun
    • Journal of Applied Biological Chemistry
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    • v.60 no.3
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    • pp.271-277
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    • 2017
  • Ethyl formate has been used for the control of insect pests by fumigation. However, there were not many reports to show its target site of fumigant toxicity on insect pests since its first use in the agricultural industry. In the present study, we showed the presumable target sites of ethyl formate fumigation in insect pests using Myzus persicae nymphs. After ethyl formate fumigation, the nymphs of this species were collected and the changes at the biochemical and molecular level were determined. The activity of cytochrome c oxidase (COX) was approximately two-fold higher after ethyl formate fumigation. In addition, the expression levels of acetylcholinesterase (AChE) decreased gradually with increasing ethyl formate concentration. These two findings suggested that COX and AChE might be the major target sites of ethyl formate fumigation. In addition to these results, the analysis of lipid content using MALDI-TOF MS/MS identified 9 phospholipids differently generated 2-fold higher in the ethyl formate-treated nymphs than that in the control nymphs, thereby leading to changes in cell membrane composition in M. persicae nymphs. Therefore, the ethyl formate fumigation caused lethal effects on M. persicae nymphs by changing COX activity, AChE gene expression, and phospholipid production.

Characterization of Thermal Degradation of Polytrimethylene Terephthalate by MALDI-TOF Mass Spectrometry

  • Jang, Sung-Woo;Yang, Eun-Kyung;Jin, Sung-Il;Cho, Young-Dal;Choe, Eun-Kyung;Park, Chan-Ryang
    • Bulletin of the Korean Chemical Society
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    • v.33 no.3
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    • pp.833-838
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    • 2012
  • The thermal degradation products of polytrimethylene terephthalate (PTT) obtained by heating the sample in the temperature range of $250-360^{\circ}C$ under non-oxidative conditions was characterized using MALDI-TOF (matrix assisted laser desorption/ionization) mass spectrometry. The structures of the degradation products were determined and the relative compositions were estimated. The MALDI-TOF mass spectra of the thermally degraded PTT sample showed three main series of oligomer products with different end groups, which were carboxyl/carboxyl, carboxyl/allyl, and allyl/allyl. In contrast to the thermal degradation of polyethylene terephthalate (PET), the oligomers containing terephthalic anhydrides were not detected, whereas the formation of oligomers containing the unsaturated allyl ester group was confirmed by mass assignment. From these results, it was concluded that the thermal degradation of PTT proceeds exclusively through the ${\beta}$-CH hydrogen transfer mechanism, which is in accordance with the proposed reaction mechanism for the thermal degradation of polybutylene terephthalate (PBT).

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.

Postharvest Biological Control of Colletotrichum acutatum on Apple by Bacillus subtilis HM1 and the Structural Identification of Antagonists

  • Kim, Hae-Min;Lee, Kui-Jae;Chae, Jong-Chan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1954-1959
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    • 2015
  • Bacillus subtilis HM1 was isolated from the rhizosphere region of halophytes for its antifungal activity against Colletotrichum acutatum, the causative agent of anthracnose. Treatment of postharvest apples with the cell culture or with a cell-free culture supernatant reduced disease severity 80.7% and 69.4%, respectively. Both treatments also exhibited antifungal activity against various phytopathogenic fungi in vitro. The antifungal substances were purified and analyzed by acid precipitation, gel filtration, high-performance liquid chromatography, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Three compounds were identified as fengycin, iturin, and surfactin. The MALDI-TOF/TOF mass spectrum revealed the presence of cyclized fengycin homologs A and B, which were distinguishable on the basis of the presence of either alanine or valine, respectively, at position 6 of the peptide sequence. In addition, the cyclized structure of fengycin was shown to play a critical role in antifungal activity.

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.

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.