• Title/Summary/Keyword: MALDI-TOF-MS

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Comparative Proteome Analysis of Two Antagonist Bacillus subtilis Strains

  • Zhang, C.X.;Zhao, X.;Han, F.;Yang, M.F.;Chen, H.;Chida, T.;Shen, S.H.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.351-357
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    • 2009
  • Natural wild-type strains of Bacillus subtilis are extensively used in agriculture as biocontrol agents for plants. This study examined two antagonist B. subtilis strains, KB-1111 and KB-1122, and the results illustrated that KB-1122 was a more potent inhibitor of the indicator pathogen than KB-1111. Thus, to investigate the intrinsic differences between the two antagonist strains under normal culture conditions, samples of KB-1111 and KB-1122 were analyzed using MALDI-TOF-MS. The main differences were related to 20 abundant intracellular and 17 extracellular proteins. When searching the NCBI database, a number of the differentially expressed proteins were identified, including 11 cellular proteins and 10 secretory proteins. Among these proteins, class III stress-response-related ATPase, aconitate hydratase, alpha-amylase precursor, and a secretory protein, endo-l, 4-beta-glucanase, were differentially expressed by the two strains. These results are useful to comprehend the intrinsic differences between the antagonism of KB-1111 and KB-1122.

Assessment of Neuronal Cell-Based Cytotoxicity of Neurotoxins from an Estuarine Nemertean in the Han River Estuary

  • Kwon, Yeo Seon;Min, Seul Ki;Yeon, Seung Ju;Hwang, Jin ha;Hong, Jae-Sang;Shin, Hwa Sung
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.725-730
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    • 2017
  • A heteronemertean, Yininemertes pratensis, was collected in Han River Estuary, South Korea. This estuarine nemertean has been known by the local fishermen for harmful effects to the glass eels, juveniles of Japanese eel Anguilla japonica, migrating to fresh water. The present study confirmed the neurotoxic effects of this heteronemertean ribbon worm at the cellular level. Derivative types of neurotoxic tetrodotoxin (TTX), 5,11-dideoxy TTX (m/z 288) and 11-norTTX-6(S)-01 (m/z 305.97), were identified through HPLC and MALDI-TOF MS. However, significant neurotoxicity was confirmed in the fraction containing an undefined molecule corresponding to the 291.1 (m/z) peak, when tested in rat primary astrocytes and dorsal ganglion cells. This study is the first to report neurotoxins of the estuarine nemertean, fairly abundant in the Han River estuary, and suggests the long-term monitoring of population dynamics and surveillance of the toxicity in this river estuary.

Tropomyosin and triosephosphate isomerase are upregulated proteins affecting Ginseng treatments in chicken muscle

  • Jung, Kie-Chul;Choi, Kang-Duk;Jang, Byoung-Gui;Sang, Byung-Don;Lee, Jun-Heon
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2004.11a
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    • pp.21-22
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    • 2004
  • The present study was aimed to investigate proteome affected by Panax ginseng extracts in chicken muscles. More than 300 protein spots were detected on silver staining gels. Among them. four protein spots were distinctively up-regulated by Panax ginseng treatments. The up-regulated proteins were finally identified as tropomyosin (2 spots), triosephosphate isomerase, and one unknown protein. Based on the known functions of the identified proteins. they are highly related to the muscle development and enhanced immunity in chicken. These proteins can give valuable information of biochemical roles for Panax ginseng in chicken meats.

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Chemical Characterization of Neutral Extracts Prepared by Treating Pinus radiata Bark with Sodium Bicarbonate

  • MUN, Ji Sun;KIM, Hwan Chul;MUN, Sung Phil
    • Journal of the Korean Wood Science and Technology
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    • v.48 no.6
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    • pp.878-887
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    • 2020
  • A neutral extract (NE), that is soluble in cold water and has excellent antioxidant activity, from Pinus radiata pine bark was prepared by sodium bicarbonate treatment, and its chemical characteristics were investigated. NE was prepared by treating P. radiata bark with 0.8% NaHCO3 aqueous solution with a 5 : 1 liquor-to-bark ratio at boiling temperature for 1 h, resulting in 44% yield and final pH of 6.66. The yield of NE was 11% higher than that of the hot water extract (HWE) due to the increase in the solubility of polyphenols, the main component in the bark, by NaHCO3 treatment. NE was characterized through FT-IR, NMR, and MALDI TOF MS analyses. The results indicated that NE is mostly composed of proanthocyanidins (PAs) consisting of procyanidin (PC) units. The acetylated neutral extract (Ac-NE) had weight average molecular weight (${\bar{M}}w$) of 5,300 Da. The Ac-NE had wide molecular weight distribution and its polydispersity (${\bar{M}}w/{\bar{M}}n$) was 6 times higher than that of pure PA. The antioxidant activity of NE was determined by 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and showed that NE had comparable antioxidant activity with pure PA.

Proteome Analysis of Waito-c Rice Seedlings Treated with Culture Fluid of Gibberellin-producing Fungus, Fusarium proliferatum KGL0401

  • Rim, Soon-Ok;Lee, Jin-Hyung;Hwang, Seon-Kap;Suh, Seok-Jong;Lee, Jin-Man;Rhee, In-Koo;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1990-1994
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    • 2006
  • Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited a high GA productivity. A gas chromatography-mass spectrometry (GC-MS) analysis of extracts of the culture fluid of F proliferatum KGL0401 also revealed the presence of $GA_1$, $GA_3$, $GA_4$, $GA_7$, $GA_{20}$, and $GA_{24}$. Therefore, the present study conducted a proteome analysis of waito-c rice treated with the culture fluid of the isolated F proliferatum KGL0401 to identify the protein expression triggered by the GA-containing culture fluid. The results revealed the overexpression of 180 protein spots in the sample treated with the culture fluid. Among them, 75 induced proteins were selected and analyzed by MALDI-TOF (matrix-assisted laser desorption-iorrization time-of-flight) mass spectrometry, followed by database searching, and 51 proteins were identified.

Assembly Mechanism of [$Fe_2S_2$] Cluster in Ferredoxin from Acidithiobacillus ferrooxidans

  • Chen, Qian;Mo, Hongyu;Tang, Lin;Du, Juan;Qin, Fang;Zeng, Jia
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.124-128
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    • 2011
  • Ferredoxin is a typical iron-sulfur protein that is ubiquitous in biological redox systems. This study investigates the in vitro assembly of a [$Fe_2S_2$] cluster in the ferredoxin from Acidithiobacillus ferrooxidans in the presence of three scaffold proteins: IscA, IscS, and IscU. The spectra and MALDI-TOF MS results for the reconstituted ferredoxin confirm that the iron-sulfur cluster was correctly assembled in the protein. The inactivation of cysteine desulfurase by L-allylglycine completely blocked any [$Fe_2S_2$] cluster assembly in the ferredoxin in E. coli, confirming that cysteine desulfurase is an essential component for iron-sulfur cluster assembly. The present results also provide strong evidence that [$Fe_2S_2$] cluster assembly in ferredoxin follows the AUS pathway.

Purification and Characterization of Lipase Inhibitor from Desmodium oxyphyllum DC. (도둑놈의 갈고리의 지질분해효소 저해물질의 정제 및 특성)

  • Lee, Jong-Kug;Kang, Min-Gu;Baek, Seung-Ye;Ahn, Young-Geun;Lee, Jong-Soo
    • The Korean Journal of Food And Nutrition
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    • v.24 no.3
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    • pp.291-294
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    • 2011
  • A lipase inhibitor from Desmodium oxyphyllum DC. was purified by methanol extraction, systematic solvent extraction, silica gel column chromatography, $C_{18}$ solid phase extraction chromatography and RP-HPLC. We obtained the purified lipase inhibitor with 182 ng($IC_{50}$) of lipase inhibitory activity for a 0.06% yield. Its molecular weight was estimated to be 655.37 Da from an instrumental analysis of MALDI-TOF-MS and it was identified copper-3,5-dibromo-2-hydroxybenzoic acid ($C_{14}H_8Br_4CuO_6$) by $^1H$, $^{13}C$ NMR analysis.

Pituitary Adenoma Biomarkers Identified Using Proteomic Fingerprint Technology

  • Zhou, Kai-Yu;Jin, Hang-Huang;Bai, Zhi-Qiang;Liu, Chi-Bo
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4093-4095
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    • 2012
  • Objective: To determine whether pituitary adenomas can be diagnosed by identifying protein biomarkers in the serum. Methods: We compared serum proteins from 65 pituitary adenoma patients and 90 healthy donors using proteomic fingerprint technology combining magnetic beads with matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Results: A total of 42 M/Z peaks were identified as related to pituitary adenoma (P<0.01). A diagnostic model established based on three biomarkers (3382.0, 4601.9, 9191.2) showed that the sensitivity of diagnosing pituitary adenoma was 90.0% and the specificity was 88.3%. The model was further tested by blind analysis showing that the sensitivity was 88.0% and the specificity was 83.3%. Conclusions: These results suggest that proteomic fingerprint technology can be used to identify pituitary adenoma biomarkers and the model based on three biomarkers (3382.0, 4601.9, 9191.2) provides a powerful and reliable method for diagnosing pituitary adenoma.

Description of 17 unrecorded bacterial species isolated from freshwater showing antibiotic resistance in Korea

  • Baek, Kiwoon;Kim, Eui-Jin;Han, Ji-Hye;Choi, Ahyoung
    • Korean Journal of Environmental Biology
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    • v.38 no.2
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    • pp.289-298
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    • 2020
  • As part of the research program "2018 Rapid screening and identification of freshwater microorganisms using MALDI-TOF/MS library" freshwater samples were collected from a branch of the Nakdong River. Almost 300 antibiotic-resistant bacterial strains were isolated from freshwater samples and subsequently identified by 16S rRNA gene sequencing. Seventeen strains among the isolates shared high 16S rRNA gene sequence similarity (>99.0%) with known species that were not previously recorded in Korea, and each of the isolates also formed a robust phylogenetic clade with the closest species. These species were phylogenetically diverse, belonging to four phyla, seven classes, 10 orders, and 13 genera. At the genus and class level, the previously unrecorded species belonged to Rhodovarius, Xanthobacter, and Shinella of the class Alphaproteobacteria; Ottowia, Simplicispira, and Zoogloea of Betaproteobacteria; Pseudomonas, Acinetobacter, and Shewanella of Gammaproteobacteria; Arcobacter of Epsilonproteobacteria; Sphingobacterium of Sphingobacteriia; Trichococcus of Bacilli; and Leucobacter of Actinobacteria. The previously unrecorded species were further characterized by examining their gram-staining, colony and cell morphology, biochemical properties, and phylogenetic position.

In Vitro Glycosylation of Peptide (RKDVY) and RNase A by PNGase F

  • Park, Su-Jin;Lee, Ji-Youn;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.191-195
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    • 2003
  • The in vitro glycosylation of pentapeptide (Arg-Lys-Asp-Val-Tyr; RKDVY) and RNase A was carried out using PNGase F (peptide-N-glycosidase F), and the results were analyzed using MALDI-TOF-MS. Aminated N,N-diretyl chitobiose was used as the sugar in the glycosylation reaction, and the amination yield of N,N'-diacetyl chitobiose was about $60\%$. To reduce the water activity and shift the reaction equilibrium to a reverse reaction, 1,4-dioxane or ethylene glycol was used as the organic solvent in the enzymatic glycosylation. A certain extent of nonenzymatic glycosylaton, known as the Maillard reaction, was also observed, which occurs on an arginine or lysine residue when the length of tie sugar residue is one or two. However, the extent of glycosylation was much higher in the enzymatic reaction, indicating that PNGase F can be effectively used to produce glycopeptides and glycoproteins in vitro.