• Title/Summary/Keyword: MALDI-TOF mass spectrometry

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The study for the synthesis and analysis of metal chelated protein (단백질의 메탈 킬레이션 화합물 제조 및 확인에 관한 연구)

  • Kim, Sung-Ho;Nam, Hae-Seon;Lee, Yoon-Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.8 no.5
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    • pp.1273-1278
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    • 2007
  • In living system, various types of metal chelated protein show their intrinsic biological activities. In order to investigate the chelation reaction between protein and metal, zinc chelated peptide was systhesized by the peptide, hydrolyzed protein, with zinc. And simple analytical method fot the chelation reaction was developed using MALDI-TOF mass spectrometry.

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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.

Rapid Identification of Vibrio Species Isolated from the Southern Coastal Regions of Korea by MALDI-TOF Mass Spectrometry and Comparison of MALDI Sample Preparation Methods

  • Cho, Youngjae;Kim, Eiseul;Han, Sun-Kyung;Yang, Seung-Min;Kim, Mi-ju;Kim, Hyun-Joong;Kim, Chang-Gyeom;Choo, Dong-Won;Kim, Young-Rok;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1593-1601
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    • 2017
  • Vibrio species are generally recognized as pathogens predominant in seafood along coastal areas. The food industry has sought to develop efficient microbial detection methods. Owing to the limits of conventional methods, this study aimed to establish a rapid identification method for Vibrio isolated from Korea, based on matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Four different preparation procedures were compared to determine the appropriate means to pretreat Vibrio species, using 17 isolates and five reference strains. Extended direct transfer and full formic acid extraction methods using bacterial colonies on agar plates revealed very low identification rates. Formic acid and trifluoroacetic acid (TFA) extractions using bacterial broth cultures were also performed. All Vibrio isolates and reference strains prepared by TFA extraction were successfully identified to the species level (17/22, 77.3%) and to the genus level (5/22, 22.7%). Thus, TFA extraction was considered the most appropriate method to pretreat Vibrio species for MALDI-TOF MS. The remaining 33 isolates and two reference strains were prepared by TFA extraction and analyzed by MALDI-TOF MS. Overall, 50 isolates were identified to the species level (40/50, 80%) and to the genus level (10/50, 20%). All isolates were identified as 43 V. alginolyticus, six V. parahaemolyticus, and one V. vulnificus species. V. alginolyticus and V. parahaemolyticus were isolated from fish offal (87.5% and 12.5%, respectively), seawater (91.3%, 8.7%), and shellfish (62.5%, 37.5%), whereas V. alginolyticus and V. vulnificus were isolated from sediment (90.9% and 9.1%, respectively). This study established a reliable system of MALDI-TOF MS preparation and analysis for Vibrio identification.

Enrichment of Peptides using Novel C8-functionalized Magnetic Nanoparticles for Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometric Analysis

  • Song, Sun-Mi;Yang, Hyo-Jik;Kim, Jin-Hee;Shin, Seong-Jae;Park, Eun-Hye;Kim, Jeong-Kwon
    • Mass Spectrometry Letters
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    • v.2 no.2
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    • pp.53-56
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    • 2011
  • [ $C_8$ ]functionalized magnetic nanoparticles were synthesized by coating magnetic $Fe_3O_4$ nanoparticles with silicaamine groups using 3-aminopropyltriethoxysilane and by subsequently modifying the amine groups with chloro(dimethyl)octylsilane to produce octyl groups on the surface of the MNPs. The $C_8$-functionalized MNPs were used to enrich peptides from tryptic protein digests of myoglobin and ${\alpha}$-casein. The enriched peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). MALDI-MS was also used to investigate desalting of the $C_8$-functionalized MNPs. Sample solutions were prepared in 1.0 M NaCl, and the successful removal of salt was observed. Enrichment with $C_8$-functionalized MNPs was very effective for separating and concentrating tryptic peptides.

Effective Identification of Ochrobactrum anthropi Isolated from Clinical Specimens (임상검체에서 분리된 Ochrobactrum anthropi의 효과적인 동정)

  • Ko, Hyun-Mi;Jo, Jun-Hyeon;Baek, Hae-Gyeong
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.221-228
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    • 2020
  • Ochrobactrum anthropi is a non-fermentative oxidative gram-negative bacillus that produces oxidase. Distinguishing a mixed culture with non-fermenting bacteria having a similar appearance and oxidase-positive is difficult, and there is a limit to accurate identification with a biochemical identification system. This paper proposes that the Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Platform (MALDI-TOF) method is useful for classifying bacteria that are difficult to identify using biochemical testing methods. As a result of analyzing five cases of O. anthropi examined using MicroScan, it took 6.5 days to the final report, which was 3.5 days more than the 3.0 days of E. coli. The pus sample in patient 5 was a mixed infection with Achromobacter xylosoxidans, and it took 11.3 days because of multiple subculture and retests. Four patients were over 60 years old with an underlying disease, and the possibility of opportunistic and nosocomial infections could not be excluded. Among them, samples collected after 92 days of hospitalization were resistant to imipenem and meropenem. Therefore, an examination using the MALDI-TOF method will be useful for the rapid and adequate treatment of patients with difficult identification, such as O. anthropi.

MALDI-TOF Analysis of Polyhexamethylene Guanidine (PHMG) Oligomers Used as a Commercial Antibacterial Humidifier Disinfectant

  • Hwang, Hyo Jin;Nam, Jungjoo;Yang, Sung Ik;Kwon, Jung-Hwan;Oh, Han Bin
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1708-1714
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    • 2013
  • Polyhexamethylene guanidine (PHMG) polymers used as an active ingredient in an antibacterial humidifier disinfectant were reported to cause harm to the human health when inhaled, although physical contact with this material was known to present low toxicity to humans. It is therefore necessary to develop an optimal analysis method which enables detection and analysis of PHMG polymers. MALDI-TOF investigations of PHMG are performed with a variety of matrices, and it is found that CHCA and 2,5-DHB are excellent matrices which well reflects the polymer population even at high mass. For the provided PHMG sample, the number-average ($M_n$) and weight-average ($M_w$) molecular masses were determined to be 744.8 and 810.7, respectively, when the CHCA was used as a matrix. The rank of the matrices in terms of averaged molecular weight was CHCA ~2,5-DHB > 5-NSA > DHAP, THAP > ATT > IAA ~ super-DHB ~ HABA. In addition, PSD of the PHMG oligomer ions exhibited a few unique fragmenation characteristics. The formation of a- and c-type fragments was the major fragmentation pathway, and the 25-Da loss peaks generally accompanied a- and c-type fragments.

Comparative Proteomic Analyses of the Yeast Saccharomyces cerevisiae KNU5377 Strain Against Menadione-Induced Oxidative Stress

  • Kim, Il-Sup;Yun, Hae-Sun;Jin, In-Gnyol
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.207-217
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    • 2007
  • The Saccharomyces0 cerevisiae KNU5377 strain, which was isolated from spoilage in nature, has the ability to convert biomass to alcohol at high temperatures and it can resist against various stresses [18, 19]. In order to understand the defense mechanisms of the KNU5377 strain under menadione (MD) as oxidative stress, we used several techniques for study: peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) followed by two-dimensional (2D) gel electrophoresis, liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. Among the 35 proteins identified by MALDI-TOF MS, 19 proteins including Sod1p, Sod2p, Tsa1p, and Ahp1p were induced under stress condition, while 16 proteins were augmented under normal condition. In particular, five proteins, Sod1p, Sod2p, Ahp1p, Rib3p, Yaf9p, and Mnt1p, were induced in only stressed cells. By LC-ESI-MS/MS analysis, 37 proteins were identified in normal cells and 49 proteins were confirmed in the stressed cells. Among the identified proteins, 32 proteins were found in both cells. Five proteins including Yel047cp and Met6p were only upregulated in the normal cells, whereas 17 proteins including Abp1P and Sam1p were elevated in the stressed cells. It was interesting that highly hypothetical proteins such as Ynl281wp, Ygr279cp, Ypl273wp, Ykl133cp, and Ykr074wp were only expressed in the stressed cells. SELDI-TOF analysis using the SAX2 and WCX2 chips showed that highly multiple-specific protein patterns were reproducibly detected in ranges from 2.9 to 27.0 kDa both under normal and stress conditions. Therefore, induction of antioxidant proteins, hypothetical proteins, and low molecular weight proteins were revealed by different proteomic techniques. These results suggest that comparative analyses using proteomics might contribute to elucidate the defense mechanisms of KNU5377 under MD stress.

Differential Proteome Analysis of Breast and Thigh Muscles between Korean Native Chickens and Commercial Broilers

  • Liu, Xian De;Jayasena, Dinesh D.;Jung, Yeon-Kuk;Jung, Samooel;Kang, Bo-Seok;Heo, Kang-Nyeong;Lee, Jun-Heon;Jo, Cheo-Run
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.6
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    • pp.895-902
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    • 2012
  • The Korean native chickens (Woorimotdak$^{TM}$, KNC) and commercial broilers (Ross, CB) show obvious differences in meat flavor after cooking. To understand the contribution of protein and peptide for meat flavor, 2-dimensional (2-D) gel electrophoresis and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry was performed. A total of 16 protein spots were differentially expressed in the breast and thigh meat between the two breeds. A total of seven protein spots were represented by different levels between KNC and CB for breast meat. Among them three protein spots (TU39149, TU40162 and TU39598) showed increases in their expressions in KNC while other four protein spots (BU40125, BU40119, BU40029 and BU39904) showed increases in CB. All nine protein spots that were represented by different levels between KNC and CB for thigh meat showed increases in their expression in KNC. Phosphoglucomutase 1 (PGM 1), myosin heavy chain (MyHC), heat shock protein B1 (HSP27), cytochrome c reductase (Enzyme Q), Glyoxylase 1, DNA methyltransferase 3B (DNA MTase 3) were identified as the main protein spots by MALDI-TOF mass spectrometry. These results can provide valuable basic information for understanding the molecular mechanism responsible for breed specific differences in meat quality, especially the meat flavour.

Multidrug-resistant Staphylococci Found on Book Surfaces in East London Libraries

  • Idris, Adi;Cutler, Ron R
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.354-357
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    • 2017
  • There is an increase in the presence of drug-resistant staphylococci outside of the nosocomial and healthcare setting. Although the presence of staphylococci has been studied in several public spaces, nothing is known on the presence of staphylococci in public libraries. Book surfaces from public libraries in the East London area, United Kingdom were swabbed and cultured and identity of the isolates determined by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS). Seven different staphylococcal species were identified by MALDI-TOF-MS analysis. This short study provides evidence of the presence of multidrug-resistant staphylococci in public libraries in the East London area.