• Title/Summary/Keyword: Bacillus strain

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Isolation and Its Optimal Culture Condition for High Agarase-Producing Mutant (한천분해효소의 고생산성 변이주의 분리 및 최적배양조건)

  • 황선희;하순득;김봉조;김학주;공재열
    • KSBB Journal
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    • v.14 no.3
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    • pp.351-357
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    • 1999
  • A marine bacterium Bacillus cereus ASK202, agarase producing strain, was treated with some mutagenic agents, ultraviolte(UV), 1-methyl-3-nitro-1-nitrosoguanidine(NTG), and ethyl methane sulfonate(EMS), several times for the increasing of the agarase production After mutagen treatment, we isolated one mutant strain treated with NTG showed the highest stability and agarase productivity and named as Bacillus cereus ASK202-N3. This Bacillus cereus ASK202-N3 strain was well grown in the modified marine medium containing 0.5%(w/v) agar, 0.3%(w/v) yeast extract, and 5.0%(w/v) NaCl, and the optimal initial pH, temperature and culture time were 7.8, $25^{\circ}C$ and 32h, respectively. In the optimal culture conditions, the agarase production was increased to 5.3 fold(850units/L) compared to that of the wild type.

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Isolation and Characterization of Bacillus thuringiensis Strain BT-209 producing Cuboidal $\delta$ -endotoxin crystals

  • Jung, Yong-Chul;Kim, Sung-uk;Son, Kwang-Hee;Lee, Hyung-Hoan;Bok, Song-Hae
    • Journal of Microbiology and Biotechnology
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    • v.5 no.3
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    • pp.138-142
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    • 1995
  • Bacillus thuringiensis strain BT-209 was isolated from a soybean grain dust sample in Korea. The strain BT-209 produced two different sizes of cuboidal crystals and one spore in the cell. In the biochemical characterization, the strain BT-209 showed negative reactions on the production of urease, and the utilization of citrate and sucrose. Examination of its antibiotic resistance revealed that while the strain BT-209 showed higher sensitivity than B. thuringiensis subsp. kurstaki HD-1 to ampicillin, bacitracin, chlortetracycline, gentamycin, neomycin, penicillin G, tetracycline and tobramycin, it was more resistant to methicillin than B. thuringiensis subsp. kurstaki HD-1. The $\delta$-endotoxin crystal of strain BT-209 consisted of three proteins with apparent molecular weights of appoximately 148, 135 and 62 kDa on a 10% SDS-PAGE. The strain BT-209 had at least eight different plasmids with sizes of 4.1, 5.2, 6.3, 8.6, 14.6, 24.5, 67.6 and 77.6 Kb. The strain BT-209 showed strong lethalities of 70% and 87% against Bombyx mori and Hyphantria cunea larvae. at 72 h, respectively.

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Isolation and Enzyme Production of a Xylanase-producing Strain, Bacillus sp. AMX-4. (Xylanase를 생산하는 Bacillus sp. AMX-4 균주의 분리와 효소 생산성)

  • 윤기홍;설숙자;조효찬;이미성;최준호;조기행
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.123-128
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    • 2002
  • A bacterium producing the extracellular xylanase was isolated from soil and has been identified as a Bacillus sp. strain. The isolate, named Bacillus sp. AMX-4, was shown to be similar to B. subtilis strain on the basis of its chemical compositions. The xylanase of culture supernatant was most active at 50℃ and pH 6.0. The additional carbon sources including monosaccharides, disaccharides, wheat bran, and rice straw increased the enzyme productivity. Especially, the maximum xylanase productivity was reached 29.2 units/ml in LB medium supplemented with 1.5% (w/v) xylose, which was 16-folds more than that in LB medium. As the results of investigating the effects of xylose on cell growth and xylanase productivity of Bacillus sp. AMX-4, increase of xylanase production was owing to the induction of xylanase biosynthesis. It was also found that the enzyme production was in association with the growth of Bacillus sp. AMX-4.

Purification of the Three-subunit, Recombinant Bacillus pasteurii Urease Expressed in Escherichia coli

  • Lee, Ji Hyun;Sang Dal Kim;Mann Hyung Lee
    • Journal of Microbiology and Biotechnology
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    • v.6 no.1
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    • pp.26-29
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    • 1996
  • The genes coding for the urease of alkalophilic Bacillus pasteurii have been previously cloned and recently sequenced. (You, J. H., B. H. Song, J. H. Kim, M. H. Lee, and S. D. Kim (1995) Molecules and Cells 5, 359-369.) The recombinant Bacillus pasteurii urease expressed in an E. coli HB101 strain was purified 31.2 fold by using combinations of anion-exchange and hydrophobic chromatography followed by Mono-Q chromatography on a FPLC. In spite of the presence of three discrete structural peptide genes in the Bacillus pasteurii urease gene cluster, only one or two enzyme subunits have been observed to date. Here we report for the first time that the recombinant Bacillus pasteurii urease expressed in a E. coli strain consists of three distinct subunits. One large subunit was estimated to be of $M_r$=65, 200 and the two small-subunit peptides are of $M_r$=14, 500 and $M_r$=13, 700, respectively.

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Isolation of Thermostable ${\alpha}$-Amylase Hyperproducing Bacillus sp. No. 32H417 and Some Properties of the Enzyme (耐熱性 ${\alpha}$-Amylase 高 生産性 Bacillus sp. No. 32H417의 分離 및 酵素 特性)

  • Kim, Moo-Sung;O, Pyong-Su
    • Microbiology and Biotechnology Letters
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    • v.19 no.2
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    • pp.122-127
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    • 1991
  • A bacterial strain NO. 32 which produced thermostable ${\alpha}$-amylase was isolated from soil and identified to genus of Bacillus. To enhance ${\alpha}$-amylase productivity, a successive mutation of Bacillus sp. No. 32 was attempted with treatment of N-methyl-N'-nitro-N-nitrosoguanidine (NTG). The resulting mutant, Bacillus sp. No. 32H417, which is risistant to refampicin and deficient in spore formation, produced about 90-fold high level of ${\alpha}$-amylase when compared with parental strain. The properties of the enzyme for thermostability were investigated. The optimal temperature and pH for enzyme reaction were 95$^{\circ}C$ and pH6.5, respectively, in the presence of 0.3mM $Ca^{2+}$ as an effective stabilizer.

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Isolation and Characterization of Bacillus sp. P16 Producing Extracellular Chitosanase (키토산분해효소를 생산 분비하는 Bacillus sp. P16의 선발 및 특성)

  • Jung, Mi-Ra;Jo, Yoo-Young;Chi, Yeon-Tae;Park, Ro-Dong
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.369-374
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    • 1997
  • An endochitosanase-producing bacterium was isolated from soil and identified as a strain of Bacillus sp. The isolate was gram positive, rod shape $(0.4-0.6{\times}1.6-2.2{\mu}m)$, endospore-forming, catalase positive, and mobility positive, and grown at pH 4.5-11.0 and upto $42^{\circ}C$ in the medium containing 2% NaCl. RAPD analysis of the DNA purified from the strain was also performed, and the chitosanase-producing strain was named as Bacillus sp. P16. The culture supernatant of the strain showed strong liquefaction activity and rapidly decreased viscosity of chitosan solution. By TLC and HPLC, chitooligosaccharides of DP 2-7 were separated and identified from the enzyme hydrolyzates of chitosan. The chitosanase from Bacillus sp. P16 was thus regarded as an endo-splitting type.

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Isolation and Characterization of Thermophilic Microorganism Producing Starch-hydrolyze Enzyme (한국 토양으로부터 전분가수분해효소를 생산하는 고온성 균주의 선별과 동정)

  • Choi, Wonseok;Bai, Dong-Hoon
    • Food Engineering Progress
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    • v.14 no.1
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    • pp.7-13
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    • 2010
  • A thermophilic microorganism, which is able to hydrolyze starch, was isolated from soil and compost in Korea. It was Gram-positive, rod-shaped, catalase positive, nonmotile, glucose and mannitol fermentative, xylose oxidative, and spore forming microorganism. It also has an ability to hydrolyze casein and gelatin. The color of colony was yellowish white. The sequence of 16S rDNA of strain 2719 showed 99.5% sequence homology with the sequence of 16S rDNA of Bacillus thermoglucosidasius. On the basis of biochemical and physiological properties and phylogenetic analysis, the isolated strain was named as Bacillus thermoglucosidasius 2719.

Isolation and Characterization of Bacillus sp. Producing Broad-Spectrum Antibiotics Against Human and Plant Pathogenic Fungi

  • Chen, Na;Jin, Min;Qu, Hong-Mei;Chen, Zhi-Qiang;Chen, Zhao-Li;Qiu, Zhi-Gang;Wang, Xin-Wei;Li, Jun-Wen
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.256-263
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    • 2012
  • A strain of bacterium producing antifungal antibiotic was isolated and identification of the strain was attempted. We could identify the bacterium as being a Bacillus sp., based on morphological observation, physiological characteristics, and 16S rDNA sequence analysis, thus leading us to designate the strain as Bacillus sp. AH-E-1. The strain showed potent antibiotic activity against phytopathogenic and human pathogenic fungi by inducing mycelial distortion and swelling and inhibiting spore germination. The antibiotic metabolite produced by the strain demonstrated excellent thermal and pH (2-11) stability, but was labile to autoclaving. From these results, we could find a broader antifungal activity of Bacillus genus. Isolation and characterization of the active agent produced by the strain are under progress.

Characterization and Organic Hydrocarbons Degradation Potential of Euryhaline Marine Microorganism, Bacillus sp. EBW4 Isolated from Polychaete (Perinereis aibuhitensis) (갯지렁이(Perinereis aibuhitensis)에서 분리한 광염성 해양 미생물 Bacillus sp. EBW4의 특성 및 유기물 분해능 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Lee, Dong-Heon;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.38-45
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    • 2013
  • In this study, euryhaline marine microorganism, Bacillus sp. strain EBW4 isolated from polychaete (Perinereis aibuhitensis) of Suncheon Bay was physiologically, biochemically and genetically characterized. Based on 16S rRNA sequence, EBW14 was found to share 98.25% similarity with Bacillus hemicentroti $JSM076093^T$, 97.96% similarity with Bacillus hwajinponensis SW-$72^T$ and 96.28% similarity with B. algicoa $KMM3737^T$, respectively. The temperature range for the growth of strain EBW4 was $4-40^{\circ}C$, NaCl concentration range 0-17% and pH range pH 5-9, revealing that EBW4 was euryhaline bacterium. Major fatty acids in strain EBW4 were composed of anteiso $C_{15:0}$ (48.2%), iso $C_{16:0}$ (12.1%), anteiso $C_{17:0}$ (11.6%) and iso $C_{14:0}$ (9.4%). EBW4 was found to have DNase, amylase, protease and lipase for the degradation of macromolecules such as DNA, carbohydrates, proteins, lipids, etc. The enzyme activities of alkaline phosphatase, esterase (C4), leucine arylamidase and ${\alpha}$-chymotrypsin were also found in strain EBW4. Analysis of the biodegradation ability of EBW4 for organic hydrocarbons under different salinity conditions using synthetic water waste revealed that EBW4 exhibited the ability to degrade organic hydrocarbons very quickly, suggesting strain EBW4 may be a good candidate for the application to various industries.

Production and Characteristics of Lytic Enzyme against Streptococcus mutans Cell Wall from Alkalophilic Bacillus sp. 4830 (호알카리성 Bacillus sp. 4830이 생산하는 Streptococcus mutans 세포벽 분해효소의 분리와 특성)

  • Kim, Yun-Keun;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1143-1149
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    • 2003
  • To elucidate a method of preventing dental caries, strains producing lytic enzymes were isolated and their characteristics were investigated. Among 5,00 alkalophilic strains isolated from soil, 22 strains showed lytic activity against Streptococcus mutans. Strain No. 4830, with the highest lytic activity, was selected for further study. Strain 4830 showed 94% sequence homology with the 16S rDNA sequence of Bacillus alcalophilus, but it was concluded to be different from Bacillus alcalophilus because of its biochemical characteristics. The strain was named Bacillus sp. 4830. The lytic enzyme from Bacillus sp. 4830 was purified by ethanol precipitation and CM-agarose column chromatography. The molecular weight of the lytic enzyme was determined to be 28 kDa by SDS-PAGE. The lytic enzyme was stable between pH 5.0 and pH 11 and up to $40^{\circ}C$. The optimal pH and temperature for the lytic activity was 9.0 and $50^{\circ}C$, respectively.