• Title/Summary/Keyword: Bacillus identification

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Reliable Identification of Bacillus cereus Group Species Using Low Mass Biomarkers by MALDI-TOF MS

  • Ha, Miyoung;Jo, Hyeon-Ju;Choi, Eun-Kyeong;Kim, Yangsun;Kim, Junsung;Cho, Hyeon-Jong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.887-896
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    • 2019
  • Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based pathogen identification relies on the ribosomal protein spectra provided in the proprietary database. Although these mass spectra can discern various pathogens at species level, the spectra-based method still has limitations in identifying closely-related microbial species. In this study, to overcome the limits of the current MALDI-TOF MS identification method using ribosomal protein spectra, we applied MALDI-TOF MS of low-mass profiling to the identification of two genetically related Bacillus species, the food-borne pathogen Bacillus cereus, and the insect pathogen Bacillus thuringiensis. The mass spectra of small molecules from 17 type strains of two bacilli were compared to the morphological, biochemical, and genetic identification methods of pathogens. The specific mass peaks in the low-mass range (m/z 500-3,000) successfully identified various closely-related strains belonging to these two reference species. The intensity profiles of the MALDI-TOF mass spectra clearly revealed the differences between the two genetically-related species at strain level. We suggest that small molecules with low molecular weight, 714.2 and 906.5 m/z can be potential mass biomarkers used for reliable identification of B. cereus and B. thuringiensis.

Screening and Identification of the Fibrinolytic Bacterial Strain from Jeot-Gal, Salt-fermented Fish (젓갈류로부터 혈전용해 균주의 분리 및 동정)

  • Jang, Young-Ryeol;Kim, Won-Keuk;Kwon, Ik-Boo;Lee, Hyun-Yong
    • Korean Journal of Food Science and Technology
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    • v.30 no.3
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    • pp.655-659
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    • 1998
  • Bacterial strain showing the strong fibrinolytic activity (2.04 plasmin unit) was screened from Jeot-Gal, Korean salt-fermented fish collected from various region. For the identification, when the strain was characterized morphologically, culturally, and biochemically, it was identified to Bacillus pumilus. And, when the fatty acids composition of the strain was analyzed, it was identified to Bacillus atropheus. Finally, the 16S rRNA partial sequence (V3 region) showed that the fibrinolytic stain screened from Jeot-Gal was identified as Bacillus subtilis. So, we named it Bacillus subtilis KJ-48.

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Microbiological Identification of Medical Probiotic Bisspan Strain (의약용 프로바이오틱 비스판균의 미생물학적 동정)

  • 전경동;이광호;김원석;백현동
    • Microbiology and Biotechnology Letters
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    • v.28 no.2
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    • pp.124-127
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    • 2000
  • Beneficial bacteria, which have been used for medical purpose and for medicines for treating intestinal disorders, include strains of Bifidobacterium sp., Lactobacillus sp., Enterococcus sp., Clostridium butyricum, Lactobacillus sporogenes, Bacillus subtilis, Bacillus polyfermenticus and the like. Bacillus polyfermenticuss SCD with is commonly called as Bispan strain has been appropriately used for the treatment of long-term intestinal disorders, since the live strains in the form of active endospores can successfully reach the target intestine. In this study, the identification and characterization of Bispan strain was done using SEM observation, API 50CHB kits, isoprenoid quinone analysis, and fatty acid analysis. These results suggest that Bispan strain is very similar to Bacillus subtilis.

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Isolation and Identification of Microorganisms in Korean Traditional Soybean Paste and Soybean Sauce (전통 된장 및 간장의 숙성기간별 생육 미생물의 분리 및 동정)

  • 유승구;조원희;강수민;이선희
    • Microbiology and Biotechnology Letters
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    • v.27 no.2
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    • pp.113-117
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    • 1999
  • As a basic study for quality improvement of Korean soybean paste and soybean sauce, we investigated on microflora of soybean paste and soybean sauce fermentation. Major Microorganisms were isolated from the sample pastes and sauces, and identified systematically. Selected Microorganisms were identified by MIS whole cell fatty acid analysis by gas chromatography. Identification results showed that Bacillus licheniformis, bacillus pumilis and Bacillus subtilis were dominant in soybean paste and Staphylococcus vitulus, Bacillus subtilis, Bacillus pumilus, and Lactobacillus fermentum were dominant in soybean sauce. It seemed that these Microorganisms played an important role in soybean paste and soybean sauce fermentation and could be used for the further studies such as protease and amylase activities.

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Identification Based on Computational Analysis of rpoB Sequence of Bacillus anthracis and Closely Related Species (Bacillus anthracis와 그 유연종의 rpoB 유전자 컴퓨터 분석을 통한 동정)

  • Kim, Kyu-Kwang;Kim, Han-Bok
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.333-338
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    • 2008
  • Computational analysis of partial rpoB gene sequence (777 bp) was done in this study to identify B. anthracis and its closely related species B. cereus and B. thuringiensis. Sequence data including 17 B. anthracis strains, 9 B. cereus strains, and 7 B. thuringiensis strains were obtained by searching databases. Those sequences were aligned and used for other computational analysis. B. anthracis strains were identificated by in silico restriction enzyme digestion. B. cereus and B. thuringiensis were not segregated by this method. Those sequencing and BLAST search were required to distinguish the two. In actual identification tests, B. anthracis strains could be identified by PCR-RFLP, and B. cereus and B. thuringiensis strains were distinguished by BLAST search with reliable e-value. In this study fast and accurate method for identifying three Bacillus species, and flow chart of identification were developed.

The studies on microbe isolated from the cocoon in Korea. (Isolation and identification of bacteria) (한국산 잠견에서 분리된 미생물에 관한 연구 제 1보 잠견에서 분리된 Bacteria의 분리동정)

  • 이상원;이철준
    • Journal of Sericultural and Entomological Science
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    • v.7
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    • pp.53-63
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    • 1967
  • In order to identify the bacteria living on the cocoons in Korea, the isolated bacterias' morphological. cultural and physiological characters has been determined through the detailed study. The second aim of this experiment was to protect against the bacteria which damage silk protein during storage. 1. The twelve strains of the bacteria were isolated and identified in the cocoons produced in Korea. The results of the identification are as the following. No 1, No 8; Bacillus subtilis variation No 2, ; Bacillus stearothermophilus No 3, ; Bacillus circulans No 5, No 6; Bacillus thuringiensis No 7, No 11; Bacillus brevis No 12, No l0; Bacillus cereus variation

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Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

  • Jeong, Young-Su;Lee, Jonghee;Kim, Seong-Joo
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2635-2639
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    • 2013
  • The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field.

Isolation and identification of Bacillus sp, GS Producing Xylanase (Xylanase생산성 Bacillus sp. GS의 분리 및 동정)

  • An, Jun-Bae;Park, Heon-Guk;Lee, Gye-Ho
    • The Korean Journal of Food And Nutrition
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    • v.7 no.1
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    • pp.8-15
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    • 1994
  • To utilize hemicellulosic biomass efficiently, the microorganism producing xylanase was isolated from fermented sawdust. It was a Gram positive, aerobic and rod shape bacterium. It had endospore and secreted strong hydrolases, such as amylase and protease. Through morphological, cultural and physiological tests, it was identified as Bacillus sp. GS. To increase the productivity of xylanase from Bacillus sp. GS, the enzyme production medium was optimized. The composition of the medium and incubation conditions were like follows xylan 1.25%, yeast extract 0.1%, NaN030.2%, K2HP04 0.1%, MgSO4 0.02%, mineral salt 0.005%, pH 6.5, incubation temperature 37$^{\circ}C$.

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Isolation and Identification of Novel Alkalophilic Bacillus alkalophishaggy JY-827 with Anticaries microbe Streptococcus mutans. (치아 우식 미생물 Streptococcus mutans 에 대해 활균활성을 갖는 신규 호알칼리성 Bacillus alkalophilshaggy JY-827의 분리 및 동정)

  • 전주연;류일환;이상욱;이갑상
    • Microbiology and Biotechnology Letters
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    • v.28 no.5
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    • pp.243-250
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    • 2000
  • The study was performed to investigated the excellent microbial anticaries substance which is more effective than the chlorohexidine in the dental caries treatment. For the screening of alkaliphilic microorganism, more than 1200 bacterial strains were isolated from sea soil sample. A typ-ical strain which produced the most excellent antimicrobial substance was selected. The strain was identified novel alkalophilic Bacillus sp. through the results of morphological, biochemical and chemotaxonomical characteristics and 16S rDNA sequencing and designated as Bacillus alkalophilshaggy JY-827.

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Discrimination of Bacillus anthracis from Bacillus cereus Group Using KHT5 Marker (KHT5 마커를 사용한 Bacillus cereus 그룹에서 Bacillus anthracis의 구별)

  • 김형태;김성주;채영규
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.40-44
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    • 2003
  • Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. In order to develop a DNA marker specific for Bacillus anthracis and to discriminate this species from Bacillus cereus group, we applied the randomly amplified polymorphic DNA (RAPD)-PCR technique to a collection of 29 strains of the genus Bacillus, including 22 species of the B. cereus group. A 709-bp RAPD marker (KHT5) specific for B. anthracis was obtained from B. anthracis BAK. The PCR product of internal primer set from the KHT5 fragment distinguished B. anthracis from the other species of the B. cereus group.