• Title/Summary/Keyword: Bacillus Bacillus licheniformis

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Isolation of a Bacillus licheniformis DK42 Producing Cellulase and Xylanase, and Properties of the Enzymes (Cellulase 및 Xylanase를 분비하는 Bacillus licheniformis DK42의 분리 및 효소 특성)

  • Kim, Min Jeong;Lim, Soo Jin;Kang, Dae-Kyung
    • Journal of Animal Science and Technology
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    • v.50 no.3
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    • pp.429-436
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    • 2008
  • A bacterium producing cold-active cellulase and xylanase was isolated from pig feces. The isolate, DK42 strain, was found to be the Gram-positive, non-motile, catalase-positive, and spore-forming stain. Under an electron microscope, the cells were observed to be rod-shaped. The isolate was identified as Bacillus licheniformis DK42 on the basis of morphological and biochemical properties as well as 16S rRNA gene sequences. The characterization of crude cellulase and xylanase from B. licheniformis DK42 was investigated. Cellulase exhibited an optimum temperature and pH at 45℃ and 6.0, whereas xylanase exhibited an optimum temperature and pH at 55℃ and 6.0. Especially cellulase maintained approx. 50% of its maximum activity even at 10℃, indicating that it is cold-active. Both cellulase and xylanase were stable after 2hr at 35℃, whereas they lost their activities after 30min at 65℃.

The Poly-γ-ᴅ-Glutamic Acid Capsule of Bacillus licheniformis, a Surrogate of Bacillus anthracis Capsule Induces Interferon-Gamma Production in NK Cells through Interactions with Macrophages

  • Lee, Hae-Ri;Jeon, Jun Ho;Rhie, Gi-Eun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.1032-1037
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    • 2017
  • The poly-${\gamma}$-$\small{D}$-glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis, provides protection of the bacterium from phagocytosis and allows its unimpeded growth in the host. We investigated crosstalk between murine natural killer (NK) cells and macrophages stimulated with the PGA capsule of Bacillus licheniformis, a surrogate of the B. anthracis capsule. PGA induced interferon-gamma production from NK cells cultured with macrophages. This effect was dependent on macrophage-derived IL-12 and cell-cell contact interaction with macrophages through NK cell receptor NKG2D and its ligand RAE-1. The results showed that PGA could enhance NK cell activation by inducing IL-12 production in macrophages and a contact-dependent crosstalk with macrophages.

Increased Production of γ-Aminobutyric Acid from Brewer's Spent Grain through Bacillus Fermentation

  • Tao Kim;Sojeong Heo;Hong-Eun Na;Gawon Lee;Jong-Hoon Lee;Ji-Yeon Kim;Do-Won Jeong
    • Journal of Microbiology and Biotechnology
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    • v.33 no.4
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    • pp.527-532
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    • 2023
  • Brewer's spent grain (BSG) is a waste product of the beer industry, and γ-aminobutyric acid (GABA) is a physiologically active substance important for brain and neuron physiology. In this study, we used the bacterial strains Bacillus velezensis DMB06 and B. licheniformis 0DA23-1, respectively, to ferment BSG and produce GABA. The GABA biosynthesis pathways were identified through genomic analysis of the genomes of both strains. We then inoculated the strains into BSG to determine changes in pH, acidity, reducing sugar content, amino-type nitrogen content, and GABA production, which was approximately doubled in BSG inoculated with Bacillus compared to that in uninoculated BSG; however, no significant difference was observed in GABA production between the two bacterial strains. These results provide the experimental basis for expanding the use of BSG by demonstrating the potential gain in increasing GABA production from a waste resource.

Cloning of the Cellulase Gene and Characterization of the Enzyme from a Plant Growth Promoting Rhizobacterium, Bacillus licheniformis K11 (고추역병 방제능이 있는 식물성장촉진 균주 Bacillus licheniformis K11의 cellulase 유전자의 cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.95-100
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    • 2007
  • The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

Toxin Gene Analysis of Bacillus cereus and Bacillus thuringiensis Isolated from Cooked Rice (쌀밥에서 분리한 Bacillus cereus와 Bacillus thuringiensis의 독소유전자 분석)

  • Jeon, Jong-Hyuk;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.42 no.3
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    • pp.361-367
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    • 2010
  • Bacterial contamination of cooked rice was analyzed to evaluate the microbial safety. Thirty raw rice samples were collected in Korea and cooked in an electric rice cooker. Mesophilic aerobe, food-poisoning Bacillus cereus group, and their toxin genes were determined on cooked rice. The percentage of total mesophilic aerobe based on 1-3 log CFU/g was 27% among the samples. Bacillus spp. in MYP selective medium was similar to the number of mesophilic aerobe, whileas Bacillus spp. was detected in most samples after enrichment. Thirty-seven isolates from 30 cooked rices were identified as B. thuringiensis, B. cereus, B. valismortis, B. pumilus, B. coagulans, B. licheniformis, Geobacillus stearothermophilus, and Brevibacillus laterosporus. Twenty isolates (54%), more than half of the isolates, were B. thuringiensis while nine (27%) were identified as B. cereus. All B. thuringiensis isolates possessed non-hemolytic toxin genes and interestingly, seven B. cereus among nine isolates possessed emetic toxin genes. More B. thuringiensis was present on the cooked rice than B. cereus and most B. cereus possessed emetic toxin genes rather than diarrheal toxin genes. Therefore, food-borne outbreak due to B.cereus on the cooked rice kept at room temperature might be examples of emetic food-poisoning.

Studies on the Microflora and Enzymes Influencing on Korea Native Kochuzang (Red Pepper Soybean Paste) Aging (재래식(在來式) 고추장 숙성(熟成)에 미치는 미생물(微生物) 및 그 효소(酵素)에 관(關)한 연구(硏究))

  • Lee, Ke-Ho;Lee, Myo-Sook;Park, Sung-O
    • Applied Biological Chemistry
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    • v.19 no.2
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    • pp.82-92
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    • 1976
  • The study was carried out to investigate the changes of the various chemical components and the microflora during the aging period of Korean navive Kochuzang. (Red pepper soybean paste) Korean native maeju loaves were separated into surface and inner parts. Three kinds of Korean native Kochuzang were prepared from surface part, inner part, and ordinary of maeju. The selection and the indentification of the high enzyme producing strains from the microflora and characteristics of their enzymes were studied. I. The changes of the various chemical components during the aging period of Kochuzang. 1) The changes of pH in the 3 kinds of Kochuzang displayed rapid decrease for the first 10 days after preparing and gradual curve of decrease until 60 days, but slight increase for the next 30 days. The pH of the surface part Kochuzang was lower than that of inner part or ordinary Kochuzang. 2) The total acid contents in the 3 kinds of Kochuzang showed gradual increase until the 60 days but it slowly reduced after this time. 3) The total nitrogen contents in the 3 kind of Kochuzang showed gradual inerease up to the 60 days, but slight decrease after this time. 4) The changes of trichloroacetic acid soluble nitrogen in the 3 kinds of Kochuzang showed a remarkable increase for the first 10 days, however gradual increase after this time. 5) The increase of amino nitrogen contents in the 3 kinds of Kochuzang seemed to be remarkable until the first 30 days, however to be less remarkable after this time. 6) The contents of reducing sugar in the 3 kinds of Kochuzang showed remarkable increase until the first 50 days and it slowly reduced after this time. II. The changes of microflora during the aging period of Kochuzang. 1) Aerobic, anaerobic bacteria and mold in the 3 kinds of Kochuzang were increased until the first 30 to 40 days, but they were reduced after this time. 2) No yeast in the three kinds of Kochuzang appeared until the first 20 days. Yeast were proved to grow, when the pH value was decreased below 5.4 after the 30 days. Yeasts in the surface part and ordinary Kochuzang were gradually increased and those in the inner part Kochuzang were decreased as aging. III. The selection and identification of high amylase and protease producing strains from the microflora during the aging period of Kochuzang. 1) The amylase and protease highly producing strains from microflora were identified as Bacillus subtilis-P, Bacillus subtilis-G, Bacillus licheniformis-K, Aspergillus oryzae-B. 2) Amylase activity of Aspergillus oryzae-B was highest among the strains and the strains in order of the higher activity to the lower one were Bacillus subtilis-P Bacillus licheniformis-K, Bacillus subtilis-G. Protease activities of Aspergillus oryzae-B and Bacillus subtilis-P were about the same and the strains in order of the higher activity to the lower one were Bacillus licheniformis-K, Bacillus subtilis-G. 3) Amylase activity was inhibited more than protease activity was with NaCl concentration. Amylase activity was inhibited by 45 to 65 percent and protease activity by 40 to 46 percent at the concentration of 15 percent NaCl, which was the average concentration of NaCl in Kochuzang.

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Isolation, Identification, and Characterization of Bacillus spp. from the Traditionally fermented Cheonggukjangs in the Gyeonggi and the Gangwon Provinces (경기.강원 지역의 전통발효 청국장으로부터 Bacillus균주의 분리 동정 및 특성 분석)

  • Lee, Nam-Keun;Jeon, Eun-Hee;Lee, Hyo-Jin;Cho, Il-Jae;Hahm, Young-Tae
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.276-280
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    • 2006
  • Traditionally fermented Cheonggukjangs were collected from Gyeonggi and Gangwon provinces and 22 strains were isolated and identified by using 165 rDNA sequences. Most of the identified strains were Bacillus subtilis and B. licheniformis, B. subtilis and B. licheniformis are dominant in the Gyeonggi area and B. licheniformis in the Gangwon area. In the growth pattern of the isolated strains, the duration of lag phase was generally 5 to 7 hours and stationary phase was reached after 23 to 40 hours of incubation. Total cell populations at the stationary phase were between $1{\times}10^6\;CFU/ml$ and $5{\times}10^7\;CFU/ml$. The fermenting ability of carbohydrates of isolates showed some differences among the regions. The isolated strains from Yong-In, Gyeonggi showed higher fermenting abilities with D-xylose, xylitol, D-tagatose and Methyl-$\alpha$-D-mannopyranoside. D-lactose, D-tagatose, D-xylose, Methyl-$\alpha$-D-mannopyranoside, amygdalin, arbutin, esculin and 2-keto-gluconate were well fermented with the An-Seong's strains; L-rhamnose, inositol, D-mannitol, D-sorbitol, celibiose and gluconate with the Kawang-Ju's stains; and D-lactose with the Odaesan's strains.

Isolation of Bacillus sp. Producing Xylanase and Cellulase and Optimization of Medium Conditions for Its Production. (Xylanase, Cellulase의 생산성이 높은 Bacillus sp.의 분리 및 효소생산을 위한 배지조건의 최적화)

  • 정원형;양시용;송민동;하종규;김창원
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.383-388
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    • 2003
  • A bacterium producing the extracellular xylanase and CMCase was isolated from soil and has been identified as Bacillus sp. The isolate, named Bacillus sp. A-7, was shown to be very similar to Bacillus licheniformis on the basis of its biochemical and physiological properties. The maximum xylanase and CMCase production were obtained when 2.0% (w/v) glucose and 0.3% (w/v) yeast extract were used as carbon source and nitrogen source, respectively. The best mineral conditions for xylanase and CMCase production were 0.1%(w/v) $CaC1_2$. Among the various feedstuffs, 1.0%(w/v) soybean meal was selected for the best xylanase and CMCase production.

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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A Study on the Extraction of Alkaline Protease from Bacillus licheniformis Fermentation Broth using Reverse Micelle (역미셀을 이용한 Bacillus licheniformis 발효액으로 부터 알카리성 단백질 분해효소의 추출에 관한 연구)

  • 권성필;구윤모홍성안
    • KSBB Journal
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    • v.6 no.1
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    • pp.105-109
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    • 1991
  • In separating alkaline protease from the bacteria (Bacillus licheniformis) fermentation broth using reverse micelle, effects of various factors;ionic strength, pH and surfactant concentration, on separation efficiency were studied. KCl controls the ionic strength. The lower KCl concentration was in the feed solution, the more protein and activity was recovered. The higher KCl concentration was in the stripping solution, the more protein and activity was recovered. Using sodium-di-2-ethylhexyl sulfosuccinate(Aerosol-OT or AOT) as a surfactant, the higher AOT concentration in the solvent, the more activity and protein were recovered. 0.1N NaOH and IN HCl were used to adjust pH. Maximum recovery of protein mass and activity were obtained at feed solution of pH 5.3. Maximum activity was recovered at stripping solution of pH 7.5

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