• Title/Summary/Keyword: Bacillus identification

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Isolation and Identification of Bacteria Producing a Soybean Milk Clotting Enzyme (두유 응고효소 생산균의 분리 및 동정)

  • 하덕모;이철우
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.109-114
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    • 1989
  • Seventeen bacterial strains producing an extracellular soybean milk clotting enzyme were Isolated from 150 soil samples, and identified as Bacillus cereus(8 strains), Bacillus pumilus(8 strains) and Bacillus licheniformis (1 strain). Among them, Bacillus pumilus strain 118 and Bacillus licheniformis strain 192 showed relatively high soybean milk clotting activity. The coagulability of enzymes from these strains decreased as the pH of soybean milk was increased from 6.0 to 7.0. The optimum temperature for soybean milk clotting activity was $65^{\circ}C$.

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Immunological Characterization of Bacillus thuringiensis Antigens (Bacillus thuringiensis 항원들의 면역학적 분석)

  • Jung, Jae-Deuk;Park, Jung-Sun;Jo, Young-Soo;Hong, Soon-Bok;Lee, Hyung-Hoan;Cho, Myung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.110-117
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    • 1995
  • This study was carried out to immunologically characterize Bacillus thuringiensis (B.t) antigens. Protein patterns of ultrasonicated- antigens of B. thuringiensis subspecies using SDS- PAGE revealed marked similarities among all the strains analyzed except for the difference between quantative variations of bands and some protein antigens. The comparison of the protein patterns showed that the protein antigen of 45 kilodalton (kd) was common in 11 strains and that the difference between B. thuringiensis subsp. canadensis and galleriae was noticed in quantative variations of bands despite of ambiguous serogrouping, suggesting a useful method for identification. All strains examined showed similar antigenic patterns in SDS-PAGE, while immunodominant bands differed in antigenic reactivity in western blot using polyclonal antibodies. Polyclonal antibody to B. thuringiensis subsp. thuringiensis and israelensis in indirect immunofluorescence assay reacted with flagella and cell surface antigens. The present study indicates that SDS-PAGE and western blot analysis may be used as tools for differentiation and identification of B. thuringiensis subspecies.

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Simultaneous Detection and Identification of Bacillus cereus Group Bacteria Using Multiplex PCR

  • Park, Si-Hong;Kim, Hyun-Joong;Kim, Jae-Hwan;Kim, Tae-Woon;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1177-1182
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    • 2007
  • Bacillus cereus group bacteria share a significant degree of genetic similarity. Thus, to differentiate and identify the Bacillus cereus group efficiently, a multiplex PCR method using the gyrB and groEL genes as diagnostic markers is suggested for simultaneous detection. The assay yielded a 400 bp amplicon for the groEL gene from all the B. cereus group bacteria, and a 253 bp amplicon from B. anthracis, 475 bp amplicon from B. cereus, 299 bp amplicon from B. thuringiensis, and 604 bp amplicon from B. mycoides for the gyrB gene. No nonspecific amplicons were observed with the DNA from 29 other pathogenic bacteria. The specificity and sensitivity of the B. cereus group identification using this multiplex PCR assay were evaluated with different kinds of food samples. In conclusion, the proposed multiplex PCR is a reliable, simple, rapid, and efficient method for the simultaneous identification of B. cereus group bacteria from food samples in a single tube.

Isolation, Identification, and Characterization of Bacillus strains from the Traditional Korean Soybean-fermented Food, Chungkookjang

  • Joo, Myeong-Hoon;Hur, Sung-Ho;Han, Yong-Soo;Kim, Ji-Yeon
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.202-210
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    • 2007
  • A total of 45 bacterial strains were isolated from the traditional Korean soybean-fermented food, Chungkookjang. Among these strains, seven strains were selected and identified based on morphological, physiological, and biochemical characteristics, as well as phylogenetic analysis using 16S rDNA sequences. All strains were Gram-positive, aerobic, motile, oxidase-positive, rod-shaped, and endospore-forming bacteria, and produced extracellular enzymes such as amylase, cellulase, lipase, protease, and xylanase. The isolates were grown in the presence of 0-11% (w/v) NaCl. Growth was optimal at pH 6-9 and at temperatures of $30-45^{\circ}C$. According to VITEK automicrobic system tests and supplementary tests, the isolates were similar to several species of the genus Bacillus. The phylogenetic analysis of seven bacterial strains based on comparisons of 16S rDNA sequences, revealed that the strains were closely related to Bacillus species. The identification of strains that produced surfactin was also carried out, based on PCR screening of the sfp gene. Among the seven isolated strains, six yielded a surfactin-positive result with PCR.

Random Amplified Polymorphic DNA-PCR Analysis for Identification of Bacillus anthracis (탄저균의 Random Amplified Polymorphic DNA-PCR 분석)

  • 김성주;박경현;김형태;조기승;김기천;최영길;박승환;이남택;채영규
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.56-60
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    • 2001
  • Molecular typing of Bacillus anthracis has been extremely difficult due to the lack of polymorphic DNA markers. Aiming to develop a DNA marker specific for Bacillus anthracis and to be able to discriminate this species from Bacillus genus, we applied the random amplified polymorphic DNA (RAPD)-PCR. We have identified B. anthracis from various Bacillus species. The analysis performed by RAPD clearly demonstrated substantial genetic variations among Bacillus species. This type of analysis is an easy, quick and highly discriminatory technique that may help in diagnosis of anthrax.

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Identification of Spoilage Bacteria Isolated from Aseptic Packaged Cooked Rice and Application of Acidic Electrolyzed Saline Solution as Water-for-Cooked Rice (무균포장밥으로부터 분리된 부패 미생물의 동정 및 전해산화수의 취반수로서의 이용 효과)

  • Jeong, Jeong-Hwan;Han, Seon-Jin;Cho, Won-Dae;Hwang, Han-Joon
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.788-793
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    • 1999
  • In this study, it was investigated that the isolation and identification of spoilage bacteria from contaminated aseptic packaged cooked rice, the potential for application of acidic electrolyzed saline solution (AESS) as water-for-cooked rice and the microbiological safety of AESS-based cooked rice. Five strains of Bacillus subtilis and a B. cereus strain among the total six isolates were partially identified by biochemical method and by Microbial Identification System (MIS). The bactericidal effect of AESS was similar as 0.1% NaOCl and 70% ethanol solution, or better than that. All of the test microorganisms except Bacillus spp. that were exposed to AESS for five seconds were destructed. The effect of AESS against Bacillus spp. was much better than that of the two solutions and all of them were destructed or inhibited on exposure for five minutes. The pH value of cooked rice prepared using AESS was in the range of 3.6 to 4.3 and was not almost changed through the storage period. Various concentrations of cell suspension of Bacillus isolates were inoculated to cooked rice, which were prepared with tap water and AESS, and stored at $37^{\circ}C$ for two weeks. The result was shown that the bacteria in tap water-based cooked rice appeared normal cell growth, while they were completely repressed in AESS-based cooked rice.

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Isolation and Identification of Pathogenic Microorganisms from Soybean Sprouts

  • Kim, Hye-Jung;Koo, Kyoung-Mo;Kim, Gi-Nahm;Lee, Dong-Sun;Paik, Hyun-Dong
    • Preventive Nutrition and Food Science
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    • v.7 no.3
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    • pp.305-309
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    • 2002
  • Raw soybean sprouts were tested for contamination with the following bacteria which have potential for pathogenesis or food spoilage : Salmonella spp., Escherichia coli O157:H7, Yersinia enterocolitica, Vibrio parahae-molyticus, Aeromonas hydrophila, Plesidomonas shigeloides, Pseudomonas aeruginosa, Staphylococcus aureus, Lis-teria monocytogenes, Bacillus cereus, Clostridium perfringens, Campylobacter jejuni, Erwinia spp., and Fusarium spp. Three of the above strains were isolated from the sprouts, and identified by morphological and biochemical methods including an API kit and ATB automated identification system. The isolate cultured in Cereus selective agar, a selective medium, was a Gram-positive, rod shaped, anaerobic spore former. The biochemical and culture tests revealed the following characteristics: catalase-positive, no growth on Simmon's citrate, NO₂ production and requirement of arginine for growth; the ATB automated identification system gave 99.8 % agreement for the identification of Bacillus cereus to the species level. The isolate cultured in Macconkey agar selective medium was Gram-negative, rod shaped and a gas former; the ATB-system gave 99.9% agreement for the identification of Aeromonas hydrophila to the species level. The isolate found in Pseudomonas isolation agar was Gram-negative, rod shaped, cytochrome oxidase-positive, a reducer of nitrates to nitrogen, and pyocyanin producer; the ATB-system gave 99.9 % agreement for the identification of Pseudomonas aeruginosa to the species level. These results indicate that the three bacteria species present in the soybean sprouts were Bacillus cereus, Aero-monas hydrophila, and Pseudomonas aeruginosa. Salmonella spp., Escherichia coli O157:H7, and Yersinia enter-ocolitica, which are associated with serious disease in humans, were not isolated from soybean sprouts examined in this study.

Screening and Identification of Bacillus sp. TS0611 from Marine Sediments for Protease Production (단백질 분해효소 생산을 위한 해양 유래 Bacillus sp. TS0611의 탐색과 동정)

  • Jang, Young-Boo;Choi, Gyeong-Lim;Hong, Yu-Mi;Choi, Jong-Duck;Choi, Yeung-Joon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.5
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    • pp.461-467
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    • 2009
  • A bacterial strain was screened and identified from sea sediments in Tongyeong coastal area in order to obtain proteases or peptidase cleaving C-terminal of glutamic acid. Strain TS0611, which showed the highest activity from 5 isolated protease producing strains, was selected and its properties investigated. Strain TS0611 was a gram-positive rod, $1.2\;{\mu}m$ in cell length, catalase positive, motility-positive, melanin-negative and grew at 15~$50^{\circ}C$, and hydrolyzed gelatin and casein. This strain was identified as Bacillus thuringiensis or Bacillus cereus based on results from the API system(API 50 CHB) which examined saccharides properties, fatty acid composition of cell wall, and 16S rRNA gene sequence.

Isolation, Identification, and Characterization of Microorganisms which Possess the Flocculating Activity to Food Wastewater (음식물류폐수에 특이적 응집제를 생성하는 미생물의 분리, 동정 및 응집특성)

  • Chung, Myung-Hee;Chung, Doo-Young;Kim, Young-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.15 no.4
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    • pp.125-130
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    • 2007
  • Six bacteria, which showed the flocculating activity to food wastewater, were isolated from various environment. These strains were identified as Bacillus pumilus, Enterobacter sp., Pantotea agglomerans, Bacillus licheniformis, and two Bacillus sps. Among them, the flocculating activities of three strains, such as Enterobacter sp.(YK102), Bacillus sp.(YK103), and Pantotea agglomerans (YK104), were eight times or more higher than that of the control strain, Zoogloea ramigera. in the test with 0.5% kaolin. In the experiment with food wastewater, Enterobacter sp.(YK102) showed the highest flocculating activity which was 2.5 times higher than that of a control strain, Pseudomonas fluorescens.

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Identification of Adenosine Deaminase Inhibitor-producing Bacterium Isolated from Soil

  • SHIN, YONG KOOK;YONG-HA PARK;JAE-DONG LEE;HONG-KI JUN
    • Journal of Microbiology and Biotechnology
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    • v.7 no.1
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    • pp.32-36
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    • 1997
  • An adenosine deaminase inhibitor-producing bacterium was isolated from soil. An isolate exhibiting high adenosine deaminase inhibitory activity, was designated J-89, and classified as a strain of Bacillus subtilis on the basis of its morphological, phenotypic characteristics, the menaquinone content and cellular fatty acid composition. To confirm the taxonomic position of the strain we need more information such as DNA-DNA homology and other chemotaxonomic characteristics. In this paper we provisionally named strain J-89 as Bacillus sp. J-89 pending further chemotaxonomic study and analysis of adenosine deaminase inhibitor.

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