• Title/Summary/Keyword: Bacillus broth

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Cloning of the MLS Antibiotics Inducible Resistance Gene and Its Control Mechanism -Inducible Resistance to MLS Antibiotics of pMB4 Plasmid Isolated from Streptococcus sp. TR-1- (MLS계 항생물질 유도 내성 유전자의 크로닝과 유전자의 조절기전 -Streptococcus sp. TR-1에서 분리한 pMB 4 Plasmid의 MLS계 항생물질 유도내성-)

  • Jeong, Soon-Hak;Kwak, Jin-Hwan;Kim, Hee-Sun;Shim, Mi-Ja;Choi, Eung-Chil;Kim, Byong-Kak
    • YAKHAK HOEJI
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    • v.34 no.2
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    • pp.139-146
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    • 1990
  • Streptococcus sp. TR-1 which has inducible resistance to MLS antibiotics was isolated from soil samples in Korea. Streptococcus sp. TR-1 was cultured in Lysis broth, then a plasmid was isolated by modified Elliker method. Bacillus subtilis UOTO277 was transformed with that plasmid. This result showed that the plasmid has the gene relating with inducible resistance to MLS antibiotics. It was named pMB4 and its size was determined about 2.4 Kb by results of digestion with various restriction enzymes. Restriction endonuclease cleavage site map of pMB4 plasmid was made by double digestion of the plasmid. pMB4 plasmid has different restriction endonuclease site map from the other plasmids that have been discovered in Streptococcus sp. so far. And it could be identified that pMB4 plasmid does not have homology with ermK of Bacillus licheniformis EMR but has homology with ermC of Staphylococcus aureus from the results of Southern hybridization.

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Isolation and Characterization of Marine Microorganisms Producing Cellulase from the Seashore of the Kyungsang Province in Korea

  • Jo, Kang-Ick;Lee, Bo-Hwa;Kim, Bo-Kyung;Jo, Hae-Young;Kim, Sung-Koo;Nam, Soo-Wan;Lee, Jin-Woo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.307-311
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    • 2005
  • Marine microorganisms to produce functional biopolymers were isolated from the seashore of the Kyungsang province. Microorganisms to hydrolyze carboxy-methyl cellulose(CMC) were cultured in marin broth and the other liquid medium that contained 2.0% (w/v) glucose, 0.25% yeast extract, 0.5% $K_2HPO_4$, 1% NaCl, 0.02% $MgSO_4{\cdot}7H_2O$ and 0.06% $(NH_4)_2SO_4$ to investigate the ability to produce carboxymethyl cellululase (CMCase) under aerobic conditions. Twelve microorganisms among them showed higher activities of CMCase than B. amyloliquefaciens DL-3, which was known as a cellulase-producing strain. The microorganism showing highest activity of CMCase in this study was identified as Bacillus subtilis subsp. subtilis with 16S rDNA partial sequencing and gyrase A partial sequencing and named as B. subtilis subsp. subtilis A-53.

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Biological Control of Meloidogyne hapla Using an Antagonistic Bacterium

  • Park, Jiyeong;Seo, Yunhee;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.30 no.3
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    • pp.288-298
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    • 2014
  • We examined the efficacy of a bacterium for biocontrol of the root-knot nematode (RKN) Meloidogyne hapla in carrot (Daucus carota subsp. sativus) and tomato (Solanum lycopersicum). Among 542 bacterial isolates from various soils and plants, the highest nematode mortality was observed for treatments with isolate C1-7, which was identified as Bacillus cereus based on cultural and morphological characteristics, the Biolog program, and 16S rRNA sequencing analyses. The population density and the nematicidal activity of B. cereus C1-7 remained high until the end of culture in brain heart infusion broth, suggesting that it may have sustainable biocontrol potential. In pot experiments, the biocontrol efficacy of B. cereus C1-7 was high, showing complete inhibition of root gall or egg mass formation by RKN in carrot and tomato plants, and subsequently reducing RKN damage and suppressing nematode population growth, respectively. Light microscopy of RKN-infected carrot root tissues treated with C1-7 showed reduced formation of gall cells and fully developed giant cells, while extensive gall cells and fully mature giant cells with prominent cell wall ingrowths formed in the untreated control plants infected with RKNs. These histopathological characteristics may be the result of residual or systemic biocontrol activity of the bacterium, which may coincide with the biocontrol efficacies of nematodes in pots. These results suggest that B. cereus C1-7 can be used as a biocontrol agent for M. hapla.

Chnracterization and Inhibitory Activity on Staphylococcus aureus of a Bacteriocin Produced by Lactobacillus plantarum KU107 (Lactobacillus plantarum KU107이 생산하는 박테리토신의 특성 및 Staphylococcus aureus 억제 작용)

  • 주관석;오세종;한경식;전우민;김세헌
    • Food Science of Animal Resources
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    • v.22 no.1
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    • pp.81-86
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    • 2002
  • A bacteriocin producing lactic acid bacteria was isolated from ground beef and the strain was identified as Lactobacillus plantarum ssp. by use of API carbohydrate fermentation pattern and physiological tests. The bacteriocin produced by L. plantarum KU107 exhibited a good spectrum of activity against foodborne pathogens including Bacillus cereus, Escherichia coli, Listeria ivanovii, Listeria monocytogenes, Pseudomonas aeruginosa, Pseudomonas chlororaphis, Staphylococcus aureus, Staphylococcus intermedius, Salmonella typhimurium and Yersinia enterocolitica. The bacteriocin was active over a wide pH range and stable of heat treatment, and inactivated by treatment with proteases. A bacteriocin from L. plantarum KU107 was effetive in reducing S. aureus in tryptic soy broth. On the ground beef containing S. aureus was added with the crude bacteriocin, S. aureus was inhibited during storage period at 4$\^{C}$.

Isolation and Identification of GABA-producing Microorganism from Chungkookjang (GABA 함량이 높은 청국장을 발효하는 균주의 분리 및 동정)

  • Mann, So-Yon;Kim, Eun-Ah;Lee, Ga-Young;Kim, Ro-Ui;Hwang, Dae-Youn;Son, Hong-Joo;Kim, Dong-Seob
    • Journal of Life Science
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    • v.23 no.1
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    • pp.102-109
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    • 2013
  • To isolate GABA-producing microorganisms, 1,500 strains were isolated from different Chungkookjang samples and screened. From these strains, 20 were selected for further analyses based on a protease and slime-producing activity test. The MC 31 strain showed the highest GABA concentration in Chungkookjang and was used in this study. MC 31 was identified as Bacillus subtilis by an API 50CHB kit and 16S rDNA sequences analysis and named as B. subtilis MC 31. B. subtilis MC 31 showed exponential growth up to 12 hours at $37^{\circ}C$ in LB broth, and it reached a stationary phase after 24 to 36 hours of incubation. B. subtilis MC 31 showed maximum GABA content at 72 hours after incubation at $40^{\circ}C$.

Preparation and Characterization of Antioxidant Peptides from Fermented Goat Placenta

  • Hou, Yinchen;Zhou, Jiejing;Liu, Wangwang;Cheng, Yongxia;Wu, Li;Yang, Gongming
    • Food Science of Animal Resources
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    • v.34 no.6
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    • pp.769-776
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    • 2014
  • The goat placenta was fermented by Bacillus subtilis and the optimal fermentation parameters of strongest antioxidant capacity of peptides were obtained using response surface methodology (RSM). The effects of fermentation time, initial pH value and glucose content on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity of the goat peptides were well fitted to a quadric equation with high determination coefficients. According to the data analysis of design expert, the strongest DPPH radical scavenging capacity value was obtained with the following conditions: content of glucose was 2.23%, initial pH value was 7.00 and fermentation time was 32.15 h. The DPPH radical scavenging capacity commonly referring antioxidant activity showed a concentration dependency and increased with increasing peptide concentration. The effects of temperature and pH were assessed to determine the stability of antioxidant peptides prepared from goat placenta. Antioxidant peptides showed good stabilities when temperature was lower than $70^{\circ}C$. However, the antioxidant peptides lost antioxidant activities rapidly under alkaline and excessive acid condition. Ultrafiltration technique was performed to separate fermentation broth with different Mw (molecular weight). It was found that peptides in the range of < 3 KDa mainly accounted for the antioxidant activities.

Optimization of Culture Medium for Novel Cell-Associated Tannase Production from Bacillus massiliensis Using Response Surface Methodology

  • Belur, Prasanna D.;Goud, Rakesh;Goudar, Dinesh C.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.199-206
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    • 2012
  • Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

Characterization of a Novel Fibrinolytic Enzyme Produced from Bacillus subtilis BK-17 (Bacillus subtilis BK-17 유래 혈전용해 효소의 특성)

  • Hyun Bek;Lim Hak-Seob;Chung Kyung Kae;Choi Yung Hyun;Choi Byung Tae;Seo Min-Jeong;Kim Ji-Eun;Ryu Eun-Ju;Huh Man Kyu;Joo Woo Hong;Jeong Young Kee
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.987-993
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    • 2005
  • A bacterium, producing a fibrinolytic enzyme, was screened from a decaying rice plant. The bacterium was identified as Bacillus subtilis by morphological, biochemical, and physiological properties and named Bacillus subtilis BK-17. The fibrinolytic enzyme (BK) was purified from supernatant of Bacillus subtilis BK-17 culture broth. The molecular weight was 31 kDa as determined by SDS-PAGE. The effect of temperature, pH, and plasminogen on the activity of the bacillokinase (BK) was analysed and the activity was compared with urokinase. The optimal temperature and pH were $50^{circ}C$ and pH 7, pH 8, respectively. The BK activity was inhibited to $45\%$, $35\%$, and $23\%$ with 1mM EDTA, $Zn^{2+}$, and $Ca^{2+}$, respectively. However, $Mg^{2+}$, $Mn^{2+}$, and $Co^{2+}$ ions did not have any significant effect on the enzyme activity The BK showed the artivity in the both plates, plasminogen-free fibrin plate and plasminogen-rich fibrin plate. The result indicates that the BK can directly act the fibrin. In comparison of fibrinolytic activity with urokinase on the fibrin plate, the BK shows about 20 folds higher activity than that of the urokinase.

Combination of Hydrophobic Filtration and Enrichment Methods for Detecting Bacillus cereus in Fresh-Cut Cabbage

  • Lee, Sujung;Choi, Yukyung;Lee, Heeyoung;Kim, Sejeong;Lee, Jeeyeon;Ha, Jimyeong;Oh, Hyemin;Lee, Yewon;Kim, Yujin;Yoon, Yohan;Lee, Soomin
    • Journal of Food Hygiene and Safety
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    • v.33 no.5
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    • pp.325-329
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    • 2018
  • This study developed a rapid detection method for Bacillus cereus in fresh-cut cabbages. Fresh-cut cabbage samples were inoculated at 1-, 2- and 3-Log CFU/g, and pathogens were enriched in tryptic soy broth containing 0.15% polymyxin B at $30^{\circ}C$, $37^{\circ}C$, and $42^{\circ}C$ to determine the detection limit and appropriate enrichment temperature for multiplex PCR detection. Enriched bacterial cells in enrichment broth were collected in a hydrophobic filter prior to DNA extraction for multiplex PCR. Filters were resuspended in distilled water, and DNA was extracted from the suspension. DNA samples were further analyzed by multiplex PCR. Detection limit of multiplex PCR was 5-Log CFU/mL. B. cereus cell counts were higher (P < 0.05) at $42^{\circ}C$ than other temperatures. Detection rate of 1-, 2-, and 3-Log CFU/g inoculated samples were 60%, 80%, and 100% after enrichment respectively. However, when enriched samples were filtered with hydrophobic membrane filter, detection rates became 100%, regardless of inoculation level. Results indicate a combination of enrichment with hydrophobic filtration improves rapid detection efficiency of B. cereus in fresh-cut cabbage by multiplex PCR.

Effect of Microbial Products Made of Bacillus stearothermophilus DL-3 on Growth of Chickens and Pigs (Bacillus stearothermophilus DL-3을 사용하여 제조한 미생물 제재가 닭과 돼지의 성장에 미치는 영향)

  • Kim Soon-Hee;Jo Kang-Ik;Lee You-Jung;Oh Ju-Sung;Jung Soon-Jae;Moon Byung-Ju;Kang Kyung-Hee;Lee Jin-Woo
    • Journal of Life Science
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    • v.16 no.1
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    • pp.22-29
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    • 2006
  • Effect of microbial products made of Bacillus stearothermophilus DL-3 on growth of chickens and pigs was investigated. Two types of microbial product were made in this study. One is the microbial product made of culture broth of B. stearothemophilus DL-3 and rice bran which named as the microbial product A. The other is the microbial product made of culture broth of B. stearothermophilus DL-3, apple pomace, soybean pomace and rice bran which named as the microbial product B. Chickens were divided into three groups and each group was fed with $100\%$ general feed, $90\%$ general feed supplemented with $10\%$ microbial product A or $90\%$ general feed supplemented with $10\%$ microbial product B. The average chicken weight of each group was $41.1{\pm}2.5g,\;41.6{\pm}3.2g\;and\;42.3{\pm}2.9g$ and those after 28 days was $547.7{\pm}91.7g,\;560.1{\pm}17.2g\;and\;562.2{\pm}32.5g$, respectively. The average weight gain for each group was 506.6 g, 518.6 g and 519.9 g, respectively, and weight increases of groups fed with $90\%$ general feeder and $10\%$ microbial product A and B were $2.4\%\;and\;2.6\%$ higher than the group fed with $100\%$ general feed. Pigs were also divided into three groups and each group was fed like chickens. The average weight of each group was $9.3{\pm}1.0kg,\;9.4{\pm}1.1kg\;and\;9.6{\pm}1.0kg$ and those after 37 days was $19.3{\pm}4.1kg,\;20.2{\pm}3.9kg\;and\;20.8{\pm}4.2kg$, respectively. The average Weight gain for each group was 10.65 kg, 10.82 kg and 11.20 kg, respectively, and weight increases of groups fed with $90\%$ general feeder and $10\%$ microbial product A and B were $1.6\%$ and $5.2\%$ higher than the group fed with $100\%$ general feed.