• Title/Summary/Keyword: Bacillus subtilis K-20

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Purification and Characterization of Fibrinolytic Enzyme Produced by Bacillus subtilis K7 Isolated from Korean Traditional Soy Sauce (한국재래간장 발효균 Bacillus subtilis K7 유래의 혈전용해 Protease의 정제 및 특성)

  • Kim, Doo-Young;Lee, Eun-Tag;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.176-182
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    • 2003
  • An alkaline fibrinolytic protease-producing bacteria was isolated front Korean traditional soy sauce and identified as Bacillus subtilis K7 from the results of analyses of its morphological and physiological properties, $API^{\circledR}$, and Biolog system. The enzyme was purified by 75% ammonium sulfate fractionation, QAE-Sephadex anion and SP-Sephadex cation exchange column chromatography and Sephadex G-100 gel filtration. The specific activity of the purified enByme was 233.9 unit/mg protein and the yield of enzyme was 3.8%. The homogeneity of the purified enzyme was confirmed by polyacrylamide gel electrophoresis. Molecular mass of the enzyme was estimated about 21,500 Da by SDS-polyacrylamide get electrophoresis and gel chromatography. The optimum temperature and pH for the enzyme activity were $40^{\circ}C$ and 9.0, respectively. The enzyme was stable in a pH range of 5.0 to 12.0, and 60% of its activity was lost on heat treatment at $50^{\circ}C$ for 20 min. The activity of the purified enzyme was inhibited by the presence of $Fe^{2+},\;Ag^{2+},\;Cu6{2+}$, iodoacetate, ethylene diamine tetraacetic acid (EDTA), and trans-1,2-diaminocycloheane-N,N,N',N'-tetraacetic acid (CDTA). The results indicates that the enzyme requires a metal ion for its enzymatic activity.

Variation of fibrinolytic enzyme activity produced Bacillus subtilis by gene cloning (유전자 cloning에 의한 Bacillus subtilis의 fibrinolytic enzyme 활성 변화)

  • 이홍석;유천권;이철수;강상모
    • Microbiology and Biotechnology Letters
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    • v.28 no.1
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    • pp.14-20
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    • 2000
  • The transformation of Bacillus subtilis K-54 and J-10 was carried out with constructed vectors containing structure and enhancer genes of aprN and prtR, to increase their fibrinolytic enzyme activity. Bands for the aprN and prtR genes were identified from B. subtilis J-10 by PCR that was carried out with the constructed primers for the genes. In addition, the gene fragments contained promoter site based on the results of analysing their nucleotide sequence. The two gene fragments, aprN and prtR, obtained by the PCR, were, then, inserted to vector such as T-vector and E.coli/Bacillus shuttle vector. The constructed vector were designated as pAPR2 (aprN), pENC2 (prtR) and pFLA1 (aprN and prtR), respectively. The constructed vector was used for transformation of the strains of B.subtilis J-10 and B. subtilis K-54 and the fribrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and the fibrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and pFLA1, resulted in the increase of fibrinolyitic enzyme activity in B. subtilis J-10 by 27.3% and 16%, respectively. However, the introduction of pENC2 to B. subtilis J-10 did not seem to induce increase of the enzyme activity. The strain of B.subtilis K-54 transformed with pENC2 showed an increased fibrinolytic activity by 5 folds compared with that of the original strain of B. subtilis K-54.

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Effect of Bacillus spp. Having Gelatin Decomposing Activity on Root-knot Nematode, Meloidogyne incognita on Pepper (젤라틴 분해력이 있는 Bacillus균의 고추 고구마뿌리혹선충에 미치는 영향)

  • Kim, Jin Han;Park, Chan Sun;Kim, Young-Ho;Kim, Yu Ri;Lee, Seung Woong;Rho, Mun Chual
    • The Korean Journal of Pesticide Science
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    • v.20 no.4
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    • pp.375-379
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    • 2016
  • This study investigated the decomposing activities on gelatin and egg sac of root-knot nematode (Meloidogyne incognita). Ten Bacillus isolates (KRB-1~10) isolated from the soils of Mt. Naejang. Among them, KRB-5, 9, and 10 showed decomposing activities, and identified as Bacillus subtilis KRB-5, Bacillus amyloliquefaciens KRB-9, and KRB-10 by 16S rRNA sequence analysis, respectively. Under pot experiments using pepper, 100-fold diluted culture broth of three isolates reduced the number of egg sac on roots collected 60 days and 90 days and increased the growth of aerial part compared with the group only treated with Meloidogyne incognita. Especially, the group treated B. subtilis KRB-5 was superior in the growth of pepper. These results suggest that the B. subtilis KRB-5 can be used to control the root-knot nematode on pepper.

Structural Identification of $Siderophore_{AH18}$ from Bacillus subtilis AH18, a Biocontrol agent of Phytophthora Blight Disease in Red-pepper (Bacillus subtilis AH18의 고추역병 방제능과 $Siderophore_{AH18}$의 구조분석)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.326-335
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    • 2008
  • The siderophore ($siderophore_{AH18}$) of Bacillus subtilis AR18 was determined to be one of catechol type and purified by using Amberlite XAD-2, Sephadex LR-20 chromatography, and reversed-phase RPLC. The $Siderophore_{AH18}$ was identified bacillibactin with its structure by GC-MS, $^1H$-NMR, and $^{13}C$-NMR. $Siderophore_{AH18}$ (bacillibactin) had been confirmed its molecular weight of 883 and chemical structure of $(2,3-dihydroxybenzoate-glycine-threonine)_3$. Purified $siderophore_{AH18}$ showed strong biocontrol ability towards the spore of Phytophthora capsici on PDA and able to effectively suppress (55%) P. capsici causing red-pepper blight in the pot in vivo test.

Characterization of Bacteriocin from Bacillus subtilis cx 1 (Bacillus subtilis cx1이 생산하는 박테리오신의 특성)

  • 김수인;장지윤;김인철;장해춘
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.50-55
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    • 2001
  • A new bacteriocin produced by Bacillus subtilis cx1, was partially purified and characterized. The bactericoin from B. subtilis cx1 was stable in the range of pH 2.5-9.5. B. subtilis csx1 retained its antimicrobial activity to long-term exposure at $-20^{\circ}C$ and $-70^{\circ}C$. However, B. subtilis cx1 was inactivated completely within 15 min over $60^{\circ}C$ and lost 50% of its antimicrobial activity within 15 min at $50^{\circ}C$, B. subtilis cx1 was inactivated by protease, trypsin, proteinase K and carboxypeptidase, which indi-cates its protein nature. Direct detection of the antimicrobial activity on Tricine -SDS-PAGE suggested an apparent molecular mass of about 9,500 dalton.

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Application of Hydrogen Peroxide on the Bacterial Control of Seaweed, Capsosiphon fulvescens (Mesaengi) (해조류 매생이(Capsosiphon fulvescens)의 저장기간 연장을 위한 과산화수소의 활용)

  • Kim, Du-Woon;Kim, Mi-Jung;Shin, Tai-Sun;Kim, Sun-Jae;Jung, Bok-Mi
    • Food Science and Preservation
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    • v.15 no.2
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    • pp.169-173
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    • 2008
  • Bacillus subtilis subsp. subtilis constitutes 90% of the total viable bacteria present on Capsosiphon fulvescens. We found that hydrogen peroxide (50 ppm) and NaOCl (50 ppm) were more effective than electrolyzed water (EW, 50ppm) against B. subtilis subsp. subtilis that was isolated from this seaweed. Relative to a control, 50 ppm hydrogen peroxide reduced the total viable population by $1.8{\pm}0.4$ log CFU/g, whereas 50 ppm EW increased the total viable population by $1.7{\pm}0.5$ log CFU/g. CFUs were evaluated following 30 days of storage at $4^{\circ}C$ using air- and vacuum-packaging. Samples treated with 50 ppm hydrogen peroxide and NaOCl showed a $1.6{\pm}0.1$-fold decrease in initial hardness ($7.9{\times}10^6dyne/cm^2$), while the samples treated with 50 ppm EW had a $2.1{\pm}0.1$-fold decrease in initial hardness ($7.9{\times}10^6dyne/cm^2$). Again, measurements were performed after storage at $4^{\circ}C$ for 20 days. This study indicates that B. subtilis subsp. subtilis is the most common contaminant in aerobically or anaerobically packaged seaweed and should therefore be the main target for quality control during long-term storage. Hydrogen peroxide and NaOCl are more effective than EW in inhibiting B. subtilis subsp. subtilis and in reducing total bacterial loads in air- and vacuum-packaged seaweed.

Chromosomal Mapping of the cdd Gene Encoding Deoxycytidine-cytidine Deaminase in Bacillus subtilis (Bacillus subtilis의 시티딘 디아미나제를 코드하는 cdd 유전자의 Chromosomal Mapping)

  • Song, Bang-Ho;Jan Neuhard
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.536-539
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    • 1988
  • A mutant of Bacillus subtilis with a defective cdd gene encoding deoxycytidine-cytidine deaminase (EC 3.5.4.5) has been characterized genetically. The genetic lesion, cdd, causing the altered deoxycytidine-cytidine deaminase was mapped at 225 min on the linkage map of B. subtilis by AR9 transduction, Transductional analysis of the cdd region established the gene order in clockwise as trp-lys-cdd-aroD. The cdd gene was linked 72% with the aroD and 20% with the lys.

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Characteristics of Antifungal Bacterium, Bacillus subtilis YS1 and It′s Mutant Induced by Gamma Radiation (온천수로부터 분리한 항진균세균의 특성 및 감마선$(Co^{60})$ 조사를 이용한 돌연변이체 유기)

  • 이영근;김재성;송인근;정혜영;장화형
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.305-311
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    • 2001
  • Antifungal bacterium, Bacillus subtilis YS1 was isolated from Yusong hot spring showed broad antifungal spectrum against 12 kinds of plant pathogenic fungi and Candida albicans, animal pathogen. From the gamma($Co^{60}$) radiation sensitivity test, $D_10$ value was 2.08 kGy and it survived above 20 kGy of radiation dose. Several mutants were induced by gamma radiation. Among them, YS1-1009 mutant showed resistance against tebuconazole of herbicide, increased activity against Botryoshaeria dothidea and ligninase activity. YS67 mutant was antifungal deficient auxotrophic mutants(trp-pro-or arg-ura-). From this results, it suggested that gamma irradiation could be useful method for mutant induction.

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Changes of Free Sugar and Free Amino Acid during the Natto Fermentation used by Bacillus subtilis S.N.U 816 (Bacillus subtilis S. N. U 816 균주를 이용한 Natto 제조중 유리당 및 유리아미노산의 변화)

  • Kim, Bok-Ran;Han, Yong-Bong;Park, Chang-Hee
    • Applied Biological Chemistry
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    • v.30 no.2
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    • pp.192-197
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    • 1987
  • Natto was produced by fermenting local soybeans Bacillus subtilis S.N.U. 816. The changes of chemical composition, enzyme activity and texture of NATTO during the fermentation were investigated. The amount of amino type and watr soluble nitrogens were increased as the fermentation progressed, although the former seemed to reach a plateau at about 20 hours of the fermentation, of the protease activity were increased until 16 hours of fermentation at which time they tended to reach plateaus. Among the inspected free sugars (fructose, glucose, sucrose, maltose), remarkable increases in the levels of fructose and glucose were observe3 after 4 hours of the fermentation. Since then their contents, however, were reduced very low as the processing went on, and sucrose contents dropped drastically to about 10% level and stayed low thereafter. Free amino acid contents of natto during 20 hours of the fermentation were or 2 times greater than those of the unfermented steamed soybean, the 24 hours ferment, respectively. Sensory evaluation revealed that 20 hours of fermentation produced the best quality products based on taste, odor, and color, considering all the data, it seems possible to conclude that the optimum of time for fermentation of natto at $42^{\circ}C$ is 20 hours.

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