• Title/Summary/Keyword: Bacillus identification

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Identification of Alkalophilic Bacillus sp. S-1013 Producing Non-Cariogenicity Sugar Fuc($1{\to}4$)gaINAc($2{\to}6$)NeuAc and Optimization of Culture Condition for Its Production (비우식성 당 Fuc($1{\to}4$)gaINAc($2{\to}6$)NeuAc를 생산하는 호알칼리성 Bacillus sp. S-1013의 동정 및 생산조건의 최적화)

  • Ryu Il-Hwan;Kim Sun-Sook;Lee Kap-Sang;Lee Eun-Sook
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.235-243
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    • 2006
  • The study was performed to identification of producing microbe Non-Cariogenicity Sugar (NCS; Fuc($1{\to}4$) gaINAc($2{\to}6$)NeuAc) with anti-caries activity, and to optimization of production condition. A typical strain which produced the NCS was identified alkalophilic Bacillus sp. S-1013 through the results of morphological, biochemical and chemotaxonomic characteristics and 16S rDNA sequencing. The optimal medium composition for the maximal production of the NCS from alkalophilic Bacillus sp. S-1013 was as follow: soluble starch 30 g, dextrin 15 g, yeast extract 5 g, peptone 10 g, $K_{2}HPO_4$ 2 g in a liter of distilled water. Optimal temperature and pH were 25 and 11.0, respectively. The highest production of NCS was shown 60 hrs cultivation using the optimal medium, and then NCS productivity and dry cell weight of culture broth increased 4.24 and 2.67 time than initial medium, respectively.

The isolation of Bacillus subtilis KYS-10 with antifungal activity against plant pathogens (식물 병원균에 대한 항진균 활성을 갖는 Bacillus subtilis KYS-10의 분리)

  • Kang, Dae-Won;Ryu, Il-Hwan;Han, Seong-Soo
    • The Korean Journal of Pesticide Science
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    • v.16 no.2
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    • pp.178-186
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    • 2012
  • This study was investigated for the purpose of the isolation and identification of antagonistic bacteria with antifungal activity against plant pathogens. This bacteria denominated Bacillus subtilis KYS-10 and the optimum growth condition were 4% sucrose, 1% yeast extract, 0.2% $K_2HPO_4$, pH 7, 150 rpm, $30^{\circ}C$, 8 day. The antifungal activities against nine plant pathogens determined inhibition zone size by diffusion methods. The results, G. zeae (scab) 70 mm and P. grisea KACC 40439 (blast), P. capsici KACC 40177 (phytophthora blight) and C. destructans KACC 41077 (root rot of ginseng) 40~43 mm, and C. gloeosporioides KACC 43520 (ripe rot), C. gloesporioides KACC 40003 (anthracnose), S. shiraiana KACC 41065 (stem rot) and S. shiraiana (mulberry sclerotial disease) 35~39 mm and F. Oxysporum KACC 44452 (bulb rot of ginseng) 28 mm. From these experiment results, author suggest that Bacillus subtilis KYS-10 would be developed as a biological control agent thorough the field experimet in the near future.

Studies on the Isolation and Identification of Bacillus sp. for the Composting of Swine Manure and the Removal of Malodorous Gases from its Liquid Compost (돈분의 퇴비화를 위한 Bacillus sp.의 분리 동정 및 그 액체 비료의 악취 제거 연구)

  • 김규동;김기연;함영태
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.154-159
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    • 2004
  • Bacillus species were isolated from swine manure to develope the microbial additive suitable for the rapid com-posting. The 3 of 4 isolated strains were identified as Bacillus cereus KD-2, B. pumilus KD-3, and B. licheni-formis KD-4. Bacillus sp. KD-1 was, however, not highly identical with any Bacillus sp. The isolated strains were analyzed their growth rates, enzyme activities, and antibacterial activities. The maximum growth tem-peratures of KD-1, KD-2, KD-3 and KD-4 were $45^{\circ}C$, $50^{\circ}C$, $53^{\circ}C$, and $55^{\circ}C$, respectively. The activities of pro-tease or amylase in mixed culture of 4 strains were similar in the range of $37^{\circ}C$ to $53^{\circ}C$ and activities of lipase in the range of $37^{\circ}C$ to $42^{\circ}C$ were twice higher than those of lipase in the range of $47^{\circ}C$ to $53^{\circ}C$. The antibacterial activity of KD-l, KD-2, or KD-3 against each other was not detected. That of KD-4 against KD-1, KD-2, or KD-3 was, however, detected. The organic compound and C/N ratio of compost fermented by the mixed culture were determined as 61.9% and 22.4%, respectively. The concentration of the ammonia gas was 12.35 mg/l in the liquid compost.

Isolation and Identification of Bacillus Strains with Antagonistic Properties against Film-forming Yeasts Overgrown in Low Salted Soybean Pastes (저염 장류에서 증식하는 산막 효모에 길항 작용을 갖는 Bacillus 균주의 분리)

  • Jeon, SaeBom;Ryu, MyeongSeon;Kim, Yong Sang;Jo, Seung Wha;Jeong, Do Yeon;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.286-291
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    • 2013
  • Soybean pastes with 8% (w/w) salinity were prepared instead of soybean paste with 14% (w/w) salinity to meet the growing demands of Korean's low sodium diet. After aging, white films had appeared on the surface of all low-salted soybean pastes [8% (w/w) salinity] unlike high-salted soybean pastes [14% (w/w) salinity]. All of eight microbes isolated from the surface film were identified as Pichia kudriavzevii. Eleven Bacillus strains with good characteristics of fermentation were isolated from traditionally fermented soybean pastes in order to preserve their unique flavors and aromas after aging, and as a result of analyzing the biochemical characteristics and 16S rRNA sequences, those were identified as B. subtilis, B. licheniformis, and B. methylotrophicus. All of the Bacillus isolates had antagonistic activities against 8 isolates of the film-forming yeasts and harbored the genes for synthesis of antimicrobial surfactants including lichenysin and/or surfactin.

Properties of Antimicrobial Substances Produced by Bacillus species Isolated from Rice Straw (볏짚에서 분리한 Bacillus 균주들이 분비하는 항균물질들의 특성들)

  • Liu, Xiaoming;Yao, Zhuang;Shim, Jae Min;Lee, Kang Wook;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.133-142
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    • 2017
  • Two hundred bacilli were isolated from rice straw, and 3 strains showing strong inhibitory activities against Bacillus cereus ATCC14579 were selected for further analyses. RSC15 was identified as B. licheniforms, and RSC26 and RSC42 were identified as B. amyloliquefaciens by molecular identification methods. The inhibitory activities were heat stable, and half of the activity was retained for 20 min at $100^{\circ}C$. SDS-PAGE analyses of the culture supernatant indicated that 2 different kinds of antibacterial substances were present with sizes below 3.5 kDa. Antibacterial substances produced by B. licheniformis RSC15 were partially purified by column chromatography, and the specific activity increased from 955.0 AU/mg to 6,400 AU/mg.

A Bacterial Strain Identified as Bacillus licheniformis using Vitek 2 Effectively Reduced NH3 Emission from Swine Manure (Vitek 2 Compact System을 이용한 Bacillus licheniformis의 동정 및 NH3 저감효과)

  • Lim, Joung-Soo;Han, Deug-Woo;Lee, Sang-Ryong;Hwang, Ok-Hwa;Kwag, Jung-Hoon;Cho, Sung-Back
    • Journal of Animal Environmental Science
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    • v.21 no.3
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    • pp.83-92
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    • 2015
  • An attempt to produce more pigs in limited spaces inevitably generalized concentrated feeding operation (CFO). As concentrated pig production practice expanded, concerns on environmental issues grow concurrently. Since odor is the concerned most among those, we attempted to develop means to tackle odor emission from livestock operations. Previously, we excavated few microorganisms from pig manure and, one of them, Bacillus licheniformis was particularly useful to handle odor problem. In this study, we conducted our investigation to further characterize Bacillus licheniformis. Strain identification was conducted using Vitek 2 compact, and the optimal temperature and pH conditions to growth B. licheniformis were searched for by analyzing turbidity on O.D 600 nm. Results of this study can be summarized as these, (1) it was re-verified that the bacterial strain that purified from pig manure was, in fact, Bacillus licheniformis, (2) the bacterial growth was highest when the temperature was kept at $30^{\circ}C$, also (3) growth rate was dependent on media pH as it was high at neutral (6, 7 and 8) but dropped when it was diverged from neutral (4, 5, 9 and 10), and (4) regarding ammonia removal efficiency, B. licheniformis recorded 64% effectiveness after 48 h incubation and reached its highest (80%) at 72 h.

Detection of plcR-papR Genes by PCR in Identifying Enterotoxin Genes-Harboring Bacillus cereus Strains (장독소 유전자 함유 Bacillus cereus 확인을 위한 독소 전사 조절 유전자 plcR-papR의 PCR 검출법)

  • Yun, Suk-Hyun;Kim, Yong-Sang;So, Soon-Ku;Jeong, Do-Yeon;Hahn, Kum-Su;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.425-429
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    • 2009
  • Identification of virulent Bacillus cereus strains was examined by PCR using primers specific for the detection of plcR-papR, which encode regulatory proteins controlling the transcription of virulence factors in B. cereus. Total 96 strains of B. cereus that carried at least one of diarrheal toxin genes including hblACD, nheABC, and cytK showed all positive PCR products, while other 48 Bacillus strains that lacked the toxin genes were plcRpapR-negative. This PCR method targeting the plcR-papR genes appears to be simple and effective in identifying the enterotoxin genes-harboring B. cereus strains.

Discrimination of Bacillus subtilis from Other Bacillus Species Using Specific Oligonucleotide Primers for the Pyruvate Carboxylase and Shikimate Dehydrogenase Genes

  • Lee, Gawon;Heo, Sojeong;Kim, Tao;Na, Hong-Eun;Park, Junghyun;Lee, Eungyo;Lee, Jong-Hoon;Jeong, Do-Won
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1011-1016
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    • 2022
  • Bacillus subtilis is a useful bacterium in the food industry with applications as a starter strain for fermented food and as a probiotic. However, it is difficult to discriminate B. subtilis from other Bacillus species because of high phenotypic and genetic similarity. In this study, we employed five previously constructed multilocus sequence typing (MLST) methods for the discrimination of B. subtilis from other Bacillus species and all five MLST assays clearly distinguished B. subtilis. Additionally, the 17 housekeeping genes used in the five MLST assays also clearly distinguished B. subtilis. The pyruvate carboxylase (pyrA) and shikimate dehydrogenase (aroE) genes were selected for the discrimination of B. subtilis because of their high number of polymorphic sites and the fact that they displayed the lowest homology among the 17 housekeeping genes. Specific primer sets for the pyrA and aroE genes were designed and PCR products were specifically amplified from B. subtilis, demonstrating the high specificity of the two housekeeping genes for B. subtilis. This species-specific PCR method provides a quick, simple, powerful, and reliable alternative to conventional methods in the detection and identification of B. subtilis.

Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus

  • Sung Wook Hong;Jong-Hui Kim;Hyun A Cha;Kun Sub Chung;Hyo Ju Bae;Won Seo Park;Jun-Sang Ham;Beom-Young Park;Mi-Hwa Oh
    • Journal of Microbiology and Biotechnology
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    • v.32 no.11
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    • pp.1462-1470
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    • 2022
  • Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0-70℃ and pH 2-10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15.