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Sample records for bacillus licheniformis system

  1. Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis.

    Science.gov (United States)

    Rachinger, Michael; Bauch, Melanie; Strittmatter, Axel; Bongaerts, Johannes; Evers, Stefan; Maurer, Karl-Heinz; Daniel, Rolf; Liebl, Wolfgang; Liesegang, Heiko; Ehrenreich, Armin

    2013-09-20

    Conjugative shuttle vectors of the pKVM series, based on an IncP transfer origin and the pMAD vector with a temperature sensitive replication were constructed to establish a markerless gene deletion protocol for Bacilli without natural competence such as the exoenzyme producer Bacillus licheniformis. The pKVM plasmids can be conjugated to strains of B. licheniformis and B. subtilis. For chromosomal gene deletion, regions flanking the target gene are fused and cloned in a pKVM vector prior to conjugative transfer from Escherichia coli to B. licheniformis. Appropriate markers on the vector backbone allow for the identification of the integration at the target locus and thereafter the vector excision, both events taking place via homologous recombination. The functionality of the deletion system was demonstrated with B. licheniformis by a markerless 939 bp in-frame deletion of the yqfD gene and the deletion of a 31 kbp genomic segment carrying a PBSX-like prophage. PMID:23916947

  2. EXPERIMENTAL-INFECTION IN MICE WITH BACILLUS-LICHENIFORMIS

    DEFF Research Database (Denmark)

    Agerholm, J.S.; Jensen, H.E.; Jensen, N.E.

    1995-01-01

    The pathogenicity of Bacillus licheniformis was assessed in normal and immunodepressed BALB/c mice. The animals were challenged intravenously with 4 x 10(7) colony forming units of B, licheniformis (ATCC 14580) and both normal and immunodepressed mice were susceptible. However, the infection was...... more severe in the immunosuppressed animals. In normal mice, lesions were restricted to the liver and kidneys, while lesions also occurred in other organs of immunodepressed mice. By crossed immunoelectrophoresis it was shown that antigens of B. licheniformis are potent immunogens, and the bacteria...

  3. Expression of alpha-amylase in Bacillus licheniformis.

    OpenAIRE

    Rothstein, D. M.; Devlin, P E; Cate, R. L.

    1986-01-01

    In Bacillus licheniformis, alpha-amylase production varied more than 100-fold depending on the presence or absence of a catabolite-repressing carbon source in the growth medium. alpha-Amylase was produced during the growth phase and not at the onset of the stationary phase. Induction of alpha-amylase correlated with synthesis of mRNA initiating at the promoter of the alpha-amylase gene.

  4. Biodegradation of malathion by Bacillus licheniformis strain ML-1

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    Khan Sara

    2016-01-01

    Full Text Available Malathion, a well-known organophosphate pesticide, has been used in agriculture over the last two decades for controlling pests of economically important crops. In the present study, a single bacterium, ML-1, was isolated by soil-enrichment technique and identified as Bacillus licheniformis on the basis of the 16S rRNA technique. The bacterium was grown in carbon-free minimal salt medium (MSM and was found to be very efficient in utilizing malathion as the sole source of carbon. Biodegradation experiments were performed in MSM without carbon source to determine the malathion degradation by the selected strain, and the residues of malathion were determined quantitatively using HPLC techniques. Bacillus licheniformis showed very promising results and efficiently consumed malathion as the sole carbon source via malathion carboxylesterase (MCE, and about 78% malathion was degraded within 5 days. The carboxylesterase activity was determined by using crude extract while using malathion as substrate, and the residues were determined by HPLC. It has been found that the MCE hydrolyzed 87% malathion within 96 h of incubation. Characterization of crude MCE revealed that the enzyme is robust in nature in terms of organic solvents, as it was found to be stable in various concentrations of ethanol and acetonitrile. Similarly, and it can work in a wide pH and temperature range. The results of this study highlighted the potential of Bacillus licheniformis strain ML-1 as a biodegrader that can be used for the bioremediation of malathion-contaminated soil.

  5. N-terminal amino acid sequence of Bacillus licheniformis alpha-amylase: comparison with Bacillus amyloliquefaciens and Bacillus subtilis Enzymes.

    OpenAIRE

    Kuhn, H; Fietzek, P P; Lampen, J. O.

    1982-01-01

    The thermostable, liquefying alpha-amylase from Bacillus licheniformis was immunologically cross-reactive with the thermolabile, liquefying alpha-amylase from Bacillus amyloliquefaciens. Their N-terminal amino acid sequences showed extensive homology with each other, but not with the saccharifying alpha-amylases of Bacillus subtilis.

  6. Stress responses of the industrial workhorse Bacillus licheniformis to osmotic challenges.

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    Rebecca Schroeter

    Full Text Available The Gram-positive endospore-forming bacterium Bacillus licheniformis can be found widely in nature and it is exploited in industrial processes for the manufacturing of antibiotics, specialty chemicals, and enzymes. Both in its varied natural habitats and in industrial settings, B. licheniformis cells will be exposed to increases in the external osmolarity, conditions that trigger water efflux, impair turgor, cause the cessation of growth, and negatively affect the productivity of cell factories in biotechnological processes. We have taken here both systems-wide and targeted physiological approaches to unravel the core of the osmostress responses of B. licheniformis. Cells were suddenly subjected to an osmotic upshift of considerable magnitude (with 1 M NaCl, and their transcriptional profile was then recorded in a time-resolved fashion on a genome-wide scale. A bioinformatics cluster analysis was used to group the osmotically up-regulated genes into categories that are functionally associated with the synthesis and import of osmostress-relieving compounds (compatible solutes, the SigB-controlled general stress response, and genes whose functional annotation suggests that salt stress triggers secondary oxidative stress responses in B. licheniformis. The data set focusing on the transcriptional profile of B. licheniformis was enriched by proteomics aimed at identifying those proteins that were accumulated by the cells through increased biosynthesis in response to osmotic stress. Furthermore, these global approaches were augmented by a set of experiments that addressed the synthesis of the compatible solutes proline and glycine betaine and assessed the growth-enhancing effects of various osmoprotectants. Combined, our data provide a blueprint of the cellular adjustment processes of B. licheniformis to both sudden and sustained osmotic stress.

  7. Nanoactivator mediated modifications in thermostable amylase from Bacillus licheniformis.

    Science.gov (United States)

    Khairnar, Rajendra S; Mahabole, Megha P; Pathak, Anupama P

    2012-12-01

    Gram-positive rod-shaped thermophilic bacteria were isolated using samples collected from terrestrial natural thermal spring located at Unkeshwar (Longitude 78.22 degree East to 78.34 degree East, Latitude 19 degree 34' North to 19 degree 40' North), District Nanded, Maharashtra State, India. The isolates were then cultivated using selective media and identified using culture-dependent techniques. One prominent isolate (UN1) exhibited high temperature stability and remarkable amylase production and was identified as Bacillus licheniformis. Amylase production was carried out in starch media and the enzyme was partially purified and characterized for optimization of pH and temperature. Amylolytic activity of the enzyme was determined. Nanoactivator-mediated modifications were carried out to enhance amylolytic activity of the partially purified amylase. Three-fold increase in catalytic efficiency of amylase was obtained after modification. PMID:23350283

  8. IMPROVING OF THE IMPREGNABILITY OF REFRACTORY SPRUCE WOOD BY BACILLUS LICHENIFORMIS PRETREATMENT

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    Sibel Yildiz,

    2011-11-01

    Full Text Available In this study it was aimed to improve impregnability of spruce (Picea orientalis L. wood with bacteria (Bacillus licheniformis A1 pretreatment, using copper/chromium/arsenic Type C (CCA-C and copper azole Type A (CBA-A. The effects of Bacillus licheniformis A1 on weight loss, copper uptake, and compression strength of samples were determined. Weight loss was slightly changed by bacterial degradation in all test groups. The best copper uptake cases were 1466 ppm for CCA-C and 2730 ppm for CBA-A. Improvement on copper uptake with bacteria pretreatment was in a range of 18 to 103% compared to control samples. Compression strength was decreased by bacterial degradation. However strength losses might be acceptable for several construction applications. Bacillus licheniformis A1 seems to have a good potential for increasing the permeability of spruce wood.

  9. STUDY REGARDING EFFICIENCY OF INDUCED GENETIC TRANSFORMATION IN BACILLUS LICHENIFORMIS WITH PLASMID DNA

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    T. VINTILĂ

    2013-12-01

    Full Text Available A strain of Bacillus licheniformis was subject to genetic transformation with plasmid vectors (pLC1 and pNC61, using electroporation technique, protoplast transformation and bivalent cations (CaCl2 mediated transformation. In the case of transformation by electroporation of Bacillus licheniformis B40, the highest number of transformed colonies (3 were obtained only after a 1,79 KV electric shock, for 2,2 milliseconds. Using this transformation technique we have obtained six kanamycin resistant transformants. The frequency of Bacillus licheniformis B40 protoplasts transformation using pLC1 and pNC61 plasmid vectors is approximately 10% (TF = 10%. As a result of pLC1 plasmid integration in Bacillus licheniformis protoplasts, six kanamycin resistant transformants were obtained. The pNC61 plasmid, which confers trimethoprim resistance, does not integrate in receiver cells by protoplast transformation. The direct genetic transformation in the presence of bivalent cations (CaCl2, mediated by pLC1 and pNC61 plasmid vectors, produce a low transformation frequency. Using this technique, we have obtained three trimethoprim resistant colonies and four kanamycin resistant colonies. The chemical way of transformation is the only technique, which realizes the integration of pNC61 in B. licheniformis B40 cells.

  10. STUDY REGARDING EFFICIENCY OF INDUCED GENETIC TRANSFORMATION IN BACILLUS LICHENIFORMIS WITH PLASMID DNA

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    VINTILĂ T.

    2007-01-01

    Full Text Available A strain of Bacillus licheniformis was subject to genetic transformation with plasmidvectors (pLC1 and pNC61, using electroporation technique, protoplasttransformation and bivalent cations (CaCl2 mediated transformation. In the case oftransformation by electroporation of Bacillus licheniformis B40, the highest numberof transformed colonies (3 were obtained only after a 1,79 KV electric shock, for 2,2milliseconds. Using this transformation technique we have obtained six kanamycinresistant transformants. The frequency of Bacillus licheniformis B40 protoplaststransformation using pLC1 and pNC61 plasmid vectors is approximately 10% (TF =10%. As a result of pLC1 plasmid integration in Bacillus licheniformis protoplasts,six kanamycin resistant transformants were obtained. The pNC61 plasmid, whichconfers trimethoprim resistance, does not integrate in receiver cells by protoplasttransformation. The direct genetic transformation in the presence of bivalent cations(CaCl2, mediated by pLC1 and pNC61 plasmid vectors, produce a lowtransformation frequency. Using this technique, we have obtained three trimethoprimresistant colonies and four kanamycin resistant colonies. The chemical way oftransformation is the only technique, which realizes the integration of pNC61 in B.licheniformis B40 cells.

  11. Control of Bacillus licheniformis spores isolated from dairy materials in yogurt production.

    Science.gov (United States)

    Tanaka, Takashi; Ito, Akiko; Kamikado, Hideaki

    2012-01-01

    To determine the effects of sporulation temperature and period on Bacillus licheniformis spore heat resistance, B. licheniformis strain No.25 spores were sporulated at 30, 37, 42, or 50°C for 11 d and at 50°C for 1.7, 4, 7, or 11 d. The heat resistance of B. licheniformis strain No.25 spores at 110°C increased with an increase in the sporulation temperature. Spores sporulated at 50°C were 1.4-fold more heat resistant than those sporulated at 30°C. Furthermore, the heat resistance of B. licheniformis strain No.25 spores at 110°C increased with an increase in the sporulation period. Spores sporulated for 11 d were 5.3-fold more heat resistant than those sporulated for 1.7 d. The heat resistance of B. licheniformis strain No.25 spores at 110°C increased with increases in the sporulation temperature and sporulation period. The results presented in this study can be applied to the pasteurization process to control B. licheniformis spores. Pasteurization at 110°C for about 60sec. is effective in controlling B. licheniformis spores isolated from dairy materials in yogurt production.

  12. Hydrolysis of Whey Protein Isolate with Bacillus licheniformis Protease: Fractionation and Identification of Aggregating Peptides

    NARCIS (Netherlands)

    Creusot, N.P.; Gruppen, H.

    2007-01-01

    The objective of this work was to identify the dominant aggregating peptides from a whey protein hydrolysate (degree of hydrolysis of 6.8%) obtained with Bacillus licheniformis protease. The aggregating peptides were fractionated with preparative reversed-phase chromatography and identified with liq

  13. Enzyme-induced aggregation of whey proteins with Bacillus licheniformis protease

    NARCIS (Netherlands)

    Creusot, N.P.

    2006-01-01

    Whey proteins are commonly used as ingredient in food. In relation with the gelation properties of whey proteins, this thesis deals with understanding the mechanism of peptide-induced aggregation of whey protein hydrolysates made with Bacillus licheniformis protease (BLP). The results show that BLP

  14. A Bacillus licheniformis pectin acetylesterase is specific for homogalacturonans acetylated at O-3

    NARCIS (Netherlands)

    Remoroza, C.A.; Wagenknecht, M.; Buchholt, H.C.; Moerschbacher, B.M.; Schols, H.A.; Gruppen, H.

    2014-01-01

    A recombinant acetylesterase from Bacillus licheniformis DSM13, belonging to carbohydrate esterase family 12, was purified and biochemically characterized. The purified enzyme, termed BliPAE, was capable of deacetylating acetylated pectins, e.g. sugar beet pectin (SBP). Contrary to its provisional a

  15. The extracellular proteome of Bacillus licheniformis grown in different media and under different nutrient starvation conditions

    NARCIS (Netherlands)

    Voigt, B; Schweder, T; Sibbald, MJJB; Albrecht, D; Ehrenreich, A; Bernhardt, J; Feesche, J; Maurer, KH; Gottschalk, G; van Dijl, JM; Hecker, M

    2006-01-01

    The now finished genome sequence of Bacillus licheniformis DSM 13 allows the prediction of the genes involved in protein secretion into the extracellular environment as well as the prediction of the proteins which are translocated. From the sequence 296 proteins were predicted to contain an N-termin

  16. OPTIMIZATION OF MEDIA CONSTITUENTS FOR THE PRODUCTION OF ALKALINE PROTEASE FROM BACILLUS LICHENIFORMIS Mohideen

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    Mohideen Askar Nawas P

    2015-07-01

    Full Text Available Production of alkaline protease by Bacillus licheniformis has been investigated under submerged fermentation. The physical and chemical parameters influencing submerged fermentation were optimized. The effect of incubation time, temperature, pH, carbon sources and nitrogen sources and additional nutrients on the production of alkaline protease was characterized. The optimum conditions for the protease production by Bacillus licheniformis were found to be at pH 9.0 and temperature at 40ºC. The outcome of carbon and inorganic nitrogen sources on protease production proved that glucose and casein were the effective medium ingredients for Bacillus licheniformis respectively. The maximum amount of protease production was recorded in medium supplemented with ammonium sulphate. Among the tested metal ions, the level of protease yield was found to be high in medium supplemented with magnesium chloride. The protease production was amplified in the presence of 1.5% sodium chloride. The extreme stability towards Triton X-100, Tween 20 and SDS was observed in Bacillus licheniformis alkaline protease.

  17. A preliminary study on the pathogenicity of Bacillus licheniformis bacteria in immunodepressed mice

    DEFF Research Database (Denmark)

    Agerholm, J.S.; Jensen, N.E.; Giese, Steen Bjørck;

    1997-01-01

    The pathogenicity of 13 strains of Bacillus licheniformis was studied in immunodepressed mice. The strains had been isolated from cases of bovine abortions (n=5), bovine feedstuffs (n=3), soil (n=l), and grain products (n=2). The origin of two strains was unknown. Groups of 10 mice were inoculated...

  18. Kinetics and thermodynamic studies of alpha amylase from bacillus licheniformis mutant

    International Nuclear Information System (INIS)

    The present investigation deals with the purification and characterization of enzyme a'-amylase from a mutant strain of Bacillus licheniformis EMS-6. A laboratory scale stirred fermentor of 7.5 L capacity was used for the enzyme production under optimal conditions. The enzyme was purified up to homogeneity level by Ammonium sulphate and ion-exchange chromatography using a fast protein liquid chromatography (FPLC) system. The specific activity of the enzyme increased 4-5 times while the yield was found to be 40.4%. The purification fold by RESOURCE-S was recorded to be 3.58. The molecular weight was found to be 55 KDa. In the present research work, the Vmax (2778 U/mg/min) and Km (8.3mg/ml) of a'-amylase were derived from the Lineweaver Burke plot. Thermodynamic parameters for soluble starch hydrolysis, Ea, AH, AS and AG of a'-amylase from B. licheniformis EMS-6 were found to be 25.14 KJ/mol, 22.53 KJ/mole, -110.95 J/mole/K and 36968 J/mole, respectively. The enzyme was stable over a pH range of 4.5-9.0 and gave pH optimum of 7.0. The pKa1 and pKa2 of ionizable groups of active site controlling Vmax, determined by Dixon plot, were 6.0 and 7.5, respectively. (author)

  19. Partial purification of a Bacillus licheniformis levansucrase producing levan with antitumor activity.

    Science.gov (United States)

    Dahech, Imen; Belghith, Karima Srih; Belghith, Hafedh; Mejdoub, Hafedh

    2012-10-01

    The extracellular fructosyltransferase (FTase) of a novel strain of Bacillus licheniformis capable of producing fructooligosaccharides (FOS) and a polysaccharide type levan was obtained and partially purified. The purification was achieved by ammonium sulfate precipitation, DEAE cellulose and gel filtration chromatographies. The enzyme was partially purified as determined by SDS-PAGE, and the specific activity reached was 67.5, representing a purification factor of 177 and yield of 40%. Levan was isolated from the cultures of B. licheniformis. The levan was composed mainly of fructose residues when analyzed by TLC after acid hydrolysis and NMR analysis. In a previous study, the levan produced exhibited a hypoglycemiant activity. The present paper deals with the study of the antitumor and anti-cytotoxic effect of levan produced by B. licheniformis strain. In the in vitro antitumor activity test of levan against some tumor cell lines, relatively the significantly high activity was observed against the HepG(2). PMID:22579870

  20. Regulation of the activity of the Bacillus licheniformis A5 glutamine synthetase.

    OpenAIRE

    Donohue, T J; Bernlohr, R W

    1981-01-01

    The regulation of glutamine synthetase activity by positive and negative effectors of enzyme activity singularly and in combinations was studied by using a homogeneous enzyme preparation from Bacillus licheniformis A5. Phosphorylribosyl pyrophosphate at concentrations greater than 2mM stimulated glutamine synthetase activity by approximately 70%. The concentration of phosphorylribosyl pyrophosphate required for half-maximal stimulation of enzyme activity was 0.4 mM. Results obtained from stud...

  1. Effect of oilseed cakes on alpha-amylase production by Bacillus licheniformis CUMC305.

    OpenAIRE

    Krishnan, T.; Chandra, A. K.

    1982-01-01

    The effects of oilseed cakes on extracellular thermostable alpha-amylase production by Bacillus licheniformis CUMC305 was investigated. Each oilseed cake was made of groundnut, mustard, sesame, linseed, coconut copra, madhuca, or cotton. alpha-Amylase production was considerably improved in all instances and varied with the oilseed cake concentration in basal medium containing peptone and beef extract. Maximum increases were effected by a low concentration (0.5 to 1.0%) of groundnut or coconu...

  2. [Maxillary sinus infection by Bacillus licheniformis: a case report from Djibouti].

    Science.gov (United States)

    Garcia Hejl, C; Sanmartin, N; Samson, T; Soler, C; Koeck, J-L

    2015-01-01

    Aerobic, spore-forming gram-positive Bacillus spp infections are rare and reported mainly in immunocompromised hosts. We report a case of acute unilateral maxillary sinusitis, caused by Bacillus licheniformis, in a 35-year-old French soldier stationed in Djibouti. It was easily identifiable due to its typical culture and resistance profile. This case is interesting for two reasons: first, it is, to our knowledge, the first case of sinusitis attributed to this microbe, and second, it has rarely been described in immunocompetent patients without altered skin or mucous membranes. PMID:26370779

  3. Fungicin M4: a narrow spectrum peptide antibiotic from Bacillus licheniformis M-4.

    Science.gov (United States)

    Lebbadi, M; Gálvez, A; Maqueda, M; Martínez-Bueno, M; Valdivia, E

    1994-07-01

    The strain Bacillus licheniformis M-4 produces a 3.4 kDa hydrophilic peptide with antifungal activity, named fungicin M4. Analysis of the purified peptide shows that it contains the amino acids Glu (8), Arg (5), Pro (4), Tyr (8), Val (3), Met (2) and Orn (4). Its inhibitory spectrum is restricted to Microsporum canis CECT 2797, Mucor mucedo CECT 2653, Mucor plumbeus CCM 443, Sporothrix schenckii CECT 2799, Bacillus megaterium and Corynebacterium glutamicum CECT 78. Fungicin M4 exerts biocidal activity on liquid cultures of Sporothrix schenckii CECT 2799.

  4. Morphological and molecular based identification of pectinase producing Bacillus licheniformis from rotten vegetable

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    Haneef Ur Rehman

    2015-12-01

    Full Text Available Pectinase catalyzed the degradation of pectin substances and has been used in various biotechnological industries. In the current study, 23 bacterial strains were isolated from rotten vegetables, soil and air. The isolated bacterial strains were qualitatively screened for pectinase production on pectin agar medium and only three strains HR 4, HR 21 and HR 23 were observed to produce extracellular pectinase. These strains were further screened quantitatively for pectinase production through submerged fermentation technology in pectin containing fermentation medium. Strain HR 4 from rotten brinjal (Solanum melongena was found to produce higher pectinase as compared to others. The maximum pectinase producing bacterial strain was identified as Bacillus licheniformis on the basis of morphological, physiological and biochemical characteristics. For further confirmation of identification, 16S rDNA sequence analysis was performed. The 16S rDNA sequences were aligned and the phylogenetic tree was constructed. The phylogenetic tree confirmed that the strain was belonging to B. licheniformis. The 16S rDNA sequences of this new strain were submitted to GenBank and designated as B. licheniformis KIBGE-IB21 with the GenBank accession number JQ 411812. The newly isolated pectinase producing B. licheniformis used apple pectin as carbon and yeast extract as nitrogen source for maximum pectinase production.

  5. High levan production by Bacillus licheniformis NS032 using ammonium chloride as the sole nitrogen source.

    Science.gov (United States)

    Kekez, B D; Gojgic-Cvijovic, G D; Jakovljevic, D M; Stefanovic Kojic, J R; Markovic, M D; Beskoski, V P; Vrvic, M M

    2015-03-01

    In this study, levan production by Bacillus licheniformis NS032 isolated from a petroleum sludge sample was investigated. High levan yield was obtained in a wide range of sucrose concentrations (up to 400 g/L) and, contrary to most levan-producing strains, using ammonium chloride as the sole N source. Interaction between sucrose, ammonium chloride, and initial pH of the medium in a low sucrose (60-200 g/L) and a high sucrose (300-400 g/L) system was analyzed by response surface methodology. According to the calculated model in the low sucrose system, maximum predicted levan yield was 47.8 g/L (sucrose 196.8 g/L, ammonium chloride 2.4 g/L, pH 7.0), while in the high sucrose system, levan yield was 99.2 g/L (sucrose 397.6 g/L, ammonium chloride 4.6 g/L, pH 7.4). In addition, protective effect of microbial levan against copper toxicity to Daphnia magna is observed for the first time. The acute toxicity (48 h EC50) of copper decreased from 0.14 to 0.44 mg/L by levan in concentration of 50 ppm. PMID:25592434

  6. Effect of oilseed cakes on alpha-amylase production by Bacillus licheniformis CUMC305.

    Science.gov (United States)

    Krishnan, T; Chandra, A K

    1982-08-01

    The effects of oilseed cakes on extracellular thermostable alpha-amylase production by Bacillus licheniformis CUMC305 was investigated. Each oilseed cake was made of groundnut, mustard, sesame, linseed, coconut copra, madhuca, or cotton. alpha-Amylase production was considerably improved in all instances and varied with the oilseed cake concentration in basal medium containing peptone and beef extract. Maximum increases were effected by a low concentration (0.5 to 1.0%) of groundnut or coconut, a high concentration (3%) of linseed or mustard, and an Rintermediate concentration (2%) of cotton, madhuca, or sesame. The oilseed cakes made of groundnut or mustard could completely replace the conventional peptone-beef extract medium as the fermentation base for the production of alpha-amylase by B. licheniformis. The addition of corn steep liquor to cotton, linseed, sesame, or madhuca cake in the medium improved alpha-amylase production. PMID:6181738

  7. The Sponge-associated Bacterium Bacillus licheniformis SAB1: A Source of Antimicrobial Compounds

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    Prabha Devi

    2010-04-01

    Full Text Available Several bacterial cultures were isolated from sponge Halichondria sp., collected from the Gujarat coast of the Indo Pacific region. These bacterial cultures were fermented in the laboratory (100 mL and the culture filtrate was assayed for antibiotic activity against 16 strains of clinical pathogens. Bacillus sp. (SAB1, the most potent of them and antagonistic to several clinically pathogenic Gram-positive, Gram-negative bacteria and the fungus Aspergillus fumigatus was chosen for further investigation. Analysis of the nucleotide sequence of the 16S rDNA gene of Bacillus sp. SAB1 showed a strong similarity (100% with the 16S rDNA gene of Bacillus licheniformis HNL09. The bioactive compounds produced by Bacillus licheniformis SAB1 (GenBank accession number: DQ071568 were identified as indole (1, 3-phenylpropionic acid (2 and a dimer 4,4′-oxybis[3-phenylpropionic acid] (3 on the basis of their Fourier Transform Infrared (FTIR, Nuclear Magnetic Resonance (NMR and Electrospray Ionization Mass Spectrometer (ESI-MS data. There is a single reference on the natural occurrence of compound 3 from the leaves of a terrestrial herb Aptenia cordifolia in the literature, so to the best of our knowledge, this is a first report of its natural occurrence from a marine source. The recovery of bacterial strains with antimicrobial activity suggests that marine-invertebrates remain a rich source for the isolation of culturable isolates capable of producing novel bioactive secondary metabolites.

  8. Plasmid-Mediated Dimethoate Degradation by Bacillus licheniformis Isolated From a Fresh Water Fish Labeo rohita

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    Manisha Deb Mandal

    2005-01-01

    Full Text Available The Bacillus licheniformis strain isolated from the intestine of Labeo rohita by an enrichment technique showed capability of utilizing dimethoate as the sole source of carbon. The bacterium rapidly utilized dimethoate beyond 0.6 mg/mL and showed prolific growth in a mineral salts medium containing 0.45 mg/mL dimethoate. The isolated B licheniformis exhibited high level of tolerance of dimethoate (3.5 mg/mL in nutrient broth, while its cured mutant did not tolerate dimethoate beyond 0.45 mg/mL and it was unable to utilize dimethoate. The wild B licheniformis strain transferred dimethoate degradation property to E coli C600 (Nar, F− strain. The transconjugant harbored a plasmid of the same molecular size (approximately 54 kb as that of the donor plasmid; the cured strain was plasmid less. Thus a single plasmid of approximately 54 kb was involved in dimethoate degradation. Genes encoding resistance to antibiotic and heavy metal were also located on the plasmid.

  9. Seeking new mutation clues from Bacillus licheniformis amylase by molecular dynamics simulations

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    Lu, Tao

    2009-07-01

    Amylase is one of the most important industrial enzymes in the world. Researchers have been searching for a highly thermal stable mutant for many years, but most focus on point mutations of one or few nitrogenous bases. According to this molecular dynamic simulation of amylase from Bacillus licheniformis (BLA), the deletion of some nitrogenous bases would be more efficacious than point mutations. The simulation reveals strong fluctuation of the BLA structure at optimum temperature. The fluctuation of the outer domains of BLA is stronger than that of the core domain. Molecular simulation provides a clue to design thermal stable amylases through deletion mutations in the outer domain.

  10. Cyclodextrin glycosyltransferase from Bacillus licheniformis: optimization of production and its properties Cyclodextrina glycosyltransferase de Bacillus licheniformis: otimização da produção e suas propriedades

    OpenAIRE

    Paulo Roberto Martins Bonilha; Vivian Menocci; Antonio José Goulart; Maria de Lourdes Teixeira de Moraes Polizeli; Rubens Monti

    2006-01-01

    Cyclodextrin glycosyltransferase (EC 2.4.1.19) is an enzyme that produces cyclodextrins from starch via an intramolecular transglycosylation reaction. An alkalophilic Bacillus strain, isolated from cassava peels, was identified as Bacillus licheniformis. CGTase production by this strain was better when potato starch was used as carbon source, followed by cassava starch and amylopectin. Glucose and amylose, on the other hand, acted as synthesis repressors. When the cultivation was supplemented...

  11. Bacillus licheniformis Isolated from Traditional Korean Food Resources Enhances the Longevity of Caenorhabditis elegans through Serotonin Signaling.

    Science.gov (United States)

    Park, Mi Ri; Oh, Sangnam; Son, Seok Jun; Park, Dong-June; Oh, Sejong; Kim, Sae Hun; Jeong, Do-Youn; Oh, Nam Su; Lee, Youngbok; Song, Minho; Kim, Younghoon

    2015-12-01

    In this study, we investigated potentially probiotic Bacillus licheniformis strains isolated from traditional Korean food sources for ability to enhance longevity using the nematode Caenorhabditis elegans as a simple in vivo animal model. We first investigated whether B. licheniformis strains were capable of modulating the lifespan of C. elegans. Among the tested strains, preconditioning with four B. licheniformis strains significantly enhanced the longevity of C. elegans. Unexpectedly, plate counting and transmission electron microscopy (TEM) results indicated that B. licheniformis strains were not more highly attached to the C. elegans intestine compared with Escherichia coli OP50 or Lactobacillus rhamnosus GG controls. In addition, qRT-PCR and an aging assay with mutant worms showed that the conditioning of B. licheniformis strain 141 directly influenced genes associated with serotonin signaling in nematodes, including tph-1 (tryptophan hydroxylase), bas-1 (serotonin- and dopamine-synthetic aromatic amino acid decarboxylase), mod-1 (serotonin-gated chloride channel), ser-1, and ser-7 (serotonin receptors) during C. elegans aging. Our findings suggest that B. licheniformis strain 141, which is isolated from traditional Korean foods, is a probiotic generally recognized as safe (GRAS) strain that enhances the lifespan of C. elegans via host serotonin signaling.

  12. Design of thermostable rhamnogalacturonan lyase mutants from Bacillus licheniformis by combination of targeted single point mutations

    DEFF Research Database (Denmark)

    da Silva, Ines Isabel Cardoso Rodrigues; Jers, Carsten; Otten, Harm;

    2014-01-01

    Rhamnogalacturonan I lyases (RGI lyases) (EC 4.2.2.-) catalyze cleavage of α-1,4 bonds between rhamnose and galacturonic acid in the backbone of pectins by β-elimination. In the present study, targeted improvement of the thermostability of a PL family 11 RGI lyase from Bacillus licheniformis (DSM......, were obtained due to additive stabilizing effects of single amino acid mutations (E434L, G55V, and G326E) compared to the wild type. The crystal structure of the B. licheniformis wild-type RGI lyase was also determined; the structural analysis corroborated that especially mutation of charged amino...

  13. Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis

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    Jin-Song Gong

    2015-12-01

    Full Text Available In the present study, we isolated a trypsin-producing strain DMN6 from the leather waste and identified it as Bacillus licheniformis through a two-step screening strategy. The trypsin activity was increased up to 140 from 20 U/mL through culture optimization. The enzyme was purified to electrophoretic homogeneity with a molecular mass of 44 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the specific activity of purified enzyme is 350 U/mg with Nα-Benzoyl-l-arginine ethylester as the substrate. The optimum temperature and pH for the trypsin are 65 °C and pH 9.0, respectively. Also, the enzyme can be significantly activated by Ba2+. This enzyme is relatively stable in alkaline environment and displays excellent activity at low temperatures. It could retain over 95% of enzyme activity after 180 min of incubation at 45 °C. The distinguished activity under low temperature and prominent stability enhance its catalytic potential. In the current work, the open reading frame was obtained with a length of 1371 nucleotides that encoded a protein of 456 amino acids. These data would warrant the B. licheniformis trypsin as a promising candidate for catalytic application in collagen preparation and leather bating through further protein engineering.

  14. Regulation of the activity of the Bacillus licheniformis A5 glutamine synthetase.

    Science.gov (United States)

    Donohue, T J; Bernlohr, R W

    1981-10-01

    The regulation of glutamine synthetase activity by positive and negative effectors of enzyme activity singularly and in combinations was studied by using a homogeneous enzyme preparation from Bacillus licheniformis A5. Phosphorylribosyl pyrophosphate at concentrations greater than 2mM stimulated glutamine synthetase activity by approximately 70%. The concentration of phosphorylribosyl pyrophosphate required for half-maximal stimulation of enzyme activity was 0.4 mM. Results obtained from studies of fractional inhibition of glutamine synthetase activity were consistent with the presence of one allosteric site for glutamine binding (apparent I0.5, 2.2mM) per active enzyme unit at a glutamate concentration of 50 mM. At a glutamate concentration of 30 mM or less, the data were consistent with the enzyme containing two binding sites for glutamine (one of which was an allosteric site with an apparent I0.5 of 0.4 mM). Bases on an analysis of the response of glutamine synthetase activity to positive and negative effectors in vitro and to the intracellular concentration of these effectors in vivo, the primary modulators of glutamine synthetase activity in B. licheniformis A5 appear to be glutamine and alanine (apparent I0.5, 5.2mM). PMID:6169702

  15. Comparative study on production of α-Amylase from Bacillus licheniformis strains

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    Dibu Divakaran

    2011-12-01

    Full Text Available Alpha amylase (α-1, 4-glucan-glucanhydrolase, EC 3.2.1.1, an extracellular enzyme, degrades α, 1-4 glucosidic linkages of starch and related substrates in an endo-fashion producing oligosaccharides including maltose, glucose and alpha limit dextrin (7. The present study deals with the production and comparative study of production of α-amylase from two strains of Bacillus licheniformis, MTCC 2617 and 2618, by using four different substrates, starch, rice, wheat and ragi powder as carbon source by submerged fermentation. The effect of varying pH and incubation temperature, activator, inhibitor, and substrate concentration was investigated on the activity of α-amylase produced by MTCC strain 2618. The results shows that the production of the α-amylase by the B.licheniformis strain MTCC 2618, using four different substrates were found to be maximum (Starch 3.64 IU/ml/minutes, Rice powder 2.93 IU/ml/minutes, Wheat powder 2.67 IU/ml/minutes, Ragi powder 2.36 IU/ml/minutes on comparing the enzyme production of two strains. It was also observed that the maximum production was found on the 3rd day (i.e. 72 hr and characterization of crude enzyme revealed that optimum activity was at pH 7 and 37ºC.

  16. Two-step purification and partial characterization of an extra cellular α-amylase from Bacillus licheniformis

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    Zare Mirakabadi, A.

    2012-11-01

    Full Text Available The aim of this study was production and partial purification of α-amylase enzyme by Bacillus licheniformis. B. Licheniformis was allowed to grow in broth culture for purpose of inducing α-amylase enzyme. Optimal conditions for amylase production by B. Licheniformis are incubation period of 120 h, temperature of 37 °C and pH 7.0. The α-amylase enzyme was purified by ion exchange chromatography on DEAE-sepharose CL-6B and sephadex G-100 gel filtration with a 19.1-fold increase in specific activity as compared to the concentrated supernatant and with a specific activity of 926.47 U/mg. The α-amylase had the highest activity at pH 7.0 and 65 °C. According to the data on native polyacrylamide gel electrophoresis, the molecular weight of the purified enzyme was 72 kDa.

  17. Study of the solubility of a modified Bacillus licheniformis alpha-amylase around the isoelectric point

    DEFF Research Database (Denmark)

    Faber, Cornilius; Hobley, Timothy John; Mollerup, Jørgen;

    2007-01-01

    The solubility of a modified recombinant Bacillus licheniformis alpha-amylase (mBLA) has been studied by batch crystallization. A semi-pure preparation was chosen containing five isoforms with pI values from 6 to 7.3 (weighted average of 6.6). Small amounts (... sodium sulfate at all pH values and increased with 0.5 mol.L-1 sodium thiocyanate at pH 7 and pH 8. The effect of anions on alpha-amylase solubility followed the Hofmeister series, and only weak evidence of reversal was seen below the isoelectric point. Cations had little effect on solubility. The sign...... and magnitude of the alpha-amylase zeta potential was determined in the presence and absence of 0.1 mol.L-1 salt. Qualitatively, zeta potential correctly predicted the different salts influence on mBLA solubility....

  18. Improving the functional expression of a Bacillus licheniformis laccase by random and site-directed mutagenesis

    Directory of Open Access Journals (Sweden)

    Schmid Rolf D

    2009-02-01

    Full Text Available Abstract Background Laccases have huge potential for biotechnological applications due to their broad substrate spectrum and wide range of reactions they are able to catalyze. These include, for example, the formation and degradation of dimers, oligomers, polymers, and ring cleavage as well as oxidation of aromatic compounds. Potential applications of laccases include detoxification of industrial effluents, decolorization of textile dyes and the synthesis of natural products by, for instance, dimerization of phenolic acids. We have recently published a report on the cloning and characterization of a CotA Bacillus licheniformis laccase, an enzyme that catalyzes dimerization of phenolic acids. However, the broad application of this laccase is limited by its low expression level of 26 mg l-1 that was achieved in Escherichia coli. To counteract this shortcoming, random and site-directed mutagenesis have been combined in order to improve functional expression and activity of CotA. Results A CotA double mutant, K316N/D500G, was constructed by combining random and site-directed mutagenesis. It can be functionally expressed at an 11.4-fold higher level than the wild-type enzyme. In addition, it is able to convert ferulic acid much faster than the wild-type enzyme (21% vs. 14% and is far more efficient in decolorizing a range of industrial dyes. The investigation of the effects of the mutations K316N and D500G showed that amino acid at position 316 had a major influence on enzyme activity and position 500 had a major influence on the expression of the laccase. Conclusion The constructed double mutant K316N/D500G of the Bacillus licheniformis CotA laccase is an appropriate candidate for biotechnological applications due to its high expression level and high activity in dimerization of phenolic acids and decolorization of industrial dyes.

  19. Identification of a new Bacillus licheniformis strain producing a bacteriocin-like substance.

    Science.gov (United States)

    Guo, Yaoqi; Yu, Zhanqiao; Xie, Jianhua; Zhang, Rijun

    2012-06-01

    The emergence of antibiotic resistance has spurred a great number of studies for development of new antimicrobials in the past decade. The purpose of this study was to screen environmental samples for Bacillus strains producing potent antimicrobial agents. A new strain, which showed strong antimicrobial activity against Staphylococcus aureus and Salmonella enterica ser. Pullorum, was isolated from soil and designated as B116. This new isolate was identified as Bacillus licheniformis by morphological, biochemical and genetic analyses. The production of bacteriocin-like substance (BLS) started at early exponential phase and achieved highest level at early stationary phase. The BLS was precipitated by ammonium sulfate and its molecular mass was determined as ∼4 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Culture supernatant of the new isolate exhibited antimicrobial activity against both Gram-positive and Gram-negative bacteria, including Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Micrococcus luteus, Escherichia coli, and Salmonella spp. The BLS was resistant to heat, acid and alkaline treatment. Activity of the BLS was totally lost after digestion by pronase and partially lost after digestion by papain and lipase. The new isolate and relevant BLS are potentially useful in food and feed applications. PMID:22752909

  20. Production of the novel two-peptide lantibiotic lichenicidin by Bacillus licheniformis DSM 13.

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    Jasmin Dischinger

    Full Text Available BACKGROUND: Lantibiotics are small microbial peptide antibiotics that are characterized by the presence of the thioether amino acids lanthionine and methyllanthionine. Lantibiotics possess structural genes which encode inactive prepeptides. During maturation, the prepeptide undergoes posttranslational modifications including the introduction of rare amino acids as lanthionine and methyllanthione as well as the proteolytic removal of the leader. The structural gene (lanA as well as the other genes which are involved in lantibiotic modification (lanM, lanB, lanC, lanP, regulation (lanR, lanK, export (lanT(P and immunity (lanEFG are organized in biosynthetic gene clusters. METHODOLOGY/PRINCIPAL FINDINGS: Sequence comparisons in the NCBI database showed that Bacillus licheniformis DSM 13 harbours a putative lantibiotic gene cluster which comprises two structural genes (licA1, licA2 and two modification enzymes (licM1, licM2 in addition to 10 ORFs that show sequence similarities to proteins involved in lantibiotic production. A heat labile antimicrobial activity was detected in the culture supernatant and a heat stabile activity was present in the isopropanol cell wash extract of this strain. In agar well diffusion assays both fractions exhibited slightly different activity spectra against Gram-positive bacteria. In order to demonstrate the connection between the lantibiotic gene cluster and one of the antibacterial activities, two Bacillus licheniformis DSM 13 mutant strains harbouring insertions in the structural genes of the modification enzymes licM1 and licM2 were constructed. These strains were characterized by a loss of activity in the isopropanol extract and substractive MALDI-TOF predicted masses of 3020.6 Da and 3250.6 Da for the active peptides. CONCLUSIONS/SIGNIFICANCE: In conclusion, B. licheniformis DSM 13 produces an antimicrobial substance that represents the two-peptide lantibiotic lichenicidin and that shows activity against a wide

  1. Ability of a solid state fermentation technique to significantly minimize catabolic repression of. alpha. -amylase production by Bacillus licheniformis M27

    Energy Technology Data Exchange (ETDEWEB)

    Ramesh, M.V.; Lonsane, B.K. (Central Food Technological Research Inst., Mysore (India). Fermentation Technology and Bioengineering Discipline)

    1991-08-01

    The production of {alpha}-amylase by Bacillus licheniformis M27 in submerged fermentation was completely inhibited due to catabolic repression in medium containing 1% glucose. In contrast, the enzyme production in a solid state fermentation system was 19,550 units/ml extract even when the medium contained 15% glucose. The peak in enzyme titre was, however, shifted from 48 to 72 h. The ability of the solid state fermentation system to significantly overcome catabolic repression was not known earlier and is probably conferred by various physico-chemical factors and culture conditions specific to the system. (orig.).

  2. Production of Levan by Bacillus licheniformis for Use as a Soil Sealant in Earthen Manure Storage Structures

    Directory of Open Access Journals (Sweden)

    Abdel E. Ghaly

    2007-01-01

    Full Text Available Manure application is not permitted on frozen land in Canada and therefore, manure management and storage are the primary issues facing the agri-food industry. Low-cost, effective and environmentally safe earthen manure storage (EMS facilities will lower costs and help make the livestock industry more competitive and efficient. The goal of this study was to develop a biological sealing technology for earthen manure storages. The results showed that it is feasible to use a growing culture of Bacillus licheniformis to produce a non viscous water insoluble levan. Levan can only be produced by Bacillus licheniformis during the growth mode. No levan was produced during the death phase. About 0. 36 g of levan was produced per gram of sucrose which is 91. 1% of theoretical yield. The polymer can be used as a plugging agent to plug the pores of high permeability soils. From the biological and biochemical characteristics of the Bacillus licheniformis, it appears that the organism is capable of producing levan from sucrose under most field and soil conditions. As a soil organism, Bacillus licheniformis should be able to compete with most common soil species such as Arthrobacter and Bacillus. The bacteria could be grown either in the non-polysaccharide producing mode or in the polysaccharide producing mode. The first would permit distribution of the bacteria to the lower soil layers but would delay the production of the polysaccharide due to the lag period required to produce the enzyme (levansucrase. Upon production of levan, pore spaces would close and hence, the hydraulic conductivity would be substantially reduced.

  3. Beneficial Effect of Sugar Osmolytes on the Refolding of Guanidine Hydrochloride-Denatured Trehalose-6-phosphate Hydrolase from Bacillus licheniformis

    OpenAIRE

    Jiau-Hua Chen; Meng-Chun Chi; Min-Guan Lin; Long-Liu Lin; Tzu-Fan Wang

    2015-01-01

    The influence of three sugar osmolytes on the refolding of guanidine hydrochloride- (GdnHCl-) denatured trehalose-6-phosphate hydrolase of Bacillus licheniformis (BlTreA) was studied by circular dichroism (CD) spectra, fluorescence emission spectra, and the recovery of enzymatic activity. These experimental results clearly indicated that sorbitol, sucrose, and trehalose at a concentration of 0.75 M improved the refolding yields of GdnHCl-denatured  BlTreA, probably due to the fact that these ...

  4. Cloning and characterization of the glutamate dehydrogenase gene in Bacillus licheniformis

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The gdhA genes of IRC-3 GDH-strain and IRC-8 GDH+ strain were cloned,and they both successfully complemented the nutritional lesion of an E.coli glutamate auxotroph,Q100 GDH-.However,the gdhA gene from the mutant IRC-8 GDH+ strain failed to complement the glutamate deficiency of the wild type strain IRC-3.The gdhA genes of the wild type and mutant origin were sequenced separately.No nucleotide difference was detected between them.Further investigations indicated that the gdhA genes were actively expressed in both the wild type and the mutant.Additionally,no GDH inhibitor was found in the wild type strain IRC-3.It is thus proposed that the inactivity of GDH in wild type is the result of the deficiency at the post-translational level of the gdhA expression.Examination of the deduced amino acid sequence of Bacillus licheniformis GDH revealed the presence of the motifs characteristic of the familyⅠ-type hexameric protein,while the GDH of Bacillus subtilis belongs to family II.

  5. Cloning and characterization of the glutamate dehydrogenase gene in Bacillus licheniformis

    Institute of Scientific and Technical Information of China (English)

    朱冰; 俞冠翘; 朱家璧; 沈善炯

    2000-01-01

    The gdhA genes of IRC-3 GDH strain and IRC-8 GDH+ strain were cloned, and they both successfully complemented the nutritional lesion of an E. coli glutamate auxotroph, Q100 GDH". However, the gdhA gene from the mutant IRC-8 GDH+ strain failed to complement the glutamate deficiency of the wild type strain IRC-3. The gdhA genes of the wild type and mutant origin were sequenced separately. No nucleotide difference was detected between them. Further investigations indicated that the gdhA genes were actively expressed in both the wild type and the mutant. Additionally, no GDH inhibitor was found in the wild type strain IRC-3. It is thus proposed that the inactivity of GDH in wild type is the result of the deficiency at the post-translational level of the gdhA expression. Examination of the deduced amino acid sequence of Bacillus licheniformis GDH revealed the presence of the motifs characteristic of the family I -type hexameric protein, while the GDH of Bacillus subtilis belongs to family II.

  6. Anti-biofilm activity of an exopolysaccharide from a sponge-associated strain of Bacillus licheniformis

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    Cordone Angela

    2011-09-01

    Full Text Available Abstract Background Secondary metabolites ranging from furanone to exo-polysaccharides have been suggested to have anti-biofilm activity in various recent studies. Among these, Escherichia coli group II capsular polysaccharides were shown to inhibit biofilm formation of a wide range of organisms and more recently marine Vibrio sp. were found to secrete complex exopolysaccharides having the potential for broad-spectrum biofilm inhibition and disruption. Results In this study we report that a newly identified ca. 1800 kDa polysaccharide having simple monomeric units of α-D-galactopyranosyl-(1→2-glycerol-phosphate exerts an anti-biofilm activity against a number of both pathogenic and non-pathogenic strains without bactericidal effects. This polysaccharide was extracted from a Bacillus licheniformis strain associated with the marine organism Spongia officinalis. The mechanism of action of this compound is most likely independent from quorum sensing, as its structure is unrelated to any of the so far known quorum sensing molecules. In our experiments we also found that treatment of abiotic surfaces with our polysaccharide reduced the initial adhesion and biofilm development of strains such as Escherichia coli PHL628 and Pseudomonas fluorescens. Conclusion The polysaccharide isolated from sponge-associated B. licheniformis has several features that provide a tool for better exploration of novel anti-biofilm compounds. Inhibiting biofilm formation of a wide range of bacteria without affecting their growth appears to represent a special feature of the polysaccharide described in this report. Further research on such surface-active compounds might help developing new classes of anti-biofilm molecules with broad spectrum activity and more in general will allow exploring of new functions for bacterial polysaccharides in the environment.

  7. ABILITY OF BACTERIAL CONSORTIUM: Bacillus coagulans, Bacilus licheniformis, Bacillus pumilus, Bacillus subtilis, Nitrosomonas sp. and Pseudomonas putida IN BIOREMEDIATION OF WASTE WATER IN CISIRUNG WASTE WATER TREATMENT PLANT

    Directory of Open Access Journals (Sweden)

    Ratu SAFITRI

    2015-10-01

    Full Text Available This study was conducted in order to determine the ability of bacterial consortium: Bacillus coagulans, Bacilus licheniformis, Bacillus pumilus, Bacillus subtilis, Nitrosomonas sp., and Pseudomonas putida in bioremediation of wastewater origin Cisirung WWTP. This study uses an experimental method completely randomized design (CRD, which consists of two treatment factors (8x8 factorial design. The first factor is a consortium of bacteria (K, consisting of 8 level factors (k1, k2, k3, k4, k5, k6, k7, and k8. The second factor is the time (T, consisting of a 7 level factors (t0, t1, t2, t3, t4, t5, t6, and t7. Test parameters consist of BOD (Biochemical Oxygen Demand, COD (Chemical Oxygen Demand, TSS (Total Suspended Solid, Ammonia and Population of Microbes during bioremediation. Data were analyzed by ANOVA, followed by Duncan test. The results of this study showed that the consortium of Bacillus pumilus, Bacillus subtilis, Bacillus coagulans, Nitrosomonas sp., and Pseudomonas putida with inoculum concentration of 5% (k6 is a consortium of the most effective in reducing BOD 71.93%, 64.30% COD, TSS 94.85%, and 88.58% of ammonia.

  8. Isolation and physico-chemical characterization of an antifungal and antibacterial peptide produced by Bacillus licheniformis A12.

    Science.gov (United States)

    Gálvez, A; Maqueda, M; Martínez-Bueno, M; Lebbadi, M; Valdivia, E

    1993-07-01

    An antifungal substance named peptide A12-C has been purified to homogeneity from supernatants of sporulated cultures of Bacillus licheniformis A12. It consists of a 0.77-kDa hydrophilic peptide containing two residues of Glu and one of Arg, Ala, Pro, Tyr and Orn. No fatty acids, phosphorus or carbohydrates have been detected. Peptide A12-C is active on several fungi (Microsporum canis CECT 2797, Mucor mucedo CECT 2653, M. plumbeus (CCM F 443, Sporothrix schenckii CECT 2799 and Trichophyton mentagrophytes CECT 2793) and bacteria (Bacillus megaterium, Corynebacterium glutamicum, Sarcina and Mycobacterium), although the latter are less sensitive.

  9. Systematic mutagenesis method for enhanced production of bacitracin by Bacillus licheniformis mutant strain UV-MN-HN-6

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    Muhammad Nauman Aftab

    2012-03-01

    Full Text Available The purpose of the current study was intended to obtain the enhanced production of bacitracin by Bacillus licheniformis through random mutagenesis and optimization of various parameters. Several isolates of Bacillus licheniformis were isolated from local habitat and isolate designated as GP-35 produced maximum bacitracin production (14±0.72 IU ml-1. Bacitracin production of Bacillus licheniformis GP-35 was increased to 23±0.69 IU ml-1 after treatment with ultraviolet (UV radiations. Similarly, treatment of vegetative cells of GP-35 with chemicals like N-methyl N'-nitro N-nitroso guanidine (MNNG and Nitrous acid (HNO2 increased the bacitracin production to a level of 31±1.35 IU ml-1 and 27±0.89 IU ml-1 respectively. Treatment of isolate GP-35 with combined effect of UV and chemical treatment yield significantly higher titers of bacitracin with maximum bacitracin production of 41.6±0.92 IU ml-1. Production of bacitracin was further enhanced (59.1±1.35 IU ml-1 by optimization of different parameters like phosphate sources, organic acids as well as temperature and pH. An increase of 4.22 fold in the production of bacitracin after mutagenesis and optimization of various parameters was achieved in comparison to wild type. Mutant strain was highly stable and produced consistent yield of bacitracin even after 15 generations. On the basis of kinetic variables, notably Yp/s (IU/g substrate, Yp/x (IU/g cells, Yx/s (g/g, Yp/s, mutant strain B. licheniformis UV-MN-HN-6 was found to be a hyperproducer of bacitracin.

  10. A RETROSPECTIVE STUDY OF BOVINE ABORTIONS ASSOCIATED WITH BACILLUS-LICHENIFORMIS

    DEFF Research Database (Denmark)

    Agerholm, J.S.; Krogh, H.V.; Jensen, H.E.

    1995-01-01

    established due to the lack of sufficient materials, or the isolation of the bacterium was considered to be a result of contamination. In four cases concomitant infections with B, licheniformis and bovine virus diarrhoea virus were present. Abortions caused by B. licheniformis were predominantly seen during...... isolations, especially from the placenta, lungs, and abomasal contents, combined with the histological findings points to B, licheniformis abortions as being of haematogenous origin with subsequent transplacental spread to the fetus....

  11. Production and characterization of keratinase of a feather-degrading Bacillus licheniformis PWD-1.

    Science.gov (United States)

    Cheng, S W; Hu, H M; Shen, S W; Takagi, H; Asano, M; Tsai, Y C

    1995-12-01

    The keratinase produced by Bacillus licheniformis PWD-1 was induced by feather powder. Maximal enzyme production could be achieved by culturing in a medium containing 1% hammer-milled feather powder (100 mesh) at 45 degrees C for 30 h. Maximal growth of PWD-1 was achieved at 50 degrees C, and maximal enzyme induction was at 45 degrees C. The molecular mass and isoelectric point of this enzyme were 31.4 kDa and 8.5, respectively. This enzyme was stable from pH 5 to 12. The optimal reaction pHs for feather powder and casein were 8.5 and 10.5 to 11.5, respectively. The optimal reaction temperature was 50 degrees C to 55 degrees C. The relative activity of this enzyme toward casein, feather powder, keratin, elastin, and collagen was 100:52:41:18:7, and 100:56:32:3 for Suc-AAPL-pNA, Suc-AAPF-pNA, Suc-AAPM-pNA, and Suc-AAVA-pNA (Suc, succinyl; pNA, p-nitrophenylanilide).

  12. Extracellular polysaccharide production in Bacillus licheniformis SVD1 and its immunomodulatory effect

    Directory of Open Access Journals (Sweden)

    J. Susan van Dyk

    2012-11-01

    Full Text Available Bacillus licheniformis SVD1 exhibited highest production of three different polysaccharides when sucrose was used as the carbon source for polysaccharide production and yeast extract was used as the nitrogen source. Polysaccharides were characterized using size exclusion chromatography (SEC, thin layer chromatography (TLC, gas chromatography with mass spectrometry (GCMS, and Fourier Transform Infrared (FTIR analysis. Field emission scanning electron microscopy (FESEM and transmission electron microscopy (TEM were used to examine the topography of the cells and polysaccharides. The cell-associated polysaccharides were composed of galactose, while two different polysaccharides were present in the extracellular medium, one of 2,000 kDa (EPS1, consisting of fructose monomers and identified as a levan with (2→6-linkages and (1→2-branching linkages. The other extracellular polysaccharide (EPS2 consisted of mannose and galactose and had a range of sizes as identified through SEC. All three polysaccharides displayed an immune modulatory effect as measured using Interleukin 6 (IL6 and tumor necrosis factor alpha (TNFα.

  13. Enhanced production of alkaline protease by a mutant of Bacillus licheniformis N-2 for dehairing

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    Muhammad Nadeem

    2010-10-01

    Full Text Available The purpose of the present investigations was to improve the yield of alkaline protease for leather dehairing by subjecting the indigenous proteolytic strain Bacillus licheniformis N-2 to various mutagenic treatments viz. UV irradiations, NTG (N-methyl-N-nitro-N-nitrosoguinidine and MMS (methyl methane sulfonate. After screening on skim milk agar plates, a total of nine positive mutants were selected for shake flask experiments. Among these, the best proteolytic mutant designated as UV-9 showed 1.4 fold higher alkaline protease activity in preoptimized growth medium than the parent strain. The fermentation profile and kinetic parameters such u(h-1, Yp/s, Yp/x, Yx/s, q s, Qs, q p and Qp also indicated the superiority of the selected mutant UV-9 for alkaline protease production over the parent strain and rest of the mutants. The dehairing capability of mutant UV-9 alkaline protease was analyzed by soaking goat skin pieces for different time intervals (3-15 h at 40 º C. A complete dehairing without degradation of collagen was achieved after 12 h, indicating its commercial exploitation in leather industry.

  14. In Vitro Antibiofilm Activity of an Exopolysaccharide from the Marine Thermophilic Bacillus licheniformis T14.

    Science.gov (United States)

    Spanò, Antonio; Laganà, Pasqualina; Visalli, Giuseppa; Maugeri, Teresa L; Gugliandolo, Concetta

    2016-05-01

    The development of antibiofilm strategies is of major interest in contrasting bacterial pathogenic biofilms. A novel fructose and fucose rich exopolysaccharide (EPS1-T14) produced by the recently described thermophilic Bacillus licheniformis T14, isolated from a shallow hydrothermal vent of Panarea Island (Eolian Island, Italy), was evaluated for its effects on biofilm formation by multiresistant clinical strains of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. The antibiofilm activity of EPS1-T14 was assessed by microtiter plate assays and visualized by confocal laser scanning microscopic images. EPS1-T14, with molecular weight of 1000 kDa, reduced biofilm formation on abiotic surfaces without affecting bacterial vitality. The novel EPS1-T14 is a water-soluble, noncytotoxic exopolymer able to prevent biofilm formation and its use may represent a promising therapeutic strategy for combating bacterial biofilm-associated infections. EPS1-T14 as antiadhesive biomolecule could be useful for novel prospective in medical and nonmedical applications. PMID:26750122

  15. Functional characterization of a Na(+)-coupled dicarboxylate transporter from Bacillus licheniformis.

    Science.gov (United States)

    Strickler, Melodie A; Hall, Jason A; Gaiko, Olga; Pajor, Ana M

    2009-12-01

    The Na(+)-coupled dicarboxylate transporter, SdcL, from Bacillus licheniformis is a member of the divalent anion/Na(+) symporter (DASS) family that includes the bacterial Na(+)/dicarboxylate cotransporter SdcS (from Staphyloccocus aureus) and the mammalian Na(+)/dicarboxylate cotransporters, NaDC1 and NaDC3. The transport properties of SdcL produced in Escherichia coli are similar to those of its prokaryotic and eukaryotic counterparts, involving the Na(+)-dependent transport of dicarboxylates such as succinate or malate across the cytoplasmic membrane with a K(m) of approximately 6 microM. SdcL may also transport aspartate, alpha-ketoglutarate and oxaloacetate with low affinity. The cotransport of Na(+) and dicarboxylate by SdcL has an apparent stoichiometry of 2:1, and a K(0.5) for Na(+) of 0.9 mM. Our findings represent the characterization of another prokaryotic protein of the DASS family with transport properties similar to its eukaryotic counterparts, but with a broader substrate specificity than other prokaryotic DASS family members. The broader range of substrates carried by SdcL may provide insight into domains of the protein that allow a more flexible or larger substrate binding pocket.

  16. Statistical Approach for Optimization of Physiochemical Requirements on Alkaline Protease Production from Bacillus licheniformis NCIM 2042

    Directory of Open Access Journals (Sweden)

    Biswanath Bhunia

    2012-01-01

    Full Text Available The optimization of physiochemical parameters for alkaline protease production using Bacillus licheniformis NCIM 2042 were carried out by Plackett-Burman design and response surface methodology (RSM. The model was validated experimentally and the maximum protease production was found 315.28 U using optimum culture conditions. The protease was purified using ammonium sulphate (60% precipitation technique. The HPLC analysis of dialyzed sample showed that the retention time is 1.84 min with 73.5% purity. This enzyme retained more than 92% of its initial activity after preincubation for 30 min at 37∘C in the presence of 25% v/v DMSO, methanol, ethanol, ACN, 2-propanol, benzene, toluene, and hexane. In addition, partially purified enzyme showed remarkable stability for 60 min at room temperature, in the presence of anionic detergent (Tween-80 and Triton X-100, surfactant (SDS, bleaching agent (sodium perborate and hydrogen peroxide, and anti-redeposition agents (Na2CMC, Na2CO3. Purified enzyme containing 10% w/v PEG 4000 showed better thermal, surfactant, and local detergent stability.

  17. Covalent Immobilization of Bacillus licheniformis γ-Glutamyl Transpeptidase on Aldehyde-Functionalized Magnetic Nanoparticles

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    Meng-Chun Chi

    2013-02-01

    Full Text Available This work presents the synthesis and use of surface-modified iron oxide nanoparticles for the covalent immobilization of Bacillus licheniformis γ-glutamyl transpeptidase (BlGGT. Magnetic nanoparticles were prepared by an alkaline solution of divalent and trivalent iron ions, and they were subsequently treated with 3-aminopropyltriethoxysilane (APES to obtain the aminosilane-coated nanoparticles. The functional group on the particle surface and the amino group of BlGGT was then cross-linked using glutaraldehyde as the coupling reagent. The loading capacity of the prepared nanoparticles for BlGGT was 34.2 mg/g support, corresponding to 52.4% recovery of the initial activity. Monographs of transmission electron microscopy revealed that the synthesized nanoparticles had a mean diameter of 15.1 ± 3.7 nm, and the covalent cross-linking of the enzyme did not significantly change their particle size. Fourier transform infrared spectroscopy confirmed the immobilization of BlGGT on the magnetic nanoparticles. The chemical and kinetic behaviors of immobilized BlGGT are mostly consistent with those of the free enzyme. The immobilized enzyme could be recycled ten times with 36.2% retention of the initial activity and had a comparable stability respective to free enzyme during the storage period of 30 days. Collectively, the straightforward synthesis of aldehyde-functionalized nanoparticles and the efficiency of enzyme immobilization offer wide perspectives for the practical use of surface-bound BlGGT.

  18. Characterization and Application of Biosurfactant Produced by Bacillus licheniformis R2.

    Science.gov (United States)

    Joshi, Sanket J; Geetha, S J; Desai, Anjana J

    2015-09-01

    The biosurfactant produced by Bacillus licheniformis R2 was characterized and studied for enhancing the heavy crude oil recovery at 80 °C in coreflood experiments. The strain was found to be nonpathogenic and produced biosurfactant, reducing the surface tension of medium from 70 to 28 mN/m with 1.1 g/l yield. The biosurfactant was quite stable during exposure to elevated temperatures (85 °C for 90 days), high salinity (10 % NaCl), and a wide range of pH (5-12) for 10 days. It was characterized as lipopeptide similar to lichenysin-A, with a critical micelle concentration of about 19.4 mg/l. The efficiency of crude biosurfactant for enhanced oil recovery by core flood studies revealed it to recovering additional 37.1 % oil from Berea sandstone cores at 80 °C. The results are indicative of the potential for the development of lipopeptide biosurfactant-based ex situ microbial enhanced heavy oil recovery from depleting oil fields with extreme temperatures.

  19. Cloning, Sequencing, and In Silico Analysis of β-Propeller Phytase Bacillus licheniformis Strain PB-13

    Directory of Open Access Journals (Sweden)

    Vinod Kumar

    2014-01-01

    Full Text Available β-Propeller phytases (BPPhy are widely distributed in nature and play a major role in phytate-phosphorus cycling. In the present study, a BPPhy gene from Bacillus licheniformis strain was expressed in E. coli with a phytase activity of 1.15 U/mL and specific activity of 0.92 U/mg proteins. The expressed enzyme represented a full length ORF “PhyPB13” of 381 amino acid residues and differs by 3 residues from the closest similar existing BPPhy sequences. The PhyPB13 sequence was characterized in silico using various bioinformatic tools to better understand structural, functional, and evolutionary aspects of BPPhy class by multiple sequence alignment and homology search, phylogenetic tree construction, variation in biochemical features, and distribution of motifs and superfamilies. In all sequences, conserved sites were observed toward their N-terminus and C-terminus. Cysteine was not present in the sequence. Overall, three major clusters were observed in phylogenetic tree with variation in biophysical characteristics. A total of 10 motifs were reported with motif “1” observed in all 44 protein sequences and might be used for diversity and expression analysis of BPPhy enzymes. This study revealed important sequence features of BPPhy and pave a way for determining catalytic mechanism and selection of phytase with desirable characteristics.

  20. Efficient production of 2,3-butanediol from corn stover hydrolysate by using a thermophilic Bacillus licheniformis strain.

    Science.gov (United States)

    Li, Lixiang; Li, Kun; Wang, Kai; Chen, Chao; Gao, Chao; Ma, Cuiqing; Xu, Ping

    2014-10-01

    In this study, a thermophilic Bacillus licheniformis strain X10 was newly isolated for 2,3-butanediol (2,3-BD) production from lignocellulosic hydrolysate. Strain X10 could utilize glucose and xylose simultaneously without carbon catabolite repression. In addition, strain X10 possesses high tolerance to fermentation inhibitors including furfural, vanillin, formic acid, and acetic acid. In a fed-batch fermentation, 74.0g/L of 2,3-BD was obtained from corn stover hydrolysate, with a productivity of 2.1g/Lh and a yield of 94.6%. Thus, this thermophilic B. licheniformis strain is a candidate for the development of efficient industrial production of 2,3-BD from corn stover hydrolysate.

  1. Efficient production of 2,3-butanediol from corn stover hydrolysate by using a thermophilic Bacillus licheniformis strain.

    Science.gov (United States)

    Li, Lixiang; Li, Kun; Wang, Kai; Chen, Chao; Gao, Chao; Ma, Cuiqing; Xu, Ping

    2014-10-01

    In this study, a thermophilic Bacillus licheniformis strain X10 was newly isolated for 2,3-butanediol (2,3-BD) production from lignocellulosic hydrolysate. Strain X10 could utilize glucose and xylose simultaneously without carbon catabolite repression. In addition, strain X10 possesses high tolerance to fermentation inhibitors including furfural, vanillin, formic acid, and acetic acid. In a fed-batch fermentation, 74.0g/L of 2,3-BD was obtained from corn stover hydrolysate, with a productivity of 2.1g/Lh and a yield of 94.6%. Thus, this thermophilic B. licheniformis strain is a candidate for the development of efficient industrial production of 2,3-BD from corn stover hydrolysate. PMID:25151068

  2. Characterization of Biosurfactant Produced by Bacillus licheniformis TT42 Having Potential for Enhanced Oil Recovery.

    Science.gov (United States)

    Suthar, Harish; Nerurkar, Anuradha

    2016-09-01

    Bacillus licheniformis TT42 produced a low-molecular weight anionic biosurfactant that reduced the surface tension of water from 72 to 27 mN/m and the interfacial tension from 12 to 0.05 mN/m against crude oil. We have earlier reported significant enhancement in oil recovery in laboratory sand pack columns and core flood studies, by biosurfactant-TT42 compared to standard strain, Bacillus mojavensis JF2. In the context of this application of the biosurfactant-TT42, its characterization was deemed important. In the preliminary studies, the biosurfactant-TT42 was found to be functionally stable at under conditions of temperature, pH, and salinity generally prevalent in oil reservoirs. Furthermore, the purified biosurfactant-TT42 was found to have a CMC of 22 mg/l. A newly developed activity staining TLC method was used for the purification of biosurfactant-TT42. Structural characterization of biosurfactant-TT42 using TLC, Fourier transform infrared spectroscopy (FTIR), GC-MS, and matrix-assisted laser desorption ionization time of flight (MALDI-TOF)/TOF suggested that it was a mixture of lipopeptide species, all having a common hydrophilic cyclic heptapeptide head with the sequence, Gln-Leu/Ileu-Leu/Ileu-Val-Asp-Leu/Ileu-Leu/Ileu linked to hydrophobic tails of different lengths of 3β-OH-fatty acids bearing 1043, 1057 and 1071 Da molecular weight, where 3β-OH-C19 fatty acid was predominant. This is the longest chain length of fatty acids reported in a lipopeptide.

  3. Disruption of microbial biofilms by an extracellular protein isolated from epibiotic tropical marine strain of Bacillus licheniformis.

    Directory of Open Access Journals (Sweden)

    Devendra H Dusane

    Full Text Available BACKGROUND: Marine epibiotic bacteria produce bioactive compounds effective against microbial biofilms. The study examines antibiofilm ability of a protein obtained from a tropical marine strain of Bacillus licheniformis D1. METHODOLOGY/PRINCIPAL FINDINGS: B. licheniformis strain D1 isolated from the surface of green mussel, Perna viridis showed antimicrobial activity against pathogenic Candida albicans BH, Pseudomonas aeruginosa PAO1 and biofouling Bacillus pumilus TiO1 cultures. The antimicrobial activity was lost after treatment with trypsin and proteinase K. The protein was purified by ultrafiltration and size-exclusion chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF analysis revealed the antimicrobial agent to be a 14 kDa protein designated as BL-DZ1. The protein was stable at 75°C for 30 min and over a pH range of 3.0 to 11.0. The sequence alignment of the MALDI-fingerprint showed homology with the NCBI entry for a hypothetical protein (BL00275 derived from B. licheniformis ATCC 14580 with the accession number gi52082584. The protein showed minimum inhibitory concentration (MIC value of 1.6 µg/ml against C. albicans. Against both P. aeruginosa and B. pumilus the MIC was 3.12 µg/ml. The protein inhibited microbial growth, decreased biofilm formation and dispersed pre-formed biofilms of the representative cultures in polystyrene microtiter plates and on glass surfaces. CONCLUSION/SIGNIFICANCE: We isolated a protein from a tropical marine strain of B. licheniformis, assigned a function to the hypothetical protein entry in the NCBI database and described its application as a potential antibiofilm agent.

  4. Comparative evaluation of Bacillus licheniformis 5A5 and Aloe variegata milk-clotting enzymes

    Directory of Open Access Journals (Sweden)

    S. A. Ahmed

    2012-03-01

    Full Text Available The properties of a milk clotting enzyme (MCE produced by bacteria (Bacillus licheniformis 5A5 were investigated and compared to those of rennet extracted from a plant (Aloe variegata. Production of MCE by B. licheniformis 5A5 was better in static than in shaken cultures. Maximum activity (98.3 and 160.3 U/ml of clotting was obtained at 75ºC and 80ºC with bacterial and plant rennet, respectively. In the absence of substrate, the clotting activity of Aloe MCE was found to be less sensitive to heat inactivation up to 80ºC for 75 min, retaining 63.8% of its activity, while bacterial MCE was completely inhibited. CaCl2 stimulated milk clotting activity (MCA up to 2% and 1.5% for bacterial and plant enzymes. NaCl inhibited MCA for both enzymes, even at low concentration (1%. Plant MCE was more sensitive to NaCl at 3% concentration it retained 30.2% of its activity, whereas bacterial MCE retained 64.1%. Increasing skim milk concentration caused a significant increase in MCA up to 6% for both enzymes. Mn2+ stimulated the activity of bacterial and plant enzymes to 158.6 and 177.9%, respectively. EDTA and PMSF increased the activity of plant MCE by 34.4 and 41.1%, respectively, which is higher than those for the bacterial MCE (19.1 and 20.9%. Some natural materials activated MCE, the highest activation of bacterial MCE (128.1% was obtained in the presence of Fenugreek (with acid extraction. However Lupine Giza 1 (with neutral extraction gave the highest activation of plant MCE (137.9%. All extracts from Neem plant increased MCA at range from 105.6% to 136.4%. Plant MCE exhibited much better stability when stored at room temperature (25-30ºC for 30 days, retaining 51.2% of its activity. Bacterial MCE was highly stabile when stored under freezing (-18ºC, retaining 100% of its activity after 30 days. Moreover, bacterial MCE was highly tolerant to repeated freezing and thawing without loss of activity for 8 months.

  5. Codon Optimization Significantly Improves the Expression Level of α-Amylase Gene from Bacillus licheniformis in Pichia pastoris

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    Jian-Rong Wang

    2015-01-01

    Full Text Available α-Amylase as an important industrial enzyme has been widely used in starch processing, detergent, and paper industries. To improve expression efficiency of recombinant α-amylase from Bacillus licheniformis (B. licheniformis, the α-amylase gene from B. licheniformis was optimized according to the codon usage of Pichia pastoris (P. pastoris and expressed in P. pastoris. Totally, the codons encoding 305 amino acids were optimized in which a total of 328 nucleotides were changed and the G+C content was increased from 47.6 to 49.2%. The recombinants were cultured in 96-deep-well microplates and screened by a new plate assay method. Compared with the wild-type gene, the optimized gene is expressed at a significantly higher level in P. pastoris after methanol induction for 168 h in 5- and 50-L bioreactor with the maximum activity of 8100 and 11000 U/mL, which was 2.31- and 2.62-fold higher than that by wild-type gene. The improved expression level makes the enzyme a good candidate for α-amylase production in industrial use.

  6. Bacillus licheniformis Contains Two More PerR-Like Proteins in Addition to PerR, Fur, and Zur Orthologues.

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    Jung-Hoon Kim

    Full Text Available The ferric uptake regulator (Fur family proteins include sensors of Fe (Fur, Zn (Zur, and peroxide (PerR. Among Fur family proteins, Fur and Zur are ubiquitous in most prokaryotic organisms, whereas PerR exists mainly in Gram positive bacteria as a functional homologue of OxyR. Gram positive bacteria such as Bacillus subtilis, Listeria monocytogenes and Staphylococcus aureus encode three Fur family proteins: Fur, Zur, and PerR. In this study, we identified five Fur family proteins from B. licheniformis: two novel PerR-like proteins (BL00690 and BL00950 in addition to Fur (BL05249, Zur (BL03703, and PerR (BL00075 homologues. Our data indicate that all of the five B. licheniformis Fur homologues contain a structural Zn2+ site composed of four cysteine residues like many other Fur family proteins. Furthermore, we provide evidence that the PerR-like proteins (BL00690 and BL00950 as well as PerRBL (BL00075, but not FurBL (BL05249 and ZurBL (BL03703, can sense H2O2 by histidine oxidation with different sensitivity. We also show that PerR2 (BL00690 has a PerR-like repressor activity for PerR-regulated genes in vivo. Taken together, our results suggest that B. licheniformis contains three PerR subfamily proteins which can sense H2O2 by histidine oxidation not by cysteine oxidation, in addition to Fur and Zur.

  7. Optimization and partial characterization of culture conditions for the production of alkaline protease from Bacillus licheniformis P003

    OpenAIRE

    Sarker, Palash Kumar; Talukdar, Saimon Ahmad; Deb, Promita; Sayem, SM Abu; Mohsina, Kaniz

    2013-01-01

    Proteolytic enzymes have occupied a pivotal position for their practical applications. The present study was carried out under shake flask conditions for the production of alkaline protease from Bacillus licheniformis P003 in basal medium containing glucose, peptone, K2HPO4, MgSO4 and Na2CO3 at pH 10. The effect of culture conditions and medium components for maximum production of alkaline protease was investigated using one factor constant at a time method along with its characterization. Ma...

  8. Supplementation of Carbohydrate to Enhance the α-amylase Production by Bacillus licheniformis ATCC 6346 in Presence of Seed Cakes

    OpenAIRE

    Vengadaramana, A.; Vasanthy, A.; Balakumar, S

    2012-01-01

    Aims: The effect of carbohydrate and amino acids on the production of a-amylase by Bacillus licheniformis ATCC 6346 was investigated. Methodology and results: To find out the influence of carbohydrate the total carbohydrate content of the medium containing different concentration (2-18 g/L) of defatted seed cake powder of sesamum and mustard containing medium was kept constant by the addition of soluble starch separately. The highest a-amylase activity obtained in the medium containing 18g/L ...

  9. High expression level of levansucrase from Bacillus licheniformis RN-01 and synthesis of levan nanoparticles.

    Science.gov (United States)

    Nakapong, Santhana; Pichyangkura, Rath; Ito, Kazuo; Iizuka, Masaru; Pongsawasdi, Piamsook

    2013-03-01

    LsRN from Bacillus licheniformis was cloned and expressed in Escherichia coli. From a 1793 bp genomic sequence, the lsRN gene was found to be composed of a single 1446 bp ORF with a putative promoter consensus boxes and a ribosome-binding site. This ORF was predicted to encode for 482 amino acid residues. The LsRN was constitutively expressed at a relatively high level without sucrose induction. The enzyme was highly purified and an apparent size of 52 kDa with an optimum temperature and pH of 50 °C and 6.0 were determined. The wide range of M(w) of levan (1-600 kDa) was synthesized in a controlled reaction with two variable parameters: temperature and ionic strength. At high temperature (50 °C), LsRN synthesized high M(w) levan (612 kDa) as a major product while at low temperature (30 °C), low M(w) levan (11 kDa) was mainly synthesized. When 0.5M NaCl was added into the reaction, the major products at both temperatures were of the size 11 kDa. Moreover we report for the first time, an enzymatic synthesis of levan nanoparticles (NPs) by a single step reaction. The LsRN synthesized levan NPs as agglomerate with average particle size of 50 nm. The encapsulation of O-acetyl-α-tocopherol was carried out to demonstrate the applicable use of levan NPs. PMID:23219733

  10. Combined effects of dietary fructooligosaccharide and Bacillus licheniformis on innate immunity, antioxidant capability and disease resistance of triangular bream (Megalobrama terminalis).

    Science.gov (United States)

    Zhang, Chun-Nuan; Li, Xiang-Fei; Xu, Wei-Na; Jiang, Guang-Zhen; Lu, Kang-Le; Wang, Li-Na; Liu, Wen-Bin

    2013-11-01

    This study was conducted to investigate the effects of fructooligosaccharide (FOS) and Bacillus licheniformis (B. licheniformis) and their interaction on innate immunity, antioxidant capability and disease resistance of triangular bream Megalobrama terminalis (average initial weight 30.5 ± 0.5 g). Nine experimental diets were formulated to contain three FOS levels (0, 0.3% and 0.6%) and three B. licheniformis levels (0, 1 × 10(7), 5 × 10(7) CFU g(-1)) according to a 3 × 3 factorial design. At the end of the 8-week feeding trial, fish were challenged by Aeromonas hydrophila (A. hydrophila) and survival rate was recorded for the next 7 days. The results showed that leucocyte counts, alternative complement activity as well as total serum protein and globulin contents all increased significantly (P 0.05) was observed in these parameters in terms of dietary FOS levels. Both plasma alkaline phosphatase and phenoloxidase activities were significantly (P FOS levels with the highest values observed in fish fed 0.6 and 0.3% FOS, respectively. Both immunoglobulin M content and liver superoxide dismutase (SOD) activity were significantly affected (P > 0.05) by both FOS and B. licheniformis. Liver catalase, glutathione peroxidase as well as plasma SOD activities of fish fed 1 × 10(7) CFU g(-1)B. licheniformis were all significantly (P 0.05) by either FOS levels or B. licheniformis contents, whereas a significant (P FOS and 1 × 10(7) CFU g(-1)B. licheniformis. The results of this study indicated that dietary FOS and B. licheniformis could significantly enhance the innate immunity and antioxidant capability of triangular bream, as well as improve its disease resistance. The best combination of these two prebiotics and/or probiotics was 0.3% FOS and 1 × 10(7) CFU g(-1)B. licheniformis. PMID:23932988

  11. Microbial reduction of [Co(III)–EDTA]{sup −} by Bacillus licheniformis SPB-2 strain isolated from a solar salt pan

    Energy Technology Data Exchange (ETDEWEB)

    Paraneeiswaran, Arunachalam [Departartment of Biotechnology, Pondicherry University, Puducherry (India); Shukla, Sudhir K. [Biofouling and Biofilm Processes Section, Water and Steam Chemistry Division, BARC Facilities, Kalpakkam 603102 (India); Homi Bhabha National Institute, Mumbai 400094 (India); Prashanth, K. [Departartment of Biotechnology, Pondicherry University, Puducherry (India); Rao, T. Subba, E-mail: subbarao@igcar.gov.in [Biofouling and Biofilm Processes Section, Water and Steam Chemistry Division, BARC Facilities, Kalpakkam 603102 (India); Homi Bhabha National Institute, Mumbai 400094 (India)

    2015-02-11

    Graphical abstract: - Highlights: • Bacillus licheniformis SPB-2 was used in the bioremediation of [Co(III)–EDTA]{sup −}. • The bacterial biomass adsorbed the Co–EDTA complex after its reduction. • [Co(III)–EDTA]{sup −} complex showed Bacillus spore inducing property. • B. licheniformis SPB-2 showed significantly radio-tolerance (D{sub 10} = 250 Gy). - Abstract: Naturally stressed habitats are known to be repositories for novel microorganisms with potential bioremediation applications. In this study, we isolated a [Co(III)–EDTA]{sup −} reducing bacterium Bacillus licheniformis SPB-2 from a solar salt pan that is exposed to constant cycles of hydration and desiccation in nature. [Co(III)–EDTA]{sup −} generated during nuclear waste management process is difficult to remove from the waste due to its high stability and solubility. It is reduced form i.e. [Co(II)–EDTA]{sup 2−} is less stable though it is toxic. This study showed that B. licheniformis SPB-2 reduced 1 mM [Co(III)–EDTA]{sup −} in 14 days when grown in a batch mode. However, subsequent cycles showed an increase in the reduction activity, which was observed up to four cycles. Interestingly, the present study also showed that [Co(III)–EDTA]{sup −} acted as an inducer for B. licheniformis SPB-2 spore germination. Vegetative cells germinated from the spores were found to be involved in [Co(III)–EDTA]{sup −} reduction. More detailed investigations showed that after [Co(III)–EDTA]{sup −} reduction, i.e. [Co(II)–EDTA]{sup 2−} complex was removed by B. licheniformis SPB-2 from the bulk liquid by adsorption phenomenon. The bacterium showed a D{sub 10} value (radiation dose required to kill 90% cells) of ∼250 Gray (Gy), which signifies the potential use of B. licheniformis SPB-2 for bioremediation of moderately active nuclear waste.

  12. Enhanced production of elastase by Bacillus licheniformis ZJUEL31410: optimization of cultivation conditions using response surface methodology

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Sequential methodology based on the application of three types of experimental designs was used to optimize the fermentation conditions for elastase production from mutant strain ZJUEL31410 of Bacillus licheniformis in shaking flask cultures. The optimal cultivation conditions stimulating the maximal elastase production consist of 220 r/min shaking speed, 25 h fermentation time, 5% (v/v) inoculums volume, 25 ml medium volume in 250 ml Erlenmeyer flask and 18 h seed age. Under the optimized conditions, the predicted maximal elastase activity was 495 U/ml. The application of response surface methodology resulted in a significant enhancement in elastase production. The effects of other factors such as elastin and the growth factor (corn steep flour) on elastase production and cell growth were also investigated in the current study. The elastin had no significant effect on enzyme-improved production. It is still not clear whether the elastin plays a role as a nitrogen source or not. Corn steep flour was verified to be the best and required factor for elastase production and cell growth by Bacillus licheniformis ZJUEL31410.

  13. Draft Genome Sequence of Bacillus licheniformis CG-B52, a Highly Virulent Bacterium of Pacific White Shrimp (Litopenaeus vannamei), Isolated from a Colombian Caribbean Aquaculture Outbreak.

    Science.gov (United States)

    Gálvez, Eric J C; Carrillo-Castro, Katerine; Zárate, Lina; Güiza, Linda; Pieper, Dietmar H; García-Bonilla, Erika; Salazar, Marcela; Junca, Howard

    2016-01-01

    Bacillus licheniformis strain CG-B52 was isolated as the etiological agent producing a self-limited outbreak of high mortalities in commercial Litopenaeus vannamei culture ponds on the Colombian Caribbean coast in 2005. Here, we report its draft genome and three novel extrachromosomal elements that it harbors. PMID:27174263

  14. Cloning, expression, purification, crystallization and preliminary X-ray characterization of allantoinase from Bacillus licheniformis ATCC 14580.

    Science.gov (United States)

    Conejero-Muriel, Mayte; Martínez-Gómez, Ana Isabel; Martínez-Rodríguez, Sergio; Gavira, Jose A

    2014-11-01

    Allantoinase, a member of the amidohydrolase superfamily, exists in a wide variety of organisms, including bacteria, fungi, plants and a few animals, such as fishes and amphibians. Allantoinase catalyzes the reversible hydrolysis of allantoin into allantoate by hydrolytic cleavage of the N1-C2 amide bond of the five-membered hydantoin ring. Allantoinase from Bacillus licheniformis (AllBali) presents an inverted enantioselectivity towards allantoin (R-enantioselective), which is a distinguishable feature that is not observed for other allantoinases. In this work, B. licheniformis ATCC 14580 allantoinase (AllBali) containing a C-terminal His6 tag was overproduced in Escherichia coli and purified to homogeneity. Crystals of AllBali were obtained by the vapour-diffusion method using 0.1 M potassium thiocyanate, 20%(w/v) polyethylene glycol 3350 as a crystallization solution. X-ray diffraction data were collected to a resolution of 3.5 Å with an Rmerge of 29.2% from a crystal belonging to space group P12₁1, with unit-cell parameters a=54.93, b=164.74, c=106.89 Å, β=98.49°. There are four molecules in the asymmetric unit with a solvent content of 47% as estimated from the Matthews coefficient (VM=2.34 Å3 Da(-1)). PMID:25372819

  15. Characterization and dye decolorization ability of an alkaline resistant and organic solvents tolerant laccase from Bacillus licheniformis LS04.

    Science.gov (United States)

    Lu, Lei; Zhao, Min; Wang, Tian-Nv; Zhao, Li-Yan; Du, Mei-Hui; Li, Tai-Lun; Li, De-Bin

    2012-07-01

    A new bacterial strain exhibiting laccase activity was isolated from forest soil and was identified as Bacillus licheniformis LS04. The spore laccase of B. licheniformis LS04 demonstrated a broad pH range for catalyzing substrates. It was quite stable at high temperature and alkaline pH. There was no loss of laccase activity after 10 days incubation at pH 9.0, and about 16% of the initial activity was detected after 10h at 80°C. In addition, the spore laccase was tolerant towards 1M of NaCl and 30% of organic solvents. Reactive black 5, reactive blue 19 and indigo carmine were decolorized by the spore laccase in the absence of mediator. Meanwhile, the decolorization process was efficiently promoted when acetosyringone was present, with more than 80% of color removal in 1h at pH 6.6 or 9.0. The unusual properties indicated a high potential in industrial applications for this novel spore laccase. PMID:21868217

  16. Supplementation of Carbohydrate to Enhance the α-amylase Production by Bacillus licheniformis ATCC 6346 in Presence of Seed Cakes

    Directory of Open Access Journals (Sweden)

    Vengadaramana, A.

    2012-01-01

    Full Text Available Aims: The effect of carbohydrate and amino acids on the production of a-amylase by Bacillus licheniformis ATCC 6346 was investigated. Methodology and results: To find out the influence of carbohydrate the total carbohydrate content of the medium containing different concentration (2-18 g/L of defatted seed cake powder of sesamum and mustard containing medium was kept constant by the addition of soluble starch separately. The highest a-amylase activity obtained in the medium containing 18g/L mustard (59.11+b1.48 U/mL and sesamum seed cake powder (55.23+b1.55 U/mL. The results indicated that under these conditions the carbohydrate content had no effect on the production of a-amylase. Effect of amino acids (0.2g/L of glycine, methionine, proline, lysine, leucine, threonine, serine, arginine, alanine, glutamic acid, tryptophan, glutamine, asparagine, histidine, valine, phenylalanine, isoleucine and mixture of amino acids on the production of a-amylase in fermentation medium was investigated. Among the different amino acids supplemented, eight amino acids improved the a-amylase production but casaminoacids slightly inhibited the enzyme production. In presence of tryptophan highest enzyme activity was obtained than control. Conclusion, significance and impact of study: In these study amino acids especially tryptophan takes part in a particular role rather than carbohydrate in the production of a-amylase from B. licheniformis ATCC 6346.

  17. Purification and characterization of biosurfactant produced by Bacillus licheniformis Y-1 and its application in remediation of petroleum contaminated soil.

    Science.gov (United States)

    Liu, Boqun; Liu, Jinpeng; Ju, Meiting; Li, Xiaojing; Yu, Qilin

    2016-06-15

    In our previous research, a petroleum degrading bacteria strain Bacillus licheniformis Y-1 was obtained in Dagang Oilfield which had the capability of producing biosurfactant. This biosurfactant was isolated and purified in this work. The biosurfactant produced by strain Y-1 had the capability to decrease the surface tension of water from 74.66 to 27.26mN/m, with the critical micelle concentration (CMC) of 40mg/L. The biosurfactant performed not only excellent stabilities against pH, temperature and salinity, but also great emulsifying activities to different kinds of oil, especially the crude oil. According to the results of FT-IR spectrum and (1)H NMR spectrum detection, the surfactant was determined to be a cyclic lipopeptide. Furthermore, through the addition of surfactant, the effect of petroleum contaminated soil remediation by fungi got a significant improvement. PMID:27114088

  18. Utjecaj sastava podloge na komercijalnu proizvodnju alkalne proteaze s pomoću soja Bacillus licheniformis N-2

    OpenAIRE

    Nadeem, Muhammad; Qazi, Javed Iqbal; Baig, Shahjahan; Syed, Qurat-ul-Ain

    2008-01-01

    U radu je istražena proizvodnja proteaze s pomoću alkalofilnog soja bakterije Bacillus licheniformis N-2 u 50 mL podloge sastava (u g/L): glukoza 10,0; sojina sačma 10,0; K2HPO4 3,0; MgSO4·7H2O 0,5; NaCl 0,5 i CaCl2·2H2O 0,5; pH=10. Dodatkom raznih izvora ugljika i dušika u obliku finih praškastih organskih, anorganskih i nemasnih hranjiva, odabran je prikladan supstrat za proizvodnju alkalne proteaze. Najviše alkalne proteaze (677,64 U/mL) proizvedeno je u podlozi s glukozom, a nešto manje s...

  19. A New Diketopiperazine, Cyclo(D-trans-Hyp-L-Leu) from a Kenyan Bacterium Bacillus licheniformis LB 8CT.

    Science.gov (United States)

    Lee, Seoung Rak; Beemelmanns, Christine; Tsuma, Leah M M; Clardy, Jon; Cao, Shugeng; Kim, Ki Hyun

    2016-04-01

    Bacterially-produced small molecules demonstrate a wide range of structural and functional diversity. A new diketopiperazine, cyclo(D-trans-Hyp-L-Leu) (1), and five other known diketopiperazines (2-6), were isolated and purified from the fermented broth of a Kenyan bacterium Bacillus licheniformis LB 8CT. The structure of 1 was elucidated by a combination of extensive spectroscopic analyses, including 2D NMR and HR-MS, and the absolute configuration was determined by a combination of NOESY analysis and Marfey's method. The known compounds were identified as cyclo(D-cis-Hyp-L-Leu) (2), cyclo(D-cis-Hyp-L-Phe) (3), cyclo(D-Pro-L-Tyr) (4), cyclo-(D-Trp-L-Leu) (5), and cyclo(L-Tyr-Gly) (6) by comparison of their spectroscopic and physical data with reported values. Compounds 1-6 were tested for antifungal and antimicrobial properties.

  20. Induction of Drought Stress Resistance by Multi-Functional PGPR Bacillus licheniformis K11 in Pepper.

    Science.gov (United States)

    Lim, Jong-Hui; Kim, Sang-Dal

    2013-06-01

    Drought stress is one of the major yield affecting factor for pepper plant. The effects of PGPRs were analyzed in relation with drought resistance. The PGPRs inoculated pepper plants tolerate the drought stress and survived as compared to non-inoculated pepper plants that died after 15 days of drought stress. Variations in protein and RNA accumulation patterns of inoculated and non-inoculated pepper plants subjected to drought conditions for 10 days were confirmed by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and differential display PCR (DD-PCR), respectively. A total of six differentially expressed stress proteins were identified in the treated pepper plants by 2D-PAGE. Among the stress proteins, specific genes of Cadhn, VA, sHSP and CaPR-10 showed more than a 1.5-fold expressed in amount in B. licheniformis K11-treated drought pepper compared to untreated drought pepper. The changes in proteins and gene expression patterns were attributed to the B. licheniformis K11. Accordingly, auxin and ACC deaminase producing PGPR B. licheniformis K11 could reduce drought stress in drought affected regions without the need for overusing agrochemicals and chemical fertilizer. These results will contribute to the development of a microbial agent for organic farming by PGPR. PMID:25288947

  1. Induction of Drought Stress Resistance by Multi-Functional PGPR Bacillus licheniformis K11 in Pepper

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    Jong-Hui Lim

    2013-06-01

    Full Text Available Drought stress is one of the major yield affecting factor for pepper plant. The effects of PGPRs were analyzed in relation with drought resistance. The PGPRs inoculated pepper plants tolerate the drought stress and survived as compared to non-inoculated pepper plants that died after 15 days of drought stress. Variations in protein and RNA accumulation patterns of inoculated and non-inoculated pepper plants subjected to drought conditions for 10 days were confirmed by two dimensional polyacrylamide gel electrophoresis (2D-PAGE and differential display PCR (DD-PCR, respectively. A total of six differentially expressed stress proteins were identified in the treated pepper plants by 2D-PAGE. Among the stress proteins, specific genes of Cadhn, VA, sHSP and CaPR-10 showed more than a 1.5-fold expressed in amount in B. licheniformis K11-treated drought pepper compared to untreated drought pepper. The changes in proteins and gene expression patterns were attributed to the B. licheniformis K11. Accordingly, auxin and ACC deaminase producing PGPR B. licheniformis K11 could reduce drought stress in drought affected regions without the need for overusing agrochemicals and chemical fertilizer. These results will contribute to the development of a microbial agent for organic farming by PGPR.

  2. Bacillus licheniformis proteases as high value added products from fermentation of wastewater sludge: pre-treatment of sludge to increase the performance of the process.

    Science.gov (United States)

    Drouin, M; Lai, C K; Tyagi, R D; Surampalli, R Y

    2008-01-01

    Wastewater sludge is a complex raw material that can support growth and protease production by Bacillus licheniformis. In this study, sludge was treated by different thermo-alkaline pre-treatment methods and subjected to Bacillus licheniformis fermentation in bench scale fermentors under controlled conditions. Thermo-alkaline treatment was found to be an effective pre-treatment process in order to enhance the proteolytic activity. Among the different pre-treated sludges tested, a mixture of raw and hydrolysed sludge caused an increase of 15% in the protease activity, as compared to the untreated sludge. The benefit of hydrolysis has been attributed to a better oxygen transfer due to decrease in media viscosity and to an increase in nutrient availability. Foam formation was a major concern during fermentation with hydrolysed sludge. The studies showed that addition of a chemical anti-foaming agent (polypropylene glycol) during fermentation to control foam could negatively influence the protease production by increasing the viscosity of sludge.

  3. Purification and partial characterization of bacillocin 490, a novel bacteriocin produced by a thermophilic strain of Bacillus licheniformis

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    De Felice Maurilio

    2002-04-01

    Full Text Available Abstract Background Applications of bacteriocins as food preservatives have been so far limited, principally because of their low antimicrobial activity in foods. Nisin is the only bacteriocin of significant use, but applications are restricted principally because of its very low activity at neutral or alkaline pH. Thus the isolation of new bacteriocins active in foods is desirable. Results We isolated a Bacillus licheniformis thermophilic strain producing a bacteriocin with some novel features, named here bacillocin 490. This bacteriocin was inactivated by pronase E and proteinase K and was active against closely related Bacillus spp. both in aerobic and in anaerobic conditions. Bactericidal activity was kept during storage at 4°C and was remarkably stable in a wide pH range. The bacteriocin was partially purified by elution after adhesion to cells of the food-isolated strain Bacillus smithii and had a rather low mass (2 KDa. Antimicrobial activity against B. smithii was observed also when this organism was grown in water buffalo milk. Conclusions Bacillocin 490 is a novel candidate as a food anti-microbial agent since it displays its activity in milk, is stable to heat treatment and during storage, is active in a wide pH range and has bactericidal activity also at high temperature. These features may allow the use of bacillocin 490 during processes performed at high temperature and as a complementary antimicrobial agent of nisin against some Bacillus spp. in non-acidic foods. The small size suggests its use on solid foods.

  4. Comparative Study on Biochemical Properties and Antioxidative Activity of Cuttlefish (Sepia officinalis) Protein Hydrolysates Produced by Alcalase and Bacillus licheniformis NH1 Proteases

    OpenAIRE

    Balti, Rafik; Bougatef, Ali; El Hadj Ali, Nedra; Ktari, Naourez; Jellouli, Kemel; Nedjar-Arroume, Naima; Dhulster, Pascal; Nasri, Moncef

    2011-01-01

    Antioxidative activities and biochemical properties of protein hydrolysates prepared from cuttlefish (Sepia officinalis) using Alcalase 2.4 L and Bacillus licheniformis NH1 proteases with different degrees of hydrolysis (DH) were determined. For the biochemical properties, hydrolysis by both enzymes increased protein solubility to above 75% over a wide pH range. The antioxidant activities of cuttlefish protein hydrolysates (CPHs) increase with increasing DH. In addition, all CPHs exhibited an...

  5. Raw agro-industrial orange peel waste as a low cost effective inducer for alkaline polygalacturonase production from Bacillus licheniformis SHG10

    OpenAIRE

    Embaby, Amira M.; Masoud, Aliaa A; Marey, Heba S.; Shaban, Nadia Z; Ghonaim, Tayssir M

    2014-01-01

    The current study underlines biotechnological valorization of the accumulated and the non-efficiently utilized agro-industrial orange peel waste to produce polygalacturonase (PGase), an industrially important enzyme with augmented demands in enzymes markets, from Bacillus licheniformis SHG10. Sequential statistical optimization of PGase production was performed through one variable at a time (OVAT) approach, Plackett-Burman (PB) and response surface methodology (RSM). The impact of introducti...

  6. Expression of the Acid Pullulanase in Bacillus licheniformis%酸性普鲁兰酶基因在地衣芽孢杆菌中的表达

    Institute of Scientific and Technical Information of China (English)

    谢银珠; 沈微; 王正祥

    2011-01-01

    According to the sequence of pullulanase gene from Bacillus deramificans ( NCBI accession number:AX203843 ), the gene encoding mature peptide of pulluanase was synthesized and designated pulA. The puIA was amplified by the method of PCR and cloned into the expression vector pHY - WZX, yielding hybrid plasmid pHY-WZX- pulA. Subsequently, pHY- WZX -pulA was introduced into Bacillus licheniformis B60608. Active pullulanase was expressed by recombiant B. licheniformis and secreted into medium. Culture condition of recombinant B. licheniformis were optimized for production of pullulanase. The optimized medium consists of 2 % cotton seed protein and 8 % of glycerol.%根据Genbank公布的来源于Bacillus deramificans的普鲁兰酶基因突变体序列(AX203843)合成普鲁兰酶成熟肽基因.将该基因插入芽孢杆菌分泌型表达载体pHY-WZX,重组质粒转化地衣芽孢杆菌B60608,重组地衣芽孢杆菌实现普鲁兰酶分泌表达.对重组菌产普鲁兰酶的条件进行优化,以含2%药媒和8%甘油的培养基最适合普鲁兰酶表达.

  7. Cyclodextrin glycosyltransferase from Bacillus licheniformis: optimization of production and its properties Cyclodextrina glycosyltransferase de Bacillus licheniformis: otimização da produção e suas propriedades

    Directory of Open Access Journals (Sweden)

    Paulo Roberto Martins Bonilha

    2006-09-01

    Full Text Available Cyclodextrin glycosyltransferase (EC 2.4.1.19 is an enzyme that produces cyclodextrins from starch via an intramolecular transglycosylation reaction. An alkalophilic Bacillus strain, isolated from cassava peels, was identified as Bacillus licheniformis. CGTase production by this strain was better when potato starch was used as carbon source, followed by cassava starch and amylopectin. Glucose and amylose, on the other hand, acted as synthesis repressors. When the cultivation was supplemented with sodium ions and had the pH adjusted between 6.0 and 9.0, the microorganism maintained the growth and enzyme production capacity. This data is interesting because it contradicts the concept that alkalophilic microorganisms do not grow in this pH range. After ultrafiltration-centrifugation, one protein of 85.2 kDa with CGTase activity was isolated. This protein was identified in plates with starch and phenolphthalein. Determination of the optimum temperature showed higher activities at 25ºC and 55ºC, indicating the possible presence of more than one CGTase in the culture filtrate. Km and Vmax values were 1.77 mg/mL and 0.0263 U/mg protein, respectively, using potato starch as substrate.Ciclodextrina glicosiltransferase (EC 2.4.1.19 é uma enzima que produz ciclodextrinas a partir de amido via transglicosilação intramolecular. Uma cepa de Bacillus alcalofílico, isolada de cascas de mandioca, foi identificada como Bacillus licheniformis. A produção de CGTase por esta cepa foi melhor quando amido de batata foi utilizado como fonte de carbono, seguido por amido de mandioca e amilopectina. Glicose e amilose, por outro lado, atuaram como repressor de síntese desta enzima. Quando o cultivo foi suplementado com íons sódio e teve o pH ajustado entre 6,0 e 9,0, o microrganismo manteve a capacidade de crescimento e de produção da enzima. Este dado é interessante pois contraria o conceito de que microrganismos alcalofílicos não apresentam crescimento

  8. Purification and characterization of an alkaline protease from Bacillus licheniformis UV-9 for detergent formulations

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    Muhammad Nadeem

    2013-04-01

    Full Text Available Alkaline protease produced by mutant strain B. licheniformis UV-9 was purified and characterized for its exploitationin detergent formulation. The enzyme was purified to homogeneity by employing ammonium sulphate precipitation andsephadex G-100 gel filtration chromatography with a 36.83 fold increase in specific activity and 11% recovery. The molecularweight of the protease was found to be 36.12 kDa by SDS-PAGE. The Km and Vmax values exhibited by purified proteasewere 5 mg/ml and 61.58ìM/ml/min, respectively, using casein as substrate. The enzyme exhibited highest activity at pH 11 andtemperature 60°C. Stability studies showed that the enzyme retained higher than 80% residual activity in the pH and temperature ranges of 8 to 11 and 30 to 50°C, respectively. However, in the presence of 10 mM Ca2+ ions the enzyme tained morethan 90% of its residual activity at pH 11 and temperature 60°C. Phenyl methyl sulphonyl fluoride (PMSF completelyinhibited the enzyme activity suggesting that it was serine protease. Among metal ions, the Mg2+ and Ca2+ ions enhancedactivity up to 128% and 145%, respectively. The purified enzyme showed extreme stability towards various surfactantssuch as Tween-20, Tween- 45, Tween-65 and Triton X-45. In addition, the enzyme also exhibited more than 100% residualactivity in the presence of oxidizing agents, H2O2 and sodium perborate. These biochemical properties indicate the potentialuse of B. licheniformis UV-9 enzyme in laundry detergents.

  9. Extracellular Ribonuclease from Bacillus licheniformis (Balifase), a New Member of the N1/T1 RNase Superfamily

    Science.gov (United States)

    Nadyrova, Alsu; Ulyanova, Vera; Ilinskaya, Olga

    2016-01-01

    The N1/T1 RNase superfamily comprises enzymes with well-established antitumor effects, such as ribotoxins secreted by fungi, primarily by Aspergillus and Penicillium species, and bacterial RNase secreted by B. pumilus (binase) and B. amyloliquefaciens (barnase). RNase is regarded as an alternative to classical chemotherapeutic agents due to its selective cytotoxicity towards tumor cells. New RNase with a high degree of structural similarity with binase (73%) and barnase (74%) was isolated and purified from Bacillus licheniformis (balifase, calculated molecular weight 12421.9 Da, pI 8.91). The protein sample with enzymatic activity of 1.5 × 106 units/A280 was obtained. The physicochemical properties of balifase are similar to those of barnase. However, in terms of its gene organization and promoter activity, balifase is closer to binase. The unique feature of balifase gene organization consists in the fact that genes of RNase and its inhibitor are located in one operon. Similarly to biosynthesis of binase, balifase synthesis is induced under phosphate starvation; however, in contrast to binase, balifase does not form dimers under natural conditions. We propose that the highest stability of balifase among analyzed RNase types allows the protein to retain its structure without oligomerization. PMID:27656652

  10. Some Investigations on Protease Enzyme Production Kinetics Using Bacillus licheniformis BBRC 100053 and Effects of Inhibitors on Protease Activity

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    Zahra Ghobadi Nejad

    2014-01-01

    Full Text Available Due to great commercial application of protease, it is necessary to study kinetic characterization of this enzyme in order to improve design of enzymatic reactors. In this study, mathematical modeling of protease enzyme production kinetics which is derived from Bacillus licheniformis BBRC 100053 was studied (at 37°C, pH 10 after 73 h in stationary phase, and 150 rpm. The aim of the present paper was to determine the best kinetic model and kinetic parameters for production of protease and calculating Ki (inhibition constant of different inhibitors to find the most effective one. The kinetic parameters Km (Michaelis-Menten constant and Vm (maximum rate were calculated 0.626 mM and 0.0523 mM/min. According to the experimental results, using DFP (diisopropyl fluorophosphate and PMSF (phenylmethanesulfonyl fluoride as inhibitors almost 50% of the enzyme activity could be inhibited when their concentrations were 0.525 and 0.541 mM, respectively. Ki for DFP and PMSF were 0.46 and 0.56 mM, respectively. Kinetic analysis showed that the Lineweaver-Burk model was the best fitting model for protease production kinetics DFP was more effective than PMSF and both of them should be covered in the group of noncompetitive inhibitors.

  11. Levan-type fructooligosaccharide production using Bacillus licheniformis RN-01 levansucrase Y246S immobilized on chitosan beads

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    Surawut Sangmanee

    2016-06-01

    Full Text Available Bacillus licheniformis RN-01 levansucrase Y246S (LsRN-Y246S was immobilized by covalently linking onto chitosan, Sepabead EC-EP, and Sepabead EC-HFA, beads. The stability of immobilized LsRN-Y246S was found to be the highest with chitosan beads, retaining more than 70% activity after 13 weeks storage at 4 oC, and 68% activity after 12 hours incubation at 40°C. LsRN-Y246S immobilized on chitosan beads withstands sucrose concentrations up to 70% (w/v, retaining over 85% of its activity, significantly better than LsRN-Y246S immobilized on others supporting matrices. LsRN-Y246S immobilized on chitosan showed a 2.4 fold increase in activity in the presence of Mn2+, and gave slight protection against deactivation by of Cu2+, Zn2+, Fe3+, SDS and EDTA. A maximum of 8.36 g and an average of 7.35 g LFOS yield at least up to DP 11 can be produced from 25 g of sucrose, during five production cycles. We have demonstrated that LFOS can be effectively produced by chitosan immobilized LsRN-Y246S and purified.

  12. Overcoming hydrolysis of raw corn starch under industrial conditions with Bacillus licheniformis ATCC 9945a α-amylase.

    Science.gov (United States)

    Šokarda Slavić, Marinela; Pešić, Milja; Vujčić, Zoran; Božić, Nataša

    2016-03-01

    α-Amylase from Bacillus licheniformis ATCC 9945a (BliAmy) was proven to be very efficient in hydrolysis of granular starch below the temperature of gelatinization. By applying two-stage feeding strategy to achieve high-cell-density cultivation of Escherichia coli and extracellular production of BliAmy, total of 250.5 U/mL (i.e. 0.7 g/L) of enzyme was obtained. Thermostability of amylase was exploited to simplify purification. The hydrolysis of concentrated raw starch was optimized using response surface methodology. Regardless of raw starch concentration tested (20, 25, 30 %), BliAmy was very effective, achieving the final hydrolysis degree of 91 % for the hydrolysis of 30 % starch suspension after 24 h. The major A-type crystalline structure and amorphous domains of the starch granule were degraded at the same rates, while amylose-lipid complexes were not degraded. BliAmy presents interesting performances on highly concentrated solid starch and could be of value for starch-consuming industries while response surface methodology (RSM) could be efficiently applied for the optimization of the hydrolysis.

  13. Overcoming hydrolysis of raw corn starch under industrial conditions with Bacillus licheniformis ATCC 9945a α-amylase.

    Science.gov (United States)

    Šokarda Slavić, Marinela; Pešić, Milja; Vujčić, Zoran; Božić, Nataša

    2016-03-01

    α-Amylase from Bacillus licheniformis ATCC 9945a (BliAmy) was proven to be very efficient in hydrolysis of granular starch below the temperature of gelatinization. By applying two-stage feeding strategy to achieve high-cell-density cultivation of Escherichia coli and extracellular production of BliAmy, total of 250.5 U/mL (i.e. 0.7 g/L) of enzyme was obtained. Thermostability of amylase was exploited to simplify purification. The hydrolysis of concentrated raw starch was optimized using response surface methodology. Regardless of raw starch concentration tested (20, 25, 30 %), BliAmy was very effective, achieving the final hydrolysis degree of 91 % for the hydrolysis of 30 % starch suspension after 24 h. The major A-type crystalline structure and amorphous domains of the starch granule were degraded at the same rates, while amylose-lipid complexes were not degraded. BliAmy presents interesting performances on highly concentrated solid starch and could be of value for starch-consuming industries while response surface methodology (RSM) could be efficiently applied for the optimization of the hydrolysis. PMID:26545758

  14. Extracellular Ribonuclease from Bacillus licheniformis (Balifase, a New Member of the N1/T1 RNase Superfamily

    Directory of Open Access Journals (Sweden)

    Yulia Sokurenko

    2016-01-01

    Full Text Available The N1/T1 RNase superfamily comprises enzymes with well-established antitumor effects, such as ribotoxins secreted by fungi, primarily by Aspergillus and Penicillium species, and bacterial RNase secreted by B. pumilus (binase and B. amyloliquefaciens (barnase. RNase is regarded as an alternative to classical chemotherapeutic agents due to its selective cytotoxicity towards tumor cells. New RNase with a high degree of structural similarity with binase (73% and barnase (74% was isolated and purified from Bacillus licheniformis (balifase, calculated molecular weight 12421.9 Da, pI 8.91. The protein sample with enzymatic activity of 1.5 × 106 units/A280 was obtained. The physicochemical properties of balifase are similar to those of barnase. However, in terms of its gene organization and promoter activity, balifase is closer to binase. The unique feature of balifase gene organization consists in the fact that genes of RNase and its inhibitor are located in one operon. Similarly to biosynthesis of binase, balifase synthesis is induced under phosphate starvation; however, in contrast to binase, balifase does not form dimers under natural conditions. We propose that the highest stability of balifase among analyzed RNase types allows the protein to retain its structure without oligomerization.

  15. Effect of synbiotics between Bacillus licheniformis and yeast extract on growth, hematological and biochemical indices of the Nile tilapia (Oreochromis niloticus)

    OpenAIRE

    M.S. Hassaan; M.A. Soltan; M.M.R. Ghonemy

    2014-01-01

    Twelve practical diets were formulated to contain four levels of Bacillus licheniformis (0.0, 0.24 × 106, 0.48 × 106 and 0.96 × 106 CFU g−1), respectively, with three yeast extract levels (0%, 0.5% and 1%), respectively. Each diet was randomly assigned to duplicate groups of 50 Nile tilapia (Oreochromis niloticus) (5.99 ± 0.03 g) in 24 concrete ponds (0.5 m3 and 1.25 m depth) for 12 weeks. Increasing dietary B. licheniformis levels in O. niloticus and yeast extract levels significantly (P ...

  16. Production, Purification, and Characterization of Thermostable α-Amylase Produced by Bacillus licheniformis Isolate AI20

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    Yasser R. Abdel-Fattah

    2013-01-01

    Full Text Available An optimization strategy, based on statistical experimental design, is employed to enhance the production of thermostable α-amylase by a thermotolerant B. licheniformis AI20 isolate. Using one variant at time (OVAT method, starch, yeast extract, and CaCl2 were observed to influence the enzyme production significantly. Thereafter, the response surface methodology (RSM was adopted to acquire the best process conditions among the selected variables, where a three-level Box-Behnken design was employed to create a polynomial quadratic model correlating the relationship between the three variables and α-amylase activity. The optimal combination of the major constituents of media for α-amylase production was 1.0% starch, 0.75% yeast extract, and 0.02% CaCl2. The predicted optimum α-amylase activity was 384 U/mL/min, which is two folds more than the basal medium conditions. The produced α-amylase was purified through various chromatographic techniques. The estimated enzyme molecular mass was 55 kDa and the α-amylase had an optimal temperature and pH of 60–80°C and 6–7.5, respectively. Values of Vmax and Km for the purified enzyme were 454 mU/mg and 0.709 mg/mL. The α-amylase enzyme showed great stability against different solvents. Additionally, the enzyme activity was slightly inhibited by detergents, sodium dodecyl sulphate (SDS, or chelating agents such as EDTA and EGTA. On the other hand, great enzyme stability against different divalent metal ions was observed at 0.1 mM concentration, but 10 mM of Cu2+ or Zn2+ reduced the enzyme activity by 25 and 55%, respectively.

  17. Cloning, Expression, and Purification of Xylanase Gene from Bacillus licheniformis for Use in Saccharification of Plant Biomass.

    Science.gov (United States)

    Zafar, Asma; Aftab, Muhammad Nauman; Din, Zia Ud; Aftab, Saima; Iqbal, Irfana; Shahid, Anam; Tahir, Arifa; Haq, Ikram Ul

    2016-01-01

    The xylanase gene (xynA) of Bacillus licheniformis 9945A was cloned and expressed in Escherichia coli BL21(DE3) using pET-22b(+) as an expression vector. The recombinant xylanase enzyme was purified by ammonium sulfate precipitation, followed by single-step immobilized metal ion affinity chromatography with a 57.58-fold purification having 138.2 U/mg specific activity and recovery of 70.08 %. Molecular weight of the purified xylanase, 23 kDa, was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable for up to 70 °C with a broad pH range of 4-9 pH units. The enzyme activity was increased in the presence of metal ions especially Ca(+2) and decreased in the presence of EDTA, indicating that the xylanase was a metalloenzyme. However, an addition of 1-4 % Tween 80, β-mercaptoethanol, and DTT resulted in the increase of enzyme activity by 51, 52, and 5 %, respectively. Organic solvents with a concentration of 10-40 % slightly decreased the enzyme activity. The xylanase enzyme possesses the ability of bioconversion of plant biomasses like wheat straw, rice straw, and sugarcane bagasse. Among the different tested biomasses, the highest saccharification percentage was observed with 1 % sugarcane bagasse after 72 h of incubation at 50 °C with 20 units of enzyme. The results suggest that recombinant xylanase can be used in the bioconversion of natural biomasses into simple sugars which could be further used for the production of biofuel. PMID:26438315

  18. Effects of acetic acid and arginine on pH elevation and growth of Bacillus licheniformis in an acidified cucumber juice medium.

    Science.gov (United States)

    Yang, Zhenquan; Meng, Xia; Breidt, Frederick; Dean, Lisa L; Arritt, Fletcher M

    2015-04-01

    Bacillus licheniformis has been shown to cause pH elevation in tomato products having an initial pH below 4.6 and metabiotic effects that can lead to the growth of pathogenic bacteria. Because of this, the organism poses a potential risk to acidified vegetable products; however, little is known about the growth and metabolism of this organism in these products. To clarify the mechanisms of pH change and growth of B. licheniformis in vegetable broth under acidic conditions, a cucumber juice medium representative of a noninhibitory vegetable broth was used to monitor changes in pH, cell growth, and catabolism of sugars and amino acids. For initial pH values between pH 4.1 to 6.0, pH changes resulted from both fermentation of sugar (lowering pH) and ammonia production (raising pH). An initial pH elevation occurred, with starting pH values of pH 4.1 to 4.9 under both aerobic and anaerobic conditions, and was apparently mediated by the arginine deiminase reaction of B. licheniformis. This initial pH elevation was prevented if 5 mM or greater acetic acid was present in the brine at the same pH. In laboratory media, under favorable conditions for growth, data indicated that growth of the organism was inhibited at pH 4.6 with protonated acetic acid concentrations of 10 to 20 mM, corresponding to 25 to 50 mM total acetic acid; however, growth inhibition required greater than 300 mM citric acid (10-fold excess of the amount in processed tomato products) products under similar conditions. The data indicate that growth and pH increase by B. licheniformis may be inhibited by the acetic acid present in most commercial acidified vegetable products but not by the citric acid in many tomato products.

  19. DEGRADACIÓN DEL ALDRÍN POR Bacillus licheniformis, AISLADO DEL AGUA Y SEDIMENTO DE LA CIENAGA GRANDE DE SANTA MARTA

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    Sánchez Díaz Granados José Gregorio

    2012-04-01

    Full Text Available Con el objeto de apoyar la utilización de los microorganismos como alternativa para la degradación de contaminantes orgánicos persistentes, se aisló la bacteria Bacillus licheniformis, a partir de muestras de sedimento y agua del complejo lagunar de la Ciénaga Grande de santa Marta (CGSM, Caribe colombiano; capaz de tolerar y degradar en condiciones aerobias el plaguicida organoclorado aldrín. Se realizó un bioensayo en el que se expuso al B. licheniformis a una concentración de 60ng/L de aldrín, durante un período de 30 días se evaluó la capacidad degradadora de la bacteria sobre el organoclorado. La identificación y aislamiento de B. licheniformis, se realizó a través de caracterización macroscópica y microscópica y pruebas bioquímicas (sistema BBL Crystal y la determinación de las concentraciones de aldrín con la técnica de cromatografía de gases. Los resultaron mostraron que B. licheniformis posee capacidad degradadora de un 24% del aldrín y que los factores como la exposición a la luz solar y la volatilización influyen considerablemente en la degradación del organoclorado con una reducción adicional de 31%.

  20. Positions of Trp codons in the leader peptide-coding region of the at operon influence anti-trap synthesis and trp operon expression in Bacillus licheniformis.

    Science.gov (United States)

    Levitin, Anastasia; Yanofsky, Charles

    2010-03-01

    Tryptophan, phenylalanine, tyrosine, and several other metabolites are all synthesized from a common precursor, chorismic acid. Since tryptophan is a product of an energetically expensive biosynthetic pathway, bacteria have developed sensing mechanisms to downregulate synthesis of the enzymes of tryptophan formation when synthesis of the amino acid is not needed. In Bacillus subtilis and some other Gram-positive bacteria, trp operon expression is regulated by two proteins, TRAP (the tryptophan-activated RNA binding protein) and AT (the anti-TRAP protein). TRAP is activated by bound tryptophan, and AT synthesis is increased upon accumulation of uncharged tRNA(Trp). Tryptophan-activated TRAP binds to trp operon leader RNA, generating a terminator structure that promotes transcription termination. AT binds to tryptophan-activated TRAP, inhibiting its RNA binding ability. In B. subtilis, AT synthesis is upregulated both transcriptionally and translationally in response to the accumulation of uncharged tRNA(Trp). In this paper, we focus on explaining the differences in organization and regulatory functions of the at operon's leader peptide-coding region, rtpLP, of B. subtilis and Bacillus licheniformis. Our objective was to correlate the greater growth sensitivity of B. licheniformis to tryptophan starvation with the spacing of the three Trp codons in its at operon leader peptide-coding region. Our findings suggest that the Trp codon location in rtpLP of B. licheniformis is designed to allow a mild charged-tRNA(Trp) deficiency to expose the Shine-Dalgarno sequence and start codon for the AT protein, leading to increased AT synthesis. PMID:20061467

  1. Bacillus licheniformis proteases as high value added products from fermentation of wastewater sludge: pre-treatment of sludge to increase the performance of the process.

    Science.gov (United States)

    Drouin, M; Lai, C K; Tyagi, R D; Surampalli, R Y

    2008-01-01

    Wastewater sludge is a complex raw material that can support growth and protease production by Bacillus licheniformis. In this study, sludge was treated by different thermo-alkaline pre-treatment methods and subjected to Bacillus licheniformis fermentation in bench scale fermentors under controlled conditions. Thermo-alkaline treatment was found to be an effective pre-treatment process in order to enhance the proteolytic activity. Among the different pre-treated sludges tested, a mixture of raw and hydrolysed sludge caused an increase of 15% in the protease activity, as compared to the untreated sludge. The benefit of hydrolysis has been attributed to a better oxygen transfer due to decrease in media viscosity and to an increase in nutrient availability. Foam formation was a major concern during fermentation with hydrolysed sludge. The studies showed that addition of a chemical anti-foaming agent (polypropylene glycol) during fermentation to control foam could negatively influence the protease production by increasing the viscosity of sludge. PMID:18309222

  2. Production and estimation of alkaline protease by immobilized Bacillus licheniformis isolated from poultry farm soil of 24 Parganas and its reusability

    Directory of Open Access Journals (Sweden)

    Shamba Chatterjee

    2015-01-01

    Full Text Available Microbial alkaline protease has become an important industrial and commercial biotech product in the recent years and exerts major applications in food, textile, detergent, and pharmaceutical industries. By immobilization of microbes in different entrapment matrices, the enzyme produced can be more stable, pure, continuous, and can be reused which in turn modulates the enzyme production in an economical manner. There have been reports in support of calcium alginate and corn cab as excellent matrices for immobilization of Bacillus subtilis and Bacillus licheniformis, respectively. This study has been carried out using calcium alginate, κ-carrageenan, agar-agar, polyacrylamide gel, and gelatin which emphasizes not only on enzyme activity of immobilized whole cells by different entrapment matrices but also on their efficiency with respect to their reusability as first attempt. Gelatin was found to be the best matrix among all with highest enzyme activity (517 U/ml at 24 h incubation point and also showed efficiency when reused.

  3. Optimization and partial characterization of culture conditions for the production of alkaline protease from Bacillus licheniformis P003.

    Science.gov (United States)

    Sarker, Palash Kumar; Talukdar, Saimon Ahmad; Deb, Promita; Sayem, Sm Abu; Mohsina, Kaniz

    2013-01-01

    Proteolytic enzymes have occupied a pivotal position for their practical applications. The present study was carried out under shake flask conditions for the production of alkaline protease from Bacillus licheniformis P003 in basal medium containing glucose, peptone, K2HPO4, MgSO4 and Na2CO3 at pH 10. The effect of culture conditions and medium components for maximum production of alkaline protease was investigated using one factor constant at a time method along with its characterization. Maximum level of enzyme production was obtained after 48h of incubation with 2% inoculum size at 42°C, under continuous agitation at 150 rpm, in growth medium of pH 9. Highest enzyme production was obtained using 1% rice flour as carbon source and 0.8% beef extract as organic nitrogen source. Results indicated that single organic nitrogen source alone was more suitable than using in combinations and there was no significant positive effect of adding inorganic nitrogen sources in basal medium. After optimization of the parameters, enzyme production was increased about 20 fold than that of in basal medium. The crude enzyme was highly active at pH 10 and stable from pH 7-11. The enzyme showed highest activity (100%) at 50°C, and retained 78% relative activity at 70°C. Stability studies showed that the enzyme retained 75% of its initial activity after heating at 60°C for 1h. The enzyme retained about 66% and 46% of its initial activity after 28 days of storage at 4°C and room temperature (25°C) respectively. Mn(2+) and Mg(2+) increased the residual activity of the enzyme, whereas Fe(2+) moderately inhibited its residual activity. When pre-incubated with Tween-20, Tween-80, SDS and H2O2, each at 0.5% concentration, the enzyme showed increased residual activity. These characteristics may make the enzyme suitable for several industrial applications, especially in leather industries. PMID:24133650

  4. Effect of synbiotics between Bacillus licheniformis and yeast extract on growth, hematological and biochemical indices of the Nile tilapia (Oreochromis niloticus

    Directory of Open Access Journals (Sweden)

    M.S. Hassaan

    2014-01-01

    Full Text Available Twelve practical diets were formulated to contain four levels of Bacillus licheniformis (0.0, 0.24 × 106, 0.48 × 106 and 0.96 × 106 CFU g−1, respectively, with three yeast extract levels (0%, 0.5% and 1%, respectively. Each diet was randomly assigned to duplicate groups of 50 Nile tilapia (Oreochromis niloticus (5.99 ± 0.03 g in 24 concrete ponds (0.5 m3 and 1.25 m depth for 12 weeks. Increasing dietary B. licheniformis levels in O. niloticus and yeast extract levels significantly (P < 0.01 improved growth performance and nutrient utilization. Supplementation of the experimental diets with, 0.48 × 106 CFU/g−1 and 1.0% yeast extract showed the best nutrient utilization compared to other treatments. All probiotic levels significantly (P < 0.01 increased chemical composition (P < 0.05 compared to the control group, while increasing yeast extract did not significantly alter chemical composition. Hematological indices, total protein and albumin of O. niloticus significantly increased while aspartate aminotransferase and alanine aminotransferase significantly (P < 0.01 decreased with an increase in B. licheniformis level up to 0.48 × 106 CFU g−1. Increasing levels of yeast extract had no effect on hematological parameters and the diets supplemented with 0.48 × 106 CFU g−1 and 0.5% yeast extract showed the highest hematological values.

  5. Raw agro-industrial orange peel waste as a low cost effective inducer for alkaline polygalacturonase production from Bacillus licheniformis SHG10.

    Science.gov (United States)

    Embaby, Amira M; Masoud, Aliaa A; Marey, Heba S; Shaban, Nadia Z; Ghonaim, Tayssir M

    2014-01-01

    The current study underlines biotechnological valorization of the accumulated and the non-efficiently utilized agro-industrial orange peel waste to produce polygalacturonase (PGase), an industrially important enzyme with augmented demands in enzymes markets, from Bacillus licheniformis SHG10. Sequential statistical optimization of PGase production was performed through one variable at a time (OVAT) approach, Plackett-Burman (PB) and response surface methodology (RSM). The impact of introduction of six raw agro-industrial wastes (orange, lemon, banana, pomegranate, artichoke peel wastes and wheat bran) and other synthetic carbon sources separately into the fermentation broth on PGase productivity was studied through OVAT approach. Orange peel waste as sole raw carbon source in basal medium proved to be the best PGase inducer. It promoted PGase productivity with relative specific activity of 166% comparable with the effect imposed by synthetic citrus pectin as a reference inducer. Three key determinants (orange peel waste, pH of the production medium and incubation temperature) had RSM optimal levels of 1.76% (w/v), 8.0 and 37.8°C, respectively along with maximal PGase level (2.69 μg galacturonic acid. min(-1). mg(-1)) within 48 hrs. Moreover, SHG10 PGase exhibited activity over a wide range of pH (3-11) and an optimal activity at 50°C. Data greatly encourage pilot scale PGase production from B. licheniformis SHG10. PMID:25077057

  6. Evaluación de la nisina como sustancia inactivadora de Bacillus licheniformis en el extracto líquido de café

    Directory of Open Access Journals (Sweden)

    Leidy Sierra L.

    2013-10-01

    Full Text Available Objetivo. Evaluar el efecto de la nisina en la inactivación de Bacillus licheniformis en el extracto líquido de café. Materiales y métodos. Se evaluó la acción de la nisina sobre Bacillus licheniformis en extractos líquidos de café variando su concentración, tiempo de incubación, concentración de solidos solubles (grados Brix y la concentración bacteriana contaminante. Resultados. Se observó que la concentración de nisina, para obtener un efecto inhibitorio del 55%, sin alterar las propiedades fisicoquímicas y sensoriales del producto, es 500 UI/ml que corresponden a 12.5 mg/L. Además, se determinó que la concentración de nisina 1000 UI/ml puede actuar satisfactoriamente en poblaciones bacterianas menores de 5x104 UFC/ml en un período de 48 horas. Con relación al efecto de concentración de sólidos solubles en la inactivación del microorganismo, no se encontraron diferencias significativas para un rango entre 15 y 45°Brix. Conclusiones. A partir de este estudio se puede concluir que la nisina puede ser usada como preservante del extracto de líquido de café sin afectar sensorialmente el producto, teniendo en cuenta la concentración bacteriana contaminante y el tiempo de incubación.

  7. Optimization of Culture Medium and Fermentation Conditions of Bacillus licheniformis M109%地衣芽孢杆菌M109高密度发酵条件的优化

    Institute of Scientific and Technical Information of China (English)

    付维来; 杜建涛; 刘鹏; 王安如

    2012-01-01

    Optimization of Bacillus licheniformis M109 culture medium in this research based on the orthogonal test, carbon source, nitrogen source and carbon nitrogen ratio were determine as necessary for fermentation medium of Bacillus licheniformis M109. In order to enhance Bacillus licheniformis M109 fermentation level, growth curve of Bacillus licheniformis M109 and trend of Ph, dissolved oxygen(DO) were studied. Though studying the fermentation process of growth, the optimal Ph and inoculation amount were determined. The results found that DO was a key factor that limited the growth of Bacillus licheniformis M109. It was studied regulation rotate speed of stirring to improve dissolved oxygen and the fermentation density in culture medium. Another study found that Bacillus licheniformis M109 fermentation level could be improved by adding fresh medium. Through optimization of culture medium and conditions, the fermentation level of Bacillus licheniformis M109 incneased to 1. 2×1010 CFU/Ml from the initial 1. 0×109 CFU/Ml and the rate of bacillus arrived to 88%.%试验旨在筛选一株地衣芽孢杆菌M109(Bacillus licheniformis M109)进行培养基和发酵条件的优化.采用单因素试验方法筛选出最佳碳源和氮源的培养基,并利用正交试验方法确定其最佳的碳氮比.为了继续提高地衣芽孢杆菌M109的发酵水平,研究其在发酵过程中的生长曲线、pH和溶氧(DO)水平的变化曲线,通过对发酵过程中生长参数的测定,优化了地衣芽孢杆菌M109最佳的接种量和最适pH.结果发现,溶氧限制是地衣芽孢杆菌M109生长的关键因素;在限定通风量的条件下,通过调节搅拌转速的方法来提高培养基中的溶氧水平,提高发酵密度;通过流加培养基的方法也能提高地衣芽孢杆菌M109的发酵水平.因此,通过对培养基和发酵条件的优化,使地衣芽孢杆菌M109的发酵水平由最初的1.0×109 CFU/mL提高到1.2×1010CFU/mL,芽孢形成率为88%.

  8. Anticorrosion/antifouling properties of bacterial spore-loaded sol-gel type coating for mild steel in saline marine condition: a case of thermophilic strain of Bacillus licheniformis

    OpenAIRE

    Eduok, Ubong; Suleiman, Rami; Gittens, Jeanette; Khaled, Mazen; Smith, Thomas J.; Akid, Robert; El Ali, Bassam; Khalil, Amjad

    2015-01-01

    This work reports the performance of a sol-gel type coating encapsulated with biofilm of inoculums of protective thermophilic strain of Bacillus licheniformis endospores isolated from the Gazan hot springs- Saudi Arabia for the inhibition of marine fouling and corrosion protection of S36-grade mild steel in 3.5 wt% NaCl medium. In order to improve its anticorrosion properties, the hybrid sol-gel coating is further doped with zinc molybdate (MOLY) and zinc aluminum polyphosphate (Z...

  9. 地衣芽孢杆菌1.934培养条件的优化%Optimization of Culture Condition for Bacillus licheniformis 1.934 Production

    Institute of Scientific and Technical Information of China (English)

    于淑玉; 张光明; 万甜

    2012-01-01

    Bacillus licheniformis is very important as a soil microorganism to increase the availability and uptake of mineral nutrients for plants. In this study, the effect of culture condition on Bacillus licheniformis 1. 934 production was investigated. On the basis of single factor experiments, a central composite design was used to optimize the prime factors. The results were analyzed by response surface. Analysis results show that the optimal culture condition is as follows; temperature 35℃,flask shaking speed 150r/min,amount of inoculation 3. 6%,pH 7. 5. Bacillus licheniformis 1. 934 grew well under this condition, incubated for 20h,the highest amylase activity(12. 7U /mL ) can be obtained;incubated for 27h,the highest cell number can be gotten.%研究了生物肥功能菌——地衣芽胞杆菌1.934 (Bacillus licheniformis)培养条件对菌体生长量的影响.采用了单因素实验和响应曲面法(RSM)设计实验和分析数据.获得了菌体摇瓶培养的最适条件:培养温度35℃,转速为150r/min,接种量为3.6%,pH为7.5.地衣芽孢杆菌在最适条件下培养约20h,淀粉酶的酶活最高,活力可达12.7U/mL培养液.培养约27h得到最大的菌体收益.

  10. Characterization of a salt-tolerant aminopeptidase from marine Bacillus licheniformis SWJS33 that improves hydrolysis and debittering efficiency for soy protein isolate.

    Science.gov (United States)

    Lei, Fenfen; Zhao, Qiangzhong; Sun-Waterhouse, Dongxiao; Zhao, Mouming

    2017-01-01

    An aminopeptidase was isolated from the marine Bacillus licheniformis SWJS33 (BLAP) and purified. According to the tandem mass spectrometry, the enzyme displayed 11% amino acid identity with the aminopeptidase from Bacillus (gi|496687392). BLAP exhibited maximum activity at 60°C and pH 8.0-8.5 and had a molecular mass of 100kDa. The presence of NaCl enabled 50% improvement of enzyme activity with 10-15% NaCl being the best. The observed inactivation by EDTA and bestatin and activation by Co(2+) and Ag(+) indicated that the obtained enzyme was a metalloaminopeptidase. Such an aminopeptidase could further improve the hydrolysis degree of soy protein isolate hydrolysates catalyzed by papain, Alcalase 2.4L or Flavourzyme 500MG from 8.5%, 9.5% or 14.4-18.8%, 18.7% or 20.1%, respectively, while decreasing the bitter intensity score of the SPI hydrolysates catalyzed by Alcalase 2.4L from 3.6 to 0.4. PMID:27507484

  11. Study of HMG-CoA Reductase Inhibition Activity of the Hydrolyzed Product of Snakehead Fish (Channa striata) Skin Collagen with 50 kDa Collagenase from Bacillus licheniformis F11.4.

    Science.gov (United States)

    Virginia, Agnes; Rachmawati, Heni; Riani, Catur; Retnoningrum, Debbie S

    2016-01-01

    Bioactive peptides produced from enzymatic hydrolysis fibrous protein have been proven to have several biological activities. Previous study showed that the hydrolysis product of snakehead fish skin collagen with 26 kDa collagenase from Bacillus licheniformis F11.4 showed HMG-CoA (HMGR) inhibition activity. The aim of this research was to determine the ability of the hydrolysis product produced from snakehead fish skin collagen hydrolysed by 50 kDa collagenase from B. licheniformis F11.4 in inhibiting HMGR activity. Snakehead fish skin collagen was extracted using an acid method and collagenase was produced from B. licheniformis F11.4 using half-strength Luria Bertani (LB) medium containing 5% collagen. Crude collagenase was concentrated and fractionated using the DEAE Sephadex A-25 column eluted with increasing gradient concentrations of NaCl. Collagen, collagenase, and fractions were analyzed using SDS-PAGE and collagenolytic activity was analyzed by the zymography method. Collagenase with 50 kDa molecular weight presented in fraction one was used to hydrolyze the collagen. The reaction was done in 18 hours at 50°C. The hydrolysis product using 3.51 μg collagen and 9 ng collagenase showed 25.8% inhibition activity against pravastatin. This work shows for the first time that the hydrolysis product of snakehead fish skin collagen and 50 kDa collagenase from B. licheniformis F11.4 has potential as an anticholesterol agent. PMID:27110500

  12. Study of HMG-CoA Reductase Inhibition Activity of the Hydrolyzed Product of Snakehead Fish (Channa striata) Skin Collagen with 50 kDa Collagenase from Bacillus licheniformis F11.4.

    Science.gov (United States)

    Virginia, Agnes; Rachmawati, Heni; Riani, Catur; Retnoningrum, Debbie S

    2016-01-01

    Bioactive peptides produced from enzymatic hydrolysis fibrous protein have been proven to have several biological activities. Previous study showed that the hydrolysis product of snakehead fish skin collagen with 26 kDa collagenase from Bacillus licheniformis F11.4 showed HMG-CoA (HMGR) inhibition activity. The aim of this research was to determine the ability of the hydrolysis product produced from snakehead fish skin collagen hydrolysed by 50 kDa collagenase from B. licheniformis F11.4 in inhibiting HMGR activity. Snakehead fish skin collagen was extracted using an acid method and collagenase was produced from B. licheniformis F11.4 using half-strength Luria Bertani (LB) medium containing 5% collagen. Crude collagenase was concentrated and fractionated using the DEAE Sephadex A-25 column eluted with increasing gradient concentrations of NaCl. Collagen, collagenase, and fractions were analyzed using SDS-PAGE and collagenolytic activity was analyzed by the zymography method. Collagenase with 50 kDa molecular weight presented in fraction one was used to hydrolyze the collagen. The reaction was done in 18 hours at 50°C. The hydrolysis product using 3.51 μg collagen and 9 ng collagenase showed 25.8% inhibition activity against pravastatin. This work shows for the first time that the hydrolysis product of snakehead fish skin collagen and 50 kDa collagenase from B. licheniformis F11.4 has potential as an anticholesterol agent.

  13. Solid State Fermentation of a Raw Starch Digesting Alkaline Alpha-Amylase from Bacillus licheniformis RT7PE1 and Its Characteristics

    Directory of Open Access Journals (Sweden)

    Romana Tabassum

    2014-01-01

    Full Text Available The thermodynamic and kinetic properties of solids state raw starch digesting alpha amylase from newly isolated Bacillus licheniformis RT7PE1 strain were studied. The kinetic values Qp, Yp/s, Yp/X, and qp were proved to be best with 15% wheat bran. The molecular weight of purified enzyme was 112 kDa. The apparent Km and Vmax values for starch were 3.4 mg mL−1 and 19.5 IU mg−1 protein, respectively. The optimum temperature and pH for α-amylase were 55°C, 9.8. The half-life of enzyme at 95°C was 17h. The activation and denaturation activation energies were 45.2 and 41.2 kJ mol−1, respectively. Both enthalpies (ΔH∗ and entropies of activation (ΔS∗ for denaturation of α-amylase were lower than those reported for other thermostable α-amylases.

  14. Comparative Study on Biochemical Properties and Antioxidative Activity of Cuttlefish (Sepia officinalis Protein Hydrolysates Produced by Alcalase and Bacillus licheniformis NH1 Proteases

    Directory of Open Access Journals (Sweden)

    Rafik Balti

    2011-01-01

    Full Text Available Antioxidative activities and biochemical properties of protein hydrolysates prepared from cuttlefish (Sepia officinalis using Alcalase 2.4 L and Bacillus licheniformis NH1 proteases with different degrees of hydrolysis (DH were determined. For the biochemical properties, hydrolysis by both enzymes increased protein solubility to above 75% over a wide pH range. The antioxidant activities of cuttlefish protein hydrolysates (CPHs increase with increasing DH. In addition, all CPHs exhibited antioxidative activity in a concentration-dependent manner. NH1-CPHs generally showed greater antioxidative activity than Alcalase protein hydrolysates (P<0.05 as indicated by the higher 1,1-diphenyl-1-picryhydrazyl (DPPH radical scavenging activity and ferrous chelating activity. Both Alcalase and NH1 protein hydrolysates were able to retard lipid peroxidation and β-carotene-linoleic acid oxidation. Alcalase-CPH (DH = 12.5% and NH1-CPH (DH = 15% contained 75.36% and 80.11% protein, respectively, with histidine and arginine as the major amino acids, followed by glutamic acid/glutamine, serine, lysine, and leucine. In addition, CPHs have a high percentage of essential amino acids made up 48.85% and 50.04%. Cuttlefish muscle protein hydrolysates had a high nutritional value and could be used as supplement to poorly balanced dietary proteins.

  15. Isolation, Characterization and investing the Industrial Applications of Thermostable and Solvent Tolerant Serine Protease from Hot Spring Isolated Thermophililic Bacillus licheniformis U1

    Directory of Open Access Journals (Sweden)

    Pravin Dudhagara

    2014-03-01

    Full Text Available Protease is the largest selling enzyme in the world due to its various applications in the making of detergent, food and leather, meat tenderisation and pharmaceutical industries. The aim of the study is to isolate and identify thermophilic Bacillus licheniformis U1 strains for thermostable protease production. The partial purified enzyme was characterized under different conditions using Anson-Hagihara’s method. Casein as a substrate in the concentration of 0.6 % w/v optimum for enzyme activity and tolerant up to 2.0% casein concentration. An optimum enzyme activity was reported at pH 7 and decreased with increasing in pH, while temperature optimum was found at 50 °C. The enzyme was stable at 40 °C to 50 °C for half an hour and nearly 50% residual activity was indicated at 60 °C. NaCl was not required for catalysis. Stability of enzymes in the presence of various organic solvents and different detergents was remarkable. The enzyme was stable up to 3 days into various solvents and slowly denatured with prolonged incubation. The result of the washing performance with detergent was clearly indicated. Moreover the removal of blood stains and dehairing in goat skin suggests the crucial application in the commercial production at large scale.

  16. 根际接种芽孢杆菌对辣椒和黄瓜壮苗形成的作用%Effects of Rhizosphere Inoculation with Bacillus licheniformis and Bacillus polymyxa on Pepper and Cucumber Seedling Development

    Institute of Scientific and Technical Information of China (English)

    常冬梅; 张志刚; 尚庆茂

    2010-01-01

    将地衣芽孢杆菌(Bacillus licheniformis)、多粘芽孢杆菌(Bacillus polymyxa)悬浮液注射接种至辣椒、黄瓜幼苗根际,分析对幼苗生长发育及其相关生理指标的影响.结果表明:辣椒苗期根际接种芽孢杆菌后,显著提高了幼苗净光合速率和根系活力,促进了矿质元素的吸收积累,增产幅度29.3%~33.3%,多粘芽孢杆菌的接种效果明显优于地衣芽孢杆菌;黄瓜幼苗根际接种芽孢杆菌后,幼苗根系活力降低,进而抑制了矿质元素吸收和产量形成.

  17. Construcción de un vector para la integración cromosomal de un gen de fitasa de Bacillus licheniformis

    Directory of Open Access Journals (Sweden)

    Maria Teresa Fernández

    2011-07-01

    Full Text Available Las fitasas son una clase especial de fosfatasas que catalizan la hidrólisis secuencial del fitato. La incapacidad de las plantas para utilizar el fósforo a partir de los fitatos del suelo es debido a la baja actividad de fitasas en sus raíces. Los microorganismos del suelo juegan un importante papel en los procesos que afectan la trans- formación de los compuestos fosforados. Muchos de ellos pueden solubilizar el fósforo a partir de los fitatos, mediante la liberación de fitasas. Este proceso permite la movilización del fósforo hacia las plantas y un mejor aprovechamiento de este nutriente. Sin embargo, muchas bacterias carecen de los genes que codifican para estas enzimas, lo que disminuye la disponibilidad de este elemento en el suelo. Una alternativa es mejorar las rizobacterias en cuanto a su capacidad de solubilizar los fitatos del suelo, mediante la transformación genética. En este trabajo el gen phyL de Bacillus licheniformis fue clonado en el vector de liberación suicida pJMT6 (vector derivado del sistema pUT/mini Tn5. La construcción recombinante que contiene un marcador de selección no antibiótico, fue transformada en Escherichia coli CC118λpir. Un clon transformante (F16 fue seleccionado y posteriormente caracterizado. Estos resultados constituyen un primer paso para desarrollar rizobacterias promotoras del crecimiento mejoradas en cuanto a la producción de actividad fitasa recombinante, como alternativa para reducir la contaminación ambiental y mejorar la productividad de los cultivos.

  18. GH53 Endo-Beta-1,4-Galactanase from a Newly Isolated Bacillus licheniformis CBMAI 1609 as an Enzymatic Cocktail Supplement for Biomass Saccharification.

    Science.gov (United States)

    de Lima, Evandro Antonio; Machado, Carla Botelho; Zanphorlin, Letícia Maria; Ward, Richard John; Sato, Hélia Harumi; Ruller, Roberto

    2016-06-01

    Galactanases (endo-β-1,4-galactanases-EC 3.2.1.89) catalyze the hydrolysis of β-1,4 galactosidic bonds in arabinogalactan and galactan side chains found in type I rhamnogalacturan. The aim of this work was to understand the catalytic function, biophysical properties, and use of a recombinant GH53 endo-beta-1,4-galactanase for commercial cocktail supplementation. The nucleotide sequence of the endo-β-1,4-galactanase from Bacillus licheniformis CBMAI 1609 (Bl1609Gal) was cloned and expressed in Escherichia coli, and the biochemical and biophysical properties of the enzyme were characterized. The optimum pH range and temperature of Bl1609Gal activity were 6.5-8 and 40 °C, respectively. Furthermore, Bl1609Gal showed remarkable pH stability, retaining more than 75 % activity even after 24 h of incubation at pH 4-10. The enzyme was thermostable, retaining nearly 100 % activity after 1-h incubation at pH 7.0 at 25-45 °C. The enzymatic efficiency (K cat /K m ) against potato galactan under optimum conditions was 241.2 s(-1) mg(-1) mL. Capillary zone electrophoresis demonstrated that the pattern of galactan hydrolysis by Bl1609Gal was consistent with that of endogalactanases. Supplementation of the commercial cocktail ACCELLERASE(®)1500 with recombinant Bl1609Gal increased hydrolysis of pretreated sugarcane bagasse by 25 %. PMID:26879978

  19. Optimization of the growth conditions for amylase production by bacillus licheniformis 208 isolated from local hotsprings of karachi

    International Nuclear Information System (INIS)

    Studies on the optimum conditions for the production of extracellular amylase were carried out with a newly isolated strain of Bacillus 208 from the hotsprings in Karachi. The optimum temperature, initial medium pH and incubation period for amylase production were 50 degree C, 7.0 and 24 hrs respectively. Furthermore, cells when grown in the complex media showed high amylase production compared to the minimal medium. Effect of different carbon sources revealed that soluble starch (1%) increased the amylase yield significantly. Peptone (as nitrogen source) gave higher yield as compared to other nitrogen sources tested. Under optimized conditions, the organism entered the stationary phase after 12 hrs and amylase production was observed to be maximum at 24th hrs of cultivation. Enzyme production regulation is influenced by catabolite repression. Reduction in enzyme production was observed in the presence of EDTA while addition of tween 20 and CaCl/sub 2/ helped to enhance the enzyme production. (author)

  20. Predicción de la transferencia de masa en cultivos no-newtonianos de Bacillus licheniformis

    Directory of Open Access Journals (Sweden)

    Mariela Rizo-Porro

    2015-01-01

    Full Text Available El objetivo de este trabajo es desarrollar un modelocapaz de predecir el valor del coeficiente volumétrico detransferencia de oxígeno (kLa en cultivos deBacilluslicheniformis,en los cuales se utiliza la sacarosa comosustrato limitante. El proceso se realiza en cuatro eta-pas: la selección de expresiones que correlacionen el kLacon la viscosidad del fluido; la determinación de lascaracterísticas reológicas de un cultivo de esta bacteriacon sobreexpresión de un exopolisacárido (EPS; ladeterminación experimental del kLa en diferentes con-diciones de viscosidad, velocidad de agitación y flujo deaire y por último; la selección de la correlación quemejor representa los resultados experimentales. Comoresultado del trabajo desarrollado se obtuvo que elmodelo de Oswald de Waele representa el comporta-miento pseudoplástico de un cultivo deBacillus licheni-formisen las condiciones estudiadas y se identificaronlos coeficientes de la correlación propuesta la cualrepresenta exitosamente el 99,45 % de los datos experi-mentales. Los valores de kLa experimental fueron obte-nidos entre 0,0056 y 0,981 s-1.

  1. Mutante espontâneo de Bacillus licheniformis bloqueado no estágio I da esporogênese, possuidor de metabolismo respiratório aumentado A spontaneous mutant of Bacillus licheniformis with increased respiratory metabolism, blocked in stage I of sporogenesis

    Directory of Open Access Journals (Sweden)

    Leon Rabinovitch

    1976-01-01

    Full Text Available Um mutante espontâneo de Bacillus licheniformis, derivado da amostra esporogênica 2390, foi estudado com vistas ao reconhecimento do estágio da evolução para esporo em que o mesmo se encontrava bloqueado. Eletronmicrografias sugeriram que as células desse mutante, colhidas durante a fase estacionária da curva de crescimento, não ultrapassaram o estágio I da esporogênese (i.e., permaneceram com o nucleóide disposto como filamento axial, enquanto a produção de antibiótico (bacitracina e a atividade proteolítica foram francamente detectadas. A linhagem mutante, designada Spolp-72, nas condições experimentais empregadas não biossintetizou esporos por estarvação em solução de sais inorgãnicos, mas evidenciou uma frequência de esporulação menor que 10*-7, após crescimento vegetativo em meio de cultura favorável á esporogênese. A amostra Spolp-72 externa um crescimento vegetativo inicial restringido, quando comparada com a amostra 2390, enquanto que, inversamente, sua atividade respiratória é significativamente mais elevada. Este último comportamento foi confirmado no presente trabalho, contrastando, nesse particular, com outros tipos de mutantes de esporulação já descritos, os quais se encontram bloqueados nos primeiros estágios da via esporogenética.A spontaneous mutant strain derived from the sporogenic B. licheniformis 2390 was studied with a view to determining at what developmental stage toward sporulation it was blocked. Electronmicrographs suggested that the mutant cells harvested during the stationary phase of the growth curve were unable to go beyond stage I of sporogenesis (i. e., their nucleoid remained as an axial filament. On the other hand, antibiotic production (bacitracin and proteolytic activity were easily detected. Under the present experimental conditions the mutant strain, named Spolp-72, did not synthesize spores by starvation in a solution of inorganic salts, in contrast with the parental

  2. Synthesis and Physicochemical Characterization of D-Tagatose-1-phosphate: The Substrate of the Tagatose-1-Phosphate Kinase TagK in the PTS-mediated D-Tagatose Catabolic Pathway of Bacillus licheniformis

    Science.gov (United States)

    Van der Heiden, Edwige; Delmarcelle, Michaël; Simon, Patricia; Counson, Melody; Galleni, Moreno; Freedberg, Darón I.; Thompson, John; Joris, Bernard; Battistel, Marcos D.

    2015-01-01

    We report the first enzymatic synthesis of D-tagatose-1-phosphate (Tag-1P) by the multi-component PEP-dependent:tag-PTS present in tagatose-grown cells of Klebsiella pneumoniae. Physicochemical characterization by 31P and 1H NMR spectroscopy reveals that, in solution, this derivative is primarily in the pyranose form. Tag-1P was used to characterize the putative tagatose-1-phosphate kinase (TagK) of the Bacillus licheniformis PTS-mediated D-Tagatose catabolic Pathway (Bli-TagP). For this purpose, a soluble protein fusion was obtained with the 6 His-tagged trigger factor (TFHis6) of Escherichia coli. The active fusion enzyme was named TagK-TFHis6. Tag-1P and D-fructose-1-phosphate (Fru-1P) are substrates for the TagK-TFHis6 enzyme, whereas the isomeric derivatives D-tagatose-6-phosphate (Tag-6P) and D-fructose-6-phosphate (Fru-6P) are inhibitors. Studies of catalytic efficiency (kcat/Km) reveal that the enzyme specificity is markedly in favor of Tag-1P as substrate. Importantly, we show in vivo that the transfer of the phosphate moiety from PEP to the B. licheniformis tagatose-specific enzyme II (EIITag) in E.coli is inefficient. The capability of the PTS general cytoplasmic components of B. subtilis, HPr and EI, to restore the phosphate transfer is demonstrated. PMID:26159072

  3. 高湿度对地衣芽胞杆菌胶囊质量的影响%Influence of High-Humidity on the Quality of the Bacillus lichenifor-mis Capsules

    Institute of Scientific and Technical Information of China (English)

    王丽华

    2014-01-01

    目的:探讨环境湿度对微生态制剂质量的影响。方法:以微生态制剂地衣芽胞杆菌胶囊为研究对象,在模拟的高湿度环境中研究了其干燥失重、活菌数和崩解时限的变化。结果:在相对湿度为80%的环境中放置48 h后,药品的活菌数、干燥失重、崩解时限均有显著变化。结论:高湿对益生菌制剂质量有很大影响,该类制剂应严格遵循药品贮存和使用的条件,并注意密闭包装。%Objective: To study the influence of environment humidity to the probiotics. Methods: Taking Bacil-lus licheniformis capsule as investigated subject, the changes of drying loss, survival number and disintegration time limit under a model high-humidity condition were observed. Results: Compared with normal conditions, the live bacteria number of the Bacillus licheniformis capsules were reduced significantly after 48 h on 80% humidity environment. Conclusion: The high humidity has significant impact on probiotic product. So probiotic product should be instrict accordance with the storage and use condition, and keep in tight packs.

  4. 地衣芽孢杆菌产Levan果聚糖发酵条件的优化%Optimization of Fermentation Conditions for Production of Levan by Bacillus licheniformis 8-37-0-1

    Institute of Scientific and Technical Information of China (English)

    陆娟; 肖敏; 卢丽丽

    2011-01-01

    通过单因素试验(培养基用水、碳源、氮源、培养温度和培养基初始pH值)和正交试验对地农芽孢杆菌(Bacillus licheniformis)8-37-0-1发酵产生Levan果聚糖的培养基组成及培养条件进行优化,采用苯酚-硫酸法测定多糖含量.结果表明:以蔗糖100g/L、牛肉膏1.0g/L、酵母粉0.6g/L、K2HPO4 3.0g/L、KH2PO4 3.0g/L、NaCl 1.0g/L、MgSO4·7H2O 0.2g/L、FeSO4·7H2O 0.001g/L,自来水配制,培养基初始pH5.0,30℃培养8-37-0-1菌株24h,Levan果聚糖产量达到最高值41.7g/L,约是未优化时的5.0倍.

  5. In silico modeling of the type 2 IDI enzymes of Bacillus licheniformis, Pseudomonas stutzeri, Streptococcus pyogenes, and Staphylococcus aureus for virtual screening of potential inhibitors of this therapeutic target.

    Science.gov (United States)

    Torktaz, Ibrahim; Shahbani Zahiri, Hossein; Akbari Noghabi, Kambiz

    2013-02-01

    Isopentenyl diphosphate isomerase is an essential enzyme in those living organisms such as pathogenic strains of Streptococcus and Staphylococcus genera which rely on the Mevalonate pathway for the production of isoprenoids. The pathogens contain type 2 IDI in contrast to human that contains type 1 IDI. Therefore, the type 2 IDI may be a potential target for the therapy of some infectious diseases. In the current study, a virtual screening by docking was performed among 2000 chemicals from CoCoCo library to find a specific inhibitor for type 2 IDIs. To this end, the structures of the type 2 IDIs of Bacillus licheniformis, Pseudomonas stutzeri, Streptococcus pyogenes, and Staphylococcus aureus were molded using comparative modeling and Hidden Markov Model (HMM) based prediction. The predicted models were evaluated based on Q-mean and Prosa score. Molegro Virtual Docker with MolDock scoring function was used for measuring the binding affinity of the found inhibitor to the active site of the models. Also the inhibition effect of the compound was virtually tested on the crystallography-solved structures of the Sulfolobus shibatae and Thermus thermophilus type 2 IDIs as well as the Escherichia coli type 1 IDI. Finally, the inhibition effect of the found inhibitor was virtually tested on the human type 1 IDI. Interestingly, the results suggest that the inhibitor efficiently binds to and inhibits the bacterial IDIs especially the type 2 IDIs of pathogens while it is not inhibiting the human IDI. PMID:23280415

  6. Effect analysis of Bacillus subtilis two (Live) enteric-coated capsules combined with Bacillus licheniformis capsule in the treatment of antibiotic associated diarrhea%枯草杆菌二联活菌肠溶胶囊联合地衣芽胞杆菌活菌胶囊治疗老年抗生素相关性腹泻的效果分析

    Institute of Scientific and Technical Information of China (English)

    袁宏伟

    2015-01-01

    Objective To explore the clinical effect of Bacillus subtilis two (Live) enteric-coated capsules combined with Bacillus licheniformis capsule in the treatment of antibiotic associated diarrhea. Methods 120 patients with an-tibiotic associated diarrhea in our hospital from February 2013 to February 2014 were selected,and were divided into three groups based on random number table,Bacillus subtilis two (Live) enteric-coated capsules group,Bacillus licheni-formis capsule group,and joint application of Bacillus subtilis two (Live) enteric-coated capsules and Bacillus licheni-formis capsule group.The therapeutic effects among three groups were compared. Results The cure rate of joint applica-tion of Bacillus subtilis two (Live) enteric-coated capsules and Bacillus licheniformis capsule group was higher than that of Bacillus subtilis two (Live) enteric-coated capsules group and Bacillus licheniformis capsule group,the differ-ence was significant (χ2=8.26,P=0.02).The number of diarrhea in healed patients of joint application of Bacillus subtilis two (Live) enteric-coated capsules and Bacillus licheniformis capsule group was less than that of Bacillus subtilis two (Live) enteric-coated capsules group and Bacillus licheniformis capsule group,the difference was significant (F=91.03, P=0.00). Conclusion Both Bacillus subtilis two (Live) enteric-coated capsules and Bacillus licheniformis capsule are classified into probiotics,and have some effect on treating senile associated diarrhea caused by antibiotics.Joint applica-tion of the two drugs displays remarkable effect on treating antibiotic associated diarrhea,and plays a certain assistant role in clinical treatment.%目的:探讨枯草杆菌二联活菌肠溶胶囊联合地衣芽胞杆菌活菌胶囊对老年抗生素相关性腹泻的临床疗效。方法收集2013年2月~2014年2月本院老年抗生素相关性腹泻患者120例,根据随机数字表法分为枯草杆菌二联活菌肠溶胶囊组、地衣芽胞杆

  7. Mathematical modeling of maize starch liquefaction catalyzed by α-amylases from Bacillus licheniformis: effect of calcium, pH and temperature.

    Science.gov (United States)

    Presečki, Ana Vrsalović; Blažević, Zvjezdana Findrik; Vasić-Rački, Ðurđa

    2013-01-01

    The first step of starch hydrolysis, i.e. liquefaction has been studied in this work. Two commercial α-amylases from Bacilllus licheniformis, known as Termamyl and Liquozyme have been used for this purpose. Using starch as the substrate, kinetics of both enzymes has been determined at optimal pH and temperature (pH 7, T = 80 °C) and at 65 °C and pH 5.5. Michaelis-Menten model with uncompetitive product inhibition was used to describe enzyme kinetics. Mathematical models were developed and validated in the repetitive batch and fed-batch reactor. Enzyme inactivation was described by the two-step inactivation model. All experiments were performed with and without calcium ions. The activities of both tested amylases are approximately one hundred times higher at 80 °C than at 65 °C. Lower inactivation rates of enzymes were noticed in the experiments performed at 65 °C without the addition of calcium than in the experiments at 80 °C. Calcium ions in the reaction medium significantly enhance amylase stability at 80 °C and pH 7. At other process conditions (65 °C and pH 5.5) a weaker calcium stabilizing effect was detected.

  8. Resistance to antimicrobials and acid and bile tolerance of Bacillus spp isolated from Bikalga, fermented seeds of Hibiscus sabdariffa

    DEFF Research Database (Denmark)

    Compaore, Clarisse S.; Jensen, Lars Bogø; Diawara, Brehima;

    2013-01-01

    In the aim of selecting starter cultures, thirteen species of Bacillus spp. including six Bacillus subtilis ssp. subtilis, four Bacillus licheniformis and three Bacillus amyloliquefaciens ssp. plantarum isolated from traditional Bikalga were investigated. The study included, for all isolates, gen...

  9. Evaluation of effect of oral applying of Bacillus licheniformis CH 200: DSM 5749 and Bacillus subtilis CH 201: DSM 4750 on development and composition of red-eared slider’s (Trachemys scripta elegans intestinal microflora based on water quality changes in aquaterrariums

    Directory of Open Access Journals (Sweden)

    Mateusz Rawski

    2012-09-01

    Full Text Available Frequent cases of bacterial infections caused by contact with semi-aquatic turtles and water from their thanks had been reported in US, Japan and many other countries. In Poland the topic of microbiological hazard associated with reptile keeping is very rarely discussed. Even less is mentioned about eventual impact of probiotics as factors limiting the colonization and presence of harmful Enterobacteriaceae. The aim of the study was to investigate the microbiological threat related to turtles and tortoises; evaluating the use of probiotics to reduce intestinal pathogenic microflora of red-eared sliders, which is potentially hazardous to humans. The results of the study suggest that tank water is the potential reservoir of pathogenic microorganisms. Oral application of probiotics containing the Bacillus licheniformis CH 200: DSM 5749 and Bacillus subtilis CH 201: DSM 4750 resulted in building a probiotic population in turtle gastrointestinal tract. It affected the water microflora in aquaterrariums by increasing the total number of bacteria, including lactic acid bacteria. Reduction of the presence and quantity of Escherichia coli serotype O157:H7 seems also to be relevant.

  10. Effects of Bacillus licheniformis on Production Performance of Pregnancy and Lactation Sows and Ammonia Concentration in Piggeries%地衣芽孢杆菌对妊娠及哺乳母猪生产性能及猪舍氨气浓度的影响

    Institute of Scientific and Technical Information of China (English)

    郭丽华; 索成; 刘海涛

    2013-01-01

    [Objective] The aim was to explore effects of Bacillus licheniformis on production performance of pregnancy and lactation sows and the ammonia concentration in piggeries. [Method] Landrace × Lange sows before 30 d pregnancy were fed with basic feed added inocula and power of B. licheniformis, and effects of B. licheniformis on production performance of pregnancy and lactation sows and the ammonia concentration in piggeries were investigated. [Result] Compared with control,B. licheniformis could reduce the constipation ratio of sows,the mortality ratio of piglets and ammonia concentration in piggeries significantly. It could also improve the daily feed of sows and the diarrhea ratio of piglets significantly. [Conclusion] B. licheniformis can increase the production performance of sows and piglets effectively, improve the environment of piggeries and reduce environmental pollution.%[目的]探讨地衣芽孢杆菌菌剂和菌粉对妊娠及哺乳母猪生产性能和猪舍氨气浓度的影响.[方法]选用产前30 d的妊娠母猪(长×大)为试验对象,通过在基础日粮中分别添加地衣芽孢杆菌菌剂和菌粉,考察其对妊娠及哺乳母猪生产性能和猪舍氨气浓度的影响.[结果]与对照组相比,饲喂地衣芽孢杆菌后能显著降低母猪便秘率、仔猪腹泻率及猪舍氨气浓度,且对母猪采食量及仔猪死淘率等也有明显改善.[结论]在饲料中添加地衣芽孢杆菌能有效提高母猪及仔猪的生长性能,改善猪舍环境,降低污染.

  11. Genotyping of B. licheniformis based on a novel multi-locus sequence typing (MLST scheme

    Directory of Open Access Journals (Sweden)

    Madslien Elisabeth H

    2012-10-01

    Full Text Available Abstract Background Bacillus licheniformis has for many years been used in the industrial production of enzymes, antibiotics and detergents. However, as a producer of dormant heat-resistant endospores B. licheniformis might contaminate semi-preserved foods. The aim of this study was to establish a robust and novel genotyping scheme for B. licheniformis in order to reveal the evolutionary history of 53 strains of this species. Furthermore, the genotyping scheme was also investigated for its use to detect food-contaminating strains. Results A multi-locus sequence typing (MLST scheme, based on the sequence of six house-keeping genes (adk, ccpA, recF, rpoB, spo0A and sucC of 53 B. licheniformis strains from different sources was established. The result of the MLST analysis supported previous findings of two different subgroups (lineages within this species, named “A” and “B” Statistical analysis of the MLST data indicated a higher rate of recombination within group “A”. Food isolates were widely dispersed in the MLST tree and could not be distinguished from the other strains. However, the food contaminating strain B. licheniformis NVH1032, represented by a unique sequence type (ST8, was distantly related to all other strains. Conclusions In this study, a novel and robust genotyping scheme for B. licheniformis was established, separating the species into two subgroups. This scheme could be used for further studies of evolution and population genetics in B. licheniformis.

  12. STUDY ON THE CHARACTERISTICS OF THE AMMONIA-NIROGEN AND RESIDUAL FEEDS DEGRADATION IN AQUATIC WATER BY BACILLUS LICHENIFORMIS%地衣芽孢杆菌对养殖水体氨氮、残饵降解特性研究

    Institute of Scientific and Technical Information of China (English)

    张庆华; 封永辉; 王娟; 郭婧; 张永华; 高建忠; 宋增福

    2011-01-01

    It is well known that ammonia-nitrogen is one of the toxic factors to affect the growth performances and increase the susceptibility of the aquatic animal in the culture ponds water.With the development of aquaculture, the intensive and high-density culture aggravate the serious pollution problem of the ammonium nitrogen; however, the traditional physical and chemical methods to remove the ammonia-nitrogen will be cut down for the secondary pollution step by step.Meanwhile, the biological methods to remove ammonia-nitrogen and bioremediation have attracted much more attentions from the scientists for the characteristics of high-efficiency, low cost, no residue and environmental friendly,which become more and more popular in the practices.The present studies were focused on the Bacillus subtitles, Bacillus cereus and Sporolactobacillus.Large yellow croaker (Pseudosciaena crocea) is one of the traditional Chinese Four Seafood that is the offshore main economic fishes, which culture widely in the China eastern offshore.In the present experiment, a Bacillus licheniformis strain X3914 as a potential probiotics was isolated from gastrointestinal tract of healthy large yellow croaker (Pseudosciaena crocea) cultured in Xiangshan, Zhejiang Province.The aim of the study is to explore the degradation rules of ammonia-nitrogen, protein and starch of residual feeds by Bacillus licheniformis strain X3914, which probably offers the guidance to the practices in aquaculture.In the present experiment, the ability to remove the ammonia-nitrogen of the Bacillus licheniformis strain X3914 was tested in the simulated waste water, the results indicated that the growth of the strain X3914 was synchronized with the degradation of ammonia-nitrogen and degradation rate of ammonia-nitrogen reached to 36.2% in 24h.Furthermore,Ammonia-nitrogen containing the ammonium ions, and ammonia molecules, as the toxicity of ammonium ions was influenced significantly by environmental factors, such as

  13. Comparison of different Bacillus subtilis expression systems.

    Science.gov (United States)

    Vavrová, Ludmila; Muchová, Katarína; Barák, Imrich

    2010-11-01

    Bacillus subtilis is considered to have great potential as a host for the production and secretion of recombinant proteins. Many different expression systems have been developed for B. subtilis. Here we compare two widely used expression systems, the IPTG-inducible derivative of spac system (hyper-spank) and the xylose-inducible (xyl) to the SURE (subtilin-regulated gene expression) system. Western blot analysis of the membrane protein SpoIISA together with its protein partner SpoIISB showed that the highest expression level of this complex is obtained using the SURE system. Measurement of β-galactosidase activities of the promoter-lacZ fusions in individual expression systems confirmed that the P(spaS) promoter of the SURE system is the strongest of those compared, although the induction/repression ratio reached only 1.84. Based on these results, we conclude that the SURE system is the most efficient of these three B. subtilis expression systems in terms of the amount of expressed product. Remarkably, the yield of the SpoIISA-SpoIISB complex obtained from B. subtilis was comparable to that normally obtained from the Escherichia coli arabinose-inducible expression system. PMID:20863884

  14. Comparative analysis of two-component signal transduction systems of Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis

    NARCIS (Netherlands)

    Been, M.W.H.J. de; Francke, C.; Moezelaar, R.; Abee, T.; Siezen, R.J.

    2006-01-01

    Members of the Bacillus cereus group are ubiquitously present in the environment and can adapt to a wide range of environmental fluctuations. In bacteria, these adaptive responses are generally mediated by two-component signal transduction systems (TCSs), which consist of a histidine kinase (HK) and

  15. 牛源地衣芽胞杆菌诱导小鼠临床乳房炎模型中热休克蛋白和核因子-κB表达上调%Up-regulated expression of heat shock protein 70 and NF-κB in mouse mastitis model established by artificial infection of Bacillus licheniformis from bovine mastitis

    Institute of Scientific and Technical Information of China (English)

    甘露; 徐君; 李姝; 程跃; 蔡亚非

    2012-01-01

    Objective: To further understand how the immune pathogenesis of cow mastitis occurs and look for better control methods to provide data for basic study. Methods: In this study the main mastitis pathogens were isolated and identified by the traditional bacterial culture methods and molecular methods. Bacillus licheniformis was selected from the main pathogens. Then mouse mammary gland was infected by Bacillus licheniformis. HE staining was done to observe the change of the mouse mammary gland after infection and immunohistochemical staining to detect the expression of heat shock protein 70 (HSP70) and NF-κB in those slices. Results: Symptoms of inflammation appeared in the mouse mammary gland after infection. Histopathologic examination of mammary gland revealed that numbers of polymorphonuclear neutrophils were present in alveoli. The expression of HSP70 and NF-κB in the experimental group was significantly increased compared with the saline group and the blank group. Conclusion:The results showed that Bacillus licheniformis was suitable for the replica of mastitis model in mouse. An increase in the expression of HSP70 and NF-κB in the experimental group explained their participation in the pathological process after bacterial infection.%目的:为进一步认识奶牛乳腺炎发生的免疫病理机制和寻找更理想的防治方法提供基础研究数据.方法:从采集回来的奶牛奶样中用传统细菌培养方法和分子学方法分离鉴定主要病原菌,挑选其中地衣芽胞杆菌感染小鼠乳腺组织,H-E染色分析感染后小鼠乳腺组织变化,免疫组织化学方法分析热休克蛋白70(HSP70)和核因子-kB(NF-kB)在感染小鼠体内的表达.结果:感染后小鼠乳腺出现明显的炎症症状,组织病理学观察显示腺泡腔内有大量嗜中性粒细胞浸润.实验组中HSP70和NF-kB表达量比生理盐水组和空白组均有增加.结论:成功通过地衣芽胞杆菌诱发建立实验性乳房炎小

  16. SYNTHESIS AND PROCESSING OF ESCHERICHIA-COLI TEM-BETA-LACTAMASE AND BACILLUS-LICHENIFORMIS ALPHA-AMYLASE IN ESCHERICHIA-COLI : THE ROLE OF SIGNAL PEPTIDASE-I

    NARCIS (Netherlands)

    van Dijl, J M; SMITH, H; BRON, S; VENEMA, G

    1988-01-01

    A mutant of Escherichia coli, in which signal peptidase I synthesis can be regulated, was constructed. The mutant was used to study the effects of signal peptidase I limitation on the synthesis and efficiency of processing of two proteins: the periplasmic E. coli TEM-beta-lactamase and Bacillus lich

  17. Biosynthesis of poly(g-L-glutamic acid) and comparative studies on the biodegradability of the g- and a-enantiomeric forms of the poly (glutamic acid) by Bacillus Licheniformis NCIMB 11709

    OpenAIRE

    Marqués Calvo, M. Soledad; Bou Serra, Jordi; Cerdà Cuéllar, Marta

    2013-01-01

    Poly (y-glutamic acid), a novel polyanionic and multifunctional macromolecule synthesized by Bacillus species, has attracted considerable attention because of its eco-friendly, biodegradable and biocompatible characteristics. Recently, its application in a wide range of fields such as food, agriculture, medicine, hygiene, cosmetics and the environment has been explored. This book discusses the chemistry, food sources and health benefits of glutamic acid.

  18. 胡椒经地衣芽孢杆菌发酵脱皮过程中的主要酶系及pH值变化%Dynamic Changes in Main Enzyme Activities and pH during Pepper(Piper nigrum L.) Decortication by Bacillus licheniformis Fermentation

    Institute of Scientific and Technical Information of China (English)

    熊海波; 侯源源; 刘四新; 苗子健; 李从发

    2011-01-01

    采用地衣芽孢杆菌进行静置液态发酵对胡椒进行脱皮,与生产实践中传统水沤法脱皮对比,研究二者发酵脱皮过程中的果胶酶、木聚糖酶、纤维素酶和发酵液pH值的动态变化。结果表明:在发酵脱皮过程中,两种方法的聚半乳糖醛酸酶(PG)和果胶裂解酶(PL)变化规律相似,都出现两次酶活力高峰;而纤维素酶活力都很低,果胶酯酶(PE)酶活力都比较高;在传统水沤法中木聚糖酶活力出现两个高峰,而在发酵法中木聚糖酶活力在脱皮后期逐渐上升;pH值变化总趋势也相似,脱皮完成时pH值均在5~5.5之间。由此推知,胡椒鲜果发酵脱皮过程中果胶酶系%Static liquid-state Bacillus licheniformis fermentation was used for pepper decortication and compared with traditional water retting.Meanwhile,the dynamic changes of pectinases,xylanase,cellulase and pH during pepper decortication by the two methods were explored.The results showed that the changes of polygalacturonase(PG) and pectin lyase(PL) revealed a similar pattern with two activity peaks during both decortication processes.However,cellulase activity was low and the pectin esterase(PE) activity remained high.Similar pH changes were observed during both decortication processes pH was between 5 and 5.5 at the end of decortication.Xylanase activity showed two peaks in traditional water retting method and a gradual increase in the late stage of Bacillus licheniformis fermentation.Therefore,pectinase might play an important role during pepper decortication.In the early stage of decortication,PG and PL first acted on pectin substances and damaged the pectin complex structure,and then PE hydrolyzed pectin molecules to form pectic acid accompanied with pH decrease.Further,xylanase activity increased gradually in the late stage of decortication and as a result,hemicellulose was rapidly depolymerized.Therefore,pepper decortication was completed under the

  19. The Bacillus cereus spoIIS programmed cell death system

    Directory of Open Access Journals (Sweden)

    Jana eMelnicakova

    2015-08-01

    Full Text Available Programmed cell death in bacteria is generally associated with two¬ component toxin antitoxin systems. The SpoIIS toxin-antitoxin system, consisting of a membrane bound SpoIISA toxin and a small, cytosolic antitoxin SpoIISB, was originally identified in Bacillus subtilis. In this work we describe the Bacillus cereus SpoIIS system which is a three-component system, harbouring an additional gene spoIISC. Its protein product serves as an antitoxin, and similarly as SpoIISB, is able to bind SpoIISA and abolish its toxic effect. Our results indicate that SpoIISC seems to be present not only in B. cereus but also in other Bacilli containing a SpoIIS toxin antitoxin system. In addition, we show that B. cereus SpoIISA can form higher oligomers and we discuss the possible role of this multimerization for the protein’s toxic function.

  20. Respiratory systems of the Bacillus cereus mother cell and forespore.

    OpenAIRE

    Escamilla, J E; R. Ramírez; Del-Arenal, P; Aranda, A.

    1986-01-01

    The respiratory systems of the mother cells and forespores of Bacillus cereus were compared throughout the maturation stages (III to VI) of sporulation. The results indicated that both cell compartments contain the same assortment of oxidoreductases and cytochromes. However membrane fractions from young forespores were clearly distinct from those of the mother cell, i.e., lower content of cytochrome aa3, lower cytochrome c oxidase activity, higher concentration of cytochrome o, and a lower se...

  1. Effect of supplemental Bacillus culture on rumen fermentation and performance in dairy cattle

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Two parts were involved in this experiment. In experiment 1, 32 Chinese Holstein cows with relatively similar body condition, lactation number and days in milk were selected. The cows were assigned in a randomized complete block design trial to determine the effect of supplemental Bacillus cultures to diet on production performance in dairy cattle. Four treatments, i.e., Bacillus licheniformis (strain number 1.813) group, Bacillus subtilis (strain number 1.1086) group, Bacillus cereus var. mycoides (strain number 1.260) group and control group. Each treatment had eight replicates, each replicate had one cow, 50 g per head per day. Results showed that Bacillus licheniformis group increased the milk yield (P0.05). In experiment 2, 3 Chinese Holstein cows with permanent fistulas were used. 3×3 Latin squares were assigned to three diets: Bacillus lincheniformis culture, Bacillus subtilis culture and control. Bacillus licheniformis culture increased total rumen microorganism (P0.05), increased the rate of acetic acid to propionic acid (P>0.05). Bacillus licheniformis culture decreased the methane production (P>0.05).

  2. Role of enzymes of homologous recombination in illegitimate plasmid recombination in Bacillus subtilis

    NARCIS (Netherlands)

    Meima, R; Haijema, BJ; Haan, GJ; Venema, G; Bron, S

    1997-01-01

    The structural stability of plasmid pGP1, which encodes a fusion between the penicillinase gene (penP) of Bacillus licheniformis and the Escherichia coli lacZ gene, was investigated in Bacillus subtilis strains expressing mutated subunits of the ATP-dependent nuclease, AddAB, and strains lacking the

  3. The expression of a plasmid-specified exported protein causes structural plasmid instability in Bacillus subtilis

    NARCIS (Netherlands)

    Cordes, C.; Meima, R; Twiest, B; Kazemier, B; Venema, G; vanDijl, JM; Bron, S

    1996-01-01

    The rolling-circle plasmid pGP1 was used to study the effects of the expression of a plasmid-specified exported protein on structural plasmid stability in Bacillus subtilis. pGP1 contains a fusion between the Bacillus licheniformis penP gene, encoding a C-terminally truncated penicillinase, and the

  4. Probiotic effects on cobia Rachycentron canadum larvae reared in a recirculating aquaculture system

    OpenAIRE

    M. Angélica Garrido-Pereira; Michael Schwarz; Brendan Delbos; Ricardo V. Rodrigues; Luis Romano; Luís Sampaio

    2014-01-01

    Cobia (Rachycentron canadum) is a marine finfish with good potential for mariculture. This study analyzes the effects of probiotic Bacillus spp. on the performance of cobia larvae reared in a recirculating aquaculture system (RAS). Larvae were stocked into two independent RAS for 26 days after hatching. One of the systems (Probiotic treatment) received the addition of a commercial probiotic consisting of B. subtilis, B. licheniformis and B. pumilus directly into the water and by live feed. Su...

  5. Systems biology of recombinant protein production using Bacillus megaterium.

    Science.gov (United States)

    Biedendieck, Rebekka; Borgmeier, Claudia; Bunk, Boyke; Stammen, Simon; Scherling, Christian; Meinhardt, Friedhelm; Wittmann, Christoph; Jahn, Dieter

    2011-01-01

    The Gram-negative bacterium Escherichia coli is the most widely used production host for recombinant proteins in both academia and industry. The Gram-positive bacterium Bacillus megaterium represents an increasingly used alternative for high yield intra- and extracellular protein synthesis. During the past two decades, multiple tools including gene expression plasmids and production strains have been developed. Introduction of free replicating and integrative plasmids into B. megaterium is possible via protoplasts transformation or transconjugation. Using His(6)- and StrepII affinity tags, the intra- or extracellular produced proteins can easily be purified in one-step procedures. Different gene expression systems based on the xylose controlled promoter P(xylA) and various phage RNA polymerase (T7, SP6, K1E) driven systems enable B. megaterium to produce up to 1.25g of recombinant protein per liter. Biomass concentrations of up to 80g/l can be achieved by high cell density cultivations in bioreactors. Gene knockouts and gene replacements in B. megaterium are possible via an optimized gene disruption system. For a safe application in industry, sporulation and protease-deficient as well as UV-sensitive mutants are available. With the help of the recently published B. megaterium genome sequence, it is possible to characterize bottle necks in the protein production process via systems biology approaches based on transcriptome, proteome, metabolome, and fluxome data. The bioinformatical platform (Megabac, http://www.megabac.tu-bs.de) integrates obtained theoretical and experimental data. PMID:21943898

  6. Resistance to antimicrobials and acid and bile tolerance of Bacillus spp isolated from Bikalga, fermented seeds of Hibiscus sabdariffa

    OpenAIRE

    Compaore, Clarisse S.; Jensen, Lars Bogø; Diawara, Brehima; Ouedraogo, Georges A.; Jakobsen, Mogens; Ouoba, Labia I. I.

    2013-01-01

    In the aim of selecting starter cultures, thirteen species of Bacillus spp. including six Bacillus subtilis ssp. subtilis, four Bacillus licheniformis and three Bacillus amyloliquefaciens ssp. plantarum isolated from traditional Bikalga were investigated. The study included, for all isolates, genes, determination of minimal inhibitory concentration (MIC) for 24 antimicrobials and detection of resistance by PCR using specific primers. The isolates were also examined for their resistance to pH ...

  7. Production of amylolytic enzymes by bacillus spp

    International Nuclear Information System (INIS)

    Sixty six bacteria and twenty fungi were isolated from various sources. These varied from rotten fruites to local drinks and soil samples from different parts of Sudan. On the basis of index of amylolytic activity, forty one bacteria and twelve fungi were found to hydrolyse strach. The best ten strach hydrolysing isolates were identified all as bacilli (Bacillus licheniformis SUD-K1, SUD-K2, SUD-K4, SUD-O, SUD-SRW, SUD-BRW, SUD-By, Bacillus subtilis SUD-K3, and Bacillus circulans SUD-D and SUD-K7). Their amylase productivity was studied with respect to temperature and time. Amylolytic activity was measured by spectrophotometer, the highest activity was produced in around 24 hours of growth in all; six of which gave the highest amylase activity at 50 deg C and the rest at 45C. Based on the thermal production six isolates were chosen for further investigation. These were Bacillus licheniformis SUD-K1, SUD-K2, SUD-K4, SUD-O, Bacillus subtilis SUD-K3 and Bacillus circulans SUD-K7. The inclusion of strach and Mg++ ions in the culture medium gave the highest enzyme yield. The Ph 9.0 was found to be the optimum for amylase production for all isolates except Bacillus subtilis SUD-K3 which had an optimum at pH 7.0. Three isolates (Bacillus licheniformis SUD-K1, SUD-K4 and SUD-O recorded highestamylase production in a medium supplemented with peptone while the rest (Bacillus licheniformis SUD-K2, Bacillus subtilis SUD-K3 and Bacillus circulans SUD-K7) gave highest amylase productivity in a medium supplemented with malt extract. Four isolates (Bacillus licheniformis SUD-K1 and Bacillus subtilis SUD-K3 gave maximum amylase production in a medium containing 0.5% soluble strach while the rest (gave maximum amylase production at 2%. Soluble strach was found to be best substrate among the different carbon sources tested. The maximum temperature for amylase activity ranged from 60-70 deg C and 1% strach concentration was optimum for all isolates. Addition of different metal ions

  8. Siderophore-mediated iron acquisition systems in Bacillus cereus: identification of receptors for anthrax virulence-associated petrobactin†a

    OpenAIRE

    Zawadzka, Anna M.; Abergel, Rebecca J.; Nichiporuk, Rita; Andersen, Ulla N.; Raymond, Kenneth N.

    2009-01-01

    During growth under iron limitation, Bacillus cereus and Bacillus anthracis, two human pathogens from the Bacillus cereus group of Gram-positive bacteria, secrete two siderophores, bacillibactin (BB) and petrobactin (PB), for iron acquisition via membrane-associated substrate-binding proteins (SBPs) and other ABC transporter components. Since PB is associated with virulence traits in B. anthracis, the PB-mediated iron uptake system presents a potential target for antimicrobial therapies; its ...

  9. The essential YycFG two-component system controls cell wall metabolism in Bacillus subtilis

    DEFF Research Database (Denmark)

    Bisicchia, Paola; Noone, David; Lioliou, Efthimia;

    2007-01-01

    Adaptation of bacteria to the prevailing environmental and nutritional conditions is often mediated by two-component signal transduction systems (TCS). The Bacillus subtilis YycFG TCS has attracted special attention as it is essential for viability and its regulon is poorly defined. Here we show...

  10. Bacillus pumilus reveals a remarkably high resistance to hydrogen peroxide provoked oxidative stress

    NARCIS (Netherlands)

    Handtke, S.; Schroeter, R.; Jurgen, B.; Methling, K.; Schluter, R.; Albrecht, D.; Hijum, S.A.F.T. van; Bongaerts, J.; Maurer, K.H.; Lalk, M.; Schweder, T.; Hecker, M.; Voigt, B.

    2014-01-01

    Bacillus pumilus is characterized by a higher oxidative stress resistance than other comparable industrially relevant Bacilli such as B. subtilis or B. licheniformis. In this study the response of B. pumilus to oxidative stress was investigated during a treatment with high concentrations of hydrogen

  11. The ComP-ComA Quorum System Is Essential For “Trojan horse” Like Pathogenesis in Bacillus nematocida

    OpenAIRE

    Xidan Deng; Yunxia Tian; Qiuhong Niu; Xiao'e Xu; Hui Shi; Hanbo Zhang; Lianming Liang; Keqin Zhang; Xiaowei Huang

    2013-01-01

    Bacillus nematocida B16 has been shown to use "Trojan horse" mechanism in pathogenesis that has characteristics of "social" behavior. The ComP-ComA system, a conserved quorum sensing system in the genus Bacillus, functions in many physiological processes including competence development, lipopeptide antibiotic surfactin production, degradative enzyme production and even some unknown functions. Here we investigated the requirement of ComP-ComA system in B. nematocida B16 for its pathogenicity ...

  12. Efflux-mediated multidrug resistance in Bacillus subtilis: similarities and dissimilarities with the mammalian system.

    OpenAIRE

    Neyfakh, A A; Bidnenko, V E; L. B. CHEN

    1991-01-01

    Bacillus subtilis cells selected for their resistance to rhodamine 6G demonstrated a multidrug-resistance (MDR) phenotype resembling that of mammalian MDR cells. Like MDR in mammalian cells, MDR in bacteria was mediated by the efflux of the drugs from the cells. The bacterial multidrug efflux system transported similar drugs and was sensitive to similar inhibitors as the mammalian multidrug transporter, P-glycoprotein. The gene coding for the bacterial multidrug transporter, like the P-glycop...

  13. The role of DNA restriction-modification systems in the biology of Bacillus anthracis

    Directory of Open Access Journals (Sweden)

    Ramakrishnan eSitaraman

    2016-01-01

    Full Text Available Restriction-modification (R-M systems are widespread among prokaryotes and, depending on their type, may be viewed as selfish genetic elements that persist as toxin-antitoxin modules or as cellular defense systems against phage infection. Studies in the last decade have made it amply clear that these two options do not exhaust the list of possible biological roles for R-M systems. Their presence in a cell may also have a bearing on other processes such as horizontal gene transfer and gene regulation. From genome sequencing and experimental data, we know that Bacillus anthracis encodes at least three methylation-dependent (typeIV restriction endonucleases, and an orphan DNA methyltransferase. In this article, we first present an outline of our current knowledge of R-M systems in Bacillus anthracis. Based on available DNA sequence data, and on our current understanding of the functions of similar genes in other systems, we conclude with hypotheses on the possible roles of the three restriction endonucleases and the orphan DNA methyltransferase.

  14. The sodium/proton antiport system in a newly isolated alkalophilic Bacillus sp.

    OpenAIRE

    Kitada, M; Onda, K.; Horikoshi, K

    1989-01-01

    The pH homeostasis and the sodium/proton antiport system have been studied in the newly isolated alkalophilic Bacillus sp. strain N-6, which could grow on media in a pH range from 7 to 10, and in its nonalkalophilic mutant. After a quick shift in external pH from 8 to 10 by the addition of Na2CO3, the delta pH (inside acid) in the cells of strain N-6 was immediately established, and the pH homeostatic state was maintained for more than 20 min in an alkaline environment. However, under the sam...

  15. 一株地衣芽胞杆菌S1-1对赭曲霉毒素A的吸附和降解研究%Adsorption and Degradation of Ochratoxin A by Bacillus licheniformis S1-1

    Institute of Scientific and Technical Information of China (English)

    师磊; 梁志宏; 徐诗涵; 郑浩; 黄昆仑

    2013-01-01

    赭曲霉毒素A(ochratoxin A,OTA)是一种由曲霉(Aspergillus spp.)和青霉(Penicillium spp.)等丝状真菌产生的次级代谢产物,主要污染谷物、葡萄(Vitisvinifera)、大豆(Glycine max)、咖啡(Coffea arabica)及其相关产品.动物实验表明,OTA具有肾毒性、肝毒性,致癌、致畸和致突变性,减少或消除食品及其原料中的OTA对国民食品安全至关重要.本研究以从动物粪便分离的芽胞杆菌(Bacillus spp.)为材料研究OTA的生物脱毒.结果显示,1株Bacillus spp.S1-1既能吸附又能降解OTA:活菌和高温灭活菌(121℃,20 min)均能吸附OTA,OTA浓度为6 μg/mL时,薄层层析(thin layer chromatography,TLC)测定24 h后高温灭活菌(121℃,20 min)的吸附量(80%)高于活菌(60%);菌液上清能降解OTA,OTA浓度为6.2 μg/mL时,高效液相色谱(high-performance liquid chromatograpy,HPLC)测定24 h降解率为98%,没有降解产物产生.S1-1在发霉玉米中OTA的降解率为35.0%.16S rRNA序列比对初步确定S1-1为地衣芽胞杆菌(B.licheniforms).本研究首次获得了1株既能吸附又能降解OTA的B.licheniforms,为OTA的生物脱毒提供了新材料.

  16. Functional characterization of WalRK: A two-component signal transduction system from Bacillus anthracis

    Directory of Open Access Journals (Sweden)

    Alisha Dhiman

    2014-01-01

    Full Text Available Two-component signal transduction systems (TCS, consisting of a sensor histidine protein kinase and its cognate response regulator, are an important mode of environmental sensing in bacteria. Additionally, they have been found to regulate virulence determinants in several pathogens. Bacillus anthracis, the causative agent of anthrax and a bioterrorism agent, harbours 41 pairs of TCS. However, their role in its pathogenicity has remained largely unexplored. Here, we show that WalRK of B. anthracis forms a functional TCS which exhibits some species-specific functions. Biochemical studies showed that domain variants of WalK, the histidine kinase, exhibit classical properties of autophosphorylation and phosphotransfer to its cognate response regulator WalR. Interestingly, these domain variants also show phosphatase activity towards phosphorylated WalR, thereby making WalK a bifunctional histidine kinase/phosphatase. An in silico regulon determination approach, using a consensus binding sequence from Bacillus subtilis, provided a list of 30 genes that could form a putative WalR regulon in B. anthracis. Further, electrophoretic mobility shift assay was used to show direct binding of purified WalR to the upstream regions of three putative regulon candidates, an S-layer protein EA1, a cell division ABC transporter FtsE and a sporulation histidine kinase KinB3. Our work lends insight into the species-specific functions and mode of action of B. anthracis WalRK.

  17. Friction and Adhesion Forces of Bacillus thuringiensis Spores on Planar Surfaces in Atmospheric Systems

    Energy Technology Data Exchange (ETDEWEB)

    Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Tsouris, Costas [ORNL

    2011-01-01

    The kinetic friction force and the adhesion force of Bacillus thuringiensis spores on planar surfaces in atmospheric systems were studied using atomic force microscopy. The influence of relative humidity (RH) on these forces varied for different surface properties including hydrophobicity, roughness, and surface charge. The friction force of the spore was greater on a rougher surface than on mica, which is atomically flat. As RH increases, the friction force of the spores decreases on mica whereas it increases on rough surfaces. The influence of RH on the interaction forces between hydrophobic surfaces is not as strong as for hydrophilic surfaces. The friction force of the spore is linear to the sum of the adhesion force and normal load on the hydrophobic surface. The poorly defined surface structure of the spore and the adsorption of contaminants from the surrounding atmosphere are believed to cause a discrepancy between the calculated and measured adhesion forces.

  18. Investigation and Analysis of Bacillus in Yogurt Production Environment%酸奶生产环境中芽孢杆菌的调查与分析

    Institute of Scientific and Technical Information of China (English)

    康博燕; 牟光庆; 陈历俊; 姜铁民

    2013-01-01

    According to the studies on the microbial community of yogurt production environment,found that the Bacillus.sp was the majority bacteria.Base on physiological and biochemical identification and molecular identification on the bacillus isolate from sampling plots,found that Bacillus subtilis account for 30%,Bacillus licheniformis account for 19%,Bacillus megaterium account for 14%,The rest of the seven kinds of the bacillus account for 37%.By traceability analysis,found that there was cross contamination in the connected workshop.%通过对某厂酸奶生产环境微生物菌群分析,发现此环境中微生物以芽孢杆菌属(Bacillus.sp)的菌种居多.对各个采样点分离、纯化的芽孢杆菌进行生理生化和分子鉴定,结果为枯草芽孢杆菌(Bacillus subtilis)占所分离鉴定芽孢杆菌的30%,地衣芽孢杆菌(Bacillus licheniformis)占19%,巨大芽孢杆菌(Bacillus megaterium)占14%,其余7种菌共占37%.对它们进行溯源分析,发现有连通的车间存在交叉污染.

  19. Analysis of the Bacillus cereus SpoIIS antitoxin-toxin system reveals its three-component nature.

    Science.gov (United States)

    Melničáková, Jana; Bečárová, Zuzana; Makroczyová, Jana; Barák, Imrich

    2015-01-01

    Programmed cell death in bacteria is generally associated with two-component toxin-antitoxin systems. The SpoIIS toxin-antitoxin system, consisting of a membrane-bound SpoIISA toxin and a small, cytosolic antitoxin SpoIISB, was originally identified in Bacillus subtilis. In this work we describe the Bacillus cereus SpoIIS system which is a three-component system, harboring an additional gene spoIISC. Its protein product serves as an antitoxin, and similarly as SpoIISB, is able to bind SpoIISA and abolish its toxic effect. Our results indicate that SpoIISC seems to be present not only in B. cereus but also in other Bacilli containing a SpoIIS toxin-antitoxin system. In addition, we show that B. cereus SpoIISA can form higher oligomers and we discuss the possible role of this multimerization for the protein's toxic function.

  20. [The flotation characteristics of Bacillus cells and spores].

    Science.gov (United States)

    Stabnikova, E V; Gregirchak, N N; Taranenko, T O

    1991-01-01

    Variations in hydrophobicity of the surface of bacillary cells and their capacity to flotation in the process of batch cultivation have been studied. It is shown that hydrophobicity of the cell surface increases in the course of batch cultivation of Bacillus thuringiensis, B. licheniformis and B. megaterium. Hydrophobicity of spores of the mentioned cultures is considerably higher than that of the vegetative cells. The increase of hydrophobicity of bacillary cells positively correlated with their capacity to flotation. That is why the use of flotation for the age fractionation of bacillary cells is possible: spores are concentrated in the foam while vegetative cells remain in the culture liquid. PMID:1779906

  1. Probiotic effects on cobia Rachycentron canadum larvae reared in a recirculating aquaculture system

    Directory of Open Access Journals (Sweden)

    M. Angélica Garrido-Pereira

    2014-11-01

    Full Text Available Cobia (Rachycentron canadum is a marine finfish with good potential for mariculture. This study analyzes the effects of probiotic Bacillus spp. on the performance of cobia larvae reared in a recirculating aquaculture system (RAS. Larvae were stocked into two independent RAS for 26 days after hatching. One of the systems (Probiotic treatment received the addition of a commercial probiotic consisting of B. subtilis, B. licheniformis and B. pumilus directly into the water and by live feed. Survival, final weight and water quality were not affected by probiotics. Results showed larvae of the probiotic treatment demonstrated a greater resistance to salinity stress. Immunohistochemical analysis showed a higher expression of CD4 in probiotic treatment. These results suggest that Bacillus spp. probiotics used in RAS have a potential stimulating impact on immune system differentiation and increases salinity stress resistance of cobia larvae.

  2. Rapid inactivation of seven Bacillus spp. under simulated Mars UV irradiation

    Science.gov (United States)

    Schuerger, Andrew C.; Richards, Jeff T.; Newcombe, David A.; Venkateswaran, Kasthuri

    2006-03-01

    Seven Bacillus spp. were exposed to simulations of Mars-normal UV fluence rates in order to study the effects of UV irradiation on microbial survival. A UV illumination system was calibrated to deliver 9.78 W m -2 (35.2 kJ m -2 h -1) of UVC + UVB irradiation (200-320 nm) to microbial samples, thus creating a clear-sky simulation (0.5 optical depth) of equatorial Mars. The Bacillus spp. studied were: B. licheniformis KL-196, B. megaterium KL-197, B. nealsonii FO-092, B. pumilus FO-36B, B. pumilus SAFR-032, B. subtilis 42HS1, and B. subtilis HA101. The bacteria were prepared as thin monolayers of endospores on aluminum coupons in order to simulate contaminated spacecraft surfaces. Bacterial monolayers were exposed to Mars UV irradiation for time-steps of 0, 0.25, 0.5, 1, 5, 15, 30, 60, 120, or 180 min. The surviving endospores were then assayed with a Most Probable Numbers (MPN) procedure and with a culture-based assay that utilized a bacillus spore germination medium. Results indicated that B. pumilus SAFR-032 was the most resistant, and B. subtilis 42HS-1 and B. megaterium were the most sensitive of the seven strains exposed to martian UV fluence rates. Bacillus subtilis 42HS1 and B. megaterium were inactivated after 30 min exposure to Mars UV, while B. pumilus SAFR-032 required 180 min for full inactivation in both assays. Spores of B. pumilus SAFR-032 exhibited significantly different inactivation kinetics suggesting that this wild type isolate also was more resistant than the standard dosimetric strain, B. subtilis HA101. Although the various Bacillus spp. exhibited diverse levels of UV resistance, none were immune to UV irradiation, and, thus, all species would be expected to be inactivated on Sun-exposed spacecraft surfaces within a few tens-of-minutes to a few hours on sol 1 under clear-sky conditions on equatorial Mars. The inactivation kinetics of all seven Bacillus spp. support the conclusion that significant levels of bioload reductions are possible on

  3. Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system

    Institute of Scientific and Technical Information of China (English)

    XU Ying; HE Guo-qing; LI Jing-jun

    2005-01-01

    This paper presents the evaluation of an aqueous two-phase system (ATPS) for extracting elastase produced by Bacillus sp. EL31410. The elastase and cell partition behavior in polyethylene glycol (PEG)/salt systems was investigated. The suitable system for elastase extraction was PEG/KH2PO4-K2HPO4, in which elastase is mainly partitioned into the PEG-rich phase,while the cells remained in the other phase. The influence of defined system parameters (e.g. PEG molecular mass, pH, NaCl addition) on the partitioning behavior of elastase is described. The concentration of phase forming components, PEG and KH2PO4-K2HPO4, was optimized for elastase recovery by means of response surface methodology, and it was found that they greatly influenced extraction recovery. The optimal ATPS was 23.1% (w/w) PEG 2 000 and 11.7% (w/w) KH2PO4-K2HPO4. The predicted recovery was about 89.5%, so this process is suggested to be a rapid and convenient method for elastase extraction.

  4. Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments.

    Science.gov (United States)

    Ouoba, Labia Irene I; Thorsen, Line; Varnam, Alan H

    2008-06-10

    The ability of various species of Bacillus from fermented seeds of Parkia biglobosa known as African locust bean (Soumbala) and fermented seeds of Hibiscus sabdariffa (Bikalga) was investigated. The study included screening of the isolates by haemolysis on blood agar, detection of toxins in broth and during the fermentation of African locust bean using the Bacillus cereus Enterotoxin Reverse Passive Latex Agglutination test kit (BCET-RPLA) and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (BDEVIA). Detection of genes encoding cytotoxin K (CytK), haemolysin BL (Hbl A, Hbl C, Hbl D), non-hemolytic enterotoxin (NheA, NheB, NheC) and EM1 specific of emetic toxin producers was also investigated using PCR with single pair and multiplex primers. Of 41 isolates, 29 Bacillus belonging to the species of B. cereus, Bacillus subtilis, Bacillus licheniformis and Bacillus pumilus showed haemolysis on blood agar. Using RPLA, enterotoxin production was detected for three isolates of B. cereus in broth and all B. cereus (9) in fermented seeds. Using BDEVIA, enterotoxin production was detected in broth as well as in fermented seeds for all B. cereus isolates. None of the isolates belonging to the other Bacillus species was able to produce enterotoxins either by RPLA or BDEVIA. Nhe genes were detected in all B. cereus while Hbl and CytK genes were detected respectively in five and six B. cereus strains. A weak presence of Hbl (A, D) and CytK genes was detected in two isolates of B. subtilis and one of B. licheniformis but results were inconsistent, especially for Hbl genes. The emetic specific gene fragment EM1 was not detected in any of the isolates studied.

  5. Seleção de diferentes meios para produção de lipase a partir de Bacillis licheniformis (UCP 1014

    Directory of Open Access Journals (Sweden)

    Ladiel Luiz Pedrozo Tavares

    2011-01-01

    Full Text Available Development of alternative culture media for microbial enzyme production have been widely exploited in recent decades. The microbial lipases are extracellular enzymes produced in fermentation processes, which favors its extraction, isolation and purification. Bacillus are Gram-positive bacteria, saprophytes, of great importance in various industrial sectors. In this study, we tested three methods of production using B. licheniformis (UCP 1014 media called A, B and C. The kinetics of enzyme production occurred in orbital shaker at 150 rpm, 37 °C, for 96 hours. The collected samples were subjected to the construction of the growth curve, determination of pH and lipolytic activity. The results indicated a better growth in the middle C showing an activity of 256 U/mL, while the means A and B had values of 170 and 153 U/mL, respectively. The results showed that the middle C presented higher enzyme production in the tests.

  6. Multiple regulatory systems coordinate DNA replication with cell growth in Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Heath Murray

    2014-10-01

    Full Text Available In many bacteria the rate of DNA replication is linked with cellular physiology to ensure that genome duplication is coordinated with growth. Nutrient-mediated growth rate control of DNA replication initiation has been appreciated for decades, however the mechanism(s that connects these cell cycle activities has eluded understanding. In order to help address this fundamental question we have investigated regulation of DNA replication in the model organism Bacillus subtilis. Contrary to the prevailing view we find that changes in DnaA protein level are not sufficient to account for nutrient-mediated growth rate control of DNA replication initiation, although this regulation does require both DnaA and the endogenous replication origin. We go on to report connections between DNA replication and several essential cellular activities required for rapid bacterial growth, including respiration, central carbon metabolism, fatty acid synthesis, phospholipid synthesis, and protein synthesis. Unexpectedly, the results indicate that multiple regulatory systems are involved in coordinating DNA replication with cell physiology, with some of the regulatory systems targeting oriC while others act in a oriC-independent manner. We propose that distinct regulatory systems are utilized to control DNA replication in response to diverse physiological and chemical changes.

  7. Multigene disruption in undomesticated Bacillus subtilis ATCC 6051a using the CRISPR/Cas9 system

    Science.gov (United States)

    Zhang, Kang; Duan, Xuguo; Wu, Jing

    2016-01-01

    Bacillus subtilis ATCC 6051a is an undomesticated strain used in the industrial production of enzymes. Because it is poorly transformable, genetic manipulation in this strain requires a highly efficient genome editing method. In this study, a Streptococcus pyogenes CRISPR/Cas9 system consisting of an all-in-one knockout plasmid containing a target-specific guide RNA, cas9, and a homologous repair template was established for highly efficient gene disruption in B. subtilis ATCC 6051a. With an efficiency of 33% to 53%, this system was used to disrupt the srfC, spoIIAC, nprE, aprE and amyE genes of B. subtilis ATCC 6051a, which hamper its use in industrial fermentation. Compared with B. subtilis ATCC 6051a, the final mutant, BS5 (ΔsrfC, ΔspoIIAC, ΔnprE, ΔaprE, ΔamyE), produces much less foam during fermentation, displays greater resistant to spore formation, and secretes 2.5-fold more β-cyclodextrin glycosyltransferase into the fermentation medium. Thus, the CRISPR/Cas9 system proved to be a powerful tool for targeted genome editing in an industrially relevant, poorly transformable strain. PMID:27305971

  8. Heme sensing in Bacillus thuringiensis: a supplementary HssRS-regulated heme resistance system.

    Science.gov (United States)

    Schmidt, Rachel M; Carter, Micaela M; Chu, Michelle L; Latario, Casey J; Stadler, Sarah K; Stauff, Devin L

    2016-05-01

    Several Gram-positive pathogens scavenge host-derived heme to satisfy their nutritional iron requirement. However, heme is a toxic molecule capable of damaging the bacterial cell. Gram-positive pathogens within the phylum Firmicutes overcome heme toxicity by sensing heme through HssRS, a two-component system that regulates the heme detoxification transporter HrtAB. Here we show that heme sensing by HssRS and heme detoxification by HrtAB occur in the insect pathogen Bacillus thuringiensis We find that in B. thuringiensis, HssRS directly regulates an operon, hrmXY, encoding hypothetical membrane proteins that are not found in other Firmicutes with characterized HssRS and HrtAB systems. This novel HssRS-regulated operon or its orthologs BMB171_c3178 and BMB171_c3330 are required for maximal heme resistance. Furthermore, the activity of HrmXY is not dependent on expression of HrtAB. These results suggest that B. thuringiensis senses heme through HssRS and induces expression of separate membrane-localized systems capable of overcoming different aspects of heme toxicity.

  9. Improving green roofs and rail road greening systems using Bacillus subtilis and Lactobacillus ssp.

    Science.gov (United States)

    Grüneberg, H; Oschmann, C; Dunya, S; Ulrichs, C

    2006-01-01

    Aim of the present study was the improvement of existing methods for green roof and rail road greening systems using soil borne bacteria. Bacillus subtilis and Lactobacillus ssp. alone and in combination with vinasse applied to different growing substrates were tested. The substrates were brick chips, textile mats, mineral wool mats, and a commercial available substrate for the Swedish company VegTech. All four substrates were tested along an artificial rail track on the experimental station at Humboldt University Berlin, and partly on an existing rail track in Munich, Germany. Plants selected for the experiments belong to the genus Sedum, which is relatively tolerant to dry conditions. Inoculation of plants with bacteria had no effect on plant growth parameters and on coverage of different mobile bedding systems with Sedum plants. There was no significant difference between the various treatments in Munich. In both experiments, the addition of vinasse alone improved plant growth. Plant growth was significantly different on all substrates, whereas brick chips and the commercial roof soil was the best substrate. Brick chips are a cheap substrate which can be used for rail track greening. The results indicate that the quality of the substrate is the most important factor for remediation and greening of rail tracks and roof tops. The rapid growth of plants can be influenced by the application of vinasse as additional nutrient solution (potash (K) source) or nutrient enriched substrate.

  10. The sponge-associated bacterium Bacillus licheniformis SAB1: A source of antimicrobial compounds

    Digital Repository Service at National Institute of Oceanography (India)

    PrabhaDevi; Wahidullah, S.; Rodrigues, C.; DeSouza, L.

    Skin infection 4. B4 Salmonella typhi Typhoid 5. B5 Shigella flexineri Gastrointestinal infection 6. B6 Klebsiella sp. Urinary tract infection 7. B7 Vibrio cholerae Cholera 8. F1 Fungal pathogens Aspergillus fumigatus Skin infection 9. F2... Rhodotorula sp. Skin infection 10. F3 Candida albicans Candidiasis 11. F4 Cryptococcus neoformans Skin infection 12. F5 Aspergillus niger Skin infection 13. D1 Multi- drug resistant bacteria Streptococcus pyogenes Skin infection 14. D2 Acinetobacter sp...

  11. Structural properties of hydrolyzed high-amylose rice starch by α-amylase from Bacillus licheniformis.

    Science.gov (United States)

    Qin, Fengling; Man, Jianmin; Xu, Bin; Hu, Maozhi; Gu, Minghong; Liu, Qiaoquan; Wei, Cunxu

    2011-12-14

    High-amylose cereal starch has a great benefit on human health through its resistant starch (RS) content. Enzyme hydrolysis of native starch is very helpful in understanding the structure of starch granules and utilizing them. In this paper, native starch granules were isolated from a transgenic rice line (TRS) enriched with amylose and RS and hydrolyzed by α-amylase. Structural properties of hydrolyzed TRS starches were studied by X-ray powder diffraction, Fourier transform infrared, and differential scanning calorimetry. The A-type polymorph of TRS C-type starch was hydrolyzed faster than the B-type polymorph, but the crystallinity did not significantly change during enzyme hydrolysis. The degree of order in the external region of starch granule increased with increasing enzyme hydrolysis time. The amylose content decreased at first and then went back up during enzyme hydrolysis. The hydrolyzed starches exhibited increased onset and peak gelatinization temperatures and decreased gelatinization enthalpy on hydrolysis. These results suggested that the B-type polymorph and high amylose that formed the double helices and amylose-lipid complex increased the resistance to BAA hydrolysis. Furthermore, the spectrum results of RS from TRS native starch digested by pancreatic α-amylase and amyloglucosidase also supported the above conclusion. PMID:22059442

  12. Cloning and characterization of a novel L-arabinose isomerase from Bacillus licheniformis

    DEFF Research Database (Denmark)

    Prabhu, Ponnandy; Tiwari, Manish Kumar; Jeya, Marimuthu;

    2008-01-01

    reading frame of 1,422 bp, capable of encoding a polypeptide of 474 amino acid residues with a calculated isoelectric point of pH 4.8 and a molecular mass of 53,500 Da. The gene was overexpressed in E. coli, and the protein was purified as an active soluble form using Ni-NTA chromatography. The molecular...

  13. Anti-biofilm activity of a polysaccharide from marine sponge associated Bacillus licheniformis

    OpenAIRE

    S. M. Abu, Sayem

    2011-01-01

    Secondary metabolites ranging from furanone to exo-polysaccharides have been suggested to have anti-biofilm activity in various recent studies. Among these,Escherichia coli group II capsular polysaccharides were shown to inhibit biofilm formation in a wide range of organisms and more recently marine Vibrio sp. and Kingella kingae were found to secrete complex exopolysaccharides having the potential for broad-spectrum biofilm inhibition and disruption. In this study, a ca. 1800 kDA polysacc...

  14. Anti-biofilm activity of an exopolysaccharide from a sponge-associated strain of Bacillus licheniformis

    OpenAIRE

    Cordone Angela; Tramice Annabella; Ciavatta Letizia; Manzo Emiliano; Sayem SM; Zanfardino Anna; De Felice Maurilio; Varcamonti Mario

    2011-01-01

    Abstract Background Secondary metabolites ranging from furanone to exo-polysaccharides have been suggested to have anti-biofilm activity in various recent studies. Among these, Escherichia coli group II capsular polysaccharides were shown to inhibit biofilm formation of a wide range of organisms and more recently marine Vibrio sp. were found to secrete complex exopolysaccharides having the potential for broad-spectrum biofilm inhibition and disruption. Results In this study we report that a n...

  15. Induction of Drought Stress Resistance by Multi-Functional PGPR Bacillus licheniformis K11 in Pepper

    OpenAIRE

    Lim, Jong-Hui; Kim, Sang-Dal

    2013-01-01

    Drought stress is one of the major yield affecting factor for pepper plant. The effects of PGPRs were analyzed in relation with drought resistance. The PGPRs inoculated pepper plants tolerate the drought stress and survived as compared to non-inoculated pepper plants that died after 15 days of drought stress. Variations in protein and RNA accumulation patterns of inoculated and non-inoculated pepper plants subjected to drought conditions for 10 days were confirmed by two dimensional polyacryl...

  16. A Type III protein-RNA toxin-antitoxin system from Bacillus thuringiensis promotes plasmid retention during spore development.

    Science.gov (United States)

    Short, Francesca L; Monson, Rita E; Salmond, George P C

    2015-01-01

    Members of the Bacillus cereus sensu lato group of bacteria often contain multiple large plasmids, including those encoding virulence factors in B. anthracis. Bacillus species can develop into spores in response to stress. During sporulation the genomic content of the cell is heavily compressed, which could result in counterselection of extrachromosomal genomic elements, unless they have robust stabilization and segregation systems. Toxin-antitoxin (TA) systems are near-ubiquitous in prokaryotes and have multiple biological roles, including plasmid stabilization during vegetative growth. Here, we have shown that a Type III TA system, based on an RNA antitoxin and endoribonuclease toxin, from plasmid pAW63 in Bacillus thuringiensis serovar kurstaki HD-73 can dramatically promote plasmid retention in populations undergoing sporulation and germination, and we provide evidence that this occurs through the post-segregational killing of plasmid-free forespores. Our findings show how an extremely common genetic module can be used to ensure plasmid maintenance during stress-induced developmental transitions, with implications for plasmid dynamics in B. cereus s.l. bacteria.

  17. Efficient production of artificially designed gelatins with a Bacillus brevis system.

    Science.gov (United States)

    Kajino, T; Takahashi, H; Hirai, M; Yamada, Y

    2000-01-01

    Artificially designed gelatins comprising tandemly repeated 30-amino-acid peptide units derived from human alphaI collagen were successfully produced with a Bacillus brevis system. The DNA encoding the peptide unit was synthesized by taking into consideration the codon usage of the host cells, but no clones having a tandemly repeated gene were obtained through the above-mentioned strategy. Minirepeat genes could be selected in vivo from a mixture of every possible sequence encoding an artificial gelatin by randomly ligating the mixed sequence unit and transforming it into Escherichia coli. Larger repeat genes constructed by connecting minirepeat genes obtained by in vivo selection were also stable in the expression host cells. Gelatins derived from the eight-unit and six-unit repeat genes were extracellularly produced at the level of 0.5 g/liter and easily purified by ammonium sulfate fractionation and anion-exchange chromatography. The purified artificial gelatins had the predicted N-terminal sequences and amino acid compositions and a solgel property similar to that of the native gelatin. These results suggest that the selection of a repeat unit sequence stable in an expression host is a shortcut for the efficient production of repetitive proteins and that it can conveniently be achieved by the in vivo selection method. This study revealed the possible industrial application of artificially designed repetitive proteins. PMID:10618240

  18. Occurrence of Bacillus amyloliquefaciens as a systemic endophyte of vanilla orchids.

    Science.gov (United States)

    White, James F; Torres, Mónica S; Sullivan, Raymond F; Jabbour, Rabih E; Chen, Qiang; Tadych, Mariusz; Irizarry, Ivelisse; Bergen, Marshall S; Havkin-Frenkel, Daphna; Belanger, Faith C

    2014-11-01

    We report the occurrence of Bacillus amyloliquefaciens in vanilla orchids (Vanilla phaeantha) and cultivated hybrid vanilla (V. planifolia × V. pompona) as a systemic bacterial endophyte. We determined with light microscopy and isolations that tissues of V. phaeantha and the cultivated hybrid were infected by a bacterial endophyte and that shoot meristems and stomatal areas of stems and leaves were densely colonized. We identified the endophyte as B. amyloliquefaciens using DNA sequence data. Since additional endophyte-free plants and seed of this orchid were not available, additional studies were performed on surrogate hosts Amaranthus caudatus, Ipomoea tricolor, and I. purpurea. Plants of A. caudatus inoculated with B. amyloliquefaciens demonstrated intracellular colonization of guard cells and other epidermal cells, confirming the pattern observed in the orchids. Isolations and histological studies suggest that the bacterium may penetrate deeply into developing plant tissues in shoot meristems, forming endospores in maturing tissues. B. amyloliquefaciens produced fungal inhibitors in culture. In controlled experiments using morning glory seedlings we showed that the bacterium promoted seedling growth and reduced seedling necrosis due to pathogens. We detected the gene for phosphopantetheinyl transferase (sfp), an enzyme in the pathway for production of antifungal lipopeptides, and purified the lipopeptide "surfactin" from cultures of the bacterium. We hypothesize that B. amyloliquefaciens is a robust endophyte and defensive mutualist of vanilla orchids. Whether the symbiosis between this bacterium and its hosts can be managed to protect vanilla crops from diseases is a question that should be evaluated in future research.

  19. Occurrence of Bacillus amyloliquefaciens as a systemic endophyte of vanilla orchids.

    Science.gov (United States)

    White, James F; Torres, Mónica S; Sullivan, Raymond F; Jabbour, Rabih E; Chen, Qiang; Tadych, Mariusz; Irizarry, Ivelisse; Bergen, Marshall S; Havkin-Frenkel, Daphna; Belanger, Faith C

    2014-11-01

    We report the occurrence of Bacillus amyloliquefaciens in vanilla orchids (Vanilla phaeantha) and cultivated hybrid vanilla (V. planifolia × V. pompona) as a systemic bacterial endophyte. We determined with light microscopy and isolations that tissues of V. phaeantha and the cultivated hybrid were infected by a bacterial endophyte and that shoot meristems and stomatal areas of stems and leaves were densely colonized. We identified the endophyte as B. amyloliquefaciens using DNA sequence data. Since additional endophyte-free plants and seed of this orchid were not available, additional studies were performed on surrogate hosts Amaranthus caudatus, Ipomoea tricolor, and I. purpurea. Plants of A. caudatus inoculated with B. amyloliquefaciens demonstrated intracellular colonization of guard cells and other epidermal cells, confirming the pattern observed in the orchids. Isolations and histological studies suggest that the bacterium may penetrate deeply into developing plant tissues in shoot meristems, forming endospores in maturing tissues. B. amyloliquefaciens produced fungal inhibitors in culture. In controlled experiments using morning glory seedlings we showed that the bacterium promoted seedling growth and reduced seedling necrosis due to pathogens. We detected the gene for phosphopantetheinyl transferase (sfp), an enzyme in the pathway for production of antifungal lipopeptides, and purified the lipopeptide "surfactin" from cultures of the bacterium. We hypothesize that B. amyloliquefaciens is a robust endophyte and defensive mutualist of vanilla orchids. Whether the symbiosis between this bacterium and its hosts can be managed to protect vanilla crops from diseases is a question that should be evaluated in future research. PMID:25060609

  20. A fail-safe system for the ribosome under zinc-limiting conditions in Bacillus subtilis.

    Science.gov (United States)

    Natori, Yousuke; Nanamiya, Hideaki; Akanuma, Genki; Kosono, Saori; Kudo, Toshiaki; Ochi, Kozo; Kawamura, Fujio

    2007-01-01

    As zinc is an essential trace metal ion for all living cells, cells elaborate a variety of strategies to cope with zinc starvation. In Bacillus subtilis, genes encoding ribosomal proteins L31 and S14 are duplicated into two types: one type contains a zinc-binding motif (RpmE or RpsN), whereas the other does not (YtiA or YhzA). We have previously shown that displacement of RpmE (L31) by YtiA from already assembled ribosomes is controlled by zinc, and this replacement could contribute to zinc mobilization under zinc-limiting conditions. We propose here that the switch between the two types of S14 has a different significance. rpsN is indispensable for growth and depletion of RpsN results in defective 30S subunits. YhzA can functionally replace RpsN to allow continued ribosome assembly under zinc-limiting conditions. Unlike YtiA, YhzA appeared in the ribosome at a slower rate consistent with incorporation into newly synthesized, rather than pre-existing ribosomes. These results raise the possibility that YhzA is involved in a fail-safe system for the de novo synthesis of ribosomes under zinc-limiting conditions.

  1. Interrelationships between Bacillus sp. CHEP5 and Bradyrhizobium sp. SEMIA6144 in the induced systemic resistance against Sclerotium rolfsii and symbiosis on peanut plants

    Indian Academy of Sciences (India)

    Marí­a Soledad Figueredo; María Laura Tonellie; Tania Taurian; Jorge Angelini; Fernando Ibañez; Lucio Valetti; Vanina Muñoz; Marí­a Soledad Anzuay; Liliana Ludueña; Adriana Fabra

    2014-12-01

    Plant-growth-promoting bacteria are often used to enhance crop yield and for biological control of phytopathogens. Bacillus sp. CHEP5 is a biocontrol agent that induces systemic resistance (ISR) in Arachis hypogaea L. (peanut) against Sclerotium rolfsii, the causal agent of root and stem wilt. In this work, the effect of the co-inoculation of Bacillus sp. CHEP5 and the peanut nodulating strain Bradyrhizobium sp. SEMIA 6144 was studied on induction of both systemic resistance and nodulation processes. Bradyrhizobium sp. SEMIA 6144 did not affect the ability of Bacillus sp. CHEP5 to protect peanut plants from S. rolfsii by ISR and the priming in challenged-plants, as evidenced by an increment in phenylalanine ammonia-lyase enzyme activity. Additionally, the capacity of Bradyrhizobium sp. SEMIA 6144 to induce nodule formation in pathogen-challenged plants was improved by the presence of Bacillus sp. CHEP5.

  2. The two-component signal transduction system YvcPQ regulates the bacterial resistance to bacitracin in Bacillus thuringiensis.

    Science.gov (United States)

    Zhang, Shumeng; Li, Xinfeng; Wang, Xun; Li, Zhou; He, Jin

    2016-10-01

    YvcPQ is one of the two-component signal transduction systems that respond to specific stimuli and enable cells to adjust multiple cellular functions. It consists of a histidine kinase YvcQ and a response regulator YvcP. In this study, through searching the consensus sequence recognized by YvcP, we found four YvcP-binding motifs in the promoter regions of genes yvcR (BMB171_C4100), BMB171_C4385, kapD (BMB171_C4525) and BMB171_C4835 in Bacillus thuringiensis BMB171 which is a representative of Bacillus cereus group, and confirmed that these genes are regulated by YvcP. We compared the sequence of yvcPQ and its downstream genes in genus Bacillus, and found two different kinds of yvc locus, one was the yvcPQ-RS in B. subtilis species and the other was the yvcPQ-R-S1S2 in B. cereus group. Furthermore, we found that YvcP activates the transcription of yvcS1S2 (downstream of yvcR) to promote bacterial resistance to bacitracin and deletion of either yvcPQ operon or yvcS1S2 operon renders the bacterial cells more sensitive to bacitracin. This study enriched our understanding of both the YvcPQ's function and the mechanism of bacterial resistance to bacitracin.

  3. Bioemulsifier production by a halothermophilic Bacillus strain with potential applications in microbially enhanced oil recovery.

    Science.gov (United States)

    Dastgheib, S M M; Amoozegar, M A; Elahi, E; Asad, S; Banat, I M

    2008-02-01

    A halothermotolerant Gram-positive spore-forming bacterium was isolated from petroleum reservoirs in Iran and identified as Bacillus licheniformis sp. strain ACO1 by phenotypic characterization and 16S rRNA analysis. It showed a high capacity for bioemulsifier production and grew up to 60 degrees C with NaCl at 180 g l(-1). The optimum NaCl concentration, pH and temperature for bioemulsifier production were 4% (w/v), 8.0, and 45 degrees C, respectively. Although ACO1 did not utilize hydrocarbons, it had a high emulsifying activity (E (24) = 65 +/- 5%) on different hydrophobic substrates. Emulsification was optimal while growing on yeast extract as the sole carbon source and NaNO(3) as the nitrogen source. The efficiency of the residual oil recovery increased by 22% after in situ growth of B. licheniformis ACO1 in a sand-pack model saturated with liquid paraffin. PMID:17876532

  4. Paradoxical DNA repair and peroxide resistance gene conservation in Bacillus pumilus SAFR-032.

    Directory of Open Access Journals (Sweden)

    Jason Gioia

    Full Text Available BACKGROUND: Bacillus spores are notoriously resistant to unfavorable conditions such as UV radiation, gamma-radiation, H2O2, desiccation, chemical disinfection, or starvation. Bacillus pumilus SAFR-032 survives standard decontamination procedures of the Jet Propulsion Lab spacecraft assembly facility, and both spores and vegetative cells of this strain exhibit elevated resistance to UV radiation and H2O2 compared to other Bacillus species. PRINCIPAL FINDINGS: The genome of B. pumilus SAFR-032 was sequenced and annotated. Lists of genes relevant to DNA repair and the oxidative stress response were generated and compared to B. subtilis and B. licheniformis. Differences in conservation of genes, gene order, and protein sequences are highlighted because they potentially explain the extreme resistance phenotype of B. pumilus. The B. pumilus genome includes genes not found in B. subtilis or B. licheniformis and conserved genes with sequence divergence, but paradoxically lacks several genes that function in UV or H2O2 resistance in other Bacillus species. SIGNIFICANCE: This study identifies several candidate genes for further research into UV and H2O2 resistance. These findings will help explain the resistance of B. pumilus and are applicable to understanding sterilization survival strategies of microbes.

  5. Contamination profiles and characterisation of Bacillus species in wheat bread and raw materials for bread production.

    Science.gov (United States)

    Rosenkvist, H; Hansen, A

    1995-08-01

    The Bacillus counts in white and wholemeal wheat loaves produced without preservatives or sour dough were consistently 10(6) cfu/g after two days of storage at ambient summer temperatures (25-30 degree C). Identified species were B. subtilis (70%), B. licheniformis (24%), B. pumilus (2%) and B. cereus (2%). The dominance of B. subtilis in bread could be explained by the higher resistance to heat of this species as determined by inoculation studies. Among 14 species isolated from retail bread and wheat grains, B. subtilis was the only species associated with ropiness. Samples of raw materials, particularly bran, seeds and oat products, contained low levels (10(0) - 10(2) cfu/g) of Bacillus spores, surviving a heat treatment (100 degree C, 10 min) corresponding to a baking process. Even low spore levels in raw materials with the frequently isolated species, B. licheniformis (49%) and B. subtilis (10%), resulted in 10(7) Bacillus per g bread crumb in two days as determined by test bakings. The results indicate a need for controlling growth of Bacillus in bread.

  6. Challenges and advances in systems biology analysis of Bacillus spore physiology; molecular differences between an extreme heat resistant spore forming Bacillus subtilis food isolate and a laboratory strain

    NARCIS (Netherlands)

    Brul, S.; Beilen, J.van; Caspers, M.; O'Brien, A.; Koster, C.de; Oomes, S.; Smelt, J.; Kort, R.; Beek, A.ter

    2011-01-01

    Bacterial spore formers are prime organisms of concern in the food industry. Spores from the genus Bacillus are extremely stress resistant, most notably exemplified by high thermotolerance. This sometimes allows surviving spores to germinate and grow out to vegetative cells causing food spoilage and

  7. The ComP-ComA quorum system is essential for "Trojan horse" like pathogenesis in Bacillus nematocida.

    Directory of Open Access Journals (Sweden)

    Xidan Deng

    Full Text Available Bacillus nematocida B16 has been shown to use "Trojan horse" mechanism in pathogenesis that has characteristics of "social" behavior. The ComP-ComA system, a conserved quorum sensing system in the genus Bacillus, functions in many physiological processes including competence development, lipopeptide antibiotic surfactin production, degradative enzyme production and even some unknown functions. Here we investigated the requirement of ComP-ComA system in B. nematocida B16 for its pathogenicity against nematodes. The ΔcomP mutant displayed deficiencies in attracting and killing nematodes, due to the absence of attractive signal molecules and the decreased expressions of virulence factors, respectively. Contrarily, a complemented comP mutant at least partially resumed its pathogenicity. Our data from transcriptional analysis further confirmed that this signaling system directly or indirectly regulated the expressions of two major virulence proteases in the infection of B. nematocida B16. Bioinformatics analyses from comparative genomics also suggested that the potential target genes of transcription factor ComA were involved in the processes such as the synthesis of attractants, production of extracellular degradative enzymes and sortase, secondary metabolites biosynthesis, regulation of transcription factors, mobility, as well as transporters, most of which were different from a saprophytic relative B. subtilis 168. Therefore, our investigation firstly revealed that the participation and necessity of ComP-ComA signaling system in bacterial pathogenesis.

  8. Removal of ammonium-N from ammonium-rich sewage using an immobilized Bacillus subtilis AYC bioreactor system

    Institute of Scientific and Technical Information of China (English)

    Jingjing Xiao; Changxiong Zhu; Dongyuan Sun; Ping Guo; Yunlong Tian

    2011-01-01

    A self-design bioreactor system employing a fixed bed operation process with immobilized Bacillus subtilis AYC beads for NH4+-N removal from slightly polluted water was proposed.Polyvinyl alcohol and Na-alginate were used as a gel matrix to entrap Bacillus subtilis AYC to form the immobilized beads.The NH4+-N removal process was studied in a intermittent operation mode to examine the start-up and steady state behaviors of the immobilized AYC in the reactor.The results indicated that the reactor was in the start-up state during the first week.NH4+-N began to be steadily removal since the second week,and the nitrogen removal rate was between 84.61% and 96.19% when the hydraulic retention time (HRT) was 30 min.To apply Bacillus subtilis AYC to develop a practical nitrogen removal system and further understand its nitrogen removal ability,the bioreactor was continuously operated under different experimental perameters.The results showed that under the optimum conditions of an HRT of 20 min and DO of 3.77-5.80 mg/L,the NH4+-N removal rate reached 99.55%.The NH4+-N removal rate increased as the C/N ratio increased.However,a high C/N may cause a high residual carbon level in the effluent,therefore,the most suitable C/N ratio was 10.In addition,the results showed that the bioreactor system could remove many types of nitrogen such as NH4+-N,NO3--N and organic-N,and had a good performance for inorganic nitrogen removal from sewage.

  9. Bacillus subtilis Two-Component System Sensory Kinase DegS Is Regulated by Serine Phosphorylation in Its Input Domain

    DEFF Research Database (Denmark)

    Jers, Carsten; Kobir, Ahasanul; Søndergaard, Elsebeth Oline;

    2011-01-01

    Bacillus subtilis two-component system DegS/U is well known for the complexity of its regulation. The cytosolic sensory kinase DegS does not receive a single predominant input signal like most two-component kinases, instead it integrates a wide array of metabolic inputs that modulate its activity...... demonstrate that DegS phosphorylation can be carried out by at least two B. subtilis Hanks-type kinases in vitro, and this stimulates the phosphate transfer towards DegU. The consequences of this process were studied in vivo, using phosphomimetic (Ser76Asp) and non-phosphorylatable (Ser76Ala) mutants of Deg...

  10. Microbial System for Polysaccharide Depolymerization: Enzymatic Route for Xanthan Depolymerization by Bacillus sp. Strain GL1

    OpenAIRE

    Nankai, Hirokazu; Hashimoto, Wataru; Miki, Hikaru; Kawai, Shigeyuki; Murata, Kousaku

    1999-01-01

    An enzymatic route for the depolymerization of a heteropolysaccharide (xanthan) in Bacillus sp. strain GL1, which was closely related to Brevibacillus thermoruber, was determined by analyzing the structures of xanthan depolymerization products. The bacterium produces extracellular xanthan lyase catalyzing the cleavage of the glycosidic bond between pyruvylated mannosyl and glucuronyl residues in xanthan side chains (W. Hashimoto et al., Appl. Environ. Microbiol. 64:3765–3768, 1998). The modif...

  11. Thermodynamics and Extra-thermodynamics of Bacillus subtilis α-Amylase in Some Chromatographic Systems

    Institute of Scientific and Technical Information of China (English)

    BIAN,Liujiao; WANG,Xiaohua; YANG,Xiaoyan; LIU,Li; CHEN,Chao

    2009-01-01

    The thermodynamics and extra-thermodynamics of Bacillus subtilis a-amylase in five kinds of chromatographic systems were studied by determining their adsorption equilibrium constant KsL,enthalpy change,entropy change,free energy change and compensation temperature under different temperature areas.The results showed that on anRP.C18 reversed-phase medium,a Zn-chelated Sepharose fast-flow affinity medium and a WCX-l cation-exchange medium,their In KsL separately linearly changed with the reciprocal of absolute temperatures under two different temperature ranges of 13-30 and 30-50℃:and on PEG-400 and modified PEG-400 hydrophobic media,their ln KsL separately linearly decreased with the reciprocal of absolute temperatures under the temperature areas of 13-40 and 13-30℃.while when the temperatures were separately over 40 and 30℃,they violently decreased with the reciprocal of absolute temperatures.Through studying the relations among the enthalpy change,entropy change.free energy change and the conformational change of a-amylase under the different temperature areas,it was found that on the RP-C18 reversed-phase medium and Zn-chelated Sepharose fast-flow affinity medium under the temperature range of 30-50℃and on the WCX-l cation-exchange medium under the temperature range of 13-30℃.their adsorption procedures were driven by both the enthalpy change and the entropy change;while on the Zn-chelated Sepharose fast-flow affinity medium under the temperature range of 13-30℃.on the WCX-1cation.exchange medium under the temperature range of 30-50℃and on the PEG-400 and modified PEG-400 hydrophobic media under the temperature range of 13-65℃.their adsorption procedures were driven only by the entropy change.Finally, by comparing the compensation temperatures of a-amylase in these chromatographic sys-terns each other,it Was further found that their enthalpy changes could be compensated only with their entropy changes rising from their conformational change.

  12. Siderophore-mediated iron acquisition systems in Bacillus cereus: Identification of receptors for anthrax virulence-associated petrobactin .

    Science.gov (United States)

    Zawadzka, Anna M; Abergel, Rebecca J; Nichiporuk, Rita; Andersen, Ulla N; Raymond, Kenneth N

    2009-04-28

    During growth under iron limitation, Bacillus cereus and Bacillus anthracis, two human pathogens from the Bacillus cereus group of Gram-positive bacteria, secrete two siderophores, bacillibactin (BB) and petrobactin (PB), for iron acquisition via membrane-associated substrate-binding proteins (SBPs) and other ABC transporter components. Since PB is associated with virulence traits in B. anthracis, the PB-mediated iron uptake system presents a potential target for antimicrobial therapies; its characterization in B. cereus is described here. Separate transporters for BB, PB, and several xenosiderophores are suggested by (55)Fe-siderophore uptake studies. The PB precursor, 3,4-dihydroxybenzoic acid (3,4-DHB), and the photoproduct of FePB (FePB(nu)) also mediate iron delivery into iron-deprived cells. Putative SBPs were recombinantly expressed, and their ligand specificity and binding affinity were assessed using fluorescence spectroscopy. The noncovalent complexes of the SBPs with their respective siderophores were characterized using ESI-MS. The differences between solution phase behavior and gas phase measurements are indicative of noncovalent interactions between the siderophores and the binding sites of their respective SBPs. These studies combined with bioinformatics sequence comparison identify SBPs from five putative transporters specific for BB and enterobactin (FeuA), 3,4-DHB and PB (FatB), PB (FpuA), schizokinen (YfiY), and desferrioxamine and ferrichrome (YxeB). The two PB receptors show different substrate ranges: FatB has the highest affinity for ferric 3,4-DHB, iron-free PB, FePB, and FePB(nu), whereas FpuA is specific to only apo- and ferric PB. The biochemical characterization of these SBPs provides the first identification of the transporter candidates that most likely play a role in the B. cereus group pathogenicity.

  13. Siderophore-mediated iron acquisition systems in Bacillus cereus: identification of receptors for anthrax virulence-associated petrobactin†a

    Science.gov (United States)

    Zawadzka, Anna M.; Abergel, Rebecca J.; Nichiporuk, Rita; Andersen, Ulla N.; Raymond, Kenneth N.

    2009-01-01

    During growth under iron limitation, Bacillus cereus and Bacillus anthracis, two human pathogens from the Bacillus cereus group of Gram-positive bacteria, secrete two siderophores, bacillibactin (BB) and petrobactin (PB), for iron acquisition via membrane-associated substrate-binding proteins (SBPs) and other ABC transporter components. Since PB is associated with virulence traits in B. anthracis, the PB-mediated iron uptake system presents a potential target for antimicrobial therapies; its characterization in B. cereus is described here. Separate transporters for BB, PB, and several xenosiderophores are suggested by 55Fe-siderophore uptake studies. The PB precursor, 3,4-dihydroxybenzoic acid (3,4-DHB), and the photoproduct of FePB (FePBν) also mediate iron delivery into iron-deprived cells. Putative SBPs were recombinantly expressed, and their ligand specificity and binding affinity assessed using fluorescence spectroscopy. The noncovalent complexes of the SBPs with their respective siderophores were characterized using ESI-MS. The differences between solution phase behavior and gas phase measurements are indicative of noncovalent interactions between the siderophores and the binding sites of their respective SBPs. These studies combined with bioinformatics sequence comparison identify SBPs from five putative transporters specific for BB and enterobactin (FeuA), 3,4-DHB and PB (FatB), PB (FpuA), schizokinen (YfiY), and desferrioxamine and ferrichrome (YxeB). The two PB receptors show different substrate ranges: FatB has the highest affinity for ferric 3,4-DHB, iron-free PB, FePB, and FePBν, whereas FpuA is specific to only apo- and ferric PB. The biochemical characterization of these SBPs provides the first identification of the transporter candidates that most likely play a role in the B. cereus group pathogenicity. PMID:19254027

  14. In vivo and in vitro digestibility of plant ingredients and diets by Bacillus phytases in tilapia, Oreochromis mossambicus

    Directory of Open Access Journals (Sweden)

    Rande B. Dechavez

    2012-12-01

    Full Text Available This study aimed to evaluate four Bacillus phytases for their efficacy in making plant-baseddiets bioavailable to tilapia (Oreochromis mossambicus using in vivo digestibility measurement and todetermine the in vitro level of dephosphorylation. The four Bacillus strains used were B. pumilus , B. megaterium , B. coagulans, and B. licheniformis. Phytase activities varied between bacterial sources aswell as between feed ingredients. For the cassava leaf meal, Pi released was highest in B. pumilus andwas not significantly different from those of B. megaterium and B. licheniformis. For the soybean meal, Pirelease was in this decreasing order: B. megaterium > B. pumilus > B. coagulans > B. licheniformisphytase. For the corn meal, addition of B. licheniformis phytase to the reaction mixture resulted insignificantly the highest Pi released followed by B. coagulans phytase which was not significantly differentfrom that of B. megaterium phytase which released the lowest Pi. Pi released by B. pumilus phytase fromcorn meal was not significantly different from the lowest Pi release of B. megaterium phytase. Theapparent digestibility coefficient (ADC values for the feed dry matter (DM ranged from 86.3 to 88.3%and were not significantly different from each other (p > 0.05.

  15. Maintenance metabolism and carbon fluxes in Bacillus species

    Directory of Open Access Journals (Sweden)

    Decasper Seraina

    2008-06-01

    Full Text Available Abstract Background Selection of an appropriate host organism is crucial for the economic success of biotechnological processes. A generally important selection criterion is a low maintenance energy metabolism to reduce non-productive consumption of substrate. We here investigated, whether various bacilli that are closely related to Bacillus subtilis are potential riboflavin production hosts with low maintenance metabolism. Results While B. subtilis exhibited indeed the highest maintenance energy coefficient, B. licheniformis and B. amyloliquefaciens exhibited only statistically insignificantly reduced maintenance metabolism. Both B. pumilus and B. subtilis (natto exhibited irregular growth patterns under glucose limitation such that the maintenance metabolism could not be determined. The sole exception with significantly reduced maintenance energy requirements was the B. licheniformis strain T380B. The frequently used spo0A mutation significantly increased the maintenance metabolism of B. subtilis. At the level of 13C-detected intracellular fluxes, all investigated bacilli exhibited a significant flux through the pentose phosphate pathway, a prerequisite for efficient riboflavin production. Different from all other species, B. subtilis featured high respiratory tricarboxylic acid cycle fluxes in batch and chemostat cultures. In particular under glucose-limited conditions, this led to significant excess formation of NADPH of B. subtilis, while anabolic consumption was rather balanced with catabolic NADPH formation in the other bacilli. Conclusion Despite its successful commercial production of riboflavin, B. subtilis does not seem to be the optimal cell factory from a bioenergetic point of view. The best choice of the investigated strains is the sporulation-deficient B. licheniformis T380B strain. Beside a low maintenance energy coefficient, this strain grows robustly under different conditions and exhibits only moderate acetate overflow, hence

  16. Pitting corrosion inhibition of aluminum 2024 by Bacillus biofilms secreting polyaspartate or gamma-polyglutamate.

    Science.gov (United States)

    Ornek, D; Jayaraman, A; Syrett, B C; Hsu, C-H; Mansfeld, F B; Wood, T K

    2002-04-01

    Pitting corrosion of aluminum 2024 in Luria Bertani medium was reduced by the secretion of anionic peptides by engineered and natural Bacillus biofilms and was studied in continuous reactors using electrochemical impedance spectroscopy. Compared to sterile controls, pitting was reduced dramatically by the presence of the biofilms. The secretion of a 20 amino acid polyaspartate peptide by an engineered Bacillus subtilis WB600/pBE92-Asp biofilm slightly reduced the corrosion rate of the passive aluminum alloy at pH 6.5; however, the secretion of gamma-polyglutamate by a Bacillus licheniformis biofilm reduced the corrosion rate by 90% (compared to the B. subtilis WB600/pBE92 biofilm which did not secrete polyaspartate or gamma-polyglutamate). The corrosion potential ( E(corr)) of aluminum 2024 was increased by about 0.15-0.44 V due to the formation of B. subtilis and B. licheniformis biofilms as compared to sterile controls. The increase of E(corr) and the observed prevention of pitting indicate that the pitting potential ( E(pit)) had increased. This result and the further decrease of corrosion rates for the passive aluminum alloy suggest that the rate of the anodic metal dissolution reaction was reduced by an inhibitor produced by the biofilms. Purified gamma-polyglutamate also decreased the corrosion rates of aluminum 2024. PMID:11956749

  17. Application of nisin and pediocin against resistance and germination of Bacillus spores in sous vide products.

    Science.gov (United States)

    Cabo, M L; Torres, B; Herrera, J J R; Bernárdez, M; Pastoriza, L

    2009-03-01

    Sous vide and other mild preservation techniques are increasingly demanded by consumers. However, spores often will survive in minimally processed foods, causing both spoilage and safety problems. The main objective of the present work was to solve an industrial spoilage problem associated with two sous vide products: mushrooms and shellfish salad. Bacillus subtilis and Bacillus licheniformis predominated as the most heat-resistant organisms isolated from mushrooms and shellfish salad, respectively. The combined effects of nisin and pediocin against resistance and germination of both Bacillus species were described by empirical equations. Whereas nisin was more effective for decreasing thermal resistance of B. subtilis spores, pediocin was more effective against B. licheniformis. However, a significant positive interaction between both biopeptides for decreasing the proportion of vegetative cells resulting from thermoresistant spores was demonstrated in later experiments, thus indicating the increased efficacy of applying high concentrations of both bacteriocins. This efficacy was further demonstrated in additional challenge studies carried out at 15 degrees C in the two sous vide products: mushrooms and shellfish salad. Whereas no vegetative cells were detected after 90 days in the presence of bacteriocins, almost 100% of the population in nontreated samples of mushrooms and shellfish salad was in the vegetative state after 17 and 43 days of storage at 15 degrees C, respectively. PMID:19343939

  18. Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

    Directory of Open Access Journals (Sweden)

    Eu Jin Chung

    2015-06-01

    Full Text Available Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and YC7010T, with anti-microbial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208 and Burkholderia glumae (KACC 44022, respectively, were also suppressed effectively by drenching a bacterial suspension (10⁷ cfu/ml of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC 15859T (99.67%, Bacillus methylotrophicus KACC 13105T (99.65%, Bacillus amyloliquefaciens subsp. plantarum KACC 17177T (99.60%, and Bacillus tequilensis KACC 15944T (99.45%. The DNA-DNA relatedness value between strain YC7010T and the most closely related strain, B. siamensis KACC 15859T was 50.4±3.5%, but it was 91.5±11.0% between two strains YC7007 and YC7010T, indicating the same species. The major fatty acids of two strains were anteiso-C15:0 and iso C15:0. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the

  19. Engineering the Xylan Utilization System in Bacillus subtilis for Production of Acidic Xylooligosaccharides

    OpenAIRE

    Rhee, Mun Su; Wei, Lusha; Sawhney, Neha; Rice, John D.; St John, Franz J.; Hurlbert, Jason C.; Preston, James F.

    2014-01-01

    Xylans are the predominant polysaccharides in hemicelluloses and an important potential source of biofuels and chemicals. The ability of Bacillus subtilis subsp. subtilis strain 168 to utilize xylans has been ascribed to secreted glycoside hydrolase family 11 (GH11) and GH30 endoxylanases, encoded by the xynA and xynC genes, respectively. Both of these enzymes have been defined with respect to structure and function. In this study, the effects of deletion of the xynA and xynC genes, individua...

  20. RAZOR EX Anthrax Air Detection System for detection of Bacillus anthracis spores from aerosol collection samples: collaborative study.

    Science.gov (United States)

    Hadfield, Ted; Ryan, Valorie; Spaulding, Usha K; Clemens, Kristine M; Ota, Irene M; Brunelle, Sharon L

    2013-01-01

    The RAZOR EX Anthrax Air Detection System was validated in a collaborative study for the detection of Bacillus anthracis in aerosol collection buffer. Phosphate-buffered saline was charged with 1 mg/mL standardized dust to simulate an authentic aerosol collection sample. The dust-charged buffer was spiked with either B. anthracis Ames at 2000 spores/mL or Bacillus cereus at 20 000 spores/mL. Twelve collaborators participated in the study, with four collaborators at each of three sites. Each collaborator tested 12 replicates of B. anthracis in dust-charged buffer and 12 replicates of B. cereus in dust-charged buffer. All samples sets were randomized and blind-coded. All collaborators produced valid data sets (no collaborators displayed systematic errors) and there was only one invalid data point. After unblinding, the analysis revealed a cross-collaborator probability of detection (CPOD) of 1.00 (144 positive results from 144 replicates, 95% confidence interval 0.975-1.00) for the B. anthracis samples and a CPOD of 0.00 (0 positive results from 143 replicates, 95% confidence interval 0.00-0.0262) for the B. cereus samples. These data meet the requirements of AOAC Standard Method Performance Requirement 2010.003, developed by the Stakeholder Panel on Agent Detection Assays. PMID:23767365

  1. Control of Anthracnose Caused by Colletotrichum musae on Curcuma alismatifolia Gagnep. Using Antagonistic Bacillus spp.

    Directory of Open Access Journals (Sweden)

    Supuk Mahadtanapuk

    2007-01-01

    Full Text Available Over 400 bacterial strains, isolated from leaf surfaces of Curcuma alismatifolia Gagnep. and hot springs in the Chiang Mai province of northern Thailand, were screened in vitro for antagonistic activity against Colletotrichum musae, an anthracnose fungus. Three isolates provided greater than 75% growth inhibition of the fungus in vitro and were identified as Bacillus licheniformis, B. amyloliquefaciens and B. subtilis. Using in planta tests, B. amyloliquefaciens and B. subtilis were shown to efficiently colonize the curcuma bracts, provide a statistically significant growth suppression of C. musae over that of B. licheniformis, and all three isolates could provide 100% inhibition of conidial fungal germination. When B. licheniformis was co-inoculated in combination with either of the other two bacteria, the ability of B. amyloliquefaciens and B. subtilis to suppress the fungal disease was dramatically reduced. Both B. amyloliquefaciens and B. subtilis were found to contain an isoform of iturin A with antifungal activity against C. musae. As a preventative measure to control the spread of C. musae and reduce the severity of fungal infections, B. amyloliquefaciens could be used to inoculate curcuma flowers cost effectively and reduce the need for the toxic synthetic fungicides currently in use.

  2. Assessment of the Bacteriocinogenic Potential of Marine Bacteria Reveals Lichenicidin Production by Seaweed-Derived Bacillus spp.

    Directory of Open Access Journals (Sweden)

    Gillian E. Gardiner

    2012-10-01

    Full Text Available The objectives of this study were (1 to assess the bacteriocinogenic potential of bacteria derived mainly from seaweed, but also sand and seawater, (2 to identify at least some of the bacteriocins produced, if any and (3 to determine if they are unique to the marine environment and/or novel. Fifteen Bacillus licheniformis or pumilus isolates with antimicrobial activity against at least one of the indicator bacteria used were recovered. Some, at least, of the antimicrobials produced were bacteriocins, as they were proteinaceous and the producers displayed immunity. Screening with PCR primers for known Bacillus bacteriocins revealed that three seaweed-derived Bacillus licheniformis harbored the bli04127 gene which encodes one of the peptides of the two-peptide lantibiotic lichenicidin. Production of both lichenicidin peptides was then confirmed by mass spectrometry. This is the first definitive proof of bacteriocin production by seaweed-derived bacteria. The authors acknowledge that the bacteriocin produced has previously been discovered and is not unique to the marine environment. However, the other marine isolates likely produce novel bacteriocins, as none harboured genes for known Bacillus bacteriocins.

  3. Cooperation and the evolutionary ecology of bacterial virulence: the Bacillus cereus group as a novel study system.

    Science.gov (United States)

    Raymond, Ben; Bonsall, Michael B

    2013-08-01

    How significant is social evolution theory for the maintenance of virulence in natural populations? We assume that secreted, distantly acting virulence factors are highly likely to be cooperative public goods. Using this assumption, we discuss and critically assess the potential importance of social interactions for understanding the evolution, diversity and distribution of virulence in the Bacillus cereus group, a novel study system for microbial social biology. We conclude that dynamic equilibria in Cry toxin production, as well as strong spatial structure and population bottlenecks in hosts are the main ecological factors maintaining the cooperative secretion of virulence factors and argue that collective action has contributed to the evolution of narrow host range. Non-linearities in the benefits associated with public goods, as well as the lack of private secretion systems in the Firmicutes may also explain the prevalence and importance of distantly acting virulence factors in B. cereus and its relatives.

  4. Bacillus anthracis

    OpenAIRE

    2003-01-01

    The events of 11 September 2001 and the subsequent anthrax outbreaks have shown that the West needs to be prepared for an increasing number of terrorist attacks, which may include the use of biological warfare. Bacillus anthracis has long been considered a potential biological warfare agent, and this review will discuss the history of its use as such. It will also cover the biology of this organism and the clinical features of the three disease forms that it can produce: cutaneous, gastrointe...

  5. Bacillus anthracis

    OpenAIRE

    BOSERET, GÉRALDINE; Linden, Annick; Mainil, Jacques

    2002-01-01

    The literature describes several methods for detection of Bacillus anthracis based on application of specific bacteriophages. The following methods of pahoinpitely are used to identify the causative agent of anthrax: the reaction of bacteriophage titer growth (RBTG), the reaction of phage adsorption (RPA), fagoterapii method (FTM) and fluorescentserological method (FSM). The essence of RBTG consists in the following: if there is the researchform of bacteria presents in the test material, then...

  6. Identification of Bacillus spp. colonizing the nasal mucosa of healthy adults living in the suburban area using the matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS system

    Directory of Open Access Journals (Sweden)

    Kosikowska Urszula

    2014-09-01

    Full Text Available Bacillus spp. can be regarded as a rare component of the nasal mucosa microflora. The aim of this study was to identify Bacillus spp. from the nasal mucosa of healthy adults living in the suburban area near Lublin using the matrix-assisted laser desorptionionization time-of-flight mass spectrometry (MALDI-TOF MS system.

  7. Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates.

    Science.gov (United States)

    Celandroni, Francesco; Salvetti, Sara; Gueye, Sokhna Aissatou; Mazzantini, Diletta; Lupetti, Antonella; Senesi, Sonia; Ghelardi, Emilia

    2016-01-01

    The soil-related Bacillus and Paenibacillus species have increasingly been implicated in various human diseases. Nevertheless, their identification still poses problems in the clinical microbiology laboratory and, with the exception of Bacillus anthracis and Bacillus cereus, little is known on their pathogenicity for humans. In this study, we evaluated the use of matrix-assisted laser desorption-ionization time of flight mass spectrometry (MALDI-TOF MS) in the identification of clinical isolates of these genera and conducted genotypic and phenotypic analyses to highlight specific virulence properties. Seventy-five clinical isolates were subjected to biochemical and MALDI-TOF MS identification. 16S rDNA sequencing and supplemental tests were used to solve any discrepancies or failures in the identification results. MALDI-TOF MS significantly outperformed classical biochemical testing for correct species identification and no misidentification was obtained. One third of the collected strains belonged to the B. cereus species, but also Bacillus pumilus and Bacillus subtilis were isolated at high rate. Antimicrobial susceptibility testing showed that all the B. cereus, B. licheniformis, B. simplex, B. mycoides, Paenibacillus glucanolyticus and Paenibacillus lautus isolates are resistant to penicillin. The evaluation of toxin/enzyme secretion, toxin-encoding genes, motility, and biofilm formation revealed that B. cereus displays the highest virulence potential. However, although generally considered nonpathogenic, most of the other species were shown to swim, swarm, produce biofilms, and secrete proteases that can have a role in bacterial virulence. In conclusion, MALDI-TOF MS appears useful for fast and accurate identification of Bacillus and Paenibacillus strains whose virulence properties make them of increasing clinical relevance.

  8. Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates.

    Directory of Open Access Journals (Sweden)

    Francesco Celandroni

    Full Text Available The soil-related Bacillus and Paenibacillus species have increasingly been implicated in various human diseases. Nevertheless, their identification still poses problems in the clinical microbiology laboratory and, with the exception of Bacillus anthracis and Bacillus cereus, little is known on their pathogenicity for humans. In this study, we evaluated the use of matrix-assisted laser desorption-ionization time of flight mass spectrometry (MALDI-TOF MS in the identification of clinical isolates of these genera and conducted genotypic and phenotypic analyses to highlight specific virulence properties. Seventy-five clinical isolates were subjected to biochemical and MALDI-TOF MS identification. 16S rDNA sequencing and supplemental tests were used to solve any discrepancies or failures in the identification results. MALDI-TOF MS significantly outperformed classical biochemical testing for correct species identification and no misidentification was obtained. One third of the collected strains belonged to the B. cereus species, but also Bacillus pumilus and Bacillus subtilis were isolated at high rate. Antimicrobial susceptibility testing showed that all the B. cereus, B. licheniformis, B. simplex, B. mycoides, Paenibacillus glucanolyticus and Paenibacillus lautus isolates are resistant to penicillin. The evaluation of toxin/enzyme secretion, toxin-encoding genes, motility, and biofilm formation revealed that B. cereus displays the highest virulence potential. However, although generally considered nonpathogenic, most of the other species were shown to swim, swarm, produce biofilms, and secrete proteases that can have a role in bacterial virulence. In conclusion, MALDI-TOF MS appears useful for fast and accurate identification of Bacillus and Paenibacillus strains whose virulence properties make them of increasing clinical relevance.

  9. Identification and safety evaluation of Bacillus species occurring in high numbers during spontaneous fermentations to produce Gergoush, a traditional Sudanese bread snack

    DEFF Research Database (Denmark)

    Thorsen, Line; Abdelgadir, Warda S.; Rønsbo, Mie Hvillum;

    2011-01-01

    Gergoush is a naturally fermented Sudanese Bread snack produced in three fermentation steps (primary starter, adapted starter and final dough), followed by three baking steps for a half to one hour at above 200°C. This study examines the microbiota of two sets of fermentations performed at a trad......Gergoush is a naturally fermented Sudanese Bread snack produced in three fermentation steps (primary starter, adapted starter and final dough), followed by three baking steps for a half to one hour at above 200°C. This study examines the microbiota of two sets of fermentations performed....... cereus sensu lato, 16-27% as Bacillus licheniformis, 8-32% as Bacillus subtilis and 4-20% as Bacillus sonorensis. During the second set of fermentation trials performed in 2009, the Bacillus spp. and B. cereus occurred in numbers of between 7.7-9.9 and 6.1-7.8 log(10) CFU/g, respectively, while...

  10. Phylogeny in aid of the present and novel microbial lineages: diversity in Bacillus.

    Directory of Open Access Journals (Sweden)

    Shalini Porwal

    Full Text Available Bacillus represents microbes of high economic, medical and biodefense importance. Bacillus strain identification based on 16S rRNA sequence analyses is invariably limited to species level. Secondly, certain discrepancies exist in the segregation of Bacillus subtilis strains. In the RDP/NCBI databases, out of a total of 2611 individual 16S rDNA sequences belonging to the 175 different species of the genus Bacillus, only 1586 have been identified up to species level. 16S rRNA sequences of Bacillus anthracis (153 strains, B. cereus (211 strains, B. thuringiensis (108 strains, B. subtilis (271 strains, B. licheniformis (131 strains, B. pumilus (83 strains, B. megaterium (47 strains, B. sphaericus (42 strains, B. clausii (39 strains and B. halodurans (36 strains were considered for generating species-specific framework and probes as tools for their rapid identification. Phylogenetic segregation of 1121, 16S rDNA sequences of 10 different Bacillus species in to 89 clusters enabled us to develop a phylogenetic frame work of 34 representative sequences. Using this phylogenetic framework, 305 out of 1025, 16S rDNA sequences presently classified as Bacillus sp. could be identified up to species level. This identification was supported by 20 to 30 nucleotides long signature sequences and in silico restriction enzyme analysis specific to the 10 Bacillus species. This integrated approach resulted in identifying around 30% of Bacillus sp. up to species level and revealed that B. subtilis strains can be segregated into two phylogenetically distinct groups, such that one of them may be renamed.

  11. Characterization of Micrococcus luteus and Bacillus marisflavi Recovered from Common Dentex (Dentex dentex Larviculture System

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    T. AKAYLI

    2014-06-01

    Full Text Available In this manuscript, thirty yellow-pigmented Gram-positive bacteria were isolated from natural intestine microflora and from sea water around the marine cage of a rearing tank of common dentex (Dentex dentex, in the Aegean Sea on the Turkish coast and were characterized. Eighteen isolates were assigned to the species Micrococcus luteus, the other twelve to the species Bacillus marisflavi. Eight representative strains, six from B. marisflavi and two from M. luteus, were chosen for further 16S rDNA analyses. A pathogenicity assay for the isolated bacterial strains was carried out in rainbow trout and it evidenced absence of pathogenicity in the tested strains. The isolated strains were tested for in vitro antagonistic activity against Listonella anguillarum, a pathogen bacterium diffused in Mediterranean aquaculture and affecting various fish species. The isolated bacterial strains showed antagonistic activity against the pathogenic bacterium, suggesting a possible role of isolates as probiotics. In this study, for the first time, bacterial strains of the species B. marisflavi, known as an environmental species, were recovered in the gut microbiota of a healthy fish. The use of the isolates characterized in this study, mainly the yellow-pigmented bacterium, is suggested as possible probiotics to improve fish health, along with alternative methods of maintaining a healthy environment.

  12. High levels of DegU-P activate an Esat-6-like secretion system in Bacillus subtilis.

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    Catarina Baptista

    Full Text Available The recently discovered Type VII/Esat-6 secretion systems seem to be widespread among bacteria of the phyla Actinobacteria and Firmicutes. In some species they play an important role in pathogenic interactions with eukaryotic hosts. Several studies have predicted that the locus yukEDCByueBC of the non-pathogenic, Gram-positive bacterium Bacillus subtilis would encode an Esat-6-like secretion system (Ess. We provide here for the first time evidences for the functioning of this secretion pathway in an undomesticated B. subtilis strain. We show that YukE, a small protein with the typical features of the secretion substrates from the WXG100 superfamily is actively secreted to culture media. YukE secretion depends on intact yukDCByueBC genes, whose products share sequence or structural homology with known components of the S. aureus Ess. Biochemical characterization of YukE indicates that it exists as a dimer both in vitro and in vivo. We also show that the B. subtilis Ess essentially operates in late stationary growth phase in absolute dependence of phosphorylated DegU, the response regulator of the two-component system DegS-DegU. We present possible reasons that eventually have precluded the study of this secretion system in the B. subtilis laboratory strain 168.

  13. Synthesis of Peptide Amides using Sol-Gel Immobilized Alcalase in Batch and Continuous Reaction System

    NARCIS (Netherlands)

    Corici, L.N.; Frissen, A.E.; Zoelen, van D.J.; Eggen, I.F.; Peter, F.; Davidescu, C.; Boeriu, C.G.

    2011-01-01

    Two commercial proteases from Bacillus licheniformis (Alcalase 2.4 L FG and Alcalase 2.5 L, Type DX) were screened for the production of Z-Ala-Phe-NH2 in batch reaction. Alcalase 2.4 L FG was the most efficient enzyme for the C-terminal amidation of Z-Ala-Phe-OMe using ammonium carbamate as ammonium

  14. Bacillus amyloliquefaciens levansucrase-catalyzed the synthesis of fructooligosaccharides, oligolevan and levan in maple syrup-based reaction systems.

    Science.gov (United States)

    Li, Mengxi; Seo, Sooyoun; Karboune, Salwa

    2015-11-20

    Maple syrups with selected degree Brix (°Bx) (15, 30, 60) were investigated as reaction systems for levansucrase from Bacillus amyloliquefaciens. The enzymatic conversion of sucrose present in the maple syrup and the production of the transfructosylation products were assessed over a time course of 48h. At 30°C, the use of maple syrup 30°Bx led to the highest levansucrase activity (427.53μmol/mg protein/min), while maple syrup 66°Bx led to the highest converted sucrose concentration (1.53M). In maple syrup 30°Bx, oligolevans (1080%). In maple syrup 66°Bx, the most abundant products were oligolevans at 30°C and levans (DP≥30) at 8°C. The acceptor specificity study revealed the ability of B. amyloliquefaciens levansucrase to synthesize a variety of hetero-fructooligosaccharides (FOSs) in maple syrups 15°Bx and 30°Bx enriched with various disaccharides, with lactose being the preferred fructosyl acceptor. The current study is the first to investigate maple-syrup-based reaction systems for the synthesis of FOSs/oligolevans/levans. PMID:26344273

  15. A Novel Immunogenic Spore Coat-Associated Protein in Bacillus Anthracis: Characterization via Proteomics Approaches and a Vector-Based Vaccine System

    OpenAIRE

    Liu, Yu-Tsueng; Lin, Shwu-Bin; Huang, Cheng-Po; Huang, Chun-Ming

    2007-01-01

    New generation anthrax vaccines have been actively explored with the aim of enhancing efficacies and decreasing undesirable side effects that could be caused by licensed vaccines. Targeting novel antigens and/or eliminating the requirements for multiple needle injections and adjuvants are major objectives in the development of new anthrax vaccines. Using proteomics approaches, we identified a spore coat-associated protein (SCAP) in Bacillus anthracis. An E. coli vector-based vaccine system wa...

  16. Possible Processes for Origin of First Chemoheterotrophic Microorganisms with Modeling of Physiological Processes of Bacterium Bacillus subtilis as a Model System in 2H2O

    OpenAIRE

    Ignat Ignatov; Oleg Mosin

    2015-01-01

    We studied possible processes for origin of first chemoheterotrophic microorganisms with modeling of physiological processes of a Gram-positive chemoheterotrophic bacterium Bacillus subtilis, producer of purine ribonucleoside inosine as a model system in heavy water. The physiological influence of deuterium on the chemoheterotrophic bacterium B. subtilis was studied on a heavy water (HW) medium with a maximal concentration of 2H2O (89–90 atom% 2H). Also various suitable samples of hot mineral...

  17. Cellular fatty acid profile and H(+)-ATPase activity to assess acid tolerance of Bacillus sp. for potential probiotic functional attributes.

    Science.gov (United States)

    Shobharani, P; Halami, Prakash M

    2014-11-01

    The present study has been focused widely on comparative account of probiotic qualities of Bacillus spp. for safer usage. Initially, 170 heat resistant flora were isolated and selected for non-pathogenic cultures devoid of cytK, hblD, and nhe1 virulence genes. Subsequently, through biochemical tests along with 16S rRNA gene sequencing and fatty acid profiling, the cultures were identified as Bacillus megaterium (AR-S4), Bacillus subtilis (HR-S1), Bacillus licheniformis (Csm1-1a and HN-S1), and Bacillus flexus (CDM4-3c and CDM3-1). The selected cultures showed 70-80 % survival under simulated gastrointestinal condition which was also confirmed through H(+)-ATPase production. The amount of H(+)-ATPase increased by more than 2-fold when grown at pH 2 which support for the acid tolerance ability of Bacillus isolates. The study also examined the influence of acidic pH on cellular fatty acid composition of Bacillus spp. A remarkable shift in the fatty acid profile was observed at acidic pH through an increased amount of even numbered fatty acid (C16 and C18) in comparison with odd numbered (C15 and C17). Additionally, the cultures exhibited various probiotic functional properties. Overall, the study increases our understanding of Bacillus spp. and will allow both industries and consumers to choose for well-defined probiotic with possible health benefits. PMID:25125040

  18. Development and field testing of a mobile chlorine dioxide generation system for the decontamination of buildings contaminated with Bacillus anthracis

    Energy Technology Data Exchange (ETDEWEB)

    Wood, Joseph P., E-mail: wood.joe@epa.gov [U.S. Environmental Protection Agency, Office of Research and Development, National Homeland Security Research Center, MC-E343-06, Research Triangle Park, NC 27711 (United States); Blair Martin, G., E-mail: martin.blair@epa.gov [U.S. Environmental Protection Agency, Office of Research and Development, National Risk Management Research Laboratory, MC-E340-C, Research Triangle Park, NC 27711 (United States)

    2009-05-30

    The numerous buildings that became contaminated with Bacillus anthracis (the bacterium causing the disease anthrax) in 2001, and more recent B. anthracis - related events, point to the need to have effective decontamination technologies for buildings contaminated with biological threat agents. The U.S. Government developed a portable chlorine dioxide (ClO{sub 2}) generation system to decontaminate buildings contaminated with B. anthracis spores, and this so-called mobile decontamination trailer (MDT) prototype was tested through a series of three field trials. The first test of the MDT was conducted at Fort McClellan in Anniston, AL. during October 2004. Four test attempts occurred over two weekends; however, a number of system problems resulted in termination of the activity prior to any ClO{sub 2} introduction into the test building. After making several design enhancements and equipment changes, the MDT was subjected to a second test. During this test, extensive leak checks were made using argon and nitrogen in lieu of chlorine gas; each subsystem was checked for functionality, and the MDT was operated for 24 h. This second test demonstrated the MDT flow and control systems functioned satisfactorily, and thus it was decided to proceed to a third, more challenging field trial. In the last field test, ClO{sub 2} was generated and routed directly to the scrubber in a 12-h continuous run. Measurement of ClO{sub 2} levels at the generator outlet showed that the desired production rate was not achieved. Additionally, only one of the two scrubbers performed adequately with regard to maintaining ClO{sub 2} emissions below the limit. Numerous lessons were learned in the field trials of this ClO{sub 2} decontamination technology.

  19. Development and field testing of a mobile chlorine dioxide generation system for the decontamination of buildings contaminated with Bacillus anthracis

    International Nuclear Information System (INIS)

    The numerous buildings that became contaminated with Bacillus anthracis (the bacterium causing the disease anthrax) in 2001, and more recent B. anthracis - related events, point to the need to have effective decontamination technologies for buildings contaminated with biological threat agents. The U.S. Government developed a portable chlorine dioxide (ClO2) generation system to decontaminate buildings contaminated with B. anthracis spores, and this so-called mobile decontamination trailer (MDT) prototype was tested through a series of three field trials. The first test of the MDT was conducted at Fort McClellan in Anniston, AL. during October 2004. Four test attempts occurred over two weekends; however, a number of system problems resulted in termination of the activity prior to any ClO2 introduction into the test building. After making several design enhancements and equipment changes, the MDT was subjected to a second test. During this test, extensive leak checks were made using argon and nitrogen in lieu of chlorine gas; each subsystem was checked for functionality, and the MDT was operated for 24 h. This second test demonstrated the MDT flow and control systems functioned satisfactorily, and thus it was decided to proceed to a third, more challenging field trial. In the last field test, ClO2 was generated and routed directly to the scrubber in a 12-h continuous run. Measurement of ClO2 levels at the generator outlet showed that the desired production rate was not achieved. Additionally, only one of the two scrubbers performed adequately with regard to maintaining ClO2 emissions below the limit. Numerous lessons were learned in the field trials of this ClO2 decontamination technology.

  20. Two-component system cross-regulation integrates Bacillus anthracis response to heme and cell envelope stress.

    Directory of Open Access Journals (Sweden)

    Laura A Mike

    2014-03-01

    Full Text Available Two-component signaling systems (TCSs are one of the mechanisms that bacteria employ to sense and adapt to changes in the environment. A prototypical TCS functions as a phosphorelay from a membrane-bound sensor histidine kinase (HK to a cytoplasmic response regulator (RR that controls target gene expression. Despite significant homology in the signaling domains of HKs and RRs, TCSs are thought to typically function as linear systems with little to no cross-talk between non-cognate HK-RR pairs. Here we have identified several cell envelope acting compounds that stimulate a previously uncharacterized Bacillus anthracis TCS. Furthermore, this TCS cross-signals with the heme sensing TCS HssRS; therefore, we have named it HssRS interfacing TCS (HitRS. HssRS reciprocates cross-talk to HitRS, suggesting a link between heme toxicity and cell envelope stress. The signaling between HssRS and HitRS occurs in the parental B. anthracis strain; therefore, we classify HssRS-HitRS interactions as cross-regulation. Cross-talk between HssRS and HitRS occurs at both HK-RR and post-RR signaling junctions. Finally, HitRS also regulates a previously unstudied ABC transporter implicating this transporter in the response to cell envelope stress. This chemical biology approach to probing TCS signaling provides a new model for understanding how bacterial signaling networks are integrated to enable adaptation to complex environments such as those encountered during colonization of the vertebrate host.

  1. Determination of the Influence of Substrate Concentration on Enzyme Selectivity Using Whey Protein Isolate and Bacillus licheniformis Protease

    NARCIS (Netherlands)

    Butré, C.I.; Sforza, S.; Gruppen, H.; Wierenga, P.A.

    2014-01-01

    Increasing substrate concentration during enzymatic protein hydrolysis results in a decrease in hydrolysis rate. To test if changes in the mechanism of hydrolysis also occur, the enzyme selectivity was determined. The selectivity is defined quantitatively as the relative rate of hydrolysis of each c

  2. Competition for nitrate and glucose between Pseudomonas fluorescens and Bacillus licheniformis under continuous or fluctuating anoxic conditions

    NARCIS (Netherlands)

    Nijburg, J.W.; Gerards, S.; Laanbroek, H.J.

    1998-01-01

    The dissimilatory nitrate-reducing bacterial community in the rhizosphere of aerenchymatous plant species such as Glyceria maxima, consists of oxidative. denitrifying and fermentative nitrate-ammonifying bacteria. To study the respective ecological niches of both types of nitrate-reducing bacteria,

  3. Competition for nitrate and glucose between Pseudomonas fluorescens and Bacillus licheniformis under continuous or fluctuating anoxic conditions

    NARCIS (Netherlands)

    Nijburg, J.W.; Gerards, S.; Laanbroek, H.J.

    1998-01-01

    The dissimilatory nitrate-reducing bacterial community in the rhizosphere of aerenchymatous plant species such as Glyceria maxima, consists of oxidative, denitrifying and fermentative nitrate-ammonifying bacteria. To study the respective ecological niches of both types of nitrate-reducing bacteria,

  4. Influence of different silica derivatives in the immobilization and stabilization of a Bacillus licheniformis protease (Subtilisin Carlsberg)

    OpenAIRE

    Ferreira, L.; Ramos, M. A.; Dordick, J S; Gil, M. H.

    2003-01-01

    Alcalase 2T, a commercial preparation of Subtilisin Carlsberg, was covalent immobilized onto physiochemically characterized silica supports. The effect of mean pore diameter and surface chemistry on enzyme activity in the hydrolysis of casein has been examined. Two sets of chemically distinct silica supports were used presenting terminal amino (SAPTES) or hydroxyl groups (STESPM-pHEMA). The percentage of immobilized protein was smaller in SAPTES (31-39%) than in STESPM-pHEMA (62-71%), but pre...

  5. Sol–gel immobilization of Alcalase from Bacillus licheniformis for application in the synthesis of C-terminal peptide amides

    NARCIS (Netherlands)

    Corici, L.N.; Frissen, A.E.; Zoelen, van D.J.; Eggen, I.F.; Peter, F.; Davidescu, C.M.; Boeriu, C.G.

    2011-01-01

    Alcalase 2.4L FG, a commercial preparation of Subtilisin A, was physically entrapped in glass sol–gel matrices using alkoxysilanes of different types mixed with tetramethoxysilane (TMOS). The materials were used for catalyzing C-terminal amidation of Z-Ala-Phe-OMe in a mixture of tert-butanol/DMF. F

  6. Effect of gelatinization and hydrolysis conditions on the selectivity of starch hydrolysis with alpha-amylase from Bacillus licheniformis

    NARCIS (Netherlands)

    Baks, T.; Bruins, M.E.; Matser, A.M.; Janssen, A.E.M.; Boom, R.M.

    2008-01-01

    Enzymatic hydrolysis of starch can be used to obtain various valuable hydrolyzates with different compositions. The effects of starch pretreatment, enzyme addition point, and hydrolysis conditions on the hydrolyzate composition and reaction rate during wheat starch hydrolysis with ¿-amylase from Bac

  7. Disruption of microbial biofilms by an extracellular protein isolated from epibiotic tropical marine strain of Bacillus licheniformis

    Digital Repository Service at National Institute of Oceanography (India)

    Dusane, D.H.; Damare, S.R.; Nancharaiah, Y.V.; Ramaiah, N.; Venugopalan, V.P.; Kumar, A.R.; Zinjarde, S.S.

    incubation period, the slides were removed, rinsed twice with sterile phosphate buffer (50 mM, pH 7.0) to remove the planktonic cells and the biofilms were stained with BacLight Live/Dead stain (Molecular Probes, Eugene). The cell viability was assessed... inhibits biofilm formation of the human pathogens Escherichia coli and Staphylococcus aureus [35]. 4-phenylbutanoic acid obtained from a marine strain of B. pumilus is also reported to be effective in inhibiting bacterial biofilms [36]. Interestingly...

  8. Characteristics of bacillus strains with antifungal activity against phytopathogens

    International Nuclear Information System (INIS)

    Four bacterial isolates that showed antifungal activity against Alternaria alternata and other phytopathogens were isolates from bean rhizosphere. 16S rDNA analysis and phylogenetic relationship indicated that these isolates belong to Genus Bacillus. Isolate A1 clustered with Bacillus licheniformis while other isolates A2, A3 and A4 clustered together with B.pumilus. n-Butanol extract of these isolates strongly inhibited the growth of A. alternata while, chloroform extract of isolate A2 and ethyl acetate extract of A1,A3, and A4 inhibited the test fungus partially. All the isolates except A4 produced chitinase enzyme. None of the isolates solubilized mineral phosphate. Radiation sensitivity of isolates A1, A2, A3 and A4 were assessed and the LD99 values are determined as 0.50, 6.69, 11,60, 1.53 kGy, respectively. Mutant libraries of each isolate were prepared by exposing them to gamma radiation at their respective LD99 dose. Crude metabolite caused drastic changes on A. alternata hyphal morphology. Appearance of shrunken and collapsed hyphae could be due to the leak of cell wall or changes in membrane permeability

  9. The central nervous system as target of Bacillus anthracis toxin independent virulence in rabbits and guinea pigs.

    Directory of Open Access Journals (Sweden)

    Haim Levy

    Full Text Available Infection of the central nervous system is considered a complication of Anthrax and was reported in humans and non-human primates. Previously we have reported that Bacillus anthracis possesses a toxin-independent virulent trait that, like the toxins, is regulated by the major virulence regulator, AtxA, in the presence of pXO2. This toxin-independent lethal trait is exhibited in rabbits and Guinea pigs following significant bacteremia and organ dissemination. Various findings, including meningitis seen in humans and primates, suggested that the CNS is a possible target for this AtxA-mediated activity. In order to penetrate into the brain tissue, the bacteria have to overcome the barriers isolating the CNS from the blood stream. Taking a systematic genetic approach, we compared intracranial (IC inoculation and IV/SC inoculation for the outcome of the infection in rabbits/GP, respectively. The outstanding difference between the two models is exhibited by the encapsulated strain VollumΔpXO1, which is lethal when injected IC, but asymptomatic when inoculated IV/SC. The findings demonstrate that there is an apparent bottleneck in the ability of mutants to penetrate into the brain. Any mutant carrying either pXO1 or pXO2 will kill the host upon IC injection, but only those carrying AtxA either on pXO1 or in the chromosome in the background of pXO2 can penetrate into the brain following peripheral inoculation. The findings were corroborated by histological examination by H&E staining and immunofluorescence of rabbits' brains following IV and IC inoculations. These findings may have major implications on future research both on B. anthracis pathogenicity and on vaccine development.

  10. The central nervous system as target of Bacillus anthracis toxin independent virulence in rabbits and guinea pigs.

    Science.gov (United States)

    Levy, Haim; Glinert, Itai; Weiss, Shay; Bar-David, Elad; Sittner, Assa; Schlomovitz, Josef; Altboum, Zeev; Kobiler, David

    2014-01-01

    Infection of the central nervous system is considered a complication of Anthrax and was reported in humans and non-human primates. Previously we have reported that Bacillus anthracis possesses a toxin-independent virulent trait that, like the toxins, is regulated by the major virulence regulator, AtxA, in the presence of pXO2. This toxin-independent lethal trait is exhibited in rabbits and Guinea pigs following significant bacteremia and organ dissemination. Various findings, including meningitis seen in humans and primates, suggested that the CNS is a possible target for this AtxA-mediated activity. In order to penetrate into the brain tissue, the bacteria have to overcome the barriers isolating the CNS from the blood stream. Taking a systematic genetic approach, we compared intracranial (IC) inoculation and IV/SC inoculation for the outcome of the infection in rabbits/GP, respectively. The outstanding difference between the two models is exhibited by the encapsulated strain VollumΔpXO1, which is lethal when injected IC, but asymptomatic when inoculated IV/SC. The findings demonstrate that there is an apparent bottleneck in the ability of mutants to penetrate into the brain. Any mutant carrying either pXO1 or pXO2 will kill the host upon IC injection, but only those carrying AtxA either on pXO1 or in the chromosome in the background of pXO2 can penetrate into the brain following peripheral inoculation. The findings were corroborated by histological examination by H&E staining and immunofluorescence of rabbits' brains following IV and IC inoculations. These findings may have major implications on future research both on B. anthracis pathogenicity and on vaccine development.

  11. Construction of a novel, stable, food-grade expression system by engineering the endogenous toxin-antitoxin system in Bacillus subtilis.

    Science.gov (United States)

    Yang, Sen; Kang, Zhen; Cao, Wenlong; Du, Guocheng; Chen, Jian

    2016-02-10

    Bacillus subtilis as an important workhorse that has been widely used to produce enzymes and metabolites. To broaden its applications, especially in the food and feed industry, we constructed a novel, stable, food-grade expression system by engineering its type II toxin-antitoxin system. The expression of the toxin EndoA, encoded by the chromosomal ydcE gene, was regulated by an endogenous, xylose-inducible promoter, while the ydcD gene, which encodes the unstable antitoxin EndoB, was inserted into a food-grade vector backbone, where its expression was driven by the native, constitutive promoter PylxM. By maintaining the xylose concentration above 2.0 g L(-1), this auto-regulated expression system was absolutely stable after 100 generations. Compared with traditional antibiotic-dependent expression systems, this novel expression system resulted in greater biomass and higher titers of desired products (enzymes or metabolites). Our results demonstrate that this stable, food-grade expression system is suitable for enzyme production and pathway engineering, especially for the production of food-grade enzymes and metabolites.

  12. The CssRS two-component regulatory system controls a general secretion stress response in Bacillus subtilis

    NARCIS (Netherlands)

    Westers, Helga; Westers, L; Darmon, E.; van Dijl, Jan Maarten; Quax, Wim; Zanen, Geeske

    2006-01-01

    Bacillus species are valuable producers of industrial enzymes and biopharmaceuticals, because they can secrete large quantities of high-quality proteins directly into the growth medium. This requires the concerted action of quality control factors, such as folding catalysts and 'cleaning proteases'.

  13. 腾冲热海嗜热芽孢杆菌的分离鉴定%Isolation and Characterization of Thermophilic Bacillus from in Tengchong Rehai

    Institute of Scientific and Technical Information of China (English)

    晏爱芬; 余丽; 林连兵

    2012-01-01

    26 Thermophilic Bacillus was obtained according to the disassociation from Tengchong hot springs. And one of the strains, NHH4 is selected to analyze its morphologic characteristics, development features, nitric and carbon sources. The cytomorphology of the bacillus is nemaline with gemma, gram-positive, oxygen-loving. And the most suitable temperature for its development is 55 ℃ , the PH value is 7. 5. It can use glucose, sucrose, D-frucose, D-marvnopyranose, but can' t use maltose, lactose, D-xylose, L- rhamnose. The analysis on the phyloge-ny of 16srRNA' s gene sequence indicates that the similarity of NHH4 and Bacillus licheniformis strain N8 is 99% , so the bacillus is identified as thermophilic lichen-bacillus.%从云南腾冲热海温泉中分离得到26株嗜热芽孢杆菌菌株,对其中一株嗜热芽孢杆菌菌株NHH4进行电镜形态、生长特征、碳源、氮源等分析.该菌株细胞形态为杆状,产芽胞,革兰氏染色阳性,严格好氧,其最适生长温度为55℃,最适为pH7.5.能利用葡萄糖、蔗糖、D-果糖、D-甘露糖,不能利用麦芽糖、乳糖、D-木糖、L-鼠季糖.通过对其16S rRNA基因序列的系统发育分析表明,NHH4与Bacillus licheniformis strain N8的序列相似性为99%,将此菌株鉴定为嗜热地衣芽胞杆菌菌株.

  14. EXOPOLYSACCHARIDE PRODUCTION BY DROUGHT TOLERANT BACILLUS SPP. AND EFFECT ON SOIL AGGREGATION UNDER DROUGHT STRESS

    Directory of Open Access Journals (Sweden)

    Sandhya Vardharajula

    2014-08-01

    Full Text Available Exopolysaccharides (EPS of microbial origin with novel functionality, reproducible physico-chemical properties, are important class of polymeric materials. EPS are believed to protect bacterial cells from dessication, produce biofilms, thus enhancing the cells chances of bacterial colonizing special ecological niches. In rhizosphere, EPS are known to be useful to improve the moisture-holding capacity. Three Bacillus spp. strains identified by 16s rDNA sequence analysis as B. amyloliquefaciens strain HYD-B17; B. licheniformis strain HYTAPB18; B. subtilis strain RMPB44 were studied for the ability to tolerate matric stress and produce EPS under different water potentials. EPS production in all the three Bacillus spp strains increased with increasing water stress indicating correlation between drought stress tolerance and EPS production. Among the isolates, strain HYD-17 showed highest production of EPS. The exopolysaccharide composition of the three strains was further analyzed by HPLC. Drought stress influenced the ratio of sugars in EPS and glucose was found as major sugar in strains HYTAPB18 and RMPB44 whereas raffinose was major sugar found in strain HYD-B17. Inoculation of EPS producing Bacillus spp. strains in soil resulted in good soil aggregation under drought stress conditions at different incubation periods. This study shows that exposure to water stress conditions affects the composition and ratios of sugars in EPS produced by Bacillus spp. strains HYD-B17, HYTAPB18 and RMPB44 influencing abiotic stress tolerance of the microorganisms.

  15. Screening for Pseudomonas and Bacillus antagonistic rhizobacteria strains for the biocontrol of Fusarium wilt of chickpea

    Directory of Open Access Journals (Sweden)

    Hannane Abed

    2016-07-01

    Full Text Available The aim of this work is to study the ability of several isolates belonging to Rhizobacteria (Pseudomonas and Bacillus collected from several chickpea growing areas in Algeria, to control the mycelium growth of Fusarium oxysporum f. sp. ciceris. Interesting isolates were characterized for their morphological characteristics, physiological and biochemical activities as potential bio-control agent. Fungal inhibition tests were performed using plate assay and each isolate were tested for the production of protease, cyanide hydrogen, indole acetic acid, antifungal volatile and extracellular compound. According to API 50 CH, we are able to identify six Bacillus species (B. subtilis, B. circulans, B. lentus, B. aneurinilyticus, B. firmus, B. licheniformis; and with API 20NE test we have identified three Pseudomonas species (P. aeruginosa, P. luteola, P. fluorescens. The ability of bacterial isolates was varied in production of Protease, Gelatinase, Amylase, Cellulase, Acid Indole acetic, Lipase, Catalase and Cyanid Hydrogen. This is traduced in different rate of inhibition growth due to various extracellular compounds, where B61 (Bacillus aneurinilyticus and P39 (Pseudomonas luteola and P70 (Pseudomonas fluorescens were the most efficient with 77 and 55.5% respectively, while B39 (Bacillus firmus and P41 (Pseudomonas luteola were the most efficient by volatile compounds with 70.5 and 77.5% respectively. Our results indicate that these bacteria isolates can be used in the biocontrol of Fusarium oxysporum f. sp. ciceris.

  16. Process optimization by response surface methodology for extracellular alkaline protease production from bacillus subtilis

    International Nuclear Information System (INIS)

    Three microbial cultures Bacillus subtilis DSM 1970, Bacillus subtilis GCU-8 and Bacillus licheniformis DSM 1969 were screened for protease production by casein agar plate method. Among these Bacillus subtilis GCU-8 was found to be the most potent protease producer in wide pH range (5.0 to 8.0). Fermentation conditions were optimized for the production of alkaline protease using two statistical tools: Placket Burmen Model for linear regression study and Response Surface Model for interactive effects of significant factors on production. The alkaline protease was optimally produced after 48 hours of incubation at 37 degree C in fermentation media containing equal amounts of substrates (soybean meal and wheat bran, 7.5 g), MgSO/sub 4/ 7H/sub 2/O, 0.10 g and yeast extract 0.55 g. The protease was purified to homogeneity by salt precipitation, ion-exchange chromatography and size exclusion chromatography. The homogeneity and molecular weights were checked by SDS-PAGE. The protease was 45 KDa protein, predominantly alkaline and optimally active at pH 8.0. (author)

  17. Recognition of greater diversity of Bacillus species and related bacteria in human faeces.

    Science.gov (United States)

    Hoyles, Lesley; Honda, Harue; Logan, Niall A; Halket, Gillian; La Ragione, Roberto M; McCartney, Anne L

    2012-01-01

    In a study looking at culturable aerobic Actinobacteria associated with the human gastrointestinal tract, the vast majority of isolates obtained from dried human faeces belonged to the genus Bacillus and related bacteria. A total of 124 isolates were recovered from the faeces of 10 healthy adult donors. 16S rRNA gene sequence analyses showed the majority belonged to the families Bacillaceae (n=81) and Paenibacillaceae (n=3), with Bacillus species isolated from all donors. Isolates tentatively identified as Bacillus clausii (n=32) and Bacillus licheniformis (n=28) were recovered most frequently, with the genera Lysinibacillus, Ureibacillus, Oceanobacillus, Ornithinibacillus and Virgibacillus represented in some donors. Phenotypic data confirmed the identities of isolates belonging to well-characterized species. Representatives of the phylum Actinobacteria were recovered in much lower numbers (n=11). Many of the bacilli exhibited antimicrobial activity against one or more strains of Clostridium difficile, Clostridium perfringens, Listeria monocytogenes and Staphylococcus aureus, with some (n=12) found to have no detectable cytopathic effect on HEp-2 cells. This study has revealed greater diversity within gut-associated aerobic spore-formers than previous studies, and suggests that bacilli with potential as probiotics could be isolated from the human gut. PMID:22041546

  18. Identification, Functional Characterization and Regulon Prediction of a Novel Two Component System Comprising BAS0540-BAS0541 of Bacillus anthracis

    Science.gov (United States)

    Gopalani, Monisha; Kandari, Divya; Bhatnagar, Rakesh

    2016-01-01

    Two component systems (TCSs) can be envisaged as complex molecular devices that help the bacteria to sense its environment and respond aptly. 41 TCSs are predicted in Bacillus anthracis, a potential bioterrorism agent, of which only four have been studied so far. Thus, the intricate signaling network contributed by TCSs remains largely unmapped in B. anthracis and needs comprehensive exploration. In this study, we functionally characterized one such system composed of BAS0540 (Response regulator) and BAS0541 (Histidine kinase). BAS0540-BAS0541, the closest homolog of CiaRH of Streptococcus in B. anthracis, forms a functional TCS with BAS0541 displaying autophosphorylation and subsequent phosphotransfer to BAS0540. BAS0540 was also found to accept phosphate from physiologically relevant small molecule phosphodonors like acetyl phosphate and carbamoyl phosphate. Results of qRT-PCR and immunoblotting demonstrated that BAS0540 exhibits a constitutive expression throughout the growth of B. anthracis. Regulon prediction for BAS0540 in B. anthracis was done in silico using the consensus DNA binding sequence of CiaR of Streptococcus. The predicted regulon of BAS0540 comprised of 23 genes, which could be classified into 8 functionally diverse categories. None of the proven virulence factors were a part of the predicted regulon, an observation contrasting with the regulon of CiaRH in Streptococci. Electrophoretic mobility shift assay was used to show direct binding of purified BAS0540 to the upstream regions of 5 putative regulon candidates- BAS0540 gene itself; a gene predicted to encode cell division protein FtsA; a self–immunity gene; a RND family transporter gene and a gene encoding stress (heat) responsive protein. A significant enhancement in the DNA binding ability of BAS0540 was observed upon phosphorylation. Overexpression of response regulator BAS0540 in B. anthracis led to a prodigious increase of ~6 folds in the cell length, thereby conferring it a filamentous

  19. Decoding Biomass-Sensing Regulons of Clostridium thermocellum Alternative Sigma-I Factors in a Heterologous Bacillus subtilis Host System.

    Directory of Open Access Journals (Sweden)

    Iván Muñoz-Gutiérrez

    Full Text Available The Gram-positive, anaerobic, cellulolytic, thermophile Clostridium (Ruminiclostridium thermocellum secretes a multi-enzyme system called the cellulosome to solubilize plant cell wall polysaccharides. During the saccharolytic process, the enzymatic composition of the cellulosome is modulated according to the type of polysaccharide(s present in the environment. C. thermocellum has a set of eight alternative RNA polymerase sigma (σ factors that are activated in response to extracellular polysaccharides and share sequence similarity to the Bacillus subtilis σI factor. The aim of the present work was to demonstrate whether individual C. thermocellum σI-like factors regulate specific cellulosomal genes, focusing on C. thermocellum σI6 and σI3 factors. To search for putative σI6- and σI3-dependent promoters, bioinformatic analysis of the upstream regions of the cellulosomal genes was performed. Because of the limited genetic tools available for C. thermocellum, the functionality of the predicted σI6- and σI3-dependent promoters was studied in B. subtilis as a heterologous host. This system enabled observation of the activation of 10 predicted σI6-dependent promoters associated with the C. thermocellum genes: sigI6 (itself, Clo1313_2778, xyn11B (Clo1313_0522, xyn10D (Clo1313_0177, xyn10Z (Clo1313_2635, xyn10Y (Clo1313_1305, cel9V (Clo1313_0349, cseP (Clo1313_2188, sigI1 (Clo1313_2174, cipA (Clo1313_0627, and rsgI5 (Clo1313_0985. Additionally, we observed the activation of 4 predicted σI3-dependent promoters associated with the C. thermocellum genes: sigI3 (itself, Clo1313_1911, pl11 (Clo1313_1983, ce12 (Clo1313_0693 and cipA. Our results suggest possible regulons of σI6 and σI3 in C. thermocellum, as well as the σI6 and σI3 promoter consensus sequences. The proposed -35 and -10 promoter consensus elements of σI6 are CNNAAA and CGAA, respectively. Additionally, a less conserved CGA sequence next to the C in the -35 element and a highly

  20. Exploration of the antioxidant system and photosynthetic system of a marine algicidal Bacillus and its effect on four harmful algal bloom species.

    Science.gov (United States)

    Hou, Shaoling; Shu, Wanjiao; Tan, Shuo; Zhao, Ling; Yin, Pinghe

    2016-01-01

    A novel marine bacterium, strain B1, initially showed 96.4% algicidal activity against Phaeocystis globosa. Under this situation, 3 other harmful algal species (Skeletonema costatum, Heterosigma akashiwo, and Prorocentrum donghaiense) were chosen to study the algicidal effects of strain B1, and the algicidal activities were 91.4%, 90.7%, and 90.6%, respectively. To explore the algicidal mechanism of strain B1 on these 4 harmful algal species, the characteristics of the antioxidant system and photosynthetic system were studied. Sensitivity to strain B1 supernatant, enzyme activity, and gene expression varied with algal species, while the algicidal patterns were similar. Strain B1 supernatant increased malondialdehyde contents; decreased chlorophyll a contents; changed total antioxidant and superoxide dismutase activity; and restrained psbA, psbD, and rbcL genes expression, which eventually resulted in the algal cells death. The algicidal procedure was observed using field emission scanning electron microscopy, which indicated that algal cells were lysed and cellular substances were released. These findings suggested that the antioxidant and photosynthetic system of these 4 algal species was destroyed under strain B1 supernatant stress. This is the first report to explore and compare the mechanism of a marine Bacillus against harmful algal bloom species of covered 4 phyla. PMID:26634608

  1. A system for the directed evolution of the insecticidal protein from Bacillus thuringiensis.

    Science.gov (United States)

    Ishikawa, Hiroshi; Hoshino, Yasushi; Motoki, Yutaka; Kawahara, Takuma; Kitajima, Mika; Kitami, Madoka; Watanabe, Ayako; Bravo, Alejandra; Soberon, Mario; Honda, Atsuko; Yaoi, Katsuro; Sato, Ryoichi

    2007-06-01

    Theoretically, the activity of AB-type toxin molecules such as the insecticidal toxin (Cry toxin) from B. thuringiensis, which have one active site and two binding site, is improved in parallel with the binding affinity to its receptor. In this experiment, we tried to devise a method for the directed evolution of Cry toxins to increase the binding affinity to the insect receptor. Using a commercial T7 phage-display system, we expressed Cry1Aa toxin on the phage surface as fusions with the capsid protein 10B. These recombinant phages bound to a cadherin-like protein that is one of the Cry1Aa toxin receptors in the model target insect Bombyx mori. The apparent affinity of Cry1Aa-expressing phage for the receptor was higher than that of Cry1Ab-expressing phage. Phages expressing Cry1Aa were isolated from a mixed suspension of phages expressing Cry1Ab and concentrated by up to 130,000-fold. Finally, random mutations were made in amino acid residues 369-375 in domain 2 of Cry1Aa toxin, the mutant toxins were expressed on phages, and the resulting phage library was screened with cadherin-like protein-coated beads. As a result, phages expressing abnormal or low-affinity mutant toxins were excluded, and phages with high-affinity mutant toxins were selected. These results indicate that a method combining T7 phage display with selection using cadherin-like protein-coated magnetic beads can be used to increase the activity of easily obtained, low-activity Cry toxins from bacteria. PMID:17914188

  2. Evaluation of the FilmArray® system for detection of Bacillus anthracis, Francisella tularensis, and Yersinia pestis

    Energy Technology Data Exchange (ETDEWEB)

    Seiner, Derrick R.; Colburn, Heather A.; Baird, Cheryl L.; Bartholomew, Rachel A.; Straub, Tim M.; Victry, Kristin D.; Hutchison, Janine R.; Valentine, Nancy B.; Bruckner-Lea, Cindy J.

    2013-04-29

    To evaluate the sensitivity and specificity of the Idaho Technologies FilmArray® Biothreat Panel for the detection of Bacillus anthracis (Ba), Francisella tularensis (Ft), and Yersinia pestis (Yp) DNA, and demonstrate the detection of Ba spores. Methods and Results: DNA samples from Ba, Ft and Yp strains and near-neighbors, and live Ba spores were analyzed using the Biothreat Panel, a multiplexed PCR-based assay for 17 pathogens and toxins. Sensitivity studies with DNA suggest a limit of detection of 250 genome equivalents (GEs) per sample. Furthermore, the correct call of Ft, Yp or Bacillus species was made in 63 of 72 samples tested at 25 GE or less. With samples containing 25 Ba Sterne spores, at least one of the two possible Ba markers were identified in all samples tested. We observed no cross-reactivity with near-neighbor DNAs.

  3. Differential Effects of Linezolid and Ciprofloxacin on Toxin Production by Bacillus anthracis in an In Vitro Pharmacodynamic System

    OpenAIRE

    Louie, Arnold; VanScoy, Brian D.; Heine, Henry S.; Liu, Weiguo; Abshire, Terry; Holman, Kari; Kulawy, Robert; Brown, David L.; Drusano, George L.

    2012-01-01

    Bacillus anthracis causes anthrax. Ciprofloxacin is a gold standard for the treatment of anthrax. Previously, using the non-toxin-producing ΔSterne strain of B. anthracis, we demonstrated that linezolid was equivalent to ciprofloxacin for reducing the total (vegetative and spore) bacterial population. With ciprofloxacin therapy, the total population consisted of spores. With linezolid therapy, the population consisted primarily of vegetative bacteria. Linezolid is a protein synthesis inhibito...

  4. Diversity of bacteria of the genus Bacillus on board of international space station.

    Science.gov (United States)

    Alekhova, T A; Zakharchuk, L M; Tatarinova, N Yu; Kadnikov, V V; Mardanov, A V; Ravin, N V; Skryabin, K G

    2015-01-01

    From swabs of surfaces of equipment and air samples of the Russian segment of the International Space Station, nine strains of spore-forming bacteria of the genus Bacillus belonging to the species B. pumilus, B. licheniformis, B. subtilis, B. megaterium, and B. amyloliquefaciens were isolated. The last species of bacilli on the equipment of RS ISS was detected for the first time. For these species of bacilli, there are known strains that can be opportunistic to humans, and their metabolites can cause biodegradation of equipment and materials. B. pumilus found on ISS belongs to the group of bacteria that exhibits a particularly high resistance to adverse environmental conditions, such as dehydration, ultraviolet and gamma radiation, and chemical disinfection.

  5. Diversity of bacteria of the genus Bacillus on board of international space station.

    Science.gov (United States)

    Alekhova, T A; Zakharchuk, L M; Tatarinova, N Yu; Kadnikov, V V; Mardanov, A V; Ravin, N V; Skryabin, K G

    2015-01-01

    From swabs of surfaces of equipment and air samples of the Russian segment of the International Space Station, nine strains of spore-forming bacteria of the genus Bacillus belonging to the species B. pumilus, B. licheniformis, B. subtilis, B. megaterium, and B. amyloliquefaciens were isolated. The last species of bacilli on the equipment of RS ISS was detected for the first time. For these species of bacilli, there are known strains that can be opportunistic to humans, and their metabolites can cause biodegradation of equipment and materials. B. pumilus found on ISS belongs to the group of bacteria that exhibits a particularly high resistance to adverse environmental conditions, such as dehydration, ultraviolet and gamma radiation, and chemical disinfection. PMID:26728721

  6. Production and characterization of phytase from Bacillus spp. as feed additive in aquaculture

    Directory of Open Access Journals (Sweden)

    Rande B. Dechavez

    2011-07-01

    Full Text Available Phytases are phosphohydrolases that catalyze the release of phosphate from phytate (myo inositol hexakisphosphate, the major phosphorus (P form mostly occurring in animal feeds of plant origin. These enzymes can be supplemented in animal diets to reduce inorganic phosphorus supplementation and fecal phosphorus excretion. Four species of Bacillus namely, B. pumilus , B.megaterium , B. coagulans , and B. licheniformis were used to study the biochemical characteristics of their phytases. All the strains investigated were able to hydrolyze extracellular phytate. The activity of phytase increased markedly at the late stationary phase in all the species tested. Highest enzyme activity was found in phytase from B. megaterium after the 4th day of culture. The crude phytases from the different Bacillus strains were optimally active at pH values ranging 5.5 to 7.0 at 37 0 C and retained their activity at temperatures up to 80 0 C. The enzymes exhibited thermostability, retaining ~50 %activity at 70 0 C and were fairly stable up to pH 10. These properties indicate that the Bacillus phytases appear to be suitable for animal feed supplementation in aquaculture to improve the bioavailability of phosphorus.

  7. Studies on the toxicity of an aqueous suspension of C60 nanoparticles using a bacterium (gen. Bacillus) and an aquatic plant (Lemna gibba) as in vitro model systems.

    Science.gov (United States)

    Santos, Sandra M A; Dinis, Augusto M; Rodrigues, David M F; Peixoto, Francisco; Videira, Romeu A; Jurado, Amália S

    2013-10-15

    The increasing use of C60 nanoparticles and the diversity of their applications in industry and medicine has led to their production in a large scale. C60 release into wastewaters and the possible accumulation in the environment has raised concerns about their ecotoxicological impact. In the present study, an aqueous suspension of C60 nanoparticles was prepared and its potential toxicity studied in laboratory, using a bacterium (Bacillus stearothermophilus) and an aquatic plant (Lemna gibba) as model systems. C60 nanoparticles inhibited the growth of L. gibba, in contrast to that of the bacterium. Consistently, the ultrastructure and respiratory activity of bacterial cells were not affected by C60, but the contents of chlorophylls a and b and chloroplast oxygen production decreased considerably in L. gibba. Altogether, our results suggest that C60 aqueous dispersions must be viewed as an environmental pollutant, potentially endangering the equilibrium of aquatic ecosystems. PMID:24084257

  8. Enhanced transformation efficiency of recalcitrant Bacillus cereus and Bacillus weihenstephanensis isolates upon in vitro methylation of plasmid DNA

    NARCIS (Netherlands)

    Nierop Groot, M.N.; Nieboer, F.; Abee, T.

    2008-01-01

    Digestion patterns of chromosomal DNAs of Bacillus cereus and Bacillus weihenstephanensis strains suggest that Sau3AI-type restriction modification systems are widely present among the isolates tested. In vitro methylation of plasmid DNA was used to enhance poor plasmid transfer upon electroporation

  9. Bacillus cereus AR156 primes induced systemic resistance by suppressing miR825/825? and activating defense-related genes in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Dongdong Niu; Hongwei Zhao; Jing Xia; Chunhao Jiang; Beibei Qi; Xiaoyu Ling; Siyuan Lin; Weixiong Zhang; Jianhua Guo; Hailing Jin

    2016-01-01

    Small RNAs play an important role in plant immune responses. However, their regulatory function in induced systemic resistance (ISR) is nascent. Bacillus cereus AR156 is a plant growth-promoting rhizobacterium that induces ISR in Arabidopsis against bacterial infection. Here, by comparing small RNA profiles of Pseudomonas syringae pv. tomato (Pst) DC3000-infected Arabidopsis with and without AR156 pretreatment, we identified a group of Arabidopsis microRNAs (miRNAs) that are differentially regulated by AR156 pretreatment. miR825 and miR825? are two miRNA generated from a single miRNA gene. Northern blot analysis indicated that they were significantly downregulated in Pst DC3000-infected plants pretreated with AR156, in contrast to the plants without AR156 pretreatment. miR825 targets two ubiquitin-protein ligases, while miR825? targets toll-interleukin-like receptor (TIR)-nucleotide binding site (NBS) and leucine-rich repeat (LRR) type resistance (R) genes. The expression of these target genes negatively correlated with the expression of miR825 and miR825?. Moreover, transgenic plants showing reduced expression of miR825 and miR825? displayed enhanced resistance to Pst DC3000 infection, whereas transgenic plants overexpressing miR825 and miR825? were more susceptible. Taken together, our data indicates that Bacillus cereus AR156 pretreatment primes ISR to Pst infection by suppressing miR825 and miR825? and activating the defense related genes they targeted.

  10. Comparative genome analysis of Bacillus cereus group genomes withBacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Iain; Sorokin, Alexei; Kapatral, Vinayak; Reznik, Gary; Bhattacharya, Anamitra; Mikhailova, Natalia; Burd, Henry; Joukov, Victor; Kaznadzey, Denis; Walunas, Theresa; D' Souza, Mark; Larsen, Niels; Pusch,Gordon; Liolios, Konstantinos; Grechkin, Yuri; Lapidus, Alla; Goltsman,Eugene; Chu, Lien; Fonstein, Michael; Ehrlich, S. Dusko; Overbeek, Ross; Kyrpides, Nikos; Ivanova, Natalia

    2005-09-14

    Genome features of the Bacillus cereus group genomes (representative strains of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis sub spp israelensis) were analyzed and compared with the Bacillus subtilis genome. A core set of 1,381 protein families among the four Bacillus genomes, with an additional set of 933 families common to the B. cereus group, was identified. Differences in signal transduction pathways, membrane transporters, cell surface structures, cell wall, and S-layer proteins suggesting differences in their phenotype were identified. The B. cereus group has signal transduction systems including a tyrosine kinase related to two-component system histidine kinases from B. subtilis. A model for regulation of the stress responsive sigma factor sigmaB in the B. cereus group different from the well studied regulation in B. subtilis has been proposed. Despite a high degree of chromosomal synteny among these genomes, significant differences in cell wall and spore coat proteins that contribute to the survival and adaptation in specific hosts has been identified.

  11. Phylogenetic Analysis of 16S rDNA Sequence and PCR - RFLP of Bacillus from Fumao - flavor Daqu%福矛高温大曲中芽孢杆菌16S rDNA-RFLP及系统发育分析

    Institute of Scientific and Technical Information of China (English)

    颜林春; 张守财; 马校卫; 汤二将; 黄祖新; 陈由强

    2012-01-01

    目的:从福建建瓯黄华山酿酒有限公司高温大曲中分离出89株芽孢杆菌,通过初步筛选鉴定并进行微生物多样性研究.方法:对其16S rDNA进行PCR - RFLP分析和系统发育研究.结果:初步筛选得到的18株芽孢杆菌被HhaⅠ和MspⅠ酶切聚类分为四大组.通过系统发育分析样品中有6株Bacillus subtilis,4株Bacillus cereus,2株Bacillus sonorensis,2株Bacillus licheniformis,以及Bacillus pumilus、Bacillus oleronius、Bacillus coagulans和Bacillus thuringiensis各1株.结论:研究显示该高温大曲中可培养芽孢杆菌具有微生物多样性.

  12. Establishment of a markerless mutation delivery system in Bacillus subtilis stimulated by a double-strand break in the chromosome.

    Directory of Open Access Journals (Sweden)

    Ting Shi

    Full Text Available Bacillus subtilis has been a model for gram-positive bacteria and it has long been exploited for industrial and biotechnological applications. However, the availability of facile genetic tools for physiological analysis has generally lagged substantially behind traditional genetic models such as Escherichia coli and Saccharomyces cerevisiae. In this work, we have developed an efficient, precise and scarless method for rapid multiple genetic modifications without altering the chromosome of B. subtilis. This method employs upp gene as a counter-selectable marker, double-strand break (DSB repair caused by exogenous endonuclease I-SceI and comK overexpression for fast preparation of competent cell. Foreign dsDNA can be simply and efficiently integrated into the chromosome by double-crossover homologous recombination. The DSB repair is a potent inducement for stimulating the second intramolecular homologous recombination, which not only enhances the frequency of resolution by one to two orders of magnitude, but also selects for the resolved product. This method has been successfully and reiteratively used in B. subtilis to deliver point mutations, to generate in-frame deletions, and to construct large-scale deletions. Experimental results proved that it allowed repeated use of the selectable marker gene for multiple modifications and could be a useful technique for B. subtilis.

  13. Bacillus cereus as a nongastrointestinal pathogen

    Directory of Open Access Journals (Sweden)

    Pavani G.

    2014-02-01

    Full Text Available The potential of Bacillus cereus to cause systemic infections is of serious concern. Apart from Gastrointestinal infections, it causes respiratory tract infections, nosocomial infections, eye infections, CNS infections, cutaneous infections, endocarditis, osteomyelitis and urinary tract infections. The potential of this bacterium to cause life threatening infections has increased. Trauma is an important predisposing factor for Bacillus cereus infections. The maintenance of skin and mucous membrane integrity limits infection by this micro-organism. [Int J Res Med Sci 2014; 2(1.000: 28-30

  14. Mapping by interspecies transformation experiments of several ribosomal protein genes near the replication origin of Bacillus subtilis chromosome.

    Science.gov (United States)

    Osawa, S; Tokui, A; Saito, H

    1978-08-17

    Bacillus subtilis 168 was transformed with DNAs from B. amyloliquefaciens K or B. licheniformis IAM 11054. These two species show a considerable difference in ribosomal proteins from B. subtilis. Analyses of the transformants indicated that the genes for 16 proteins, S3, S5, S8, S12, S17, S19, BL1, BL5, BL6, BL8, BL14, BL16, BL17, BL22, BL23 and BL25 are located in the cysA-str-spc region on B. subtilis chromosome. The genes for 10 proteins, S4, S6, S13, S16, S20, BL15, BL18, BL20, BL24 and BL28 could not be found in this region in the present experiments.

  15. Inactivation of Spores of Bacillus anthracis Sterne, Bacillus cereus, and Bacillus thuringiensis subsp. israelensis by Chlorination

    OpenAIRE

    Rice, E W; Adcock, N. J.; Sivaganesan, M; Rose, L. J.

    2005-01-01

    Three species of Bacillus were evaluated as potential surrogates for Bacillus anthracis for determining the sporicidal activity of chlorination as commonly used in drinking water treatment. Spores of Bacillus thuringiensis subsp. israelensis were found to be an appropriate surrogate for spores of B. anthracis for use in chlorine inactivation studies.

  16. Surfactin production by strains of Bacillus mojavensis

    Science.gov (United States)

    Bacillus mojavensis, RRC101 is an endophytic bacterium patented for control of fungal diseases in maize and other plants. DNA fingerprint analysis of the rep-PCR fragments of 35 B. mojavensis and 4 B. subtilis strains using the Diversilab genotyping system revealed genotypic distinctive strains alon...

  17. An efficient heat-inducible Bacillus subtilis bacteriophage 105 expression and secretion system for the production of the Streptomyces clavuligerus beta-lactamase inhibitory protein (BLIP).

    Science.gov (United States)

    Liu, Hong-Bing; Chui, Ka-Shun; Chan, Chi-Leong; Tsang, Chun-Wai; Leung, Yun-Chung

    2004-03-18

    The Streptomyces clavuligerus beta-lactamase inhibitory protein (BLIP) has been shown to be a potent inhibitor of class A beta-lactamases including the Escherichia coli TEM-1 beta-lactamase (Ki = 0.6 nM). A heat-inducible BLIP expression system was constructed based on a derivative of Bacillus subtilis phage phi105. The recombinant BLIP produced by this system was secreted to the culture medium, purified to homogeneity, and fully active. We have shown that the signal peptide of BLIP functions well in B. subtilis to secrete BLIP out of the cells, which facilitates purification. The absence of a His-tag also avoids the activity and structure of BLIP being altered. An unprecedented high yield of recoverable protein in culture supernatant (3.6mg of >95% pure BLIP/l culture) was achieved by a simple purification protocol. We have developed an efficient production process in which the culture time before heat-induction was 3-4h and the culture supernatant could be collected 5h after induction. This total time of 8-9h is considered to be very short compared to that of the native S. clavuligerus culturing (60-70h). We achieved a very efficient BLIP production rate of 0.8-0.9mg/l/h. Heterologous gene expression was tightly controlled and no production of BLIP was observed before heat-induction, suggesting that cell density can be further increased to improve enzyme yield.

  18. Dietary Bacillus subtilis FPTB13 and chitin, single or combined, modulate systemic and cutaneous mucosal immunity and resistance of catla, Catla catla (Hamilton) against edwardsiellosis.

    Science.gov (United States)

    Sangma, Timothy; Kamilya, Dibyendu

    2015-12-01

    Effects of dietary administration of Bacillus subtilis FPTB13 and chitin, single or combined, on the systemic immunity, mucosal immunity and resistance of catla (Catla catla) against Edwardsiella tarda infection were investigated. The probiotic attributes of B. subtilis was tested by conducting antagonism study, safety in catla, in vitro immunomodulation and dietary immunomodulation. Results of these studies indicated the probiotic potential of the strain. From the preliminary dietary immunomodulation study, a dose of 10(9) B. subtilis cells g(-1) was selected for inclusion into diets for subsequent experiments. Experimental diets were prepared by adding B. subtilis (10(9) cells g(-1)), chitin (2%) and their combination to the basal diet. Different systemic and mucosal immunological parameters viz. oxygen radical production, myeloperoxidase content, lysozyme activity, total protein content and alkaline phosphatase activity showed significant enhancement (pdiet. B. subtilis and chitin alone also significantly elevated most of the immune responses. All the diets significantly increased the resistance of catla against E. tarda challenge. The highest post-challenge survival was observed in combined group (i.e. 63.33%). In conclusion, B. subtilis and chitin, alone or combined, had a health ameliorating effect in catla. The results also collectively suggest the usefulness of applying a combined probiotic and immunostimulant supplemented diet to achieve greater benefits. PMID:26616655

  19. Feasible Management of Southern Corn Leaf Blight via Induction of Systemic Resistance by Bacillus cereus C1L in Combination with Reduced Use of Dithiocarbamate Fungicides

    Directory of Open Access Journals (Sweden)

    Yi-Ru Lai

    2016-10-01

    Full Text Available Dithiocarbamate fungicides such as maneb and mancozeb are widely used nonsystemic protectant fungicides to control various plant fungal diseases. Dithiocarbamate fungicides should be frequently applied to achieve optimal efficacy of disease control and avoid either decline in effectiveness or wash-off from leaf surface. Dithiocarbamates are of low resistance risk but have the potential to cause human neurological diseases. The objective of this study was to develop a strategy to effectively control plant disease with reduced use of dithiocarbamtes. Southern corn leaf blight was the model pathosystem for the investigation. When corn plants were drench-treated with Bacillus cereus C1L, a rhizobacterium able to induce systemic resistance in corn plants against southern leaf blight, frequency of spraying dithiocarbamate fungicides could be decreased. The treatment of B. cereus C1L was able to protect maize from southern leaf blight while residues of dithiocarbamates on leaf surface were too low to provide sufficient protection. On the other hand, frequent sprays of mancozeb slightly but significantly reduced growth of corn plants under natural conditions. In contrast, application of B. cereus C1L can significantly promote growth of corn plants whether sprayed with mancozeb or not. Our results provide the information that plant disease can be well controlled by rhizobacteria-mediated induced systemic resistance in combination with reduced but appropriate application of dithiocarbamate fungicides just before a heavy infection period. An appropriate use of rhizobacteria can enhance plant growth and help plants overcome negative effects caused by dithiocarbamates.

  20. Feasible Management of Southern Corn Leaf Blight via Induction of Systemic Resistance by Bacillus cereus C1L in Combination with Reduced Use of Dithiocarbamate Fungicides

    Science.gov (United States)

    Lai, Yi-Ru; Lin, Pei-Yu; Chen, Chao-Ying; Huang, Chien-Jui

    2016-01-01

    Dithiocarbamate fungicides such as maneb and mancozeb are widely used nonsystemic protectant fungicides to control various plant fungal diseases. Dithiocarbamate fungicides should be frequently applied to achieve optimal efficacy of disease control and avoid either decline in effectiveness or wash-off from leaf surface. Dithiocarbamates are of low resistance risk but have the potential to cause human neurological diseases. The objective of this study was to develop a strategy to effectively control plant disease with reduced use of dithiocarbamtes. Southern corn leaf blight was the model pathosystem for the investigation. When corn plants were drench-treated with Bacillus cereus C1L, a rhizobacterium able to induce systemic resistance in corn plants against southern leaf blight, frequency of spraying dithiocarbamate fungicides could be decreased. The treatment of B. cereus C1L was able to protect maize from southern leaf blight while residues of dithiocarbamates on leaf surface were too low to provide sufficient protection. On the other hand, frequent sprays of mancozeb slightly but significantly reduced growth of corn plants under natural conditions. In contrast, application of B. cereus C1L can significantly promote growth of corn plants whether sprayed with mancozeb or not. Our results provide the information that plant disease can be well controlled by rhizobacteria-mediated induced systemic resistance in combination with reduced but appropriate application of dithiocarbamate fungicides just before a heavy infection period. An appropriate use of rhizobacteria can enhance plant growth and help plants overcome negative effects caused by dithiocarbamates. PMID:27721698

  1. Comparison of Bacillus thuringiensis and Bacillus cereus

    International Nuclear Information System (INIS)

    Bacillus cereus and Bacillus thuringiensis are closely related, spore forming soil bacteria. B. thuringiensis produces insecticidal crystal proteins during sporulation and these toxins are the most important biopesticides in the world today. Genomes of the B. thuringiensis and B. cereus strains were analysed by pulsed field gel electrophoresis after treatment of the DNA with the restriction enzyme NotI. The NotI fingerprint patterns varied both within the B. thuringiensis and the B. cereus strains. The size of the fragments varied between 15 and 1350 kb. When physical maps of the B. thuringiensis and B. cereus strains were compared, B. thuringiensis appeared to be as closely related to B. cereus as the B. cereus strains were to each other. Nine out of 12 B. thuringiensis strains and 18 out of 25 B. cereus strains produced enterotoxins. The close relationship between B. thuringiensis and B. cereus should be taken into consideration when B. thuringiensis is used as a biopesticide. (author). 10 refs, 4 figs, 1 tab

  2. Characterization of Bacillus cereus

    NARCIS (Netherlands)

    Wijnands LM; Dufrenne JB; Leusden FM; MGB

    2002-01-01

    Bacillus cereus is a ubiquitary microorganism that may cause food borne disease. Pathogenicity, however, depends on various characteristics such as the ability to form (entero)-toxin(s) that can not be detected by microbiological methods. Further characterization of pathogenic properties is not only

  3. Biodiversity in Bacillus cereus

    NARCIS (Netherlands)

    Pielaat A; Fricker M; Nauta MJ; Leusden FM van; MGB

    2006-01-01

    Experiments have been performed by different partners to identify variability in properties of Bacillus cereus strains that contribute to the extent of their virulence as part of an EU project. To this end, 100 B. cereus strains were selected and screened for biological properties, such as toxin pro

  4. Essential Bacillus subtilis genes

    DEFF Research Database (Denmark)

    Kobayashi, K.; Ehrlich, S.D.; Albertini, A.;

    2003-01-01

    To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximate to4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes we...

  5. Expression of the genes encoding the CasK/R two-component system and the DesA desaturase during Bacillus cereus cold adaptation.

    Science.gov (United States)

    Diomandé, Sara Esther; Doublet, Bénédicte; Vasaï, Florian; Guinebretière, Marie-Hélène; Broussolle, Véronique; Brillard, Julien

    2016-08-01

    Two-component systems (TCS) allow a cell to elaborate a variety of adaptive responses to environment changes. The recently discovered CasK/R TCS plays a role in the optimal unsaturation of fatty acids necessary for cold adaptation of the foodborne-pathogen Bacillus cereus Here, we showed that the promoter activity of the operon encoding this TCS was repressed during growth at low temperature in the stationary phase in the parental strain when compared to the casK/R mutant, suggesting that CasR negatively regulates the activity of its own promoter in these conditions. The promoter activity of the desA gene encoding the Δ5 fatty acid desaturase, providing unsaturated fatty acids (UFAs) required for low temperature adaptation, was repressed in the casK/R mutant grown at 12°C versus 37°C. This result suggests that CasK/R activates desA expression during B. cereus growth at low temperature, allowing an optimal unsaturation of the fatty acids. In contrast, desA expression was repressed during the lag phase at low temperature in presence of UFAs, in a CasK/R-independent manner. Our findings confirm that the involvement of this major TCS in B. cereus cold adaptation is linked to the upregulation of a fatty acid desaturase. PMID:27435329

  6. Regioselective biooxidation of (+-valencene by recombinant E. coli expressing CYP109B1 from Bacillus subtilis in a two-liquid-phase system

    Directory of Open Access Journals (Sweden)

    Schmid Rolf D

    2009-07-01

    Full Text Available Abstract Background (+-Nootkatone (4 is a high added-value compound found in grapefruit juice. Allylic oxidation of the sesquiterpene (+-valencene (1 provides an attractive route to this sought-after flavoring. So far, chemical methods to produce (+-nootkatone (4 from (+-valencene (1 involve unsafe toxic compounds, whereas several biotechnological approaches applied yield large amounts of undesirable byproducts. In the present work 125 cytochrome P450 enzymes from bacteria were tested for regioselective oxidation of (+-valencene (1 at allylic C2-position to produce (+-nootkatone (4 via cis- (2 or trans-nootkatol (3. The P450 activity was supported by the co-expression of putidaredoxin reductase (PdR and putidaredoxin (Pdx from Pseudomonas putida in Escherichia coli. Results Addressing the whole-cell system, the cytochrome CYP109B1 from Bacillus subtilis was found to catalyze the oxidation of (+-valencene (1 yielding nootkatol (2 and 3 and (+-nootkatone (4. However, when the in vivo biooxidation of (+-valencene (1 with CYP109B1 was carried out in an aqueous milieu, a number of undesired multi-oxygenated products has also been observed accounting for approximately 35% of the total product. The formation of these byproducts was significantly reduced when aqueous-organic two-liquid-phase systems with four water immiscible organic solvents – isooctane, n-octane, dodecane or hexadecane – were set up, resulting in accumulation of nootkatol (2 and 3 and (+-nootkatone (4 of up to 97% of the total product. The best productivity of 120 mg l-1 of desired products was achieved within 8 h in the system comprising 10% dodecane. Conclusion This study demonstrates that the identification of new P450s capable of producing valuable compounds can basically be achieved by screening of recombinant P450 libraries. The biphasic reaction system described in this work presents an attractive way for the production of (+-nootkatone (4, as it is safe and can easily be

  7. Structure based protein engineering of Bacillus stearothermophilus α-amylase: toward a new substrate specificity

    International Nuclear Information System (INIS)

    licheniformis crystal structure as initial model) it seems that Bacillus stearothermophilus α-amylase binding site is more complex with and insertion of 40 residues. Therefore the three dimensional structure is crucial to understand the specificity of the substrate of this enzyme which will be used to drive the design of mutation to introduce new properties for industrial purpose. (author)

  8. Effects of potential probiotic Bacillus amyloliquefaciens [corrected] FPTB16 on systemic and cutaneous mucosal immune responses and disease resistance of catla (Catla catla).

    Science.gov (United States)

    Das, Anushree; Nakhro, Khriezhato; Chowdhury, Supratim; Kamilya, Dibyendu

    2013-11-01

    Effects of dietary administration of Bacillus amyloliquefaciens FPTB16 on systemic and mucosal immunity and disease resistance of catla (Catla catla) against Edwardsiella tarda infection were evaluated in the present study. The laboratory maintained B. amyloliquefaciens was used to study antagonistic activity against fish pathogenic bacteria by agar well diffusion assay. Healthy catla were challenged by this bacterium for determination of its safety. For preparation of probiotic supplemented diet, the bacteria were added to the basal diet (control) at three different inclusion levels i.e., 1 × 10(9), 1 × 10(8) and 1 × 10(7) CFU/g diet. Fish (weight 25-30 g) were fed with these diets and various immune parameters and disease resistance study were conducted at 4 weeks and 8 weeks post-feeding. The bacterial antagonism study showed inhibition zone against E. tarda, Aeromonas hydrophila, Vibrio parahaemolyticus and V. harveyi. B. amyloliquefaciens was harmless to catla as neither mortalities nor morbidities were observed after the challenge. Study of different systemic and mucosal immunological parameters viz. superoxide anion production and nitric oxide production, myeloperoxidase content, lysozyme activity and total protein content showed significant enhancement (p Diet containing 10(8) and 10(9) CFU/g B. amyloliquefaciens significantly enhanced (p < 0.05) the resistance of catla against bacterial challenge at both time points. These results collectively suggest that B. amyloliquefaciens is a potential probiotic species and can be used in aquaculture to improve health status and disease resistance with an optimal dietary supplementation of 10(9) CFU/g.

  9. Single cell FRET analysis for the identification of optimal FRET-pairs in Bacillus subtilis using a prototype MEM-FLIM system.

    Directory of Open Access Journals (Sweden)

    Ruud G J Detert Oude Weme

    Full Text Available Protein-protein interactions can be studied in vitro, e.g. with bacterial or yeast two-hybrid systems or surface plasmon resonance. In contrast to in vitro techniques, in vivo studies of protein-protein interactions allow examination of spatial and temporal behavior of such interactions in their native environment. One approach to study protein-protein interactions in vivo is via Förster Resonance Energy Transfer (FRET. Here, FRET efficiency of selected FRET-pairs was studied at the single cell level using sensitized emission and Frequency Domain-Fluorescence Lifetime Imaging Microscopy (FD-FLIM. For FRET-FLIM, a prototype Modulated Electron-Multiplied FLIM system was used, which is, to the best of our knowledge, the first account of Frequency Domain FLIM to analyze FRET in single bacterial cells. To perform FRET-FLIM, we first determined and benchmarked the best fluorescent protein-pair for FRET in Bacillus subtilis using a novel BglBrick-compatible integration vector. We show that GFP-tagRFP is an excellent donor-acceptor pair for B. subtilis in vivo FRET studies. As a proof of concept, selected donor and acceptor fluorescent proteins were fused using a linker that contained a tobacco etch virus (TEV-protease recognition sequence. Induction of TEV-protease results in loss of FRET efficiency and increase in fluorescence lifetime. The loss of FRET efficiency after TEV induction can be followed in time in single cells via time-lapse microscopy. This work will facilitate future studies of in vivo dynamics of protein complexes in single B. subtilis cells.

  10. Taxonomy Icon Data: Bacillus subtilis [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available g Bacillus_subtilis_S.png Bacillus_subtilis_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus...+subtilis&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus+subtilis&t=NL http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Bacillus+subtilis&t=S http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Bacillus+subtilis&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=214 ...

  11. Fluorescent Amplified Fragment Length Polymorphism Analysis of Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis Isolates

    OpenAIRE

    Hill, Karen K.; Ticknor, Lawrence O.; Okinaka, Richard T.; Asay, Michelle; Blair, Heather; Bliss, Katherine A.; Laker, Mariam; Pardington, Paige E.; Richardson, Amber P.; Tonks, Melinda; Beecher, Douglas J.; Kemp, John D.; Kolstø, Anne-Brit; Wong, Amy C. Lee; Keim, Paul

    2004-01-01

    DNA from over 300 Bacillus thuringiensis, Bacillus cereus, and Bacillus anthracis isolates was analyzed by fluorescent amplified fragment length polymorphism (AFLP). B. thuringiensis and B. cereus isolates were from diverse sources and locations, including soil, clinical isolates and food products causing diarrheal and emetic outbreaks, and type strains from the American Type Culture Collection, and over 200 B. thuringiensis isolates representing 36 serovars or subspecies were from the U.S. D...

  12. Possible Processes for Origin of First Chemoheterotrophic Microorganisms with Modeling of Physiological Processes of Bacterium Bacillus subtilis as a Model System in 2H2O

    Directory of Open Access Journals (Sweden)

    Ignat Ignatov

    2015-09-01

    Full Text Available We studied possible processes for origin of first chemoheterotrophic microorganisms with modeling of physiological processes of a Gram-positive chemoheterotrophic bacterium Bacillus subtilis, producer of purine ribonucleoside inosine as a model system in heavy water. The physiological influence of deuterium on the chemoheterotrophic bacterium B. subtilis was studied on a heavy water (HW medium with a maximal concentration of 2H2O (89–90 atom% 2H. Also various suitable samples of hot mineral water and sea water derived from different sources of Bulgaria were investigated using IR- and DNES-spectroscopy. It was shown that hot alkaline mineral water with temperature from +65 0C to +95 0C and pH value from 9 to 11 is more suitable for the origination of first organic forms than other analyzed water samples. There were discussed the reactions of condensation and dehydration occurring in alkaline aqueous solutions at t = +65–95 0C and рН = 9–10, resulting in synthesis from separate molecules the larger organic molecules as short polipeptides and pyrines, as well as the possible mechanisms of the deuterium accumulation in form of H2HO in hot water. The metabolism of the bacterium B. subtilis and the resistance to deuterium was also analyzed on an evolutionary level taking into account the hydrological conditions of primodial hydrosphere and the presence of H2HO, as well as the qualitative and quantitative composition of the cellular protein, amino acids and carbohydrates on media with maximum deuterium content. It was demonstrated on the example of chemoheterotrophic bacteria that first microorganisms might have been originated in hot mineral water with Ca2+ (0.5-1.0 g/l at t = + 65-95 0C and pH = 9–11, that is more suitable for maintenance and origin of life than other analyzed water samples.

  13. Peptidoglycan metabolism is controlled by the WalRK (YycFG) and PhoPR two-component systems in phosphate-limited Bacillus subtilis cells.

    Science.gov (United States)

    Bisicchia, Paola; Lioliou, Efthimia; Noone, David; Salzberg, Letal I; Botella, Eric; Hübner, Sebastian; Devine, Kevin M

    2010-02-01

    In Bacillus subtilis, the WalRK (YycFG) two-component system controls peptidoglycan metabolism in exponentially growing cells while PhoPR controls the response to phosphate limitation. Here we examine the roles of WalRK and PhoPR in peptidoglycan metabolism in phosphate-limited cells. We show that B. subtilis cells remain viable in a phosphate-limited state for an extended period and resume growth rapidly upon phosphate addition, even in the absence of a PhoPR-mediated response. Peptidoglycan synthesis occurs in phosphate-limited wild-type cells at approximately 27% the rate of exponentially growing cells, and at approximately 18% the rate of exponentially growing cells in the absence of PhoPR. In phosphate-limited cells, the WalRK regulon genes yocH, cwlO(yvcE), lytE and ydjM are expressed in a manner that is dependent on the WalR recognition sequence and deleting these genes individually reduces the rate of peptidoglycan synthesis. We show that ydjM expression can be activated by PhoP approximately P in vitro and that PhoP occupies its promoter in phosphate-limited cells. However, iseA(yoeB) expression cannot be repressed by PhoP approximately P in vitro, but can be repressed by non-phosphorylated WalR in vitro. Therefore, we conclude that peptidoglycan metabolism is controlled by both WalRK and PhoPR in phosphate-limited B. subtilis cells. PMID:20487291

  14. OVERPRODUCTION OF THE ATP-DEPENDENT NUCLEASE ADDAB IMPROVES THE STRUCTURAL STABILITY OF A MODEL PLASMID SYSTEM IN BACILLUS-SUBTILIS

    NARCIS (Netherlands)

    MEIMA, R; HAIJEMA, BJ; VENEMA, G; BRON, S

    1995-01-01

    The effect of the ATP-dependent exonuclease AddAB complex on the structural stability of plasmid pGP1 in Bacillus subtilis was studied. Using deletion mutagenesis and gene amplification techniques, B. subtilis strains were constructed either lacking or overproducing the AddAB complex, a key enzyme i

  15. Comparative genomics of iron-transporting systems in Bacillus cereus strains and impact of iron sources on growth and biofilm formation

    NARCIS (Netherlands)

    Hayrapetyan, Hasmik; Siezen, Roland; Abee, Tjakko; Nierop Groot, Masja

    2016-01-01

    Iron is an important element for bacterial viability, however it is not readily available in most environments. We studied the ability of 20 undomesticated food isolates of Bacillus cereus and two reference strains for capacity to use different (complex) iron sources for growth and biofilm format

  16. Molecular identification and safety of Bacillus species involved in the fermentation of African oil beans (Pentaclethra macrophylla Benth) for production of Ugba.

    Science.gov (United States)

    Ahaotu, I; Anyogu, A; Njoku, O H; Odu, N N; Sutherland, J P; Ouoba, L I I

    2013-03-01

    Molecular identification of Bacillus spp. involved in the fermentation of African oil bean seeds for production of Ugba, as well as ability of the Bacillus spp. isolated to produce toxins, were investigated. Forty-nine bacteria were isolated from Ugba produced in different areas of South Eastern Nigeria and identified by phenotyping and sequencing of 16S rRNA, gyrB and rpoB genes. Genotypic diversities at interspecies and intraspecies level of the isolates were screened by PCR amplification of the 16S-23S rDNA intergenic transcribed spacer (ITS-PCR) and repetitive sequence-based PCR (rep-PCR). The ability of the bacteria to produce toxins was also investigated by detection of genes encoding production of haemolysin BL (HblA, HblC, HblD), non-haemolytic enterotoxin (NheA, NheB, NheC), cytotoxin K (CytK) and emetic toxin (EM1) using PCR with specific primers. Moreover, a Bacillus cereus Enterotoxin Reverse Passive Latex Agglutination test kit (BCET-RPLA) was used to screen ability of the isolates to produce haemolysin in broth and during fermentation of African oil bean seeds. The isolates were characterized as motile, rod-shaped, endospore forming, catalase positive, Gram-positive bacteria. They were identified as Bacillus cereus sensu lato (42), Lysinibacillus xylanilyticus (3), Bacillus clausii (1), Bacillus licheniformis (1), Bacillus subtilis (1), and Bacillus safensis (1). B. cereus was the predominant Bacillus species and was present in all samples studied. Using ITS-PCR, interspecies diversity was observed among isolates, with six clusters representing each of the pre-cited species. Rep-PCR was more discriminatory (eight clusters) and allowed further differentiation at intraspecies level for the B. cereus and L. xylanilyticus isolates with two genotypes for each species. Genes encoding production of non-haemolytic enterotoxin (NheA, NheB, NheC) and cytotoxin K (CytK) genes were detected in all B. cereus isolates, while Hbl genes (HblA, HblC, HblD) were

  17. Genome analysis shows Bacillus axarquiensis is not a later heterotypic synonym of Bacillus mojavensis; reclassification of Bacillus malacitensis and Brevibacterium halotolerans as heterotypic synonyms of Bacillus axarquiensis.

    Science.gov (United States)

    Dunlap, Christopher A; Bowman, Michael J; Schisler, David A; Rooney, Alejandro P

    2016-06-01

    Bacillus axarquiensis and Bacillus malacitensis were previously reported to be later heterotypic synonyms of Bacillus mojavensis, based primarily on DNA-DNA relatedness values. We have sequenced draft genomes of Bacillus axarquiensis NRRL B-41617T and Bacillus malacitensis NRRL B-41618T. Comparative genomics and DNA-DNA relatedness calculations showed that while Bacillus axarquiensis and Bacillus malacitensis are synonymous with each other, they are not synonymous with Bacillus mojavensis. In addition, a draft genome was completed for Brevibacterium halotolerans, a strain long suspected of being a Bacillus subtilis group member based on 16S rRNA similarities (99.8 % with Bacillus mojavensis). Comparative genomics and DNA-DNA relatedness calculations showed that Brevibacterium halotolerans is synonymous with Bacillus axarquiensis and Bacillus malacitensis. The pairwise in silico DNA-DNA hybridization values calculated in comparisons between the three conspecific strains were all greater than 92 %, which is well above the standard species threshold of 70 %. While the pairwise in silico DNA-DNA hybridization values calculated in comparisons of the three conspecific strains with Bacillus mojavensis were all less than 65 %. The combined results of our genotype and phenotype studies showed that Bacillus axarquiensis, Bacillus malacitensis and Brevibacterium halotolerans are conspecific and distinct from Bacillus mojavensis. Because the valid publication of the name Bacillus axarquiensis predates the publication of the name Bacillus malacitensis, we propose that Bacillus malacitensis be reclassified as a synonym of Bacillus axarquiensis. In addition, we propose to reclassify Brevibacterium halotolerans as a synonym of Bacillus axarquiensis. An amended description of Bacillus axarquiensis is provided.

  18. Genome analysis shows Bacillus axarquiensis is not a later heterotypic synonym of Bacillus mojavensis; reclassification of Bacillus malacitensis and Brevibacterium halotolerans as heterotypic synonyms of Bacillus axarquiensis.

    Science.gov (United States)

    Dunlap, Christopher A; Bowman, Michael J; Schisler, David A; Rooney, Alejandro P

    2016-06-01

    Bacillus axarquiensis and Bacillus malacitensis were previously reported to be later heterotypic synonyms of Bacillus mojavensis, based primarily on DNA-DNA relatedness values. We have sequenced draft genomes of Bacillus axarquiensis NRRL B-41617T and Bacillus malacitensis NRRL B-41618T. Comparative genomics and DNA-DNA relatedness calculations showed that while Bacillus axarquiensis and Bacillus malacitensis are synonymous with each other, they are not synonymous with Bacillus mojavensis. In addition, a draft genome was completed for Brevibacterium halotolerans, a strain long suspected of being a Bacillus subtilis group member based on 16S rRNA similarities (99.8 % with Bacillus mojavensis). Comparative genomics and DNA-DNA relatedness calculations showed that Brevibacterium halotolerans is synonymous with Bacillus axarquiensis and Bacillus malacitensis. The pairwise in silico DNA-DNA hybridization values calculated in comparisons between the three conspecific strains were all greater than 92 %, which is well above the standard species threshold of 70 %. While the pairwise in silico DNA-DNA hybridization values calculated in comparisons of the three conspecific strains with Bacillus mojavensis were all less than 65 %. The combined results of our genotype and phenotype studies showed that Bacillus axarquiensis, Bacillus malacitensis and Brevibacterium halotolerans are conspecific and distinct from Bacillus mojavensis. Because the valid publication of the name Bacillus axarquiensis predates the publication of the name Bacillus malacitensis, we propose that Bacillus malacitensis be reclassified as a synonym of Bacillus axarquiensis. In addition, we propose to reclassify Brevibacterium halotolerans as a synonym of Bacillus axarquiensis. An amended description of Bacillus axarquiensis is provided. PMID:27030978

  19. The Eukaryotic-Like Ser/Thr Kinase PrkC Regulates the Essential WalRK Two-Component System in Bacillus subtilis.

    Science.gov (United States)

    Libby, Elizabeth A; Goss, Lindsie A; Dworkin, Jonathan

    2015-06-01

    Most bacteria contain both eukaryotic-like Ser/Thr kinases (eSTKs) and eukaryotic-like Ser/Thr phosphatases (eSTPs). Their role in bacterial physiology is not currently well understood in large part because the conditions where the eSTKs are active are generally not known. However, all sequenced Gram-positive bacteria have a highly conserved eSTK with extracellular PASTA repeats that bind cell wall derived muropeptides. Here, we report that in the Gram-positive bacterium Bacillus subtilis, the PASTA-containing eSTK PrkC and its cognate eSTP PrpC converge with the essential WalRK two-component system to regulate WalR regulon genes involved in cell wall metabolism. By continuously monitoring gene expression throughout growth, we consistently find a large PrkC-dependent effect on expression of several different WalR regulon genes in early stationary phase, including both those that are activated by WalR (yocH) as well as those that are repressed (iseA, pdaC). We demonstrate that PrkC phosphorylates WalR in vitro and in vivo on a single Thr residue located in the receiver domain. Although the phosphorylated region of the receiver domain is highly conserved among several B. subtilis response regulators, PrkC displays specificity for WalR in vitro. Consistently, strains expressing a nonphosphorylatable WalR point mutant strongly reduce both PrkC dependent activation and repression of yocH, iseA, and pdaC. This suggests a model where the eSTK PrkC regulates the essential WalRK two-component signaling system by direct phosphorylation of WalR Thr101, resulting in the regulation of WalR regulon genes involved in cell wall metabolism in stationary phase. As both the eSTK PrkC and the essential WalRK two-component system are highly conserved in Gram-positive bacteria, these results may be applicable to further understanding the role of eSTKs in Gram-positive physiology and cell wall metabolism. PMID:26102633

  20. The Eukaryotic-Like Ser/Thr Kinase PrkC Regulates the Essential WalRK Two-Component System in Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Elizabeth A Libby

    2015-06-01

    Full Text Available Most bacteria contain both eukaryotic-like Ser/Thr kinases (eSTKs and eukaryotic-like Ser/Thr phosphatases (eSTPs. Their role in bacterial physiology is not currently well understood in large part because the conditions where the eSTKs are active are generally not known. However, all sequenced Gram-positive bacteria have a highly conserved eSTK with extracellular PASTA repeats that bind cell wall derived muropeptides. Here, we report that in the Gram-positive bacterium Bacillus subtilis, the PASTA-containing eSTK PrkC and its cognate eSTP PrpC converge with the essential WalRK two-component system to regulate WalR regulon genes involved in cell wall metabolism. By continuously monitoring gene expression throughout growth, we consistently find a large PrkC-dependent effect on expression of several different WalR regulon genes in early stationary phase, including both those that are activated by WalR (yocH as well as those that are repressed (iseA, pdaC. We demonstrate that PrkC phosphorylates WalR in vitro and in vivo on a single Thr residue located in the receiver domain. Although the phosphorylated region of the receiver domain is highly conserved among several B. subtilis response regulators, PrkC displays specificity for WalR in vitro. Consistently, strains expressing a nonphosphorylatable WalR point mutant strongly reduce both PrkC dependent activation and repression of yocH, iseA, and pdaC. This suggests a model where the eSTK PrkC regulates the essential WalRK two-component signaling system by direct phosphorylation of WalR Thr101, resulting in the regulation of WalR regulon genes involved in cell wall metabolism in stationary phase. As both the eSTK PrkC and the essential WalRK two-component system are highly conserved in Gram-positive bacteria, these results may be applicable to further understanding the role of eSTKs in Gram-positive physiology and cell wall metabolism.

  1. An efficient fusion protein system for expression ofBacillus anthracis protective antigen as immunogenic and diagnostic antigen

    Institute of Scientific and Technical Information of China (English)

    Vahid Bagheri; Hossein Motamedi; Masoud Reza Seifiabad Shapouri

    2010-01-01

    Objective:To produce high quantities of recombinant protective antigen (rPA) for human vaccine and diagnosis.Methods: ThePAgene was amplified byPCR with pXO1 plasmid as template. ThePCR product was cloned into pMAL-c2X vector using theBamHI andSalI restriction enzymes. The recombinant plasmid was transformed intoEscherichia coliDH5α strain and then screened for transformation. The expression of protective antigen was analyzed bySDS-PAGE and Western blotting after isopropyl β-D-thiogalactopyranoside(IPTG) induction.Results:The full-length PA gene (2.2kb) was cloned into pMAL vector system. The recombinant vector was confirmed by restriction enzyme andPCRanalysis. The expression of cytoplasmic maltose-binding protein-protective (MBP-P) antigen fusion protein was detected bySDS-PAGE and Western blotting, and obtained a125 kDa protein band, which was similar to expected size of fusion protein.Conclusions: This expression system can be used in the high production of rPA. After purification and immunization studies, the purified rPA may be used in the development of the human recombinant anthrax vaccine and also in diagnosis of anthrax disease.

  2. 牛栏山二锅头酒醅中芽孢杆菌分离鉴定及发酵风味分析%Identification of Bacillus from Niulanshan Erguotou fermented grain and analysis of flavor compounds in the fermentation

    Institute of Scientific and Technical Information of China (English)

    杨春霞; 廖永红; 刘峻雄; 胡建华; 胡佳音; 窦屾

    2012-01-01

    从牛栏山二锅头酒醅中分离筛选出5株产风味物质能力较好的芽孢杆菌,通过16SrDNA序列分析和构建系统发育树,5株细菌分别为地衣芽孢杆菌(Bacillus licheniformis)、蜡样芽孢杆菌(Bacillus cereus)、短小芽孢杆菌(Bacillus pumilus)和枯草芽孢杆菌(Bacillus subtilis)。分别对它们进行发酵风味分析,其发酵液经固相微萃取和GC-MS分析,并除去空白培养基中物质,地衣芽孢杆菌BL-1发酵液共检测得到14种风味物质,蜡样芽孢杆菌BC-1和短小芽孢杆菌BP-1发酵都得到12种风味物质,枯草芽孢杆菌BS-1好氧发酵共得到16种风味物质,枯草芽孢杆菌BS-2厌氧发酵共得到19种风味物质。除短小芽孢杆菌外,其他4株芽孢杆菌都含有较多数量的酯类化合物,且主要代谢风味物质都是3-羟基-2-丁酮,而短小芽孢杆菌BP-1则含有数量较多的烃类化合物,其主要风味物质是苯乙醇。%Five strains of bacillus which can produce flavor were screened from Niulanshan Erguotou fermented grain.Using the sequences analysis of 16S rDNA and phylogenetic tree construction,five strains were identified as Bacillus licheniformis,Bacillus cereus,Bacillus pumilus and Bacillus subtilis.The fermentation broth of five bacillus strains were analyzed by solid phase micro-extraction and chromatography-mass spectrometry.Removing the compounds of blank,a total of 14 flavor compounds in fermentation broth of Bacillus licheniformis BL-1,12 flavor compounds in fermentation broth of Bacillus cereus BC-1 and Bacillus pumilus BP-1,16 flavor compounds in fermentation broth of Bacillus subtilis BS-1,19 flavor compounds in fermentation broth of Bacillus subtilis BS-2.Except Bacillus pumilus,the fermentation of other four Bacillus strains mainly contained esters compound,and 3-hydroxy-2-butanone was the most important flavor compound.However,the fermentation of Bacillus pumilus BP-1 mainly comprised alkynes compound and

  3. Production of Levan by Bacillus licheniformis for Use as a Soil Sealant in Earthen Manure Storage Structures

    OpenAIRE

    Abdel E. Ghaly; F. Arab; N. S. Mahmoud; Higgins, J

    2007-01-01

    Manure application is not permitted on frozen land in Canada and therefore, manure management and storage are the primary issues facing the agri-food industry. Low-cost, effective and environmentally safe earthen manure storage (EMS) facilities will lower costs and help make the livestock industry more competitive and efficient. The goal of this study was to develop a biological sealing technology for earthen manure storages. The results showed that it is feasible to use a growing cultu...

  4. Bacillus thuringiensis (Bt)

    Science.gov (United States)

    2004-01-01

    Bacillus thuringiensis (Bt), a natural bacteria found all over the Earth, has a fairly novel way of getting rid of unwanted insects. Bt forms a protein substance (shown on the right) that is not harmful to humans, birds, fish or other vertebrates. When eaten by insect larvae the protein causes a fatal loss of appetite. For over 25 years agricultural chemical companies have relied heavily upon safe Bt pesticides. New space based research promises to give the insecticide a new dimension in effectiveness and applicability. Researchers from the Consortium for Materials Development in Space along with industrial affiliates such as Abott Labs and Pern State University flew Bt on a Space Shuttle mission in the fall of 1996. Researchers expect that the Shuttle's microgravity environment will reveal new information about the protein that will make it more effective against a wider variety of pests.

  5. Induction of systemic resistance in tomato by the autochthonous phylloplane resident Bacillus cereus Indução de resistência sistêmica em tomateiro pela bactéria autóctone residente no filoplano Bacillus cereus

    Directory of Open Access Journals (Sweden)

    Bernardo de Almeida Halfeld-Vieira

    2006-08-01

    Full Text Available The objective of this work was to verify if the induced resistance mechanism is responsible for the capacity of a phylloplane resident bacteria (Bacillus cereus, isolated from healthy tomato plants, to control several diseases of this crop. A strain of Pseudomonas syringae pv. tomato was used as the challenging pathogen. The absence of direct antibiosis of the antagonist against the pathogen, the significant increase in peroxidases activity in tomato plants exposed to the antagonist and then inoculated with the challenging pathogen, as well as the character of the protection, are evidences wich suggest that biocontrol efficiency presented by the antagonist in previous works might be due to induced systemic resistance (ISR.O objetivo deste trabalho foi investigar se o mecanismo de indução de resistência é responsável pela capacidade de um isolado bacteriano residente no filoplano (Bacillus cereus, obtido de plantas sadias de tomateiro, em controlar diversas doenças da cultura. A fim de investigar o modo de ação envolvido no controle, foi utilizado um isolado de Pseudomonas syringae pv. tomato como patógeno desafiador. A ausência do mecanismo de antibiose contra o patógeno, o aumento significativo dos níveis de peroxidases em plantas expostas ao antagonista e submetidas à inoculação do patógeno desafiador, assim como a sistemicidade da proteção, são evidências que sugerem que o mecanismo de biocontrole em questão se refere à indução de resistência sistêmica (IRS.

  6. Isolation and identification of bacillus species from soil samples in anthrax epidemic area%炭疽高发地区土壤样本中常见芽胞杆菌的分离及鉴定

    Institute of Scientific and Technical Information of China (English)

    张慧娟; 魏建春; 张恩民; 张建华

    2012-01-01

    Objective To isolate and identify bacillus species in soil samples in anthrax epidemic area and evaluate the disinfection effect and understand the distributions of common bacillus species in the area. Methods Sixty soil samples were collected from the anthrax epidemic area to isolate and identify related bacillus species with PCR assay. Results No Bacillus anthracis was identified, but 13 strains of Bacillus licheniformis, 8 strains of Bacillus subtilis, 11 strains of Bacillus pumilus and 1 strain of Bacillus cereus were identified in 33 gene fragments from 48 clones by sequencing and blast alignment. The specificities of the primers for Bacillus megatherium and Bacillus circulans were not high. Conclusion The disinfection effect in the anthrax epidemic area was good. The related bacillus species exist widely in the soil, suggesting that their identifications are needed in anthrax surveillance by specific genes amplification%目的 通过分离鉴定炭疽可疑污染土壤样本的芽胞杆菌,评价消毒效果和了解监测地区土壤中的芽胞杆菌分布情况.方法 采集炭疽监测点土壤样本60份,对炭疽芽胞杆菌和其他芽胞杆菌进行分离培养和聚合酶链反应扩增鉴定.结果 在可疑污染土壤样本中未分离到炭疽芽胞杆菌;从分离到的48个单克隆菌落中扩增到33个目的片段,经测序和blast比对,确定得到地衣芽胞杆菌13株,枯草芽胞杆菌8株,短小芽胞杆菌扩增11株,蜡样芽胞杆菌扩增到1株,巨大芽胞杆菌引物和环状芽胞杆菌引物特异性不好.结论 本研究提示该监测点炭疽疫情消毒效果可信,但需要进一步研究验证;几种芽胞杆菌在土壤中广泛存在,在炭疽监测工作中进行病原体分离时需要加以鉴别,可通过特异基因扩增来辅助检验.

  7. The Eukaryotic-Like Ser/Thr Kinase PrkC Regulates the Essential WalRK Two-Component System in Bacillus subtilis

    OpenAIRE

    Libby, Elizabeth A.; Goss, Lindsie A.; Jonathan Dworkin

    2015-01-01

    Most bacteria contain both eukaryotic-like Ser/Thr kinases (eSTKs) and eukaryotic-like Ser/Thr phosphatases (eSTPs). Their role in bacterial physiology is not currently well understood in large part because the conditions where the eSTKs are active are generally not known. However, all sequenced Gram-positive bacteria have a highly conserved eSTK with extracellular PASTA repeats that bind cell wall derived muropeptides. Here, we report that in the Gram-positive bacterium Bacillus subtilis, th...

  8. Genome Differences That Distinguish Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis

    OpenAIRE

    Radnedge, Lyndsay; Agron, Peter G.; Hill, Karen K.; Jackson, Paul J.; Ticknor, Lawrence O; Keim, Paul; Andersen, Gary L.

    2003-01-01

    The three species of the group 1 bacilli, Bacillus anthracis, B. cereus, and B. thuringiensis, are genetically very closely related. All inhabit soil habitats but exhibit different phenotypes. B. anthracis is the causative agent of anthrax and is phylogenetically monomorphic, while B. cereus and B. thuringiensis are genetically more diverse. An amplified fragment length polymorphism analysis described here demonstrates genetic diversity among a collection of non-anthrax-causing Bacillus speci...

  9. Production of Cellulases, Xylanase, Pectinase, alpha-amylase and Protease Enzymes Cocktail by Bacillus spp. and Their Mixed Cultures with Candida tropicalis and Rhodotorula glutinis under Solid State Fermentation

    International Nuclear Information System (INIS)

    A group of twelve locally isolated Bacillus species, B.megaterium (MAI and MA II), B.licheniformis (MLI and ML II); B. circulans, B. stearothermophilis, B.cereus, B.sphaericus, B. pumilus, B. laterosporus, B. coagulans and B. pantothenticus, were examined for the production of cellulases, xylanase, pectinase, alpha-amylase and protease enzymes cocktail on wheat bran under solid state fermentation (SSF). All species were found to be potent hydrolyzing enzymes producers and the superior producing species were B. megaterium MAI and B. licheniformis. On the other hand, both of them still produced highest enzyme titres when mixed with Candida tropicalis or Rhodotorula glutinis, yeast strains. The two superior bacterial strains produced the highest enzymatic activities when coculturing with C. tropicalis compared with coculturing with R. glutinis only or with both C. tropicalis and R. glutinis in combination. The inferior activities of cocultures (B. megaterinm MAI and R. glutinis) were enhanced in carboxymethyl cellulase (CMCase), filter paper cellulase (FPase), avecilase, xylanase, pectinase, -amylase and protease by gamma irradiation at dose 1.0 kGy with percent increase 8 %, 20 %, 10 %, 4 %, 31 %, 22 % and 34 %, respectively as compared with un-irradiated cocultures

  10. Genome Sequence of Bacillus endophyticus and Analysis of Its Companion Mechanism in the Ketogulonigenium vulgare-Bacillus Strain Consortium.

    Directory of Open Access Journals (Sweden)

    Nan Jia

    Full Text Available Bacillus strains have been widely used as the companion strain of Ketogulonigenium vulgare in the process of vitamin C fermentation. Different Bacillus strains generate different effects on the growth of K. vulgare and ultimately influence the productivity. First, we identified that Bacillus endophyticus Hbe603 was an appropriate strain to cooperate with K. vulgare and the product conversion rate exceeded 90% in industrial vitamin C fermentation. Here, we report the genome sequencing of the B. endophyticus Hbe603 industrial companion strain and speculate its possible advantage in the consortium. The circular chromosome of B. endophyticus Hbe603 has a size of 4.87 Mb with GC content of 36.64% and has the highest similarity with that of Bacillus megaterium among all the bacteria with complete genomes. By comparing the distribution of COGs with that of Bacillus thuringiensis, Bacillus cereus and B. megaterium, B. endophyticus has less genes related to cell envelope biogenesis and signal transduction mechanisms, and more genes related to carbohydrate transport and metabolism, energy production and conversion, as well as lipid transport and metabolism. Genome-based functional studies revealed the specific capability of B. endophyticus in sporulation, transcription regulation, environmental resistance, membrane transportation, extracellular proteins and nutrients synthesis, which would be beneficial for K. vulgare. In particular, B. endophyticus lacks the Rap-Phr signal cascade system and, in part, spore coat related proteins. In addition, it has specific pathways for vitamin B12 synthesis and sorbitol metabolism. The genome analysis of the industrial B. endophyticus will help us understand its cooperative mechanism in the K. vulgare-Bacillus strain consortium to improve the fermentation of vitamin C.

  11. Genome Sequence of Bacillus endophyticus and Analysis of Its Companion Mechanism in the Ketogulonigenium vulgare-Bacillus Strain Consortium.

    Science.gov (United States)

    Jia, Nan; Du, Jin; Ding, Ming-Zhu; Gao, Feng; Yuan, Ying-Jin

    2015-01-01

    Bacillus strains have been widely used as the companion strain of Ketogulonigenium vulgare in the process of vitamin C fermentation. Different Bacillus strains generate different effects on the growth of K. vulgare and ultimately influence the productivity. First, we identified that Bacillus endophyticus Hbe603 was an appropriate strain to cooperate with K. vulgare and the product conversion rate exceeded 90% in industrial vitamin C fermentation. Here, we report the genome sequencing of the B. endophyticus Hbe603 industrial companion strain and speculate its possible advantage in the consortium. The circular chromosome of B. endophyticus Hbe603 has a size of 4.87 Mb with GC content of 36.64% and has the highest similarity with that of Bacillus megaterium among all the bacteria with complete genomes. By comparing the distribution of COGs with that of Bacillus thuringiensis, Bacillus cereus and B. megaterium, B. endophyticus has less genes related to cell envelope biogenesis and signal transduction mechanisms, and more genes related to carbohydrate transport and metabolism, energy production and conversion, as well as lipid transport and metabolism. Genome-based functional studies revealed the specific capability of B. endophyticus in sporulation, transcription regulation, environmental resistance, membrane transportation, extracellular proteins and nutrients synthesis, which would be beneficial for K. vulgare. In particular, B. endophyticus lacks the Rap-Phr signal cascade system and, in part, spore coat related proteins. In addition, it has specific pathways for vitamin B12 synthesis and sorbitol metabolism. The genome analysis of the industrial B. endophyticus will help us understand its cooperative mechanism in the K. vulgare-Bacillus strain consortium to improve the fermentation of vitamin C.

  12. 米醋中蜡状芽孢杆菌DNA提取及其ERIC-PCR体系建立%Extraction of Bacillus cereus DNA in Vinegar and the Establishment of ERIC-PCR System

    Institute of Scientific and Technical Information of China (English)

    廖永红; 任文雅; 孙宝国; 徐瑾; 沈晗

    2011-01-01

    以蜡状芽孢杆菌(Bacillus cereus)为试验材料,比较细菌基因组DNA的提取方法.将ERIC-PCR应用于醋液分离菌的研究,对此反应体系的主要因素进行优化,最终建立了适合于此菌种的ERIC-PCR体系.采用改良的传统细菌基因组DNA提取方法,所提取的DNA质量较高,能够满足ERIC-PCR反应的需要.反应体系:25μL反应体积10×扩增缓冲液(含Mg2+)2.5 μL,20 pmol/μL ERIC-PCR引物E1 1μL,20 pmol/μL ERIC-PCR引物E2 1μL,DNA模板2μL,2.5 mmol/LdNTPs混合液2.0 μL,taq聚合酶1.1 μL,双蒸水补齐;PCR反应程序为:94℃变性3min,1个循环;94℃变性30 s,55℃退火40 s,72℃延伸1min,30个循环;72℃延伸4min.%Bacillus cereus was used as experimental materials. Hie method of genomic DNA extraction was studied in Bacillus and the conditions of ERIC-PCR. Finally, ERIC-PCR reaction system suitable for our lab was established. The results showed that high-grade genomic DNA which could meet the requirements of PCR reaction was obtained by the modified methods. The established ERIC-PCR reaction system was as follows: 10×Buffer (Mg2+) 2.5 μL, 20 pmol/μL primer E1 1 μL, 20 pmol/μL primer E2 1 μL, DNA template 2 μL, 2.5 mmol/L dNTPs 2 μL, Taq polymerase 1.1 μL, ddH2O 15.4 μL, 25 μL reaction volume. The reaction program of PCR was devised as follows: 3 min degeneration at 94 ℃ (one cycle), then 30s degeneration at 94 ℃, 40s annealing at 55 ℃, and 1 min extension at 72 ℃(30cycles), and then 4min final extension at 72 ℃.

  13. NCBI nr-aa BLAST: CBRC-TGUT-17-0007 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TGUT-17-0007 ref|YP_078884.1| cytochrome caa3 oxidase (subunit I) [Bacillus lichen...iformis ATCC 14580] ref|YP_091296.1| CtaD [Bacillus licheniformis ATCC 14580] gb|AAU23246.1| cytochrome ...caa3 oxidase (subunit I) [Bacillus licheniformis ATCC 14580] gb|AAU40603.1| CtaD [Bacillus licheniformis DSM 13] YP_078884.1 0.12 29% ...

  14. NCBI nr-aa BLAST: CBRC-AGAM-02-0116 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-AGAM-02-0116 ref|YP_081361.1| gluconate permease [Bacillus licheniformis ATCC ...14580] ref|YP_093794.1| GntP [Bacillus licheniformis ATCC 14580] gb|AAU25723.1| gluconate permease [Bacillus lichen...iformis ATCC 14580] gb|AAU43101.1| GntP [Bacillus licheniformis DSM 13] YP_081361.1 1.8 41% ...

  15. Phages Preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: Past, Present and Future

    Directory of Open Access Journals (Sweden)

    Annika Gillis

    2014-07-01

    Full Text Available Many bacteriophages (phages have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here.

  16. Phages preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: past, present and future.

    Science.gov (United States)

    Gillis, Annika; Mahillon, Jacques

    2014-07-01

    Many bacteriophages (phages) have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here. PMID:25010767

  17. The fate of Bacillus cereus in the gastrointestinal tract

    NARCIS (Netherlands)

    Pielaat A; Wijnands LM; Takumi K; Nauta MJ; Leusden FM van; MGB

    2006-01-01

    This report presents a mathematical dynamical model for the behaviour of Bacillus cereus in the gastro-intestinal tract. Biological processes and system dynamics are simultaneously incorporated in this mechanistic model. Variability in growth characteristics and physical traits of different B. cereu

  18. Discrimination of press fit candidate microorganism (Enterobacter cloacae, Bacillus licheniformis) by restriction fragment length polymorphic analysis of the 16SrRNA gene; 16S rRNA idenshi no sengen danpen kchotakei kaiseki niyoru atsunyukoho biseibutsu (Enterobacter cloacae, Bacillus licheni-formis) no shikibetsu

    Energy Technology Data Exchange (ETDEWEB)

    Fujiwara, Kazuhiro; Tanaka, Shinji; Otsuka, Makiko; Ichimura, Naoya; Yonebayashi, Eiji; Enomoto, Heiji

    1999-09-01

    In MeOH viewed as one of the improvement method for recovery of the petroleum with hope, the development of discrimination technique of press fit candidate microorganism and oil reservoir resident microorganism which exists in the test object oil reservoir was tried in order to monitor the survival situation of the microorganism which inserted in the oil reservoir under pressure. 16S rRNA amplified by the PCR using the universal primer The microorganism that it cut off the gene at restriction enzyme HhaI,MspI, AluI and inhabits oil reservoir water and oil reservoir rock in the object oil reservoir by ( necessarily TaqI ) and restriction fragment length polymorphic analysis was classified. As the result, the effectiveness of the this PCR-RFLP method was indicated the microorganism which showed RFLP pattern which is identical with the press fit candidate microorganism in the oil reservoir resident microorganism for the discrimination of the press fit candidate microorganism without existing. And, it was indicated that the this PCR-RFLP method was effective for the investigation of oil reservoir resident microbial community which can positively utilize source of nutrition inserted to oil reservoir with the press fit candidate microorganism under pressure, and it was possible to grasp oil reservoir resident microorganism to be especially considered in MEOR. (translated by NEDO)

  19. NCBI nr-aa BLAST: CBRC-DDIS-03-0023 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DDIS-03-0023 ref|YP_080913.1| Sugar transporter YwtG [Bacillus licheniformis A...TCC 14580] ref|YP_093341.1| YwtG [Bacillus licheniformis ATCC 14580] gb|AAU42648.1| YwtG [Bacillus licheniformis DSM 13] YP_080913.1 3e-51 35% ...

  20. Bacillus subtilis MreB orthologs self-organize into filamentous structures underneath the cell membrane in a heterologous cell system.

    Directory of Open Access Journals (Sweden)

    Felix Dempwolff

    Full Text Available Actin-like bacterial cytoskeletal element MreB has been shown to be essential for the maintenance of rod cell shape in many bacteria. MreB forms rapidly remodelling helical filaments underneath the cell membrane in Bacillus subtilis and in other bacterial cells, and co-localizes with its two paralogs, Mbl and MreBH. We show that MreB localizes as dynamic bundles of filaments underneath the cell membrane in Drosophila S2 Schneider cells, which become highly stable when the ATPase motif in MreB is modified. In agreement with ATP-dependent filament formation, the depletion of ATP in the cells lead to rapid dissociation of MreB filaments. Extended induction of MreB resulted in the formation of membrane protrusions, showing that like actin, MreB can exert force against the cell membrane. Mbl also formed membrane associated filaments, while MreBH formed filaments within the cytosol. When co-expressed, MreB, Mbl and MreBH built up mixed filaments underneath the cell membrane. Membrane protein RodZ localized to endosomes in S2 cells, but localized to the cell membrane when co-expressed with Mbl, showing that bacterial MreB/Mbl structures can recruit a protein to the cell membrane. Thus, MreB paralogs form a self-organizing and dynamic filamentous scaffold underneath the membrane that is able to recruit other proteins to the cell surface.

  1. Characterization of the mucosal and systemic immune response induced by Cry1Ac protein from Bacillus thuringiensis HD 73 in mice

    Directory of Open Access Journals (Sweden)

    R.I. Vázquez-Padrón

    2000-02-01

    Full Text Available The present paper describes important features of the immune response induced by the Cry1Ac protein from Bacillus thuringiensis in mice. The kinetics of induction of serum and mucosal antibodies showed an immediate production of anti-Cry1Ac IgM and IgG antibodies in serum after the first immunization with the protoxin by either the intraperitoneal or intragastric route. The antibody fraction in serum and intestinal fluids consisted mainly of IgG1. In addition, plasma cells producing anti-Cry1Ac IgG antibodies in Peyer's patches were observed using the solid-phase enzyme-linked immunospot (ELISPOT. Cry1Ac toxin administration induced a strong immune response in serum but in the small intestinal fluids only anti-Cry1Ac IgA antibodies were detected. The data obtained in the present study confirm that the Cry1Ac protoxin is a potent immunogen able to induce a specific immune response in the mucosal tissue, which has not been observed in response to most other proteins.

  2. Environmental Persistence of Bacillus anthracis and Bacillus subtilis Spores

    Science.gov (United States)

    Wood, Joseph P.; Meyer, Kathryn M.; Kelly, Thomas J.; Choi, Young W.; Rogers, James V.; Riggs, Karen B.; Willenberg, Zachary J.

    2015-01-01

    There is a lack of data for how the viability of biological agents may degrade over time in different environments. In this study, experiments were conducted to determine the persistence of Bacillus anthracis and Bacillus subtilis spores on outdoor materials with and without exposure to simulated sunlight, using ultraviolet (UV)-A/B radiation. Spores were inoculated onto glass, wood, concrete, and topsoil and recovered after periods of 2, 14, 28, and 56 days. Recovery and inactivation kinetics for the two species were assessed for each surface material and UV exposure condition. Results suggest that with exposure to UV, decay of spore viability for both Bacillus species occurs in two phases, with an initial rapid decay, followed by a slower inactivation period. The exception was with topsoil, in which there was minimal loss of spore viability in soil over 56 days, with or without UV exposure. The greatest loss in viable spore recovery occurred on glass with UV exposure, with nearly a four log10 reduction after just two days. In most cases, B. subtilis had a slower rate of decay than B. anthracis, although less B. subtilis was recovered initially. PMID:26372011

  3. FORMALDEHYDE GAS INACTIVATION OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACE MATERIALS.

    Science.gov (United States)

    Research evaluated the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface material using formaldehyde gas. Spores were dried on seven types of indoor surfaces and exposed to 1100 ppm formaldehyde gas for 10 hr. Fo...

  4. Screening of Bacillus Species with Potentials of Antibiotics Production

    Directory of Open Access Journals (Sweden)

    Faruk Adamu KUTA

    2009-07-01

    Full Text Available Sixteen soil samples were collected from different refuse dump sites in Minna, the capital Niger State, and analysed for the presence of Bacillus species. Physical-chemical analysis of the soil samples revealed the followings: PH value 6.89-8.47; moisture content 1.58 – 21.21% and temperature 27-28ºC. Using both pour plate and streak method of inoculation, total bacterial count in the soil samples ranged from 3.8×104 cfu/g 16.0×104 cfu/g. The identified Bacillus species included: Bacillus cereus (30.8%, Bacillus brevis (1.9% Bacillus polymyxa (3.8%, Bacillus lichenifomis (13.5%, Bacillus spherericus (7.7%, Bacillus mycoides (13.5%, Bacillus pumilus (7.7%, Bacillus subtilis (3.8%, Bacillus alvei (1.9%, Bacillus laterosporous (1.9%, Bacillus firmus (9.6% and Bacillus circulars (3.8%. Antibiotic production tests indicated that nine Bacillus species out of twelve isolated in this study could be used to produce antibiotics that had effect on the test organisms. However, Bacillus polymyxa, Bacillus sphaericus and Bacillus laterosporous had little or no effect on the tested organisms. This study suggests that some Bacillus species have potential to produce high quality antibiotics that can be use to control microbial growth in future.

  5. Identification and Properties of Amylase-producing Bacillus Isolated from Songhe Distiller's Yeast%宋河酒曲中产淀粉酶芽孢菌的分离鉴定及产酶特性

    Institute of Scientific and Technical Information of China (English)

    侯小歌; 王俊英; 张杰; 李学思; 李绍亮; 胡炳义

    2012-01-01

    为了有效控制宋河酒曲的制曲过程和发酵进程,对宋河酒曲中产淀粉酶芽孢菌进行了分离鉴定并研究筛选菌株的产淀粉酶特性.结果表明:从宋河酒曲中共分离到12株产淀粉酶芽孢菌,均属革兰氏阳性杆状菌,初步鉴定归为1个属5个种,即芽孢杆菌属(Bacillus)、坚硬芽孢杆菌(Bacillus firmus)、枯草芽孢杆菌( Bacillus subtilis)、地衣芽孢杆菌(Bacillus licheni formis)、凝结芽孢杆菌(Bacillus coagulans)、巨大芽孢杆菌(Bacillus megaterium),其中,坚硬芽孢杆菌是宋河大曲产淀粉酶芽孢菌的数量优势菌群,获得1株地衣芽孢杆菌SQ2为中温型高产淀粉酶菌株,其液态发酵产酶特性为37℃培养,前24 h产酶较弱,此后,酶活力迅速上升,72 h达到最大值为89 μg/(mL·min),产酶旺盛期发生在菌体成熟期和衰亡初期,产酶过程pH值先稍偏酸性后接近中性.%Amylase-producing bacilli isolated from Songhe Distiller's yeast were identified and their properties were studied to effectively control the starter-making process and fermentation process. The results showed that 12 bacillus strains which could produce amylase were isolated and identified as Bacillus firmus, Bacillus subtilis, Bacillus licheniformis, Bacillus coagulans and Bacillus megaterium , which belonged to genus of Bacillus. The B. firmus was the dominant amylase-producing bacillus in Songhe Daqu. Strain SQ2 was obtained as a high producing-amylase bacillus by determining transparent circle, its properties of producing amylase brothing under 37℃ were showed as follows: amylase-producing activity kept minimum in first fermenting 24 hours, gradually raised after 24 hours, reached primary stabilizing value to 72 hours, and decreased gently after fermenting 72 hours, the maximum value of amylase activity could reach 89 μg/(mL · min), the maximum amylase production occurred at the stage of strain maturity and decline growth. The pH value kept slightly acid

  6. The poly-γ-d-glutamic acid capsule surrogate of the Bacillus anthracis capsule induces nitric oxide production via the platelet activating factor receptor signaling pathway.

    Science.gov (United States)

    Lee, Hae-Ri; Jeon, Jun Ho; Park, Ok-Kyu; Chun, Jeong-Hoon; Park, Jungchan; Rhie, Gi-Eun

    2015-12-01

    The poly-γ-d-glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis, confers protection of the bacillus from phagocytosis and allows its unimpeded growth in the host. PGA capsules released from B. anthracis are associated with lethal toxin in the blood of experimentally infected animals and enhance the cytotoxic effect of lethal toxin on macrophages. In addition, PGA capsule itself activates macrophages and dendritic cells to produce proinflammatory cytokine such as IL-1β, indicating multiple roles of PGA capsule in anthrax pathogenesis. Here we report that PGA capsule of Bacillus licheniformis, a surrogate of B. anthracis capsule, induces production of nitric oxide (NO) in RAW264.7 cells and bone marrow-derived macrophages. NO production was induced by PGA in a dose-dependent manner and was markedly reduced by inhibitors of inducible NO synthase (iNOS), suggesting iNOS-dependent production of NO. Induction of NO production by PGA was not observed in macrophages from TLR2-deficient mice and was also substantially inhibited in RAW264.7 cells by pretreatment of TLR2 blocking antibody. Subsequently, the downstream signaling events such as ERK, JNK and p38 of MAPK pathways as well as NF-κB activation were required for PGA-induced NO production. In addition, the induced NO production was significantly suppressed by treatment with antagonists of platelet activating factor receptor (PAFR) or PAFR siRNA, and mediated through PAFR/Jak2/STAT-1 signaling pathway. These findings suggest that PGA capsule induces NO production in macrophages by triggering both TLR2 and PAFR signaling pathways which lead to activation of NF-kB and STAT-1, respectively. PMID:26350415

  7. The poly-γ-d-glutamic acid capsule surrogate of the Bacillus anthracis capsule induces nitric oxide production via the platelet activating factor receptor signaling pathway.

    Science.gov (United States)

    Lee, Hae-Ri; Jeon, Jun Ho; Park, Ok-Kyu; Chun, Jeong-Hoon; Park, Jungchan; Rhie, Gi-Eun

    2015-12-01

    The poly-γ-d-glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis, confers protection of the bacillus from phagocytosis and allows its unimpeded growth in the host. PGA capsules released from B. anthracis are associated with lethal toxin in the blood of experimentally infected animals and enhance the cytotoxic effect of lethal toxin on macrophages. In addition, PGA capsule itself activates macrophages and dendritic cells to produce proinflammatory cytokine such as IL-1β, indicating multiple roles of PGA capsule in anthrax pathogenesis. Here we report that PGA capsule of Bacillus licheniformis, a surrogate of B. anthracis capsule, induces production of nitric oxide (NO) in RAW264.7 cells and bone marrow-derived macrophages. NO production was induced by PGA in a dose-dependent manner and was markedly reduced by inhibitors of inducible NO synthase (iNOS), suggesting iNOS-dependent production of NO. Induction of NO production by PGA was not observed in macrophages from TLR2-deficient mice and was also substantially inhibited in RAW264.7 cells by pretreatment of TLR2 blocking antibody. Subsequently, the downstream signaling events such as ERK, JNK and p38 of MAPK pathways as well as NF-κB activation were required for PGA-induced NO production. In addition, the induced NO production was significantly suppressed by treatment with antagonists of platelet activating factor receptor (PAFR) or PAFR siRNA, and mediated through PAFR/Jak2/STAT-1 signaling pathway. These findings suggest that PGA capsule induces NO production in macrophages by triggering both TLR2 and PAFR signaling pathways which lead to activation of NF-kB and STAT-1, respectively.

  8. Evaluation of flocculating performance of a thermostable bioflocculant produced by marine Bacillus sp.

    Science.gov (United States)

    Okaiyeto, Kunle; Nwodo, Uchechukwu U; Mabinya, Leonard V; Okoli, Arinze S; Okoh, Anthony I

    2016-01-01

    This study assessed the bioflocculant (named MBF-W7) production potential of a bacterial isolate obtained from Algoa Bay, Eastern Cape Province of South Africa. The 16S ribosomal deoxyribonucleic acids gene sequence analysis showed 98% sequence similarity to Bacillus licheniformis strain W7. Optimum culture conditions for MBF-W7 production include 5% (v/v) inoculum size, maltose and NH4NO3 as carbon and nitrogen sources of choice, medium pH of 6 as the initial pH of the growth medium. Under these optimal conditions, maximum flocculating activity of 94.9% was attained after 72 h of cultivation. Chemical composition analyses showed that the purified MBF-W7 was a glycoprotein which was predominantly composed of polysaccharides 73.7% (w/w) and protein 6.2% (w/w). Fourier transform infrared spectroscopy revealed the presence of hydroxyl, carboxyl and amino groups as the main functional groups identified in the bioflocculant molecules. Thermogravimetric analyses showed the thermal decomposition profile of MBF-W7. Scanning electron microscopy imaging revealed that bridging played an important role in flocculation. MBF-W7 exhibited excellent flocculating activity for kaolin clay suspension at 0.2 mg/ml over a wide pH range of 3-11; with the maximal flocculation rate of 85.8% observed at pH 3 in the presence of Mn(2+). It maintained and retained high flocculating activity of over 70% after heating at 100°C for 60 min. MBF-W7 showed good turbidity removal potential (86.9%) and chemical oxygen demand reduction efficiency (75.3%) in Tyume River. The high flocculating rate of MBF-W7 makes it an attractive candidate to replace chemical flocculants utilized in water treatment. PMID:26797258

  9. Comparative genomics analysis of the companion mechanisms of Bacillus thuringiensis Bc601 and Bacillus endophyticus Hbe603 in bacterial consortium.

    Science.gov (United States)

    Jia, Nan; Ding, Ming-Zhu; Gao, Feng; Yuan, Ying-Jin

    2016-01-01

    Bacillus thuringiensis and Bacillus endophyticus both act as the companion bacteria, which cooperate with Ketogulonigenium vulgare in vitamin C two-step fermentation. Two Bacillus species have different morphologies, swarming motility and 2-keto-L-gulonic acid productivities when they co-culture with K. vulgare. Here, we report the complete genome sequencing of B. thuringiensis Bc601 and eight plasmids of B. endophyticus Hbe603, and carry out the comparative genomics analysis. Consequently, B. thuringiensis Bc601, with greater ability of response to the external environment, has been found more two-component system, sporulation coat and peptidoglycan biosynthesis related proteins than B. endophyticus Hbe603, and B. endophyticus Hbe603, with greater ability of nutrients biosynthesis, has been found more alpha-galactosidase, propanoate, glutathione and inositol phosphate metabolism, and amino acid degradation related proteins than B. thuringiensis Bc601. Different ability of swarming motility, response to the external environment and nutrients biosynthesis may reflect different companion mechanisms of two Bacillus species. Comparative genomic analysis of B. endophyticus and B. thuringiensis enables us to further understand the cooperative mechanism with K. vulgare, and facilitate the optimization of bacterial consortium. PMID:27353048

  10. Comparative genomics analysis of the companion mechanisms of Bacillus thuringiensis Bc601 and Bacillus endophyticus Hbe603 in bacterial consortium.

    Science.gov (United States)

    Jia, Nan; Ding, Ming-Zhu; Gao, Feng; Yuan, Ying-Jin

    2016-06-29

    Bacillus thuringiensis and Bacillus endophyticus both act as the companion bacteria, which cooperate with Ketogulonigenium vulgare in vitamin C two-step fermentation. Two Bacillus species have different morphologies, swarming motility and 2-keto-L-gulonic acid productivities when they co-culture with K. vulgare. Here, we report the complete genome sequencing of B. thuringiensis Bc601 and eight plasmids of B. endophyticus Hbe603, and carry out the comparative genomics analysis. Consequently, B. thuringiensis Bc601, with greater ability of response to the external environment, has been found more two-component system, sporulation coat and peptidoglycan biosynthesis related proteins than B. endophyticus Hbe603, and B. endophyticus Hbe603, with greater ability of nutrients biosynthesis, has been found more alpha-galactosidase, propanoate, glutathione and inositol phosphate metabolism, and amino acid degradation related proteins than B. thuringiensis Bc601. Different ability of swarming motility, response to the external environment and nutrients biosynthesis may reflect different companion mechanisms of two Bacillus species. Comparative genomic analysis of B. endophyticus and B. thuringiensis enables us to further understand the cooperative mechanism with K. vulgare, and facilitate the optimization of bacterial consortium.

  11. Aerobic biodegradation of Azo dye by Bacillus cohnii MTCC 3616; an obligately alkaliphilic bacterium and toxicity evaluation of metabolites by different bioassay systems.

    Science.gov (United States)

    Prasad, A S Arun; Rao, K V Bhaskara

    2013-08-01

    An obligate alkaliphilic bacterium Bacillus cohnii MTCC 3616 aerobically decolorized a textile azo dye Direct Red-22 (5,000 mg l⁻¹) with 95 % efficiency at 37 °C and pH 9 in 4 h under static conditions. The decolorization of Direct Red-22 (DR-22) was possible through a broad pH (7-11), temperature (10-45 °C), salinity (1-7 %), and dye concentration (5-10 g l⁻¹) range. Decolorization of dye was assessed by UV-vis spectrophotometer with reduction of peak intensity at 549 nm (λ(max)). Biodegradation of dye was analyzed by Fourier transform infrared spectroscopy (FTIR) and high-performance liquid chromatography (HPLC). The FTIR spectrum revealed that B. cohnii specifically targeted azo bond (N=N) at 1,614.42 cm⁻¹ to break down Direct Red-22. Formation of metabolites with different retention times in HPLC analysis further confirmed the degradation of dye. The phytotoxicity test with 5,000 mg l⁻¹ of untreated dye showed 80 % germination inhibition in Vigna mungo, 70 % in Sorghum bicolor and 80 % in Vigna radiata. No germination inhibition was noticed in all three plants by DR-22 metabolites at 5,000 mg l⁻¹. Biotoxicity test with Artemia salina proved the lethality of the azo dye at LC₅₀ of 4 and 8 % for degraded metabolites by causing death of its nauplii compared to its less toxic-degraded metabolites. Bioaccumulation of dye was observed in the mid-gut of A. salina. The cytogenotoxicity assay on the meristematic root tip cells of Allium cepa further confirmed the cytotoxic nature of azo dye (DR-22) with decrease in mitotic index (0.5 % at 500 ppm) and increase in aberrant index (4.56 %) over 4-h exposure period. Genotoxic damages (lagging chromosome, metaphase cluster, chromosome bridges, and dye accumulation in cytoplasm) were noticed at different stages of cell cycle. The degraded metabolites had negligible cytotoxic and genotoxic effects.

  12. Resistance of Bacillus amyloliquefaciens spores to melt extrusion process conditions

    OpenAIRE

    Ciera, Lucy Wanjiru; Beladjal, Lynda; Almeras, Xavier; Gheysens, Tom; Nierstrasz, Vincent; Van Langenhove, Lieva; Mertens, Johan

    2014-01-01

    With the increasing demand for functionalised textile materials, industry is focusing on research that will add novel properties to textiles. Bioactive compounds and their benefits have been and are still considered as a possible source of unique functionalities to be explored. However, incorporating bioactive compounds into textiles and their resistance to textile process parameters has not yet been studied. In this study, we developed a system to study the resistance of Bacillus amyloliquef...

  13. Historical Distribution and Molecular Diversity of Bacillus anthracis, Kazakhstan

    OpenAIRE

    Aikembayev, Alim M.; Lukhnova, Larissa; Temiraliyeva, Gulnara; Meka-Mechenko, Tatyana; Pazylov, Yerlan; Zakaryan, Sarkis; Denissov, Georgiy; Easterday, W. Ryan; Matthew N. Van Ert; Keim, Paul; Francesconi, Stephen C.; Jason K Blackburn; Hugh-Jones, Martin; Hadfield, Ted

    2010-01-01

    To map the distribution of anthrax outbreaks and strain subtypes in Kazakhstan during 1937–2005, we combined geographic information system technology and genetic analysis by using archived cultures and data. Biochemical and genetic tests confirmed the identity of 93 archived cultures in the Kazakhstan National Culture Collection as Bacillus anthracis. Multilocus variable number tandem repeat analysis genotyping identified 12 genotypes. Cluster analysis comparing these genotypes with previousl...

  14. Bacillus anthracis IsdG, a Heme-Degrading Monooxygenase

    OpenAIRE

    Skaar, Eric P.; Gaspar, Andrew H.; Schneewind, Olaf

    2006-01-01

    Bacillus anthracis, the causative agent of anthrax, utilizes hemin and hemoglobin for growth in culture, suggesting that these host molecules serve as sources for the nutrient iron during bacterial infection. Bioinformatic analyses of the B. anthracis genome revealed genes with similarity to the iron-regulated surface determinant (isd) system responsible for heme uptake in Staphylococcus aureus. We show that the protein product of one of these genes, isdG, binds hemin in a manner resembling t...

  15. The effects of mixture commercial live bakers’ yeast and probiotic bacillus on growth and feeding performance and survival rate of silver carp (Hypophthalmichthys molitrix larvae via bioencapsulated Artemia urmiana nauplii

    Directory of Open Access Journals (Sweden)

    Hossein Adineh

    2011-07-01

    Full Text Available Hypophthalmichthys molitrix (Valenciennes, 1844 larvae is an important species forfreshwater aquaculture. This study evaluated the effects of feeding a blend of probiotic bacilli bacteria(B. polymixa, B. licheniformis, B. circulans and baker´s yeast (Saccharomyces cerevisiae on growth and feeding parameters and survival rate of Silver carp larvae. Artemia urmiana (Gǘnther, 1899 nauplii is an important live food that was used as a vector to carry probiotic bacillus to digestive tract of silver carp larvae. The fish larvae were fed at a level of 10 percent body weight at 4 times a day for 30 days. Fish larvae in experimental treatments were fed A. urmiana nauplii that were enriched by blend of 50 percent of Bacillus spp. with concentrations of 1×105, 2×105, 3×105 and 4×105 CFU mL-1 and 50 percent of baker´s yeast with concentrations of 1×105, 2×105, 3×105 and 4×105 cellsmL-1 (T1, T2, T3, T4, respectively and were compared to fish larvae fed control diets of unbioencapsulated A. urmiananauplii. The experiment indicated that feeding and growth parameters in fish fed experimental treatments were significantly higher than fish fed control diets (P<0.05 but survival rate did not significantly differ. Overall, the best group was fed the highest level of yeast and probiotic.

  16. Isolation and Characterization of a New Heterotrophic Nitrifying Bacillus sp. Strain

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective To characterize the heterotrophic nitrifying bacteria. Methods The bacteria were isolated from membrane bioreactor for treating synthetic wastewater using the method newly introduced in this study. Fluorescence in situ hybridization (FISH) was used to validate the nonexistence of autotrophic ammonia oxidizers and nitrite oxidizers. Batch tests were carried out to investigate the capability of heterotrophic nitrification by the pure culture. Phylogenetic analysis of the pure culture was performed. Results A heterotrophic nitrifier, named Bacillus sp. LY, was newly isolated from the membrane bioreactor system in which the efficiency of TN removal was up to 80%. After 24-day, incubation, the removal efficiency of COD by Bacillus sp. LYwas 71.7%. The ammonium nitrogen removal rate after assimilation nearly ceased by Bacillus sp. LYwas 74.7%.The phylogenetic tree of Bacillus sp. LY and the neighbouring nitrifiers were given. Conclusions The batch test results indicate that Bacillus sp. LY can utilize the organic carbon as the source of assimilation when it grows on glucose and ammonium chloride medium accompanying the formation of oxidized-nitrogen. It also can denitrify nitrate while nitrifying. Bacillus sp. LY may become a new bacterial resource for heterotrophic nitrification and play a bioremediation role in nutrient removal.

  17. Cyclic Lipopeptide Biosynthetic Genes and Products, and Inhibitory Activity of Plant-Associated Bacillus against Phytopathogenic Bacteria.

    Directory of Open Access Journals (Sweden)

    Isabel Mora

    Full Text Available The antibacterial activity against bacterial plant pathogens and its relationships with the presence of the cyclic lipopeptide (cLP biosynthetic genes ituC (iturin, bmyB (bacillomycin, fenD (fengycin and srfAA (surfactin, and their corresponding antimicrobial peptide products have been studied in a collection of 64 strains of Bacillus spp. isolated from plant environments. The most frequent antimicrobial peptide (AMP genes were bmyB, srfAA and fenD (34-50% of isolates. Most isolates (98.4% produced surfactin isoforms, 90.6% iturins and 79.7% fengycins. The antibacterial activity was very frequent and generally intense among the collection of strains because 75% of the isolates were active against at least 6 of the 8 bacterial plant pathogens tested. Hierarchical and correspondence analysis confirmed the presence of two clearly differentiated groups. One group consisted of Bacillus strains that showed a strong antibacterial activity, presented several cLPs genes and produced several isoforms of cLPs simultaneously, mainly composed of B. subtilis and B. amyloliquefaciens, although the last one was exclusive to this group. Another group was characterized by strains with very low or none antibacterial activity, that showed one or none of the cLP genes and produced a few or none of the corresponding cLPs, and was the most heterogenous group including B. subtilis, B. licheniformis, B. megaterium, B. pumilus, B. cereus and B. thuringiensis, although the last two were exclusive to this group. This work demonstrated that the antagonistic capacity of plant-associated Bacillus against plant pathogenic bacteria is related to the presence of cLP genes and to the production of the corresponding cLPs, and it is mainly associated to the species B. subtilis and B. amyloliquefaciens. Our findings would help to increase the yield and efficiency of screening methods to obtain candidate strains to biocontrol agents with a mechanism of action relaying on the

  18. Cyclic Lipopeptide Biosynthetic Genes and Products, and Inhibitory Activity of Plant-Associated Bacillus against Phytopathogenic Bacteria.

    Science.gov (United States)

    Mora, Isabel; Cabrefiga, Jordi; Montesinos, Emilio

    2015-01-01

    The antibacterial activity against bacterial plant pathogens and its relationships with the presence of the cyclic lipopeptide (cLP) biosynthetic genes ituC (iturin), bmyB (bacillomycin), fenD (fengycin) and srfAA (surfactin), and their corresponding antimicrobial peptide products have been studied in a collection of 64 strains of Bacillus spp. isolated from plant environments. The most frequent antimicrobial peptide (AMP) genes were bmyB, srfAA and fenD (34-50% of isolates). Most isolates (98.4%) produced surfactin isoforms, 90.6% iturins and 79.7% fengycins. The antibacterial activity was very frequent and generally intense among the collection of strains because 75% of the isolates were active against at least 6 of the 8 bacterial plant pathogens tested. Hierarchical and correspondence analysis confirmed the presence of two clearly differentiated groups. One group consisted of Bacillus strains that showed a strong antibacterial activity, presented several cLPs genes and produced several isoforms of cLPs simultaneously, mainly composed of B. subtilis and B. amyloliquefaciens, although the last one was exclusive to this group. Another group was characterized by strains with very low or none antibacterial activity, that showed one or none of the cLP genes and produced a few or none of the corresponding cLPs, and was the most heterogenous group including B. subtilis, B. licheniformis, B. megaterium, B. pumilus, B. cereus and B. thuringiensis, although the last two were exclusive to this group. This work demonstrated that the antagonistic capacity of plant-associated Bacillus against plant pathogenic bacteria is related to the presence of cLP genes and to the production of the corresponding cLPs, and it is mainly associated to the species B. subtilis and B. amyloliquefaciens. Our findings would help to increase the yield and efficiency of screening methods to obtain candidate strains to biocontrol agents with a mechanism of action relaying on the production of

  19. 77 FR 33455 - Bacillus mycoides isolate J; Receipt of Application for Emergency Exemption for Use on Potato in...

    Science.gov (United States)

    2012-06-06

    ... section 18 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) (7 U.S.C. 136p), at the... action of Bacillus mycoides isolate J is to induce the plants systemic acquired resistance (SAR)...

  20. Triple fixation of Bacillus subtilis dormant spores.

    OpenAIRE

    Kozuka, S; Tochikubo, K

    1983-01-01

    A triple-fixation method with a sequential application of 5% glutaraldehyde, 1% osmium tetroxide, and 2% potassium permanganate gave superior preservation of the ultrastructure of Bacillus subtilis dormant spores with a thick spore coat.

  1. Narrow terahertz attenuation signatures in Bacillus thuringiensis.

    Science.gov (United States)

    Zhang, Weidong; Brown, Elliott R; Viveros, Leamon; Burris, Kellie P; Stewart, C Neal

    2014-10-01

    Terahertz absorption signatures from culture-cultivated Bacillus thuringiensis were measured with a THz photomixing spectrometer operating from 400 to 1200 GHz. We observe two distinct signatures centered at ∼955 and 1015 GHz, and attribute them to the optically coupled particle vibrational resonance (surface phonon-polariton) of Bacillus spores. This demonstrates the potential of the THz attenuation signatures as "fingerprints" for label-free biomolecular detection. PMID:23821459

  2. Narrow terahertz attenuation signatures in Bacillus thuringiensis.

    Science.gov (United States)

    Zhang, Weidong; Brown, Elliott R; Viveros, Leamon; Burris, Kellie P; Stewart, C Neal

    2014-10-01

    Terahertz absorption signatures from culture-cultivated Bacillus thuringiensis were measured with a THz photomixing spectrometer operating from 400 to 1200 GHz. We observe two distinct signatures centered at ∼955 and 1015 GHz, and attribute them to the optically coupled particle vibrational resonance (surface phonon-polariton) of Bacillus spores. This demonstrates the potential of the THz attenuation signatures as "fingerprints" for label-free biomolecular detection.

  3. Bacillus-shape design of polymer based drug delivery systems with janus-faced function for synergistic targeted drug delivery and more effective cancer therapy.

    Science.gov (United States)

    Cui, Fei; Lin, Jinyan; Li, Yang; Li, Yanxiu; Wu, Hongjie; Yu, Fei; Jia, Mengmeng; Yang, Xiangrui; Wu, Shichao; Xie, Liya; Ye, Shefang; Luo, Fanghong; Hou, Zhenqing

    2015-04-01

    The particle shape of the drug delivery systems had a strong impact on their in vitro and in vivo performance, but there was limited availability of techniques to produce the specific shaped drug carriers. In this article, the novel methotrexate (MTX) decorated MPEG-PLA nanobacillus (MPEG-PLA-MTX NB) was prepared by the self-assembly technique followed by the extrusion through SPG membrane with high N2 pressure for targeted drug delivery, in which Janus-like MTX was not only used as a specific anticancer drug but could also be served as a tumor-targeting ligand. The MPEG-PLA-MTX NBs demonstrated much higher in vitro and in vivo targeting efficiency compared to the MPEG-PLA-MTX nanospheres (MPEG-PLA-MTX NSs) and MPEG-PLA nanospheres (MPEG-PLA NSs). In addition, the MPEG-PLA-MTX NBs also displayed much more excellent in vitro and in vivo antitumor activity than the MPEG-PLA-MTX NSs and free MTX injection. To our knowledge, this work provided the first example of the integration of the shape design (which mediated an early phase tumor accumulation and a late-phase cell internalization) and Janus-faced function (which mediated an early phase active targeting effect and a late-phase anticancer effect) on the basis of nanoscaled drug delivery systems. The highly convergent and cooperative drug delivery strategy opens the door to more drug delivery systems with new shapes and functions for cancer therapy.

  4. Cannibalism stress response in Bacillus subtilis.

    Science.gov (United States)

    Höfler, Carolin; Heckmann, Judith; Fritsch, Anne; Popp, Philipp; Gebhard, Susanne; Fritz, Georg; Mascher, Thorsten

    2016-01-01

    When faced with carbon source limitation, the Gram-positive soil organism Bacillus subtilis initiates a survival strategy called sporulation, which leads to the formation of highly resistant endospores that allow B. subtilis to survive even long periods of starvation. In order to avoid commitment to this energy-demanding and irreversible process, B. subtilis employs another strategy called 'cannibalism' to delay sporulation as long as possible. Cannibalism involves the production and secretion of two cannibalism toxins, sporulation delaying protein (SDP) and sporulation killing factor (SKF), which are able to lyse sensitive siblings. The lysed cells are thought to then provide nutrients for the cannibals to slow down or even prevent them from entering sporulation. In this study, we uncovered the role of the cell envelope stress response (CESR), especially the Bce-like antimicrobial peptide detoxification modules, in the cannibalism stress response during the stationary phase. SDP and SKF specifically induce Bce-like systems and some extracytoplasmic function σ factors in stationary-phase cultures, but only the latter provide some degree of protection. A full Bce response is only triggered by mature toxins, and not by toxin precursors. Our study provides insights into the close relationship between stationary-phase survival and the CESR of B. subtilis. PMID:26364265

  5. Bacillus thuringiensis Conjugation in Simulated Microgravity

    Science.gov (United States)

    Beuls, Elise; van Houdt, Rob; Leys, Natalie; Dijkstra, Camelia; Larkin, Oliver; Mahillon, Jacques

    2009-10-01

    Spaceflight experiments have suggested a possible effect of microgravity on the plasmid transfer among strains of the Gram-positive Bacillus thuringiensis, as opposed to no effect recorded for Gram-negative conjugation. To investigate these potential effects in a more affordable experimental setup, three ground-based microgravity simulators were tested: the Rotating Wall Vessel (RWV), the Random Positioning Machine (RPM), and a superconducting magnet. The bacterial conjugative system consisted in biparental matings between two B. thuringiensis strains, where the transfer frequencies of the conjugative plasmid pAW63 and its ability to mobilize the nonconjugative plasmid pUB110 were assessed. Specifically, potential plasmid transfers in a 0-g position (simulated microgravity) were compared to those obtained under 1-g (normal gravity) condition in each device. Statistical analyses revealed no significant difference in the conjugative and mobilizable transfer frequencies between the three different simulated microgravitational conditions and our standard laboratory condition. These important ground-based observations emphasize the fact that, though no stimulation of plasmid transfer was observed, no inhibition was observed either. In the case of Gram-positive bacteria, this ability to exchange plasmids in weightlessness, as occurs under Earth's conditions, should be seen as particularly relevant in the scope of spread of antibiotic resistances and bacterial virulence.

  6. Purification on N and P in Aquiculturie Wastewater with a Bacteria-Alga System%菌-藻体系去除水产养殖废水中氮和磷的净化实验

    Institute of Scientific and Technical Information of China (English)

    周武旋; 赵江萍; 栗越妍

    2012-01-01

    以投加人工饲料喂养罗非鱼7天的玻璃鱼缸内的废水为样品,接种地衣芽孢杆菌(Bacillus licheniformis)、硝化细菌、月牙藻(Selenastrum reinsch)和四尾栅藻(Scenedesmus quadricanda)后于光照箱内培养,于0、12h、24h、48h、84h、120h、168h测定废水样品的pH值、溶解氧、氨氮、亚硝酸盐氮、硝酸盐氮和可溶性磷酸盐的去除率,以24h氨氮和168h可溶性磷酸盐的去除率为指标进行L3(34)正交实验,研究菌藻体系去除水产养殖废水中氮和磷的净化效果.结果表明,地衣芽孢杆菌、硝化细菌、月牙藻和四尾栅藻组成的菌-藻体系可以通过其新陈代谢过程中形成的原始共生关系有效地去除养殖水体中的氮、磷污染物.菌-藻体系去除氨氮的最佳反应时间为24h,最大去除率98%,在初始密度为5×105 cells/ml条件下,最佳菌-藻体积配比为1∶2∶2∶3,即最佳菌-藻初始密度分别为2.5×10s cell/ml、5.0×105 cell/ml、5.0×105 cell/ml、10.0×l05 cell/ml.菌-藻体系去除可溶性磷酸盐的最佳反应时间为168h,去除率100%,在初始密度为5×105 cells/ml条件下,最佳菌藻体积配比为1∶1∶3∶2,即最佳菌-藻初始密度分别为2.5×105 cell/ml、2.5× 105 cell/ml、10.0×10s cell/ml、5.0×105 cell/ml.%Using the waste water after feeding tuapia 7 days with artificial additive in the aquarium glass as samples. Bacillus licheniformis (Bacillus licheniformis ), nitrifying bacteria , crescent algae ( Selenastrum reinsch ) and four tail gate algae ( Scenedesmus quadricanda )were vaccinated and cultured in the light box. pH value,dissolved oxygen,ammonia nitrogen,nitrite nitrogen,nitrate nitrogen and soluble phosphate removal rate of the waste water samples were measured after 0,12h,24h,48h,84h,120h and 168h, respectively cultivation. ThenL9( 34) orthogonal experiments were conducted to analyze the ammonia nitrogen at 24h and the removal rate of soluble phosphate at 168h, in order to

  7. Hydrazine vapor inactivates Bacillus spores

    Science.gov (United States)

    Schubert, Wayne W.; Engler, Diane L.; Beaudet, Robert A.

    2016-05-01

    NASA policy restricts the total number of bacterial spores that can remain on a spacecraft traveling to any planetary body which might harbor life or have evidence of past life. Hydrazine, N2H4, is commonly used as a propellant on spacecraft. Hydrazine as a liquid is known to inactivate bacterial spores. We have now verified that hydrazine vapor also inactivates bacterial spores. After Bacillus atrophaeus ATCC 9372 spores deposited on stainless steel coupons were exposed to saturated hydrazine vapor in closed containers, the spores were recovered from the coupons, serially diluted, pour plated and the surviving bacterial colonies were counted. The exposure times required to reduce the spore population by a factor of ten, known as the D-value, were 4.70 ± 0.50 h at 25 °C and 2.85 ± 0.13 h at 35 °C. These inactivation rates are short enough to ensure that the bioburden of the surfaces and volumes would be negligible after prolonged exposure to hydrazine vapor. Thus, all the propellant tubing and internal tank surfaces exposed to hydrazine vapor do not contribute to the total spore count.

  8. DECONTAMINATION ASSESSMENT OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS, AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACTS USING A HYDROGEN PERIOXIDE GAS GENERATOR

    Science.gov (United States)

    Aims: To evaluate the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface materials using hydrogen peroxide gas. Methods and Results: B. anthracis, B. subtilis, and G. Stearothermophilus spores were dried on seven...

  9. Production, Secretion and Biological Activity of Bacillus cereus Enterotoxins

    Directory of Open Access Journals (Sweden)

    Sonia Senesi

    2010-06-01

    Full Text Available Bacillus cereus behaves as an opportunistic pathogen frequently causing gastrointestinal diseases, and it is increasingly recognized to be responsible for severe local or systemic infections. Pathogenicity of B. cereus mainly relies on the secretion of a wide array of toxins and enzymes and also on the ability to undergo swarming differentiation in response to surface-sensing. In this report, the pathogenicity exerted by B. cereus toxins is described with particular attention to the regulatory mechanisms of production and secretion of HBL, Nhe and CytK enterotoxins.

  10. TRANSGENIC PLANTS EXPRESSING BACILLUS THURINGIENSIS DELTA-ENDOTOXINS

    Institute of Scientific and Technical Information of China (English)

    Hua-rong,Li; BrendaOppert; KunYanZhu; RandallA.Higgins; Fang-nengHuang; LawrentL.Buschman

    2003-01-01

    Commercial varieties of transgenic Bacillus thuringiensis (Bt) plants have been developed in many countries to control target pests. Initially, the expression of native Bt genes in plants was low due to mRNA instability, improper splicing, and post-translation modifications. Subsequently, modifications of the native Bt genes greatly enhanced expression levels. This is a review of the developments that made modem high-expression transgenic Bt plants possible, with an emphasis on the reasons for the low-level expression of native Bt genes in plant systems, and the techniques that have been used to improve plant expression of Bt toxin genes.

  11. Successful Treatment of Bacillus cereus Bacteremia in a Patient with Propionic Acidemia

    Science.gov (United States)

    Aygun, Fatih; Cam, Halit

    2016-01-01

    Bacillus cereus can cause serious, life-threatening, systemic infections in immunocompromised patients. The ability of microorganism to form biofilm on biomedical devices can be responsible for catheter-related bloodstream infections. Other manifestations of severe disease are meningitis, endocarditis, osteomyelitis, and surgical and traumatic wound infections. The most common feature in true bacteremia caused by Bacillus is the presence of an intravascular catheter. Herein, we report a case of catheter-related bacteremia caused by B. cereus in a patient with propionic acidemia. PMID:27195164

  12. Successful Treatment of Bacillus cereus Bacteremia in a Patient with Propionic Acidemia.

    Science.gov (United States)

    Aygun, Fatma Deniz; Aygun, Fatih; Cam, Halit

    2016-01-01

    Bacillus cereus can cause serious, life-threatening, systemic infections in immunocompromised patients. The ability of microorganism to form biofilm on biomedical devices can be responsible for catheter-related bloodstream infections. Other manifestations of severe disease are meningitis, endocarditis, osteomyelitis, and surgical and traumatic wound infections. The most common feature in true bacteremia caused by Bacillus is the presence of an intravascular catheter. Herein, we report a case of catheter-related bacteremia caused by B. cereus in a patient with propionic acidemia. PMID:27195164

  13. Successful Treatment of Bacillus cereus Bacteremia in a Patient with Propionic Acidemia

    Directory of Open Access Journals (Sweden)

    Fatma Deniz Aygun

    2016-01-01

    Full Text Available Bacillus cereus can cause serious, life-threatening, systemic infections in immunocompromised patients. The ability of microorganism to form biofilm on biomedical devices can be responsible for catheter-related bloodstream infections. Other manifestations of severe disease are meningitis, endocarditis, osteomyelitis, and surgical and traumatic wound infections. The most common feature in true bacteremia caused by Bacillus is the presence of an intravascular catheter. Herein, we report a case of catheter-related bacteremia caused by B. cereus in a patient with propionic acidemia.

  14. 地衣芽孢杆菌总蛋白双向电泳方法的建立和优化%Establishment and optimization of two-dimensional gel electrophoresis of total proteins from Bacillus lincheniformis

    Institute of Scientific and Technical Information of China (English)

    江慎华; 陈惠; 陈静; 汪涛; 姚中平; 周英棠

    2013-01-01

    探索建立有效的地衣芽孢杆菌蛋白质组双向电泳体系,为进一步揭示地衣芽孢杆菌促进氧化葡萄糖酸杆菌产酸的作用机制奠定基础.以地衣芽孢杆菌为材料,比较蛋白质制备超声破壁时间、新型细胞裂解液、pH梯度和不同上样量对地衣芽孢杆菌蛋白双向电泳结果的影响.结果显示:采用15min超声破壁提取地衣芽孢杆菌总蛋白,选用新型蛋白质裂解,用长24cm、pH4~7的IPG胶条,在上样量为80μg进行等电聚焦,于60V 15min、120V 6h条件下进行SDS-PAGE垂直电泳,可以获得背景清晰、重复性好的双向电泳图谱.在探索出一种新型可行的新型细胞裂解液的同时,建立一套用于地衣芽孢杆菌蛋白质组分析的双向电泳方法.%A two dimensional gel electrophoresis protocol proteomic study of Bacillus lincheniformis and offer further information how Bacillus cereus stimulate the growth of Gluconobacter oxydans to produce 2-keto-L-gulonic acid (2-KLG) after entering into stationary phase was established.Different parameters,including protein preparation by different ultrasonic broken time,new type lysis,pH gradient and different sample of Bacillus lincheniformis protein,were used to evaluate the effect of two-dimensional electrophoresis of Bacillus lincheniformis proteins.Results showed that the clear background and reproducibility of two dimensional gel electrophoresis were established on the condition of using 15min of ultrasonic broken extraction,selection of protein cleavage I,pH4~7 24cm IPG strips,80 μg loading volume,isoelectric focus at 60V 15min,120V 6h.The aim of this study was not only to explore a novel cell lysate in two-dimensional gel electrophoresis,but also to establish a method of proteome analysis for two-dimensional electrophoresis of Bacillus licheniformis.

  15. BAC Library Construction and Physical Mapping of Bacillus anthracis A16R

    Institute of Scientific and Technical Information of China (English)

    Zhang Da; Zhu Houchu; Huang Liuyu

    2013-01-01

    Bacillus anthracis is an endospore-forming bacterium that causes severe inhalational anthrax, and bacillus anthracis A16R is an attenuated strain derived from Bacillus anthracis A16. The development of bacterial artificial chromosome (BAC) system has allowed the construction of large insert-size DNA libraries, and the bacterial artificial chromosomes (BACs) have become the preferred large insert cloning system for genomic analysis because such libraries are characteristically stable, high in ifdelity and easy to handle. To facilitate genome studies of this bacterium, a bacterial artiifcial chromosome library (BAC) has been established from genome DNA of Bacillus anthracis A16R. This library consisted of 9 600 clones randomly selected from more than 15 000 recombinant clones carrying inserts in the plindigoBAC-5 vectors. The mean insert size was 56 kbp, representing an approximate 12-fold genome coverage, while end sequences were obtained from 700 randomly selected clones. Sequences were compared with Bacillus anthracis Ames and Bacillus cereus ATCC 14579 Genome Project databases using the NCBI BLASTN search project. And most BLASTN results showed high identities and that the sequences’ sites could be used as STSs. To construct this physical map, Excel was used for the array of STSs and some gaps of the map were iflled up by PCR walking. Artemis-V4 was used in the construction of a genome-wide physical map with 93%genome coverage. The A16R BAC library proved to be a vital tool for the generation of a map that would not only allow the subsequent sequencing of defined areas of genome, but also provide immediate access to clones that were stable and convenient for functional genomic researches.

  16. BAC Library Construction and Physical Mapping of Bacillus anthracis A16R

    Directory of Open Access Journals (Sweden)

    Da Zhang

    2013-12-01

    Full Text Available Bacillus anthracis is an endospore-forming bacterium that causes severe inhalational anthrax, and bacillus anthracis A16R is an attenuated strain derived from Bacillus anthracis A16. The development of bacterial artificial chromosome (BAC system has allowed the construction of large insert-size DNA libraries, and the bacterial artificial chromosomes (BACs have become the preferred large insert cloning system for genomic analysis because such libraries are characteristically stable, high in fidelity and easy to handle. To facilitate genome studies of this bacterium, a bacterial artificial chromosome library (BAC has been established from genome DNA of Bacillus anthracis A16R. This library consisted of 9 600 clones randomly selected from more than 15 000 recombinant clones carrying inserts in the plindigoBAC-5 vectors. The mean insert size was 56 kbp, representing an approximate 12-fold genome coverage, while end sequences were obtained from 700 randomly selected clones. Sequences were compared with Bacillus anthracis Ames and Bacillus cereus ATCC 14579 Genome Project databases using the NCBI BLASTN search project. And most BLASTN results showed high identities and that the sequences’ sites could be used as STSs. To construct this physical map, Excel was used for the array of STSs and some gaps of the map were filled up by PCR walking. Artemis-V4 was used in the construction of a genome-wide physical map with 93% genome coverage. The A16R BAC library proved to be a vital tool for the generation of a map that would not only allow the subsequent sequencing of defined areas of genome, but also provide immediate access to clones that were stable and convenient for functional genomic researches.

  17. A positive selection vector for the analysis of structural plasmid instability in Bacillus subtilis

    NARCIS (Netherlands)

    Meima, R; Venema, G; Bron, S

    1996-01-01

    A system for the positive selection of structural plasmid rearrangements in Bacillus subtilis was developed. Random deletions removing a transcription terminator structure in the assay plasmid, designated pGP100, resulted in expression of the cat-86 gene, under control of a constitutive bacteriophag

  18. BOOK REVIEW – BACILLUS THURINGIENSIS: A CORNERSTONE OF MODERN AGRICULTURE BACILLUS THURINGIENSIS

    Science.gov (United States)

    Are you interested in the technical issues surrounding the use of Bacillus thuringiensis pesticidal traits as sprays and as plant incorporated protectants (transgenic crops)? Should the dimensions of human health, ecology, entomology, risk assessment, resistance management, and d...

  19. The effects of citrus extract (Citrox©) on the naturally occurring microflora and inoculated pathogens, Bacillus cereus and Salmonella enterica, in a model food system and the traditional Greek yogurt-based salad Tzatziki.

    Science.gov (United States)

    Tsiraki, Maria I; Savvaidis, Ioannis N

    2016-02-01

    The antimicrobial effect of citrus extract (at 1 mL/kg [TC1] and 2 mL/kg [TC2]) on the naturally occurring microflora and inoculated pathogens (Bacillus cereus and Salmonella enterica, at ca. 6 log cfu/g) in the traditional Greek yogurt-based salad Tzatziki during storage under vacuum at 4 or 10 °C was examined. We also examined the effect of citrus extract (Citrox(©)) against the two aforementioned pathogens in tryptic soy broth (TSB). Of the two treatments, TC2 yielded the lowest yeast counts, irrespective of temperature, resulting in approximately 2 (4 °C) and 3 (10 °C) log reductions on the final day of storage (70 and 30 days, respectively). Although panelists preferred the TC1-treated salad, the TC2-treated product was sensorily acceptable. Therefore, at the concentrations used, Citrox had no negative sensorial effect on the Tzatziki. During storage, the Bacillus populations in the Citrox-treated Tzatziki samples progressively decreased, showing major declines from days 12 and 28 (at 10 and 4 °C, respectively). Citrox, especially at 2 mL/kg, had a significant effect on the survival of B. cereus. S. enterica showed major declines in all untreated Tzatziki samples from day 0-70 (4 °C) and from day 0-30 (10 °C), with averages of 2.5 and 2.8 log cfu/g, respectively. The results indicate that Citrox (at 1 and 2 mL/kg) is effective, from a safety standpoint, for reducing Bacillus and Salmonella spp. in Tzatziki. In addition, 2% citrus extract also showed a higher inhibitory effect against B. cereus and S. enterica grown in TSB than 1% citrus extract.

  20. Differentiation of Bacillus anthracis from Bacillus cereus by gas chromatographic whole-cell fatty acid analysis.

    OpenAIRE

    Lawrence, D; Heitefuss, S; Seifert, H S

    1991-01-01

    Three strains of Bacillus anthracis and seven strains of Bacillus cereus were grown on complex medium and on synthetic medium. Gas chromatographic analysis of whole-cell fatty acids of strains grown on complex medium gave nearly identical fatty acid patterns. Fatty acid patterns of strains grown on synthetic medium showed a high content of branched-chain fatty acids. Significant differences between the fatty acid patterns of the two species were found. Odd iso/anteiso fatty acid ratios were a...

  1. Automated thermochemolysis reactor for detection of Bacillus anthracis endospores by gas chromatography–mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Li, Dan [Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602 (United States); Rands, Anthony D.; Losee, Scott C. [Torion Technologies, American Fork, UT 84003 (United States); Holt, Brian C. [Department of Statistics, Brigham Young University, Provo, UT 84602 (United States); Williams, John R. [Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602 (United States); Lammert, Stephen A. [Torion Technologies, American Fork, UT 84003 (United States); Robison, Richard A. [Department of Microbiology and Molecular Biology, Brigham Young University, Provo, UT 84602 (United States); Tolley, H. Dennis [Department of Statistics, Brigham Young University, Provo, UT 84602 (United States); Lee, Milton L., E-mail: milton_lee@byu.edu [Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602 (United States)

    2013-05-02

    Graphical abstract: -- Highlights: •An automated sample preparation system for Bacillus anthracis endospores was developed. •A thermochemolysis method was applied to produce and derivatize biomarkers for Bacillus anthracis detection. •The autoreactor controlled the precise delivery of reagents, and TCM reaction times and temperatures. •Solid phase microextraction was used to extract biomarkers, and GC–MS was used for final identification. •This autoreactor was successfully applied to the identification of Bacillus anthracis endospores. -- Abstract: An automated sample preparation system was developed and tested for the rapid detection of Bacillus anthracis endospores by gas chromatography–mass spectrometry (GC–MS) for eventual use in the field. This reactor is capable of automatically processing suspected bio-threat agents to release and derivatize unique chemical biomarkers by thermochemolysis (TCM). The system automatically controls the movement of sample vials from one position to another, crimping of septum caps onto the vials, precise delivery of reagents, and TCM reaction times and temperatures. The specific operations of introduction of sample vials, solid phase microextraction (SPME) sampling, injection into the GC–MS system, and ejection of used vials from the system were performed manually in this study, although they can be integrated into the automated system. Manual SPME sampling is performed by following visual and audible signal prompts for inserting the fiber into and retracting it from the sampling port. A rotating carousel design allows for simultaneous sample collection, reaction, biomarker extraction and analysis of sequential samples. Dipicolinic acid methyl ester (DPAME), 3-methyl-2-butenoic acid methyl ester (a fragment of anthrose) and two methylated sugars were used to compare the performance of the autoreactor with manual TCM. Statistical algorithms were used to construct reliable bacterial endospore signatures, and 24

  2. Automated thermochemolysis reactor for detection of Bacillus anthracis endospores by gas chromatography–mass spectrometry

    International Nuclear Information System (INIS)

    Graphical abstract: -- Highlights: •An automated sample preparation system for Bacillus anthracis endospores was developed. •A thermochemolysis method was applied to produce and derivatize biomarkers for Bacillus anthracis detection. •The autoreactor controlled the precise delivery of reagents, and TCM reaction times and temperatures. •Solid phase microextraction was used to extract biomarkers, and GC–MS was used for final identification. •This autoreactor was successfully applied to the identification of Bacillus anthracis endospores. -- Abstract: An automated sample preparation system was developed and tested for the rapid detection of Bacillus anthracis endospores by gas chromatography–mass spectrometry (GC–MS) for eventual use in the field. This reactor is capable of automatically processing suspected bio-threat agents to release and derivatize unique chemical biomarkers by thermochemolysis (TCM). The system automatically controls the movement of sample vials from one position to another, crimping of septum caps onto the vials, precise delivery of reagents, and TCM reaction times and temperatures. The specific operations of introduction of sample vials, solid phase microextraction (SPME) sampling, injection into the GC–MS system, and ejection of used vials from the system were performed manually in this study, although they can be integrated into the automated system. Manual SPME sampling is performed by following visual and audible signal prompts for inserting the fiber into and retracting it from the sampling port. A rotating carousel design allows for simultaneous sample collection, reaction, biomarker extraction and analysis of sequential samples. Dipicolinic acid methyl ester (DPAME), 3-methyl-2-butenoic acid methyl ester (a fragment of anthrose) and two methylated sugars were used to compare the performance of the autoreactor with manual TCM. Statistical algorithms were used to construct reliable bacterial endospore signatures, and 24

  3. Draft genome sequence of Bacillus endophyticus 2102.

    Science.gov (United States)

    Lee, Yong-Jik; Lee, Sang-Jae; Kim, Sun Hong; Lee, Sang Jun; Kim, Byoung-Chan; Lee, Han-Seung; Jeong, Haeyoung; Lee, Dong-Woo

    2012-10-01

    Bacillus endophyticus 2102 is an endospore-forming, plant growth-promoting rhizobacterium isolated from a hypersaline pond in South Korea. Here we present the draft sequence of B. endophyticus 2102, which is of interest because of its potential use in the industrial production of algaecides and bioplastics and for the treatment of industrial textile effluents. PMID:23012284

  4. Diagnostic Oligonucleotide Microarray Fingerprinting of Bacillus Isolates

    OpenAIRE

    Chandler, Darrell P.; Alferov, Oleg; Chernov, Boris; Daly, Don S; Golova, Julia; Perov, Alexander; Protic, Miroslava; Robison, Richard; Schipma, Matthew; White, Amanda; Willse, Alan

    2006-01-01

    A genome-independent microarray and new statistical techniques were used to genotype Bacillus strains and quantitatively compare DNA fingerprints with the known taxonomy of the genus. A synthetic DNA standard was used to understand process level variability and lead to recommended standard operating procedures for microbial forensics and clinical diagnostics.

  5. Complete Genome of Bacillus thuringiensis Myophage Spock

    OpenAIRE

    Maroun, Justin W.; Whitcher, Kelvin J.; Chamakura, Karthik R.; Kuty Everett, Gabriel F.

    2013-01-01

    Bacillus thuringiensis is a Gram-positive, sporulating soil microbe with valuable pesticide-producing properties. The study of bacteriophages of B. thuringiensis could provide new biotechnological tools for the use of this bacterium. Here, we present the complete annotated genome of Spock, a myophage of B. thuringiensis, and describe its features.

  6. Protein-Tyrosine Phosphorylation in Bacillus subtilis

    DEFF Research Database (Denmark)

    Mijakovic, Ivan; Petranovic, Dina; Bottini, N.;

    2005-01-01

    phosphorylation, indicating that this post-translational modifi cation could regulate physiological processes ranging from stress response and exopolysaccharide synthesis to DNA metabolism. Some interesting work in this fi eld was done in Bacillus subtilis , and we here present the current state of knowledge...

  7. «KING OF PROBIOTICS» BACILLUS COAGULANS IN MODERN COMBINED PROBIOTIC PREPARATIONS LAKTOVIT FORTE (FULL REVIEW

    Directory of Open Access Journals (Sweden)

    Bomko TV

    2016-03-01

    Full Text Available Bacillus coagulans has an advantage over most other bacteria used as probiotics. It occupies an intermediate position between the genera Bacillusand Lactobacillus, is a spore-forming bacteria that produce lactic acid.This bacteria in the spores form can tolerate well technology processes, resistant to antibiotics and antiseptics, does not collapse under the influence of gastric juice and bile. Getting into the duodenum, the spores germinate into vegetative forms and begin vegetation and growth, providing probiotic effects.Bacillus coagulans refers to semi-residental bacteria - performing in the human probiotic function, it passes the sporulation phase and slowly leaves the body, standing out in the faeces in the spores form. Thus, it does not violate the personal composition of intestinal microflora.Probiotic Bacillus coagulans enhances the microbiological composition of the intestine, increasing the number of obligate microorganisms and displacing pathogenic flora. Mechanisms of this action based on the lactic acid production and some bacteriocins synthesis, also on the immunomodulatory effect - stimulation of cellular and humoral immunity. The bacterial cell wall and spores are the main immunomodulatory compounds of the Bacillus coagulans.Apparently, namely Bacillus coagulans immunomodulatory properties play a crucial role in the pharmacological effects. It is now well known about the important role of immune system in the pathogenesis of many diseases; it has the clinical effect without the need for intensive growth of bacteria and intestinal colonization; even small amounts of spores are sufficient for pharmacological effect; many experimental evidences of the spore penetration into the lymphatic system and interaction with immunocompetent cells, as well as local and systemic immune effects of probiotic.In addition to this main action, Bacillus coagulans helps to digest lactose, possesses anti-inflammatory and antioxidant activity, as well

  8. Phylogenomic analysis shows that ‘Bacillus vanillea’ is a later heterotypic synonym of Bacillus siamensis.

    Science.gov (United States)

    Dunlap, Christopher A

    2015-10-01

    Bacillus vanillea’ XY18 ( = CGMCC 8629 = NCCB 100507) was isolated from cured vanilla beans and involved in the formation of vanilla aroma compounds. A draft genome of this strain was assembled and yielded a length of 3.71 Mbp with a DNA G+C content of 46.3 mol%. Comparative genomic analysis with its nearest relatives showed only minor differences between this strain and the genome of the Bacillus siamensis KCTC 13613T ( = BCC 22614T = KACC 16244T), with a calculated DNA–DNA hybridization (DDH) value of 91.2 % and an average nucleotide identity (ANI) of 98.9 %. This DDH value is well above the recommended 70 % threshold for species delineation, as well as the ANI threshold of 95 %. In addition, the results of morphological, physiological, chemotaxonomic and phylogenetic analyses indicate that the type strains of these two taxa are highly similar with phenotype coherence. A core genome multi-locus sequencing analysis was conducted for the strains and the results show that ‘Bacillus vanillea’ XY18 clusters closely to the type strain of Bacillus siamensis. Therefore, it is proposed that the species ‘Bacillus vanillea’ XY18 ( = CGMCC 8629 = NCCB 100507) should be reclassified as a later heterotypic synonym of Bacillus siamensis KCTC 13613T ( = BCC 22614T = KACC 16244T). An emended description of Bacillus siamensis is provided. PMID:26296875

  9. Investigation of biosurfactant production by Bacillus pumilus 1529 and Bacillus subtilis WPI

    Directory of Open Access Journals (Sweden)

    shila khajavi shojaei

    2016-06-01

    Full Text Available Introduction: Biosurfactants are unique amphipathic molecules with extensive application in removing organic and metal contaminants. The purpose of this study was to investigate production of biosurfactant and determine optimal conditions to produce biosurfactant by Bacillus pumilus 1529 and Bacillus subtilis WPI. Materials and methods: In this study, effect of carbon source, temperature and incubation time on biosurfactant production was evaluated. Hemolytic activity, emulsification activity, oil spreading, drop collapse, cell hydrophobicity and measurement of surface tension were used to detect biosurfactant production. Then, according to the results, the optimal conditions for biosurfactant production by and Bacillus subtilis WPI was determined. Results: In this study, both bacteria were able to produce biosurfactant at an acceptable level. Glucose, kerosene, sugarcane molasses and phenanthrene used as a sole carbon source and energy for the mentioned bacteria. Bacillus subtilis WPI produced maximum biosurfactant in the medium containing kerosene and reduced surface tension of the medium to 33.1 mN/m after 156 hours of the cultivation at 37°C. Also, the highest surface tension reduction by Bacillus pumilus 1529 occurred in the medium containing sugarcane molasses and reduce the surface tension of culture medium after 156 hours at 37°C from 50.4 to 28.83 mN/m. Discussion and conclusion: Bacillus pumilus 1529 and Bacillus subtilis WPI had high potential in production of biosurfactant and degradation of petroleum hydrocarbons and Phenanthrene. Therefore, it could be said that these bacteria had a great potential for applications in bioremediation and other environmental process.

  10. Bacteriocins synthesized by Bacillus thuringiensis: generalities and potential applications

    Science.gov (United States)

    Salazar-Marroquín, Elma Laura; Galán-Wong, Luis J.; Moreno-Medina, Víctor Ricardo; Reyes-López, Miguel Ángel; Pereyra-Alférez, Benito

    2016-01-01

    The members of the Bacillus thuringiensis group, commonly known as Bt, produce a huge number of metabolites, which show biocidal and antagonistic activity. B. thuringiensis is widely known for synthesizing Cry, Vip and Cyt proteins, active against insects and other parasporins with biocidal activity against certain types of cancerous cells. Nevertheless, B. thuringiensis also synthesizes compounds with antimicrobial activity, especially bacteriocins. Some B. thuringiensis bacteriocins resemble lantibiotics and other small linear peptides (class IIa) from the lactic acid bacteria bacteriocins classification system. Although many bacteriocins produced by Bt have been reported, there is no proper classification for them. In this work, we have grouped these based on molecular weight and functionality. Bacteriocins are small peptides synthesized by bacteria, presenting inhibitory activity against Gram-positive and Gram-negative bacteria and to a lesser extent against fungi. These molecules represent a good study model in the search for microbial control alternatives. Lactic acid bacteria produces a huge number of these types of molecules with great potential. Nonetheless, members of the Bacillus, cereus group, especially B. thuringiensis, emerge as an attractive alternative for obtaining bacteriocins showing novel activities. This review describes the potential applications of B. thuringiensis bacteriocins in the control of foodborne pathogens, environment and medical area. PMID:27340340

  11. Assessment of the effect of temperature on the carp physiology (Cyprinus carpio, L., 1758 fed with probiotics in condition of a recirculating aquaculture system

    Directory of Open Access Journals (Sweden)

    Elena BOCIOC

    2015-10-01

    Full Text Available In order to establish the effect of temperature on the physiology of the carp, special attention was awarded to leukograma and absolute number of leukocytes of carp blood. Body's reaction to stress factors action involves physiological changes, including changes in blood composition and immune mechanisms. The hematological analysis has been effectuated in the experiment that took place during 60 days, in four breeding units of 500 litters in volume each. Four kinds of variants were compared: V1-pellets with 30% crude protein, without probiotics; V2-pellets with 30% crude protein, with probiotics of 2.24×109 CFU/kg food; V3-pellets with 30% crude protein, with probiotics of 3.84×109 CFU/kg food; and V4-pellets with 30% crude protein, with probiotics of 7.04×109 CFU/kg food. We used BioPlus®2B probiotics (a mixture of Bacillus licheniformis (DSM 5749 and Bacillus subtilis (DSM 5750. The microscopic examination of blood smears, which were coloured with May-Grunewald Giemsa panoptic method (both at the beginning and at the end of each experimental stage, we found that lymphocytes were predominant in comparison with other types of leukocytes, the absolute number being: V1 73.3 – 85.9%; V2 65.3 – 84.7%; V3 67.0 – 87.5%; V4 71.3 – 88.3%. It was noticed that eosinophilic granulocytes and basophils from the fish blood, were presented in a low number (0.1–0.8%, (0.2 – 0.4% respectively, also at the beginning of the experiment and in the end of the two phases. The glucose concentration in blood registered higher values, across to normal values reported by literature for carp, also in control group (V1 – 101.8 mg/dl as well as variants with different concentrations of probiotic (V2 – 104.6 mg/dl, V3 – 102.2 mg/dl, V4 – 116.2 mg/dl. The mean values of protein level were in the normal limits for carp, between 3.9 g/dl and 4.9 g/dl, aspect which suggests that fish present a normal physiological condition. In conclusion, probiotics can

  12. Cloning and analysis of Bacillus signal peptide from soil metagenome%土壤宏基因组中芽孢杆菌信号肽的克隆及分析

    Institute of Scientific and Technical Information of China (English)

    2016-01-01

    从土壤宏基因组中筛选获得在芽孢杆菌中高效分泌重组蛋白的信号肽.通过数据分析,将预测到的信号肽DNA片段与蛋白酶基因融合,在枯草芽孢杆菌(Bacillus subtilis) WB800和地衣芽孢杆菌(Bacillus licheniformis) DL6中表达,进行胞外蛋白酶酶活测定.筛选得到25个可能来源于G+菌的信号肽序列,胞外蛋白酶活性分析的结果显示,有14个信号肽具有较好的分泌蛋白酶的能力,其中引导效果最好的信号肽(sig20)是蛋白酶自身信号肽的1.64倍.将高分泌信号肽转化到地衣芽孢杆菌中,证实其在地衣芽孢杆菌同样具有较好的引导效果.应用生物信息学与宏基因组学相结合的方法,可有效获得高效的芽孢杆菌信号肽序列,为蛋白质高效分泌表达系统的构建提供丰富的表达元件.

  13. Genetics of thermophilic bacteria. [Bacillus stearothermophilus:a2

    Energy Technology Data Exchange (ETDEWEB)

    Welker, N.E.

    1991-01-01

    Organisms adapted to high temperature have evolved a variety of unique solutions to the biochemical problems imposed by this environment. Adaptation is commonly used to describe the biochemical properties of organisms which have become adapted to their environment (genetic adaptation). It can also mean the direct response-at the cellular level-of an organism to changes in temperature (physiological adaptation). Thermophilic bacilli (strains of Bacillus stearothermophilus) can exhibit a variety of biochemical adaptations in response to changes in temperature. These include changes in the composition and stability of the membrane, metabolic potential, the transport of amino acids, regulatory mechanisms, ribose methylation of tRNA, protein thermostability, and nutritional requirements. The objectives of the research were to develop efficient and reliable genetic systems to analyze and manipulate B. Stearothermophilus, and to use these systems initiate a biochemical, molecular, and genetic investigations of genes that are required for growth at high temperature.

  14. Bacillus caldolyticus prs gene encoding phosphoribosyldiphosphate synthase

    DEFF Research Database (Denmark)

    Krath, Britta N.; Hove-Jensen, Bjarne

    1996-01-01

    The prs gene, encoding phosphoribosyl-diphosphate (PRPP) synthase, as well as the flanking DNA sequences were cloned and sequenced from the Gram-positive thermophile, Bacillus caldolyticus. Comparison with the homologous sequences from the mesophile, Bacillus subtilis, revealed a gene (gca......D) encoding N-acetylglucosamine-l-phosphate uridyltransferase upstream of prs, and a gene homologous to ctc downstream of prs. cDNA synthesis with a B. caldolyticus gcaD-prs-ctc-specified mRNA as template, followed by amplification utilising the polymerase chain reaction indicated that the three genes are co......-transcribed. Comparison of amino acid sequences revealed a high similarity among PRPP synthases across a wide phylogenetic range. An E. coli strain harbouring the B. caldolyticus prs gene in a multicopy plasmid produced PRPP synthase activity 33-fold over the activity of a haploid B. caldolyticus strain. B. caldolyticus...

  15. Computational based functional analysis of Bacillus phytases.

    Science.gov (United States)

    Verma, Anukriti; Singh, Vinay Kumar; Gaur, Smriti

    2016-02-01

    Phytase is an enzyme which catalyzes the total hydrolysis of phytate to less phosphorylated myo-inositol derivatives and inorganic phosphate and digests the undigestable phytate part present in seeds and grains and therefore provides digestible phosphorus, calcium and other mineral nutrients. Phytases are frequently added to the feed of monogastric animals so that bioavailability of phytic acid-bound phosphate increases, ultimately enhancing the nutritional value of diets. The Bacillus phytase is very suitable to be used in animal feed because of its optimum pH with excellent thermal stability. Present study is aimed to perform an in silico comparative characterization and functional analysis of phytases from Bacillus amyloliquefaciens to explore physico-chemical properties using various bio-computational tools. All proteins are acidic and thermostable and can be used as suitable candidates in the feed industry. PMID:26672917

  16. Bacillus Cereus catheter related bloodstream infection in a patient in a patient with acute lymphblastic leukemia

    Directory of Open Access Journals (Sweden)

    Lütfiye Öksüz

    2012-01-01

    Full Text Available Bacillus cereus infection is rarely associated with actual infection and for this reason single positive blood culture is usually regarded as contamination . However it may cause a number of infections, such catheter-related blood stream infections. Significant catheter-related bloodstream infections (CRBSI caused by Bacillus spp. are mainly due to B.cereus and have been predominantly reported in immunocompromised hosts1 . Catheter removal is generally advised for management of infection. In this report, catheter-related bacteremia caused by B.cereus in a patient with acute lymphoblastıc leukemia (ALL in Istanbul Medical Faculty was presented.A 44-year old man presented with fatigue, weight loss, epistaxis and high fever. A double-lumen Hickman–catheter (Bard 12.0 Fr, Round Dual Lumen was inserted by surgical cut-down to access the right subclavian vein which would be necessary for allogeneic stem cell transplantation. Three weeks later the patient presented with high fever and headache. Bacillus spp. was isolated from the cathether while blood culture obtained from the peripheral vein remained negative. The bacterial identification was confirmed as B.cereus using VITEK identification system It has been reported Bacillus cereus septicemia may be fatal in immunocompromised hosts despite broad-spectrum appropriate treatment10. Catheter removal is essential for prevention of recurrent bacteremia. Long-term cathater salvage should be reserved for appropriate patient group.

  17. Bacillus cereus Biofilms—Same, Only Different

    Science.gov (United States)

    Majed, Racha; Faille, Christine; Kallassy, Mireille; Gohar, Michel

    2016-01-01

    Bacillus cereus displays a high diversity of lifestyles and ecological niches and include beneficial as well as pathogenic strains. These strains are widespread in the environment, are found on inert as well as on living surfaces and contaminate persistently the production lines of the food industry. Biofilms are suspected to play a key role in this ubiquitous distribution and in this persistency. Indeed, B. cereus produces a variety of biofilms which differ in their architecture and mechanism of formation, possibly reflecting an adaptation to various environments. Depending on the strain, B. cereus has the ability to grow as immersed or floating biofilms, and to secrete within the biofilm a vast array of metabolites, surfactants, bacteriocins, enzymes, and toxins, all compounds susceptible to act on the biofilm itself and/or on its environment. Within the biofilm, B. cereus exists in different physiological states and is able to generate highly resistant and adhesive spores, which themselves will increase the resistance of the bacterium to antimicrobials or to cleaning procedures. Current researches show that, despite similarities with the regulation processes and effector molecules involved in the initiation and maturation of the extensively studied Bacillus subtilis biofilm, important differences exists between the two species. The present review summarizes the up to date knowledge on biofilms produced by B. cereus and by two closely related pathogens, Bacillus thuringiensis and Bacillus anthracis. Economic issues caused by B. cereus biofilms and management strategies implemented to control these biofilms are included in this review, which also discuss the ecological and functional roles of biofilms in the lifecycle of these bacterial species and explore future developments in this important research area. PMID:27458448

  18. Anthrose Biosynthetic Operon of Bacillus anthracis▿

    OpenAIRE

    Dong, Shengli; McPherson, Sylvia A.; Tan, Li; Chesnokova, Olga N.; Turnbough, Charles L.; Pritchard, David G.

    2008-01-01

    The exosporium of Bacillus anthracis spores consists of a basal layer and an external hair-like nap. The nap is composed primarily of the glycoprotein BclA, which contains a collagen-like region with multiple copies of a pentasaccharide side chain. This oligosaccharide possesses an unusual terminal sugar called anthrose, followed by three rhamnose residues and a protein-bound N-acetylgalactosamine. Based on the structure of anthrose, we proposed an enzymatic pathway for its biosynthesis. Exam...

  19. Antimicrobial Effects of Honey on Bacillus Cereus

    OpenAIRE

    This paper should be cited as: Javadzadeh M, Najafi M, Rezaei M, Dastoor M, Behzadi AS, Amiri A . [ Antimicrobial Effects of Honey on Bacillus Cereus ]. MLJ. 201 4 ; 8 ( 2 ): 55 - 61 [Article in Persian] Javadzadeh, M. (MSc; M Najafi; Rezaei, M. (MSc; Dastoor, M. (BSc; Behzadi, AS. (MSc; Amiri, A. (MSc

    2014-01-01

    Background and Objective: Honey is a healthy and nutritious food that has been used for a long time as a treatment for different diseases. One of the applied properties of honey is its antimicrobial effect, which differs between different types of honey due to variation of phenolic and antioxidant compositions. This study aimed to assess antimicrobial effect of honey on Bacillus cereus, considering its chemical properties. Material and Methods: Three samples of honey (A1 and A2 of Khorasan Ra...

  20. BACILLUS CEREUS: ISOLATION IN JENNET MILK

    Directory of Open Access Journals (Sweden)

    M.L. Scatassa

    2011-01-01

    Full Text Available Jennet milk as human food is hypoallergenic for patients affected by Cow Milk Protein Allergy and multiple food allergies. For these pathologies, jennet milk represents the best alternative to other types of milk. Therefore, jennet milk consumers are very sensible to the effects of pathogens' contaminations, and several hygienic practices during the milk production need to be adopted. During regular monitoring in one Sicilian jennet farm, Bacillus cereus in the milk was detected. In 3 bulk milk samples (maximum concentration: 1.2 x 103 ufc/ml, in 3 individual milk samples (10, 20 e 60 ufc/ml, in the milk filter (5 ufc/cm2, in the soil (maximum concentration: 1.5 x 103 ufc/g, on the hands and the gloves of two milkers, on the animal hide (from 1 to 3 ufc/cm2. No spores were detected. A total of 8 Bacillus cereus s.s. strains were analyzed for diarrhoic toxin, and 6 strains producing enterotoxins resulted. The improvement of environmental and milking hygienic conditions reduced Bacillus cereus concentration.

  1. Bioaccumulation of copper, zinc, cadmium and lead by Bacillus sp., Bacillus cereus, Bacillus sphaericus and Bacillus subtilis Bioacumulação de cobre, zinco, cádmio e chumbo por Bacillus sp., Bacillus cereus, Bacillus sphaericus e Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Antonio Carlos Augusto da Costa

    2001-03-01

    Full Text Available This work presents some results on the use of microbes from the genus Bacillus for uptake of cadmium, zinc, copper and lead ions. Maximum copper bioaccumulations were 5.6 mol/g biomass for B. sphaericus, 5.9 mol/g biomass for B. cereus and B. subtilis, and 6.4 mol/g biomass for Bacillus sp. Maximum zinc bioaccumulations were 4.3 mol/g biomass for B. sphaericus, 4.6 mol/g biomass for B. cereus, 4.8 mol/g biomass for Bacillus sp. and 5.0 mol/g biomass for B. subtilis. Maximum cadmium bioaccumulations were 8.0 mol/g biomass for B. cereus, 9.5 mol/g biomass for B. subtilis, 10.8 mol/g biomass for Bacillus sp. and 11.8 mol/g biomass for B. sphaericus. Maximum lead biomaccumulations were 0.7 mol/g biomass for B. sphaericus, 1.1 mol/g biomass for B. cereus, 1.4 mol/g biomass for Bacillus sp. and 1.8 mol/g biomass for B. subtilis. The different Bacillus strains tested presented distinct uptake capacities, and the best results were obtained for B. subtilis and B. cereus.Este trabalho apresenta resultados de acumulação dos íons metálicos cádmio, zinco, cobre e chumbo por bactérias do gênero Bacillus. A bioacumulação máxima de cobre foi 5,6 mol/g biomassa para B. sphaericus, 5,9 mol/g biomassa para B. cereus e B. subtilis, e 6,4 mol/g biomassa para Bacillus sp.. A bioacumulação máxima de zinco foi 4,3 mol/g biomassa para B. sphaericus, 4,6 mol/g biomassa para B. cereus, 4,8 mol/g biomassa para Bacillus sp. e 5,0 mol/g biomassa para B. subtilis. A bioacumulação máxima de cádmio foi 8,0 mol/g biomassa para B. cereus, 9,5 mol/g biomassa para B. subtilis, 10,8 mol/g biomassa para Bacillus sp. e 11,8 mol/g biomassa para B. sphaericus. A bioacumulação máxima de chumbo foi 0,7 mol/g biomassa para B. sphaericus, 1,1 mol/g biomassa para B. cereus, 1,4 mol/g biomassa para Bacillus sp. e 1,8 mol/g biomassa para B. subtilis. As distintas linhagens de Bacillus testadas apresentaram variáveis capacidades de carregamento de íons metálicos, sendo os

  2. Isolation of bacillus thuringiensis from different samples from Mansehra District

    International Nuclear Information System (INIS)

    The insecticidal activity of Bacillus thuringiensis has made it very interesting for the control of a variety of agricultural pests and human disease vectors. The present study is an attempt to explore the potential and diversity. of Bacillus thuringiensis. from the local environment for the control of cotton spotted bollworm (Earias sp.), a major pest of cotton. Two hundred and ninety eight samples of soil, grain dust, wild animal dung, birds dropping, decaying leaves and dead insects were collected from different ecological environments of Mansehra District yielding 438 Bacillus thuringiensis isolates that produce parasporal crystalline inclusions. In this study the soil samples were found to be the richest source for Bacillus thuringiensis. (author)

  3. Soluble Expression and Characterization of Biologically Active Bacillus anthracis Protective Antigen in Escherichia coli

    OpenAIRE

    Nagendra Suryanarayana; Vanlalhmuaka,; Bharti Mankere; Monika Verma; Kulanthaivel Thavachelvam; Urmil Tuteja

    2016-01-01

    Bacillus anthracis secretory protein protective antigen (PA) is primary candidate for subunit vaccine against anthrax. Attempts to obtain large quantity of PA from Escherichia coli expression system often result in the formation of insoluble inclusion bodies. Therefore, it is always better to produce recombinant proteins in a soluble form. In the present study, we have obtained biologically active recombinant PA in small scale E. coli shake culture system using three different expression cons...

  4. Bacillus anthracis capsule activates caspase-1 and induces interleukin-1beta release from differentiated THP-1 and human monocyte-derived dendritic cells.

    Science.gov (United States)

    Cho, Min-Hee; Ahn, Hae-Jeong; Ha, Hyun-Joon; Park, Jungchan; Chun, Jeong-Hoon; Kim, Bong-Su; Oh, Hee-Bok; Rhie, Gi-Eun

    2010-01-01

    The poly-gamma-d-glutamic acid (PGA) capsule is one of the major virulence factors of Bacillus anthracis, which causes a highly lethal infection. The antiphagocytic PGA capsule disguises the bacilli from immune surveillance and allows unimpeded growth of bacilli in the host. Recently, efforts have been made to include PGA as a component of anthrax vaccine; however, the innate immune response of PGA itself has been poorly investigated. In this study, we characterized the innate immune response elicited by PGA in the human monocytic cell line THP-1, which was differentiated into macrophages with phorbol 12-myristate 13-acetate (PMA) and human monocyte-derived dendritic cells (hMoDCs). PGA capsules were isolated from the culture supernatant of either the pXO1-cured strain of B. anthracis H9401 or B. licheniformis ATCC 9945a. PGA treatment of differentiated THP-1 cells and hMoDCs led to the specific extracellular release of interleukin-1beta (IL-1beta) in a dose-dependent manner. Evaluation of IL-1beta processing by Western blotting revealed that cleaved IL-1beta increased in THP-1 cells and hMoDCs after PGA treatment. Enhanced processing of IL-1beta directly correlated with increased activation of its upstream regulator, caspase-1, also known as IL-1beta-converting enzyme (ICE). The extracellular release of IL-1beta in response to PGA was ICE dependent, since the administration of an ICE inhibitor prior to PGA treatment blocked induction of IL-1beta. These results demonstrate that B. anthracis PGA elicits IL-1beta production through activation of ICE in PMA-differentiated THP-1 cells and hMoDCs, suggesting the potential for PGA as a therapeutic target for anthrax. PMID:19737897

  5. Biocontrol and Plant Growth Promotion Characterization of Bacillus Species Isolated from Calendula officinalis Rhizosphere.

    Science.gov (United States)

    Ait Kaki, Asma; Kacem Chaouche, Noreddine; Dehimat, Laid; Milet, Asma; Youcef-Ali, Mounia; Ongena, Marc; Thonart, Philippe

    2013-12-01

    The phenotypic and genotypic diversity of the plant growth promoting Bacillus genus have been widely investigated in the rhizosphere of various agricultural crops. However, to our knowledge this is the first report on the Bacillus species isolated from the rhizosphere of Calendula officinalis. 15 % of the isolated bacteria were screened for their important antifungal activity against Fusarium oxysporum, Botrytis cinerea, Aspergillus niger, Cladosporium cucumerinium and Alternaria alternata. The bacteria identification based on 16S r-RNA and gyrase-A genes analysis, revealed strains closely related to Bacillus amyloliquefaciens, B. velezensis, B. subtilis sub sp spizezenii and Paenibacillus polymyxa species. The electro-spray mass spectrometry coupled to liquid chromatography (ESI-LC MS) analysis showed that most of the Bacillus isolates produced the three lipopeptides families. However, the P. polymyxa (18SRTS) didn't produce any type of lipopeptides. All the tested Bacillus isolates produced cellulase but the protease activity was observed only in the B. amyloliquefaciens species (9SRTS). The Salkowsky colorimetric test showed that the screened bacteria synthesized 6-52 μg/ml of indole 3 acetic acid. These bacteria produced siderophores with more than 10 mm wide orange zones on chromazurol S. The greenhouse experiment using a naturally infested soil with Sclerotonia sclerotiorum showed that the B. amyloliquefaciens (9SRTS) had no significant (P > 0.05) effect on the pre-germination of the chickpea seeds. However, it increased the size of the chickpea plants and reduced the stem rot disease (P officinalis and other crop systems. PMID:24426149

  6. Aerobic Biodegradation of Chlorinated Hydrocarbons by Bacillus circulans WZ-12 CCTCC M 207006 under Saline Conditions

    Institute of Scientific and Technical Information of China (English)

    YU Jianming; CAI Wenji; ZHAO Shiliang; WANG Yan; CHEN Jianmeng

    2013-01-01

    A novel saline-tolerant bacterium Bacillus circulans WZ-12 was evaluated for its potential to degrade four chlorinated hydrocarbons under saline conditions.CH2Cl2 was effectively degraded by Bacillus circulans WZ-12 cells in the medium containing NaC1 concentrations ranging from 5 g·L-1 to 10 g·L-1,and the maximum degradation efficiency (85%) was achieved at NaCl concentration of 10 g·L-.Similarly,Bacillus circulans WZ-12was able to degrade CH2BtCl,C2H4Cl2,and C2H2Cl2 in the presence of 10 g NaCl per liter within 24 h.Cells of Bacillus circulans WZ-12 grown in minimal salt medium contained low levels of glycine betaine (GB),but GB levels were 3-to 5-fold higher in cells grown in media with high salt.Kinetic analysis revealed that biodegradation of the four chlorinated hydrocarbons was concentration dependent and a linear inverse correlation (R2=0.85-0.94)was observed between the rate of biodegradation (Ⅴ) and salt concentration from 5 g·L-1 to 60 g·L-1.The growing cells (in minimal salt medium) degraded approximately 50% of the CH2Cl2 within 24 h,whereas the resting cells (in physiological saline) degraded only 25% of the CH2Cl2 within 24 h and were inactive after 36 h cultivation.Biodegradation could be repeatedly performed for more than 192 h with more than 50% removal efficiency.Bacillus circulans WZ-12 grows well in an aqueous/oil system,hence,it is effective for the treatment of industrial effluents that contain chlorinated hydrocarbons with high salt concentrations.

  7. Historical distribution and molecular diversity of Bacillus anthracis, Kazakhstan.

    Science.gov (United States)

    Aikembayev, Alim M; Lukhnova, Larissa; Temiraliyeva, Gulnara; Meka-Mechenko, Tatyana; Pazylov, Yerlan; Zakaryan, Sarkis; Denissov, Georgiy; Easterday, W Ryan; Van Ert, Matthew N; Keim, Paul; Francesconi, Stephen C; Blackburn, Jason K; Hugh-Jones, Martin; Hadfield, Ted

    2010-05-01

    To map the distribution of anthrax outbreaks and strain subtypes in Kazakhstan during 1937-2005, we combined geographic information system technology and genetic analysis by using archived cultures and data. Biochemical and genetic tests confirmed the identity of 93 archived cultures in the Kazakhstan National Culture Collection as Bacillus anthracis. Multilocus variable number tandem repeat analysis genotyping identified 12 genotypes. Cluster analysis comparing these genotypes with previously published genotypes indicated that most (n = 78) isolates belonged to the previously described A1.a genetic cluster, 6 isolates belonged to the A3.b cluster, and 2 belonged to the A4 cluster. Two genotypes in the collection appeared to represent novel genetic sublineages; 1 of these isolates was from Krygystan. Our data provide a description of the historical, geographic, and genetic diversity of B. anthracis in this Central Asian region. PMID:20409368

  8. Regulation of the anaerobic metabolism in Bacillus subtilis.

    Science.gov (United States)

    Härtig, Elisabeth; Jahn, Dieter

    2012-01-01

    The Gram-positive soil bacterium Bacillus subtilis encounters changing environmental conditions in its habitat. The access to oxygen determines the mode of energy generation. A complex regulatory network is employed to switch from oxygen respiration to nitrate respiration and various fermentative processes. During adaptation, oxygen depletion is sensed by the [4Fe-4S](2+) cluster containing Fnr and the two-component regulatory system ResDE consisting of the membrane-bound histidine kinase ResE and the cytoplasmic ResD regulator. Nitric oxide is the signal recognized by NsrR. Acetate formation and decreasing pH are measured via AlsR. Finally, Rex is responding to changes in the cellular NAD(+)/NADH ration. The fine-tuned interplay of these regulators at approximately 400 target gene promoters ensures efficient adaptation of the B. subtilis physiology. PMID:23046954

  9. 艰难梭状杆菌肠毒素B在巨大芽孢杆菌中的表达和鉴定%Expression and identification of recombinant Clostridium difficile toxin B using Bacillus megaterium system

    Institute of Scientific and Technical Information of China (English)

    杨桂林; 刘威龙; 姚红艳; 周伯平; 冯汉平

    2011-01-01

    目的 获得高纯度和具有生物活性的重组肠毒素B(rTcdB).方法 以艰难梭状芽孢杆菌染色体DNA为模板,通过PCR方法扩增得到全长TcdB基因并克隆至穿梭载体pHis1522.构建的质粒经直接测序验证无误后,转化巨大芽孢杆菌原生质体,在木糖的诱导作用下进行TcdB的表达、纯化及生物活性鉴定.结果 从细菌培养液中纯化得到rTcdB,浓度达到5~10 mg/L,其相对分子质量与天然的TcdB蛋白接近,生物活性与天然的TcdB蛋白相似.结论 在巨大芽孢杆菌中成功表达了具有完整结构和活性的艰难梭状芽孢杆菌TcdB重组蛋白.%Objective To express and purify recombinant and biologically active Clostridium difficile toxin B (rTcdB). Methods The genes of TcdB were amplified by polymerase chain reaction (PCR) using chromosomal DNA from a toxigenic strain, and cloned into a shuttle vector pHis1522.The sequences of TcdB genes in the vector were verified by DNA sequencing. The construction was transformed into Bacillus megaterium protoplasts and the protein expression was driven by a xylose promoter. The purified protein was tested for biological activity. Results rTcdB was successfully purified from bacterial crude extracts. Approximately 5-10 mg of highly purified recombinant toxin was obtained from one liter of bacterial culture. The expressed rTcdB had molecular mass similar to the native toxin, and its biological activity was proved to be similar to its native counterpart after an extensive examination. Conclusion rTcdB with biological activities is successfully expressed in Bacillus megaterium.

  10. Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model

    OpenAIRE

    Lopamudra Haldar; Gandhi, D.N.

    2016-01-01

    Aim: To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. Materials and Methods: An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1) was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2) and (T3) groups received spore biomass of Bacillus coagulans B...

  11. Emetic toxin-producing strains of Bacillus cereus show distinct characteristics within the Bacillus cereus group.

    NARCIS (Netherlands)

    Carlin, Frédéric; Fricker, Martina; Pielaat, Annemarie; Heisterkamp, Simon; Shaheen, Ranad; Salonen, Mirja Salkinoja; Svensson, Birgitta; Nguyen-the, Christophe; Ehling-Schulz, Monika

    2006-01-01

    One hundred representative strains of Bacillus cereus were selected from a total collection of 372 B. cereus strains using two typing methods (RAPD and FT-IR) to investigate if emetic toxin-producing hazardous B. cereus strains possess characteristic growth and heat resistance profiles. The strains

  12. Detection of Anthrax Simulants with Microcalorimetric Spectroscopy: Bacillus subtilis and Bacillus cereus Spores

    Science.gov (United States)

    Arakawa, Edward T.; Lavrik, Nickolay V.; Datskos, Panos G.

    2003-04-01

    Recent advances in the development of ultrasensitive micromechanical thermal detectors have led to the advent of novel subfemtojoule microcalorimetric spectroscopy (CalSpec). On the basis of principles of photothermal IR spectroscopy combined with efficient thermomechanical transduction, CalSpec provides acquisition of vibrational spectra of microscopic samples and absorbates. We use CalSpec as a method of identifying nanogram quantities of biological micro-organisms. Our studies focus on Bacillus subtilis and Bacillus cereus spores as simulants for Bacillus anthracis spores. Using CalSpec, we measured IR spectra of B. subtilis and B. cereus spores present on surfaces in nanogram quantities (approximately 100 -1000 spores). The spectra acquired in the wavelength range of 690 -4000 cm-1 (2.5 -14.5 μm) contain information-rich vibrational signatures that reflect the different ratios of biochemical makeup of the micro-organisms. The distinctive features in the spectra obtained for the two types of micro-organism can be used to distinguish between the spores of the Bacillus family. As compared with conventional IR and Fourier-transform IR microscopic spectroscopy techniques, the advantages of the present technique include significantly improved sensitivity (at least a full order of magnitude), absence of expensive IR detectors, and excellent potential for miniaturization.

  13. Complete Genome Sequence of Bacillus thuringiensis Bacteriophage Smudge.

    Science.gov (United States)

    Cornell, Jessica L; Breslin, Eileen; Schuhmacher, Zachary; Himelright, Madison; Berluti, Cassandra; Boyd, Charles; Carson, Rachel; Del Gallo, Elle; Giessler, Caris; Gilliam, Benjamin; Heatherly, Catherine; Nevin, Julius; Nguyen, Bryan; Nguyen, Justin; Parada, Jocelyn; Sutterfield, Blake; Tukruni, Muruj; Temple, Louise

    2016-01-01

    Smudge, a bacteriophage enriched from soil using Bacillus thuringiensis DSM-350 as the host, had its complete genome sequenced. Smudge is a myovirus with a genome consisting of 292 genes and was identified as belonging to the C1 cluster of Bacillus phages. PMID:27540049

  14. Complete Genome of Bacillus thuringiensis Myophage BigBertha

    OpenAIRE

    Ting, Jose H.; Smyth, Trinity B.; Chamakura, Karthik R.; Kuty Everett, Gabriel F.

    2013-01-01

    BigBertha is a myophage of Bacillus thuringiensis, a widely used biocontrol agent that is active against many insect pests of plants. Here, we present the complete annotated genome of BigBertha. The genome shares 85.9% sequence identity with Bacillus cereus phage B4.

  15. Non-peptide metabolites from the genus Bacillus.

    Science.gov (United States)

    Hamdache, Ahlem; Lamarti, Ahmed; Aleu, Josefina; Collado, Isidro G

    2011-04-25

    Bacillus species produce a number of non-peptide metabolites that display a broad spectrum of activity and structurally diverse bioactive chemical structures. Biosynthetic, biological, and structural studies of these metabolites isolated from Bacillus species are reviewed. This contribution also includes a detailed study of the activity of the metabolites described, especially their role in biological control mechanisms.

  16. Quantitative immunofluorescence studies of the serology of Bacillus anthracis spores.

    OpenAIRE

    Phillips, A. P.; Martin, K L

    1983-01-01

    A fluorescein-conjugated antibody against formalin-inactivated spores of Bacillus anthracis Vollum reacted only weakly with a variety of Bacillus species in microfluorometric immunofluorescence assays. A conjugated antibody against spores of B. anthracis Sterne showed little affinity for spores of several B. anthracis isolates including B. anthracis Vollum, indicating that more than one anthrax spore serotype exists.

  17. Complete Genome Sequence of Bacillus thuringiensis Bacteriophage Smudge

    Science.gov (United States)

    Cornell, Jessica L.; Breslin, Eileen; Schuhmacher, Zachary; Himelright, Madison; Berluti, Cassandra; Boyd, Charles; Carson, Rachel; Del Gallo, Elle; Giessler, Caris; Gilliam, Benjamin; Heatherly, Catherine; Nevin, Julius; Nguyen, Bryan; Nguyen, Justin; Parada, Jocelyn; Sutterfield, Blake; Tukruni, Muruj

    2016-01-01

    Smudge, a bacteriophage enriched from soil using Bacillus thuringiensis DSM-350 as the host, had its complete genome sequenced. Smudge is a myovirus with a genome consisting of 292 genes and was identified as belonging to the C1 cluster of Bacillus phages. PMID:27540049

  18. Complete Genome Sequence of Bacillus thuringiensis Bacteriophage Smudge.

    Science.gov (United States)

    Cornell, Jessica L; Breslin, Eileen; Schuhmacher, Zachary; Himelright, Madison; Berluti, Cassandra; Boyd, Charles; Carson, Rachel; Del Gallo, Elle; Giessler, Caris; Gilliam, Benjamin; Heatherly, Catherine; Nevin, Julius; Nguyen, Bryan; Nguyen, Justin; Parada, Jocelyn; Sutterfield, Blake; Tukruni, Muruj; Temple, Louise

    2016-08-18

    Smudge, a bacteriophage enriched from soil using Bacillus thuringiensis DSM-350 as the host, had its complete genome sequenced. Smudge is a myovirus with a genome consisting of 292 genes and was identified as belonging to the C1 cluster of Bacillus phages.

  19. Genetic map of the Bacillus stearothermophilus NUB36 chromosome

    Energy Technology Data Exchange (ETDEWEB)

    Vallier, H.; Welker, N.E. (Northwestern Univ., Evanston, IL (USA))

    1990-02-01

    A circular genetic map of Bacillus stearothermophilus NUB36 was constructed by transduction with bacteriophage TP-42C and protoplast fusion. Sixty-four genes were tentatively assigned a cognate Bacillus subtilis gene based on growth response to intermediates or end products of metabolism, cross-feeding, accumulation of intermediates, or their relative order in a linkage group. Although the relative position of many genes on the Bacillus subtilis genetic map appears to be similar, some differences were detected. The tentative order of the genes in the Bacillus stearothermophilus aro region is aspB-aroBAFEC-tyra-hisH-(trp), whereas it is aspB-aroE-tyrA-hisH-(trp)-aroHBF in Bacillus subtilis. The aroA, aroC, and aroG genes in Bacillus subtilis are located in another region. The tentative order of genes in the trp operon of Bacillus stearothermophilus is trpFCDABE, whereas it is trpABFCDE in Bacillus subtilis.

  20. Secreted Expression of the Maltogenic Amylase in Bacillus subtilis%麦芽糖α-淀粉酶基因在枯草芽孢杆菌中的分泌表达

    Institute of Scientific and Technical Information of China (English)

    沈微; 俞玲; 陈献忠; 樊游; 王正祥

    2011-01-01

    The gene encoding maltogenic amylase was fused with signal peptide gene encoding amylase of Bacillus licheniformis by overlapping PCR.The fused gene(BlMa) was ligated with an expression vector of Bacillus,pHY-P43.B1Ma transforms Bacillus subtilis directly and results in the recombinant plasmid pHY-P43-BlMa.Bacillus subtilis 1A717 which is deficient in amylase was selected as expression host.The recombinant strain was designated as Bacillus subtilis / pHY-P43-BlMa.Enzyme activity assay and SDS-PAGE showed that recombinant maltogenic amylase of B.licheniformis(BlMa enzyme)expressed by B.subtilis / pHY-P43-BlMa was secreted into medium.High Performance Liquid Chromatography(HPLC) identified that major hydrolysate of starch catalyzed by BlMa enzyme was maltose.Culture condition of Bacillus subtilis / pHY-P43-BlMa in flask scale was optimized for production of BlMa enzyme,the optimized medium was: 10 % corn starch,2.5 % cotton seed protein,0.3 %(NH4)2SO4,0.03 % CaCl2 and 0.1 % NaH2PO4.The maximum enzyme activity under optimized condition reached 5.9 U/mL.%采用重叠PCR方法在麦芽糖α-淀粉酶编码基因5’端添加地衣芽孢杆菌α-淀粉酶基因信号肽编码区,获得重组基因BlMa。重组基因与芽孢杆菌表达载体pHY-P43连接后直接转化枯草芽孢杆菌,获得重组质粒pHY-P43-BlMa。枯草芽孢杆菌淀粉酶基因缺陷株1A717被用作BlMa基因表达宿主菌,重组菌命名为Bacillus subtilis/pHY-P43-BlMa。酶活检测和SDS-PAGE电泳均显示,B.subtilis/pHY-P43-BlMa表达的重组麦芽糖淀粉酶(BlMa)全部分泌到培养液中。HPLC检测表明,BlMa催化可溶性淀粉水解产物主要为麦芽糖。对B.subtilis/pHY-P43-BlMa摇瓶发酵条件进行优化。获得优化发酵培养基配方:10%玉米淀粉,2.5%药媒,0.3%(NH4)2SO4,0.03%CaCl2,0.1%NaH2PO4,在优化条件下重组菌发酵酶活为5.9 U/mL。

  1. Influence of orally fed a select mixture of Bacillus probiotics on intestinal T-cell migration in weaned MUC4 resistant pigs following Escherichia coli challenge.

    Science.gov (United States)

    Yang, Gui-Yan; Zhu, Yao-Hong; Zhang, Wei; Zhou, Dong; Zhai, Cong-Cong; Wang, Jiu-Feng

    2016-01-01

    Efficient strategies for treating enteritis caused by F4(+) enterotoxigenic Escherichia coli (ETEC)/verocytotoxigenic Escherichia coli (VTEC)/enteropathogenic E. coli (EPEC) in mucin 4 resistant (MUC4 RR; supposed to be F4ab/ac receptor-negative [F4ab/acR(-)]) pigs remain elusive. A low (3.9 × 10(8) CFU/day) or high (7.8 × 10(8) CFU/day) dose of Bacillus licheniformis and Bacillus subtilis spore mixture (BLS-mix) was orally administered to MUC4 RR piglets for 1 week before F4(+) ETEC/VTEC/EPEC challenge. Orally fed BLS-mix upregulated the expression of TLR4, NOD2, iNOS, IL-8, and IL-22 mRNAs in the small intestine of pigs challenged with E. coli. Expression of chemokine CCL28 and its receptor CCR10 mRNAs was upregulated in the jejunum of pigs pretreated with high-dose BLS-mix. Low-dose BLS-mix pretreatment induced an increase in the proportion of peripheral blood CD4(-)CD8(-) T-cell subpopulations and high-dose BLS-mix induced the expansion of CD4(-)CD8(-) T cells in the inflamed intestine. Immunostaining revealed that considerable IL-7Rα-expressing cells accumulated at the lamina propria of the inflamed intestines after E. coli challenge, even in pigs pretreated with either low- or high-dose BLS-mix, although Western blot analysis of IL-7Rα expression in the intestinal mucosa did not show any change. Our data indicate that oral administration of the probiotic BLS-mix partially ameliorates E. coli-induced enteritis through facilitating upregulation of intestinal IL-22 and IκBα expression, and preventing loss of intestinal epithelial barrier integrity via elevating ZO-1 expression. However, IL-22 also elicits an inflammatory response in inflamed intestines as a result of infection with enteropathogenic bacteria. PMID:27424033

  2. Bacillus subtilis regulatory protein GerE

    OpenAIRE

    Ducros, V M A; Brannigan, J.A.; Lewis, R J; Wilkinson, A.J.

    1998-01-01

    GerE is the latest-acting of a series of factors which regulate gene expression in the mother cell during sporulation in Bacillus. The gene encoding GerE has been cloned from B. subtilis and overexpressed in Escherichia coli. Purified GerE has been crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The small plate-like crystals belong to the monoclinic space group C2 and diffract beyond 2.2 Angstrom resolution with a synchrotron radiation X-ra...

  3. COMBINED MICROBIAL SURFACTANT-POLYMER SYSTEM FOR IMPROVED OIL MOBILITY AND CONFORMANCE CONTROL

    Energy Technology Data Exchange (ETDEWEB)

    Jorge Gabitto; Maria Barrufet

    2005-08-01

    show a significant influence of nutrient regime on alternate production of surfactants or polymers for a Bacillus licheniformis strain, NIPER 1A. The main conclusion of this work is that NIPER 1A can produce both surfactant and polymer by nutrient regime manipulation. Our experiments proved that this property leads to improved oil recovery by increasing alternatively, oil mobility and conformance control.

  4. Scale-down and parallel operation of a riboflavin production process with Bacillus subtilis

    OpenAIRE

    Knorr, Bettina

    2007-01-01

    Novel parallel bioreactor systems at a milliliter scale were recently developed for the design and improvement of biological cultivations. The objective of this work was to identify the reaction parameters that were necessary for a representative scale-down of an industrial manufacturing process to be carried out with the new technology. The process for the production of riboflavin with Bacillus subtilis, operated in a controlled fed-batch mode, served as an example for investigations in stir...

  5. Genetical and radiobiological characteristics of phage Tg13 of Bacillus thuringiensis

    International Nuclear Information System (INIS)

    The radiation-genetical aspects of interrelations between phages and cells of the spore-forming bacteria Bacillus thurin-giensis were studied. The phage Tg13 liberates C-mutants, forming transparent negative colonies, both spontaneously and under the effect of UV irradiation. UV-radiation increases reliably the level of C-mutants in the population. The phenotype of the observed mutants is, evidently, caused by the specific features of interaction in the system: preudolysogenic culture -phage Tg13

  6. Formulation and evaluation of Bacillus coagulans-loaded hypromellose mucoadhesive microspheres

    OpenAIRE

    Alli SMA

    2011-01-01

    Sk Md Athar AlliDepartment of Pharmaceutical Technology, Jadavpur University, Kolkata, West Bengal, IndiaAbstract: Development of a novel delivery system has been attempted to deliver viable probiotic cells into the gut for a prolonged period of time while maintaining high numbers of viable cells within the formulation throughout the shelf-life of the product and during the gastrointestinal transit. Core mucoadhesive microspheres of Bacillus coagulans were developed employing several grades o...

  7. Bacillus anthracis HssRS signaling to HrtAB regulates heme resistance during infection

    OpenAIRE

    Stauff, Devin L; Skaar, Eric P.

    2009-01-01

    Bacillus anthracis proliferates to high levels within vertebrate tissues during the pathogenesis of anthrax. This growth is facilitated by the acquisition of nutrient iron from host heme. However, heme acquisition can lead to the accumulation of toxic amounts of heme within B. anthracis. Here, we show that B. anthracis resists heme toxicity by sensing heme through the HssRS two-component system, which regulates expression of the heme-detoxifying transporter HrtAB. In addition, we demonstrate ...

  8. Production and Characterization of Biodiesel Using Nonedible Castor Oil by Immobilized Lipase from Bacillus aerius

    OpenAIRE

    2015-01-01

    A novel thermotolerant lipase from Bacillus aerius was immobilized on inexpensive silica gel matrix. The immobilized lipase was used for the synthesis of biodiesel using castor oil as a substrate in a solvent free system at 55°C under shaking in a chemical reactor. Several crucial parameters affecting biodiesel yield such as incubation time, temperature, substrate molar ratio, and amount of lipase were optimized. Under the optimized conditions, the highest biodiesel yield was up to 78.13%. Th...

  9. Structural and Functional Analysis of the GerD Spore Germination Protein of Bacillus Species

    OpenAIRE

    Li, Yunfeng; Jin, Kai; Ghosh, Sonali; Devarakonda, Parvathimadhavi; Carlson, Kristina; Davis, Andrew; Stewart, Kerry-Ann V.; Cammett, Elizabeth; Rossi, Patricia Pelczar; Setlow, Barbara; Lu, Min; Setlow, Peter; Hao, Bing

    2014-01-01

    Spore germination in Bacillus species represents an excellent model system with which to study the molecular mechanisms underlying the nutritional control of growth and development. Binding of specific chemical nutrients to their cognate receptors located in the spore inner membrane triggers the germination process that leads to a resumption of metabolism in spore outgrowth. Recent studies suggest that the inner membrane GerD lipoprotein plays a critical role in the receptor-mediated activati...

  10. Prosthetic Joint Infection due to Mycobacterium bovis after Intravesical Instillation of Bacillus Calmette-Guerin (BCG

    Directory of Open Access Journals (Sweden)

    Eric Gomez

    2009-01-01

    Full Text Available Intravesical instillation of Bacillus Calmette-Guerin (BCG is a treatment to prevent recurrence of superficial urothelial bladder carcinoma. Complications after bladder instillation of BCG have been reported including locally invasive and systemic infections due to dissemination of Mycobacterium bovis from the bladder. We present an uncommon case and literature review of prosthetic joint infection due to M. bovis after intravesical BCG treatment of bladder cancer.

  11. Reagent-free and portable detection of Bacillus anthracis spores using a microfluidic incubator and smartphone microscope

    Energy Technology Data Exchange (ETDEWEB)

    Hutchison, Janine R.; Erikson, Rebecca L.; Sheen, Allison M.; Ozanich, Richard M.; Kelly, Ryan T.

    2015-08-06

    Rapid, cost-effective bacterial detection systems are needed to respond to potential biothreat events. Here we report the use of smartphone-based microscopy in combination with a simple microfluidic incubation device to detect 5000 Bacillus anthracis spores in 3 hours. This field-deployable approach is compatible with real-time PCR for secondary confirmation.

  12. Growth and sporulation of Bacillus cereus ATCC 14579 under defined conditions: temporal expression of genes for key sigma factors

    NARCIS (Netherlands)

    Vries, de Y.P.; Hornstra, L.M.; Vos, de W.M.; Abee, T.

    2004-01-01

    An airlift fermentor system allowing precise regulation of pH and aeration combined with a chemically defined medium was used to study growth and sporulation of Bacillus cereus ATCC 14579. Sporulation was complete and synchronous. Expression of sigA, sigB, sigF, and sigG was monitored with real-time

  13. The Phylogeny of Bacillus cereus sensu lato.

    Science.gov (United States)

    Okinaka, Richard T; Keim, Paul

    2016-02-01

    The three main species of the Bacillus cereus sensu lato, B. cereus, B. thuringiensis, and B. anthracis, were recognized and established by the early 1900s because they each exhibited distinct phenotypic traits. B. thuringiensis isolates and their parasporal crystal proteins have long been established as a natural pesticide and insect pathogen. B. anthracis, the etiological agent for anthrax, was used by Robert Koch in the 19th century as a model to develop the germ theory of disease, and B. cereus, a common soil organism, is also an occasional opportunistic pathogen of humans. In addition to these three historical species designations, are three less-recognized and -understood species: B. mycoides, B. weihenstephanensis, and B. pseudomycoides. All of these "species" combined comprise the Bacillus cereus sensu lato group. Despite these apparently clear phenotypic definitions, early molecular approaches to separate the first three by various DNA hybridization and 16S/23S ribosomal sequence analyses led to some "confusion" because there were limited differences to differentiate between these species. These and other results have led to frequent suggestions that a taxonomic change was warranted to reclassify this group to a single species. But the pathogenic properties of B. anthracis and the biopesticide applications of B. thuringiensis appear to "have outweighed pure taxonomic considerations" and the separate species categories are still being maintained. B. cereus sensu lato represents a classic example of a now common bacterial species taxonomic quandary. PMID:26999390

  14. Fast Neutron Radiation Effects on Bacillus Subtili

    Institute of Scientific and Technical Information of China (English)

    CHEN Xiaoming; REN Zhenglong; ZHANG Jianguo; ZHENG Chun; TAN Bisheng; YANG Chengde; CHU Shijin

    2009-01-01

    To examine the sterilizing effect and mechanism of neutron radiation, Bacillus sub-tilis vat. niger, strain (ATCC 9372) spores were irradiated with the fast neutron from the Chinese fast burst reactor Ⅱ(CFBR-Ⅱ). The plate-count results indicated that the D10 value was 384.6 Gy with a neutron radiation dose rate of 7.4 Gy/min. The rudimental catalase activity of the spores declined obviously with the increase in the radiation dose. Meanwhile, under the scanning electron microscope, no visible influence of the neutron radiation on the spore configuration was detected even if the dose was increased to 4 kGy. The content and distribution of DNA double-strand breaks induced by neutron radiation at different doses were measured and quantified by pulsed-field gel electrophoresis (PFGE). Further analysis of the DNA release percentage (PR), the DNA breakage level (L), and the average molecular weight, indicated that DNA fragments were obvi-ously distributed around the 5 kb regions at different radiation doses, which suggests that some points in the DNA molecule were sensitive to neutron radiation. Both PR and L varied regularly to some extent with the increase in radiation dose. Thus neutron radiation has a high sterilization power, and can induce falling enzyme activity and DNA breakage in Bacillus subtilis spores

  15. The mutation Glu151Asp in the B-component of the Bacillus cereus non-hemolytic enterotoxin (Nhe) leads to a diverging reactivity in antibody-based detection systems.

    Science.gov (United States)

    Didier, Andrea; Jeßberger, Nadja; Krey, Victoria; Dietrich, Richard; Scherer, Siegfried; Märtlbauer, Erwin

    2015-11-09

    The ability of Bacillus cereus to cause foodborne toxicoinfections leads to increasing concerns regarding consumer protection. For the diarrhea-associated enterotoxins, the assessment of the non-hemolytic enterotoxin B (NheB) titer determined by a sandwich enzyme immunoassay (EIA) correlates best with in vitro cytotoxicity. In general, the regulation of enterotoxin expression of B. cereus is a coordinately-regulated process influenced by environmental, and probably also by host factors. As long as these factors are not completely understood, the currently-applied diagnostic procedures are based on indirect approaches to assess the potential virulence of an isolate. To date, sandwich EIA results serve as a surrogate marker to categorize isolates as either potentially low or highly toxic. Here, we report on a single amino acid exchange in the NheB sequence leading to an underestimation of the cytotoxic potential in a limited number of strains. During the screening of a large panel of B. cereus isolates, six showed uncommon features with low sandwich EIA titers despite high cytotoxicity. Sequence analysis revealed the point-mutation (Glu)151(Asp) in the potential binding region of the capture antibody. Application of this antibody also results in low titers in an indirect EIA format and shows variable detection intensities in Western-immunoblots. A commercially-available assay based on a lateral flow device detects all strains correctly as NheB producers in a qualitative manner. In conclusion, isolates showing low NheB titers should additionally be assayed in an indirect EIA or for their in vitro cytotoxicity to ensure a correct classification as either low or highly toxic.

  16. The Mutation Glu151Asp in the B-Component of the Bacillus cereus Non-Hemolytic Enterotoxin (Nhe Leads to a Diverging Reactivity in Antibody-Based Detection Systems

    Directory of Open Access Journals (Sweden)

    Andrea Didier

    2015-11-01

    Full Text Available The ability of Bacillus cereus to cause foodborne toxicoinfections leads to increasing concerns regarding consumer protection. For the diarrhea-associated enterotoxins, the assessment of the non-hemolytic enterotoxin B (NheB titer determined by a sandwich enzyme immunoassay (EIA correlates best with in vitro cytotoxicity. In general, the regulation of enterotoxin expression of B. cereus is a coordinately-regulated process influenced by environmental, and probably also by host factors. As long as these factors are not completely understood, the currently-applied diagnostic procedures are based on indirect approaches to assess the potential virulence of an isolate. To date, sandwich EIA results serve as a surrogate marker to categorize isolates as either potentially low or highly toxic. Here, we report on a single amino acid exchange in the NheB sequence leading to an underestimation of the cytotoxic potential in a limited number of strains. During the screening of a large panel of B. cereus isolates, six showed uncommon features with low sandwich EIA titers despite high cytotoxicity. Sequence analysis revealed the point-mutation Glu151Asp in the potential binding region of the capture antibody. Application of this antibody also results in low titers in an indirect EIA format and shows variable detection intensities in Western-immunoblots. A commercially-available assay based on a lateral flow device detects all strains correctly as NheB producers in a qualitative manner. In conclusion, isolates showing low NheB titers should additionally be assayed in an indirect EIA or for their in vitro cytotoxicity to ensure a correct classification as either low or highly toxic.

  17. The Mutation Glu151Asp in the B-Component of the Bacillus cereus Non-Hemolytic Enterotoxin (Nhe) Leads to a Diverging Reactivity in Antibody-Based Detection Systems

    Science.gov (United States)

    Didier, Andrea; Jeßberger, Nadja; Krey, Victoria; Dietrich, Richard; Scherer, Siegfried; Märtlbauer, Erwin

    2015-01-01

    The ability of Bacillus cereus to cause foodborne toxicoinfections leads to increasing concerns regarding consumer protection. For the diarrhea-associated enterotoxins, the assessment of the non-hemolytic enterotoxin B (NheB) titer determined by a sandwich enzyme immunoassay (EIA) correlates best with in vitro cytotoxicity. In general, the regulation of enterotoxin expression of B. cereus is a coordinately-regulated process influenced by environmental, and probably also by host factors. As long as these factors are not completely understood, the currently-applied diagnostic procedures are based on indirect approaches to assess the potential virulence of an isolate. To date, sandwich EIA results serve as a surrogate marker to categorize isolates as either potentially low or highly toxic. Here, we report on a single amino acid exchange in the NheB sequence leading to an underestimation of the cytotoxic potential in a limited number of strains. During the screening of a large panel of B. cereus isolates, six showed uncommon features with low sandwich EIA titers despite high cytotoxicity. Sequence analysis revealed the point-mutation Glu151Asp in the potential binding region of the capture antibody. Application of this antibody also results in low titers in an indirect EIA format and shows variable detection intensities in Western-immunoblots. A commercially-available assay based on a lateral flow device detects all strains correctly as NheB producers in a qualitative manner. In conclusion, isolates showing low NheB titers should additionally be assayed in an indirect EIA or for their in vitro cytotoxicity to ensure a correct classification as either low or highly toxic. PMID:26569304

  18. ISOLATION AND CHARACTERIZATION OF CRUDE OIL DEGRADING BACILLUS SPP.

    Directory of Open Access Journals (Sweden)

    A. Akhavan Sepahi, I. Dejban Golpasha, M. Emami, A. M. Nakhoda

    2008-07-01

    Full Text Available Today, application of microorganisms for removing crude oil pollution from contaminated sites as bioremediation studies, was considered by scientists because other methods such as surfactant washing and incineration lead to production of more toxic compounds and they are non-economic. Fifteen crude oil degrading bacillus spp. were isolated from contaminated sites. Two isolated showed best growth in liquid media with 1-3% (v/v crude oil and mineral salt medium, then studied for enzymatic activities on tested media. The results showed maximal increase in optical densities and total viable count concomitant with decrease in pH on fifth day of experimental period for bacillus S6. Typical generation time on mineral salt with 1% crude oil is varying between 18-20h, 25-26h respectively for bacillus S6 and S35. Total protein was monitored at determined time intervals as biodegradation indices. Increasing of protein concentration during the incubation period reveals that isolated bacillus can degrade crude oil and increase microbial biomass. These bacillus spp. reduced surface tension from 60 (mN/m to 31 and 38 (mN/m, It means that these bacillus spp. can produce sufficient surfactant and have good potential of emulsification capacity. The results demonstrated that these bacillus spp. can utilize crude oil as a carbon and energy source.

  19. Expression, purification and preliminary crystallographic characterization of FlhF from Bacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Bange, Gert; Petzold, Georg; Wild, Klemens; Sinning, Irmgard, E-mail: irmi.sinning@bzh.uni-heidelberg.de [Heidelberg University Biochemistry Centre (BZH), INF 328, 69120 Heidelberg (Germany)

    2007-05-01

    Preliminary crystallographic data are reported for the third SRP GTPase FlhF from Bacillus subtilis. The Gram-positive bacterium Bacillus subtilis contains three proteins belonging to the signal recognition particle (SRP) type GTPase family. The well characterized signal sequence-binding protein SRP54 and the SRP receptor protein FtsY are universally conserved components of the SRP system of protein transport. The third member, FlhF, has been implicated in the placement and assembly of polar flagella. This article describes the overexpression and preliminary X-ray crystallographic analysis of an FlhF fragment that corresponds to the well characterized GTPase domains in SRP54 and FtsY. Three crystal forms are reported with either GDP or GMPPNP and diffract to a resolution of about 3 Å.

  20. Occurrence and significance of Bacillus cereus and Bacillus thuringiensis in ready-to-eat food

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Ørum-Smidt, Lasse; Andersen, Sigrid R;

    2005-01-01

    had at least one gene or component involved in human diarrhoeal disease, while emetic toxin was related to only one B. cereus strain. A new observation was that 31 out of the 40 randomly selected B. cereus-like strains could be classified as Bacillus thuringiensis due to crystal production and......Among 48,901 samples of ready-to-eat food products at the Danish retail market, 0.5% had counts of Bacillus cereus-like bacteria above 10(4) cfu g(-1). The high counts were most frequently found in starchy, cooked products, but also in fresh cucumbers and tomatoes. Forty randomly selected strains....../or content of cry genes. Thus, a large proportion of the B. cereus-like organisms present in food may belong to B. thuringiensis....

  1. Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The gene coding for a thermophilic neutral protease from Bacillus stearothermophilus was expressed in Bacillus subtilis DB104, under the control of the sacB gene promoter. This was followed by either the native signal peptide sequence of this protease or the signal peptide sequence of the sacB gene. The protease was purified 3.8-fold, with a specific activity of 16530 U mg-1. As analyzed by SDS-PAGE, the molecular mass of the expressed protease was about 35 kDa, and the optimal temperature and pH of the protease were 65℃ and 7.5, respectively. Moreover, it still had about 80% activity after 1 h reaction at 65℃.

  2. Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The gene coding for a thermophilic neutral protease from Bacillus stearothermophilus was expressed in Bacillus subtilis DB104, under the control of the sacB gene promoter. This was followed by either the native signal peptide sequence of this protease or the signal peptide sequence of the sacB gene. The protease was purified 3.8-fold, with a specific activity of 16530 U mg-1. As analyzed by SDS-PAGE, the molecular mass of the expressed protease was about 35 kDa, and the optimal temperature and pH of the protease were 65℃ and 7.5, respectively. Moreover, it still had about 80% activity after 1 h reaction at 65 ℃ .

  3. An Optical Biosensor for Bacillus Cereus Spore Detection

    Science.gov (United States)

    Li, Chengquan; Tom, Harry W. K.

    2005-03-01

    We demonstrate a new transduction scheme for optical biosensing. Bacillus cereus is a pathogen that may be found in food and dairy products and is able to produce toxins and cause food poisoning. It is related to Bacillus anthracis (anthrax). A CCD array covered with micro-structured glass coverslip is used to detect the optical resonant shift due to the binding of the antigen (bacillus cereus spore) to the antibody (polyclonal antibody). This novel optical biosensor scheme has the potential for detecting 10˜100 bioagents in a single device as well as the potential to test for antigens with multiple antibody tests to avoid ``false positives.''

  4. Flow-cytometric Analysis of Bacillus anthracis Spores

    Directory of Open Access Journals (Sweden)

    D. V. Kamboj

    2006-11-01

    Full Text Available Flow-cytometric technique has been established as a powerful tool for detection andidentification of microbiological agents. Unambiguous and rapid detection of Bacillus anthracisspores has been reported using immunoglobulin G-fluorescein isothiocyanate conjugate againstlive spores. In addition to the high sensitivity, the present technique could differentiate betweenspores of closely related species, eg, Bacillus cereus and Bacillus subtilis using fluorescenceintensity. The technique can be used for detection of live as well as inactivated spores makingit more congenial for screening of suspected samples of bioterrorism.

  5. Genetic Differentiation between Sympatric Populations of Bacillus cereus and Bacillus thuringiensis

    Science.gov (United States)

    Vilas-Boas, Gislayne; Sanchis, Vincent; Lereclus, Didier; Lemos, Manoel Victor F.; Bourguet, Denis

    2002-01-01

    Little is known about genetic exchanges in natural populations of bacteria of the spore-forming Bacillus cereus group, because no population genetics studies have been performed with local sympatric populations. We isolated strains of Bacillus thuringiensis and B. cereus from small samples of soil collected at the same time from two separate geographical sites, one within the forest and the other at the edge of the forest. A total of 100 B. cereus and 98 B. thuringiensis strains were isolated and characterized by electrophoresis to determine allelic composition at nine enzymatic loci. We observed genetic differentiation between populations of B. cereus and B. thuringiensis. Populations of a given Bacillus species—B. thuringiensis or B. cereus—were genetically more similar to each other than to populations of the other Bacillus species. Hemolytic activity provided further evidence of this genetic divergence, which remained evident even if putative clones were removed from the data set. Our results suggest that the rate of gene flow was higher between strains of the same species, but that exchanges between B. cereus and B. thuringiensis were nonetheless possible. Linkage disequilibrium analysis revealed sufficient recombination for B. cereus populations to be considered panmictic units. In B. thuringiensis, the balance between clonal proliferation and recombination seemed to depend on location. Overall, our data indicate that it is not important for risk assessment purposes to determine whether B. cereus and B. thuringiensis belong to a single or two species. Assessment of the biosafety of pest control based on B. thuringiensis requires evaluation of the extent of genetic exchange between strains in realistic natural conditions. PMID:11872495

  6. Bacillus subtilis as potential producer for polyhydroxyalkanoates

    Directory of Open Access Journals (Sweden)

    Patel Sanjay KS

    2009-07-01

    Full Text Available Abstract Polyhydroxyalkanoates (PHAs are biodegradable polymers produced by microbes to overcome environmental stress. Commercial production of PHAs is limited by the high cost of production compared to conventional plastics. Another hindrance is the brittle nature and low strength of polyhydroxybutyrate (PHB, the most widely studied PHA. The needs are to produce PHAs, which have better elastomeric properties suitable for biomedical applications, preferably from inexpensive renewable sources to reduce cost. Certain unique properties of Bacillus subtilis such as lack of the toxic lipo-polysaccharides, expression of self-lysing genes on completion of PHA biosynthetic process – for easy and timely recovery, usage of biowastes as feed enable it to compete as potential candidate for commercial production of PHA.

  7. Bacillus circulans exopolysaccharide: Production, characterization and bioactivities.

    Science.gov (United States)

    Vidhyalakshmi, R; Valli, Nachiyar C; Narendra Kumar, G; Sunkar, Swetha

    2016-06-01

    A bacterium with the ability to produce appreciable amount of exopolysaccharide was isolated from slimy layer of coconut. 16S rDNA analysis identified the organism as Bacillus circulans. EPS production was observed at all stages of culture growth and reached maximum of 0.065mg/ml by 96h, which on further incubation started to decrease. Response Surface Methodology using Box Behnken design has shown the influence of sucrose which was found to be directly proportional to exopolysaccharide production with production reaching 1.09mg/ml. HPLC analysis identified the presence of glucose, mannose, fructose and verbascose and NMR analysis confirmed the presence of glucose, mannose and galactose. Even though the extracted B. circulans EPS did not show appreciable anti-bacterial or anti-fungal activity, it exhibited appreciable antioxidant, anti-inflammatory and anti-tumor activity. PMID:26902891

  8. Bacillus subtilis genome vector-based complete manipulation and reconstruction of genomic DNA for mouse transgenesis

    OpenAIRE

    Iwata, Tetsuo; Kaneko, Shinya; Shiwa, Yuh; Enomoto, Takayuki; Yoshikawa, Hirofumi; Hirota, Junji

    2013-01-01

    Background The Bacillus subtilis genome (BGM) vector is a novel cloning system for large DNA fragments, in which the entire 4.2 Mb genome of B. subtilis functions as a vector. The BGM vector system has several attractive properties, such as a large cloning capacity of over 3 Mb, stable propagation of cloned DNA and various modification strategies using RecA-mediated homologous recombination. However, genetic modifications using the BGM vector system have not been fully established, and this s...

  9. Characterization of Bacillus anthracis persistence in vivo.

    Directory of Open Access Journals (Sweden)

    Sarah A Jenkins

    Full Text Available Pulmonary exposure to Bacillus anthracis spores initiates inhalational anthrax, a life-threatening infection. It is known that dormant spores can be recovered from the lungs of infected animals months after the initial spore exposure. Consequently, a 60-day course antibiotic treatment is recommended for exposed individuals. However, there has been little information regarding details or mechanisms of spore persistence in vivo. In this study, we investigated spore persistence in a mouse model. The results indicated that weeks after intranasal inoculation with B. anthracis spores, substantial amounts of spores could be recovered from the mouse lung. Moreover, spores of B. anthracis were significantly better at persisting in the lung than spores of a non-pathogenic Bacillus subtilis strain. The majority of B. anthracis spores in the lung were tightly associated with the lung tissue, as they could not be readily removed by lavage. Immunofluorescence staining of lung sections showed that spores associated with the alveolar and airway epithelium. Confocal analysis indicated that some of the spores were inside epithelial cells. This was further confirmed by differential immunofluorescence staining of lung cells harvested from the infected lungs, suggesting that association with lung epithelial cells may provide an advantage to spore persistence in the lung. There was no or very mild inflammation in the infected lungs. Furthermore, spores were present in the lung tissue as single spores rather than in clusters. We also showed that the anthrax toxins did not play a role in persistence. Together, the results suggest that B. anthracis spores have special properties that promote their persistence in the lung, and that there may be multiple mechanisms contributing to spore persistence.

  10. Genome Sequence of Bacillus thuringiensis subsp. kurstaki Strain HD-1

    OpenAIRE

    Day, Michael; Ibrahim, Mohamed; Dyer, David; Bulla, Lee

    2014-01-01

    We report here the complete genome sequence of Bacillus thuringiensis subsp. kurstaki strain HD-1, which serves as the primary U.S. reference standard for all commercial insecticidal formulations of B. thuringiensis manufactured around the world.

  11. Effects of probiotic Bacillus species in aquaculture – An overview

    Directory of Open Access Journals (Sweden)

    Cristian-Teodor BURUIANĂ

    2014-12-01

    Full Text Available The ingestion of a large amount of certain types of beneficial bacteria can reduce the multiplication and development of pathogenic bacteria in the gut. A “probiotic” is a product that contains live microorganisms which positively influence the host intestinal microbiota by preventing the proliferation of pathogenic bacteria and promoting the growth and development of beneficial bacteria. Bacillus spp. are Gram-positive endospore-forming bacteria with beneficial effects in aquaculture industry. The dietary supplementation of Bacillus spp. in fish culture improved especially growth performance, immune response and the disease resistance of fish against pathogenic bacterial infections. The objective of the current paper is to review the recent published investigations reported in the scientific literature on the use of probiotic Bacillus spp. in aquaculture, focusing on their beneficial effects on the host. This review includes the main effects of Bacillus spp. administration in shrimp culture, carp culture, tilapia culture, and other fish culture.

  12. Two Genes Encoding Uracil Phosphoribosyltransferase Are Present in Bacillus subtilis

    DEFF Research Database (Denmark)

    Martinussen, Jan; Glaser, Philippe; Andersen, Paal S.;

    1995-01-01

    Uracil phosphoribosyltransferase (UPRTase) catalyzes the key reaction in the salvage of uracil in many microorganisms. Surprisingly, two genes encoding UPRTase activity were cloned from Bacillus subtilis by complementation of an Escherichia coli mutant. The genes were sequenced, and the putative...

  13. The Complete Genome Sequence of Bacillus thuringiensis AlHakam

    Energy Technology Data Exchange (ETDEWEB)

    Challacombe, Jean F.; Altherr, Michael R.; Xie, Gary; Bhotika,Smriti S.; Brown, Nancy; Bruce, David; Campbell, Connie S.; Campbell,Mary L.; Chen, Jin; Chertkov, Olga; Cleland, Cathy; Dimitrijevic, Mira; Doggett, Norman A.; Fawcett, John J.; Glavina, Tijana; Goodwin, Lynne A.; Green, Lance D.; Han, Cliff S.; Hill, Karen K.; Hitchcock, Penny; Jackson, Paul J.; Keim, Paul; Kewalramani, Avinash Ramesh; Longmire, Jon; Lucas, Susan; Malfatti, Stephanie; Martinez, Diego; McMurry, Kim; Meincke, Linda J.; Misra, Monica; Moseman, Bernice L.; Mundt, Mark; Munk,A. Christine; Okinaka, Richard T.; Parson-Quintana, B.; Reilly, LeePhilip; Richardson, Paul; Robinson, Donna L.; Rubin, Eddy; Saunders,Elizabeth; Tapia, Roxanne; Tesmer, Judith G.; Thayer, Nina; Thompson,Linda S.; Tice, Hope; Ticknor, Lawrence O.; Wills, Patti L.; Gilna, Paul; Brettin, Thomas S.

    2007-04-01

    Bacillus thuringiensis is an insect pathogen that is widelyused as a biopesticide (3). Here we report the finished, annotated genomesequence of B. thuringiensis Al Hakam, which was collected in Iraq by theUnited Nations Special Commission (2).

  14. BOOK REVIEW: BACILLUS THURINGIENSIS: A CORNERSTONE OF MODERN AGRICULTURE

    Science.gov (United States)

    Are you interested in the technical issues surrounding the use of Bacillus thuringiensis pesticidal traits as sprays and as plant incorporated protectants (transgenic crops)? Should the dimensions of human health, ecology, entomology, risk assessment, resistance management, and d...

  15. Antifungal activity of Bacillus sp. isolated from compost.

    Science.gov (United States)

    Czaczyk, K; Stachowiak, B; Trojanowska, K; Gulewicz, K

    2000-01-01

    Four strains of Bacillus isolated from lupine compost exhibited an antifungal activity against six plant fungal pathogens (Rhizoctonia solani, Bipolaris sorokiniana, Sclerotinia sclerotiorum, Trichothecium roseum, Fusarium solani, Fusarium oxysporum). It was significantly influenced by the composition of the cultivation media.

  16. Selection of Bacillus subtilis mutants impaired in ammonia assimilation.

    OpenAIRE

    Dean, D R; Aronson, A I

    1980-01-01

    The selection of Bacillus subtilis mutants capable of using D-histidine to fulfill a requirement for L-histidine resulted in mutants with either no glutamate synthase activity or increased amounts of an altered glutamine synthetase.

  17. Protein engineering of cyclodextrin glycosyltransferase from Bacillus circulans strain 251

    NARCIS (Netherlands)

    Penninga, Dirk

    1996-01-01

    An enormous diversity of molecular functions in living organisms is carried out by proteins. Our studies have focussed on the functional analysis of a starch-converting enzyme, cyclodextrin glycosyltransferase (CGTase) from Bacillus circulans strain 251. Zie: Summary

  18. The Adsorption Properties of Bacillus atrophaeus Spores on Single-Wall Carbon Nanotubes

    Directory of Open Access Journals (Sweden)

    P. Cortes

    2009-01-01

    Full Text Available An adsorption equilibrium and a kinetic study of Bacillus atrophaeus on Single-Wall Carbon Nanotubes (SWCNTs were here performed to provide the basis for developing biosensor devices for detecting threatening micro-organisms in water supply systems. B. atrophaeus spores and carbon nanotubes were subjected to a batch adsorption process to document their equilibria and kinetics. Here, commercial nanotubes were either studied as received or were acid-purified before adsorption experiments. The Bacillus spores appear to show higher affinity towards the purified nanotubes than to the as-received nanomaterial. The effective diffusivity of the spores onto the purified nanotubes was found to be approximately 30 percent higher than onto the as-received nanotubes. It seems that the removal of amorphous carbon from the as-received nanotubes through a purification process yielded an intimate nantoubes-spore interaction as revealed by transmission electron microscopy. Freundlich model successfully correlated the adsorption equilibrium data for the nanotubes-spore interaction. Transmission electron micrographs showed extensive contact between the Bacillus and the purified nanotubes, but the association appeared less intimate between the spores and the as-received nanotubes.

  19. Biocontrol of geosmin-producing Streptomyces spp. by two Bacillus strains from Chinese liquor.

    Science.gov (United States)

    Zhi, Yan; Wu, Qun; Du, Hai; Xu, Yan

    2016-08-16

    Streptomyces spp. producing geosmin have been regarded as the most frequent and serious microbial contamination causing earthy off-flavor in Chinese liquor. It is therefore necessary to control the Streptomyces community during liquor fermentation. Biological control, using the native microbiota present in liquor making, appears to be a better solution than chemical methods. The objective of this study was to isolate native microbiota antagonistic toward Streptomyces spp. and then to evaluate the possible action mode of the antagonists. Fourteen Bacillus strains isolated from different Daqu (the fermentation starter) showed antagonistic activity against Streptomyces sampsonii, which is one of the dominant geosmin producers. Bacillus subtilis 2-16 and Bacillus amyloliquefaciens 1-45 from Maotai Daqu significantly inhibited the growth of S. sampsonii by 57.8% and 84.3% respectively, and effectively prevented the geosmin production in the simulated fermentation experiments (inoculation ratio 1:1). To probe the biocontrol mode, the ability of strain 2-16 and 1-45 to produce antimicrobial metabolites and to reduce geosmin in the fermentation system was investigated. Antimicrobial substances were identified as lipopeptides by ultra-performance liquid chromatography tandem electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI/Q-TOF MS) and in vitro antibiotic assay. In addition, strains 2-16 and 1-45 were able to remove 45% and 15% of the geosmin respectively in the simulated solid-state fermentation. This study highlighted the potential of biocontrol, and how the use of native Bacillus species in Daqu could provide an eco-friendly method to prevent growth of Streptomyces spp. and geosmin contamination in Chinese liquor fermentation.

  20. Global microarray analysis of carbohydrate use in alkaliphilic hemicellulolytic bacterium Bacillus sp. N16-5.

    Directory of Open Access Journals (Sweden)

    Yajian Song

    Full Text Available The alkaliphilic hemicellulolytic bacterium Bacillus sp. N16-5 has a broad substrate spectrum and exhibits the capacity to utilize complex carbohydrates such as galactomannan, xylan, and pectin. In the monosaccharide mixture, sequential utilization by Bacillus sp. N16-5 was observed. Glucose appeared to be its preferential monosaccharide, followed by fructose, mannose, arabinose, xylose, and galactose. Global transcription profiles of the strain were determined separately for growth on six monosaccharides (glucose, fructose, mannose, galactose, arabinose, and xylose and four polysaccharides (galactomannan, xylan, pectin, and sodium carboxymethylcellulose using one-color microarrays. Numerous genes potentially related to polysaccharide degradation, sugar transport, and monosaccharide metabolism were found to respond to a specific substrate. Putative gene clusters for different carbohydrates were identified according to transcriptional patterns and genome annotation. Identification and analysis of these gene clusters contributed to pathway reconstruction for carbohydrate utilization in Bacillus sp. N16-5. Several genes encoding putative sugar transporters were highly expressed during growth on specific sugars, suggesting their functional roles. Two phosphoenolpyruvate-dependent phosphotransferase systems were identified as candidate transporters for mannose and fructose, and a major facilitator superfamily transporter was identified as a candidate transporter for arabinose and xylose. Five carbohydrate uptake transporter 1 family ATP-binding cassette transporters were predicted to participate in the uptake of hemicellulose and pectin degradation products. Collectively, microarray data improved the pathway reconstruction involved in carbohydrate utilization of Bacillus sp. N16-5 and revealed that the organism precisely regulates gene transcription in response to fluctuations in energy resources.

  1. Efficient transformation of Bacillus thuringiensis requires nonmethylated plasmid DNA.

    OpenAIRE

    Macaluso, A; Mettus, A M

    1991-01-01

    The transformation efficiency of Bacillus thuringiensis depends upon the source of plasmid DNA. DNA isolated from B. thuringiensis, Bacillus megaterium, or a Dam- Dcm- Escherichia coli strain efficiently transformed several B. thuringiensis strains, B. thuringiensis strains were grouped according to which B. thuringiensis backgrounds were suitable sources of DNA for transformation of other B. thuringiensis strains, suggesting that B. thuringiensis strains differ in DNA modification and restri...

  2. Regulation of cry Gene Expression in Bacillus thuringiensis

    OpenAIRE

    Chao Deng; Qi Peng; Fuping Song; Didier Lereclus

    2014-01-01

    Bacillus thuringiensis differs from the closely related Bacillus cereus group species by its ability to produce crystalline inclusions. The production of these crystals mainly results from the expression of the cry genes, from the stability of their transcripts and from the synthesis, accumulation and crystallization of large amounts of insecticidal Cry proteins. This process normally coincides with sporulation and is regulated by various factors operating at the transcriptional, post-transcr...

  3. Natural Dissemination of Bacillus anthracis Spores in Northern Canada

    OpenAIRE

    Dragon, D C; Bader, D. E.; Mitchell, J.; Woollen, N.

    2005-01-01

    Soil samples were collected from around fresh and year-old bison carcasses and areas not associated with known carcasses in Wood Buffalo National Park during an active anthrax outbreak in the summer of 2001. Sample selection with a grid provided the most complete coverage of a site. Soil samples were screened for viable Bacillus anthracis spores via selective culture, phenotypic analysis, and PCR. Bacillus anthracis spores were isolated from 28.4% of the samples. The highest concentrations of...

  4. Genetic analysis of petrobactin transport in Bacillus anthracis

    OpenAIRE

    Carlson, Paul E.; Dixon, Shandee D.; Janes, Brian K.; Carr, Katherine A.; Nusca, Tyler D.; Anderson, Erica C.; Keene, Sarra E.; Sherman, David H.; Hanna, Philip C.

    2010-01-01

    Iron acquisition mechanisms play an important role in the pathogenesis of many infectious microbes. In Bacillus anthracis, the siderophore petrobactin is required for both growth in iron depleted conditions and for full virulence of the bacterium. Here we demonstrate the roles of two putative petrobactin binding proteins FatB and FpuA (encoded by GBAA5330 and GBAA4766, respectively) in Bacillus anthracis iron acquisition and pathogenesis. Markerless deletion mutants were created using allelic...

  5. Expression of UGA-Containing Mycoplasma Genes in Bacillus subtilis

    OpenAIRE

    Kannan, T. R.; Baseman, Joel B.

    2000-01-01

    We used Bacillus subtilis to express UGA-containing Mycoplasma genes encoding the P30 adhesin (one UGA) of Mycoplasma pneumoniae and methionine sulfoxide reductase (two UGAs) of Mycoplasma genitalium. Due to natural UGA suppression, these Mycoplasma genes were expressed as full-length protein products, but at relatively low efficiency, in recombinant wild-type Bacillus. The B. subtilis-expressed Mycoplasma proteins appeared as single bands and not as multiple bands compared to expression in r...

  6. Laser-induced speckle scatter patterns in Bacillus colonies

    Directory of Open Access Journals (Sweden)

    Huisung eKim

    2014-10-01

    Full Text Available Label-free bacterial colony phenotyping technology called BARDOT (BActerial Rapid Detection using Optical scattering Technology provided successful classification of several different bacteria at the genus, species, and serovar level. Recent experiments with colonies of Bacillus species provided strikingly different characteristics of elastic light scatter (ELS patterns, which were comprised of random speckles compared to other bacteria, which are dominated by concentric rings and spokes. Since this laser-based optical sensor interrogates the whole volume of the colony, 3-D information of micro- and macro-structures are all encoded in the far-field scatter patterns. Here, we present a theoretical model explaining the underlying mechanism of the speckle formation by the colonies from Bacillus species. Except for Bacillus polymyxa, all Bacillus spp. produced random bright spots on the imaging plane, which presumably dependent on the cellular and molecular organization and content within the colony. Our scatter model-based analysis revealed that colony spread resulting in variable surface roughness can modify the wavefront of the scatter field. As the center diameter of the Bacillus spp. colony grew from 500 μm to 900 μm, average speckles area decreased 2-fold and the number of small speckles increased 7-fold. In conclusion, as Bacillus colony grows, the average speckle size in the scatter pattern decreases and the number of smaller speckle increases due to the swarming growth characteristics of bacteria within the colony.

  7. Impact of Gastrointestinal Bacillus anthracis Infection on Hepatic B Cells

    Directory of Open Access Journals (Sweden)

    Natacha Colliou

    2015-09-01

    Full Text Available Ingestion of Bacillus anthracis results in rapid gastrointestinal (GI infection, known as GI anthrax. We previously showed that during GI anthrax, there is swift deterioration of intestinal barrier function leading to translocation of gut-associated bacteria into systemic circulation. Additionally, we described dysfunction in colonic B cells. In concordance with our previous studies, here, we report early migration of the Sterne strain of B. anthracis along with other gut-resident bacteria into the infected murine liver. Additionally, despite a global decrease in the B cell population, we observed an increase in both B-1a and marginal zone (MZ-like B cells. Both of these cell types are capable of producing immunoglobulins against common pathogens and commensals, which act as a general antibody barrier before an antigen-specific antibody response. Accumulation of these cells in the liver was associated with an increase in chemokine expression. These data suggest that the presence of Sterne and other commensals in the liver trigger migration of MZ-like B cells from the spleen to the liver to neutralize systemic spread. Further research is required to evaluate the possible cause of their failure to clear the infection within the liver, including the potential role of dysfunctional mitogen-activated protein kinase (MAPK signaling.

  8. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity.

    Science.gov (United States)

    Warda, Alicja K; Siezen, Roland J; Boekhorst, Jos; Wells-Bennik, Marjon H J; de Jong, Anne; Kuipers, Oscar P; Nierop Groot, Masja N; Abee, Tjakko

    2016-01-01

    We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with 11 additional B. cereus reference genomes to provide an overview of the different types of carbohydrate transporters and utilization systems found in B. cereus strains. The combined application of API tests, defined growth media experiments and comparative genomics enabled us to link the carbohydrate utilisation capacity of 22 B. cereus strains with their genome content and in some cases to the panC phylogenetic grouping. A core set of carbohydrates including glucose, fructose, maltose, trehalose, N-acetyl-glucosamine, and ribose could be used by all strains, whereas utilisation of other carbohydrates like xylose, galactose, and lactose, and typical host-derived carbohydrates such as fucose, mannose, N-acetyl-galactosamine and inositol is limited to a subset of strains. Finally, the roles of selected carbohydrate transporters and utilisation systems in specific niches such as soil, foods and the human host are discussed. PMID:27272929

  9. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity

    Science.gov (United States)

    Warda, Alicja K.; Siezen, Roland J.; Boekhorst, Jos; Wells-Bennik, Marjon H. J.; de Jong, Anne; Kuipers, Oscar P.; Nierop Groot, Masja N.; Abee, Tjakko

    2016-01-01

    We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with 11 additional B. cereus reference genomes to provide an overview of the different types of carbohydrate transporters and utilization systems found in B. cereus strains. The combined application of API tests, defined growth media experiments and comparative genomics enabled us to link the carbohydrate utilisation capacity of 22 B. cereus strains with their genome content and in some cases to the panC phylogenetic grouping. A core set of carbohydrates including glucose, fructose, maltose, trehalose, N-acetyl-glucosamine, and ribose could be used by all strains, whereas utilisation of other carbohydrates like xylose, galactose, and lactose, and typical host-derived carbohydrates such as fucose, mannose, N-acetyl-galactosamine and inositol is limited to a subset of strains. Finally, the roles of selected carbohydrate transporters and utilisation systems in specific niches such as soil, foods and the human host are discussed. PMID:27272929

  10. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity.

    Directory of Open Access Journals (Sweden)

    Alicja K Warda

    Full Text Available We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with 11 additional B. cereus reference genomes to provide an overview of the different types of carbohydrate transporters and utilization systems found in B. cereus strains. The combined application of API tests, defined growth media experiments and comparative genomics enabled us to link the carbohydrate utilisation capacity of 22 B. cereus strains with their genome content and in some cases to the panC phylogenetic grouping. A core set of carbohydrates including glucose, fructose, maltose, trehalose, N-acetyl-glucosamine, and ribose could be used by all strains, whereas utilisation of other carbohydrates like xylose, galactose, and lactose, and typical host-derived carbohydrates such as fucose, mannose, N-acetyl-galactosamine and inositol is limited to a subset of strains. Finally, the roles of selected carbohydrate transporters and utilisation systems in specific niches such as soil, foods and the human host are discussed.

  11. A Low Dimensional Approximation For Competence In Bacillus Subtilis.

    Science.gov (United States)

    Nguyen, An; Prugel-Bennett, Adam; Dasmahapatra, Srinandan

    2016-01-01

    The behaviour of a high dimensional stochastic system described by a chemical master equation (CME) depends on many parameters, rendering explicit simulation an inefficient method for exploring the properties of such models. Capturing their behaviour by low-dimensional models makes analysis of system behaviour tractable. In this paper, we present low dimensional models for the noise-induced excitable dynamics in Bacillus subtilis, whereby a key protein ComK, which drives a complex chain of reactions leading to bacterial competence, gets expressed rapidly in large quantities (competent state) before subsiding to low levels of expression (vegetative state). These rapid reactions suggest the application of an adiabatic approximation of the dynamics of the regulatory model that, however, lead to competence durations that are incorrect by a factor of 2. We apply a modified version of an iterative functional procedure that faithfully approximates the time-course of the trajectories in terms of a two-dimensional model involving proteins ComK and ComS. Furthermore, in order to describe the bimodal bivariate marginal probability distribution obtained from the Gillespie simulations of the CME, we introduce a tunable multiplicative noise term in a two-dimensional Langevin model whose stationary state is described by the time-independent solution of the corresponding Fokker-Planck equation. PMID:27045827

  12. Transfer of the toxin protein genes of Bacillus sphaericus into Bacillus thuringiensis subsp. israelensis and their expression.

    OpenAIRE

    Bourgouin, C.; Delécluse, A; La Torre, F.; Szulmajster, J

    1990-01-01

    The genes encoding the toxic determinants of Bacillus sphaericus have been expressed in a nontoxic and a toxic strain of Bacillus thuringiensis subsp. israelensis. In both cases, the B. sphaericus toxin proteins were produced at a high level during sporulation of B. thuringiensis and accumulated as crystalline structures. B. thuringiensis transformants expressing B. sphaericus and B. thuringiensis subsp. israelensis toxins did not show a significant enhancement of toxicity against Aedes aegyp...

  13. Ground Anthrax Bacillus Refined Isolation (GABRI) method for analyzing environmental samples with low levels of Bacillus anthracis contamination

    OpenAIRE

    Fasanella, Antonio; Di Taranto, Pietro; Garofolo, Giuliano; Colao, Valeriana; Marino, Leonardo; Buonavoglia, Domenico; Pedarra, Carmine; Adone, Rosanna; Hugh-Jones, Martin

    2013-01-01

    Background In this work are reported the results of a qualitative analytical method capable of detecting Bacillus anthracis spores when they are present in very low concentration in the soil. The Ground Anthrax Bacillus Refined Isolation (GABRI) method, assessed in our laboratory, was compared with the classic method. The comparison involved artificially anthrax-contaminated soil samples (500 spores/7.5 grams soil) and naturally contaminated soil samples collected in Bangladesh during a field...

  14. Bacillus cereus in Brazilian Ultra High Temperature milk Bacillus cereus em leite UHT brasileiro

    Directory of Open Access Journals (Sweden)

    Cristiana de Paula Pacheco-Sanchez

    2007-01-01

    Full Text Available Brazilian Ultra High Temperature (UHT milk consumption has increased during the last decade from 187 to 4,200 million liters. In the continuous UHT process, milk is submitted for 2-4 s to 130-150ºC, in a continuous flow system with immediate refrigeration and aseptical packing in hermetic packages. This research had the purpose to verify the incidence of B. cereus species from the B. cereus group, in UHT milk. In 1998 high indexes of these organisms were reported, reaching 34.14% of the analyzed samples. Beyond this fact, there was the need to establish methods and processes adjusted for correct identification of B. cereus. Thus, commercial sterility tests of 6,500 UHT milk packages were investigated in two assays, after ten days incubation at 37ºC and 7ºC to germinate all possible spores and/or to recuperate injured vegetative cells followed by pH measurement. Samples (1,300 packages each from five Brazilian UHT plants of whole UHT milk processed by direct steam injection, packaged in carton were investigated for the presence of Bacillus cereus through phenotypic and genetic (PCR tests. Values of pH were different for the samples, ranging between 6.57 and 6.73. After storage of the samples, only four packages with pH measurement below the lower limit of 6.5 were found and analyzed for the presence of B. cereus. This organism was not detected in any of the samples indicating that the five Brazilian UHT milk processors control pathogenic microorganisms and it can be said that the consumption of UHT milk does not present safety problems to consumers. Fourier Transform Infrared Spectroscopy (FTIR and PCR tests were efficient and must be adopted to confirm the biochemical series for B. cereus.O consumo de leite ultra-alta temperatura (UHT brasileiro aumentou, durante a última década, de 187 milhões de litros para 4,200 milhões de litros. No processo contínuo de leite UHT o leite é submetido por 2-4 seg a 130-150ºC, em sistemas de

  15. Enterotoxigenic Bacillus spp. DNA fingerprint revealed in naturally contaminated nonfat dry milk powder using rep-PCR.

    Science.gov (United States)

    Cooper, Robin M; McKillip, John L

    2006-01-01

    Dry milk powders and functional ingredients frequently contain high levels of viable bacterial spores, some of which may result in growth of toxigenic Bacillus spp. in reconstituted and temperature-abused foods. Samples from nonfat dry milk (NFDM), infant milk formula (IMF), coffee creamer, lecithin, and cocoa powder were subjected to a short heat treatment followed by enrichment in tryptone phosphate glucose yeast extract (TPGY) broth at 32 degrees C for 12-25 hours to obtain cell densities of 10(6) CFU ml(-1). DNA was extracted using a modification of established protocol, leading to the development of an optimized method for each food system. Purified DNA was amplified by rep-PCR using extragenic sequence-targeting primers and optimized for each food. PCR fingerprints from each food were analyzed electrophoretically for banding patterns earlier correlated to that of enterotoxigenic Bacillus spp. and Bacillus cereus positive control DNA fingerprints. Reverse passive latex agglutination (RPLA) and Bacillus Diarrhoeal Enterotoxin Enzyme Linked Immunosorbent Assay (Tecra Diagnostics) confirmed the presence of HBL and NHE enterotoxin production in NFDM, Coffee creamer, infant milk formula, and two lecithin samples but not in cocoa powder. These results demonstrate the utility of rep-PCR not only as a tool for bacterial genotyping, but a unique means of quality control and hygiene monitoring in food microbiology.

  16. An inducible recA expression Bacillus subtilis genome vector for stable manipulation of large DNA fragments

    OpenAIRE

    Ogawa, Takafumi; Iwata, Tetsuo; Kaneko, Shinya; Itaya, Mitsuhiro; Hirota, Junji

    2015-01-01

    Background The Bacillus subtilis genome (BGM) vector is a novel cloning system based on the natural competence that enables B. subtilis to import extracellular DNA fragments into the cell and incorporate the recombinogenic DNA into the genome vector by homologous recombination. The BGM vector system has several attractive properties, such as a megabase cloning capacity, stable propagation of cloned DNA inserts, and various modification strategies using RecA-mediated homologous recombination. ...

  17. Bacillus sp.Treating Wastewater Containing Antimony%Bacillus sp.处理含锑废水试验研究

    Institute of Scientific and Technical Information of China (English)

    李小娇; 成应向; 龚道新; 向仁军; 王强强

    2012-01-01

    利用某芽孢杆菌属微生物(Bacillus sp.)对锑矿选矿废水进行了处理.研究微生物的接种量、作用时间、温度、体系pH值等对废水中Sb的去除效果的影响.结果表明:作用时间4d、微生物接种量为5%、处理体系pH为2、最佳处理体系温度为30℃时,效果最佳,对废水中Sb的去除率达到99.75%,处理后废水中Sb的浓度由122.21 mg/L降低至0.30 mg/L,出水Sb浓度低于湖南省地方标准排放限值0.50 mg/L.%Antimony ore dressing wastewater was treated by using a certain Bacillus microorganism (Bacillus sp.). Sb removal efficiency in mineral processing wastewater was affected by microbial inoculums size, treating time, temperature and the system pH value. Results showed that the best removal efficiency of Sb in mineral processing wastewater could reach 99.75% in 4d under the optimum conditions of 30 ℃, microbial inoculated quantity 5% and pH value 2.0. After the treatment, the concentration of Sb in wastewater was reduced from 122.21 mg/L to 0.30 mg/L, which was lower than local industrial wastewater discharge standard of 0.50 mg/L.

  18. Fluorimetric Detection of a Bacillus stearothermophilus Spore-Bound Enzyme, α-d-Glucosidase, for Rapid Indication of Flash Sterilization Failure

    OpenAIRE

    Vesley, Donald; Langholz, Ann C.; Rohlfing, Stephen R.; Foltz, William E.

    1992-01-01

    A biological indicator based on fluorimetric detection within 60 min of a Bacillus stearothermophilus spore-bound enzyme, α-d-glucosidase, has been developed. Results indicate that the enzyme survived slightly longer than spores observed after 24 h of incubation. The new system shows promise for evaluating flash sterilization cycles within 60 min compared with conventional 24-h systems.

  19. Functional analysis of 14 genes that constitute the purine catabolic pathway in Bacillus subtilis and evidence for a novel regulon controlled by the PucR transcription activator

    DEFF Research Database (Denmark)

    Schultz, Anna Charlotte; Nygaard, P.; Saxild, Hans Henrik

    2001-01-01

    The soil bacterium Bacillus subtilis has developed a highly controlled system for the utilization of a diverse array of low molecular-weight compounds as a nitrogen source when the preferred nitrogen sources, e.g., glutamate plus ammonia, are exhausted. We have identified such a system for the ut...

  20. Glyconanobiotics: Novel carbohydrated nanoparticle antibiotics for MRSA and Bacillus anthracis.

    Science.gov (United States)

    Abeylath, Sampath C; Turos, Edward; Dickey, Sonja; Lim, Daniel V

    2008-03-01

    This report describes the synthesis and evaluation of glycosylated polyacrylate nanoparticles that have covalently-bound antibiotics within their framework. The requisite glycosylated drug monomers were prepared from one of three known antibiotics, an N-sec-butylthio beta-lactam, ciprofloxacin, and a penicillin, by acylation with 3-O-acryloyl-1,2-O-isopropylidene-5,6 bis((chlorosuccinyl)oxy)-d-glucofuranose (7) or 6-O-acetyl-3-O-acryloyl-1,2-O-isopropylidene-5-(chlorosuccinyl)oxy-alpha-d-glucofuranose (10). These acrylated monomers were subjected to emulsion polymerization in a 7:3 (w:w) mixture of butyl acrylate-styrene in the presence of sodium dodecyl sulfate as surfactant (3 weight %) and potassium persulfate as a radical initiator (1 weight %). The resulting nanoparticle emulsions were characterized by dynamic light scattering and found to have similar diameters ( approximately 40 nm) and size distributions to those of our previously studied systems. Microbiological testing showed that the N-sec-butylthio beta-lactam and ciprofloxacin nanoparticles both have powerful in vitro activities against methicillin-resistant Staphylococcus aureus and Bacillus anthracis, while the penicillin-bound nanoparticles have no antimicrobial activity. This indicates the need for matching a suitable antibiotic with the nanoparticle carrier. Overall, the study shows that even relatively large, polar acrylate monomers (MW>1000 amu) can be efficiently incorporated into the nanoparticle matrix by emulsion polymerization, providing opportunities for further advances in nanomedicine. PMID:18063370

  1. The Pore-Forming Haemolysins of Bacillus Cereus: A Review

    Directory of Open Access Journals (Sweden)

    Vincent Sanchis

    2013-06-01

    Full Text Available The Bacillus cereus sensu lato group contains diverse Gram-positive spore-forming bacteria that can cause gastrointestinal diseases and severe eye infections in humans. They have also been incriminated in a multitude of other severe, and frequently fatal, clinical infections, such as osteomyelitis, septicaemia, pneumonia, liver abscess and meningitis, particularly in immuno-compromised patients and preterm neonates. The pathogenic properties of this organism are mediated by the synergistic effects of a number of virulence products that promote intestinal cell destruction and/or resistance to the host immune system. This review focuses on the pore-forming haemolysins produced by B. cereus: haemolysin I (cereolysin O, haemolysin II, haemolysin III and haemolysin IV (CytK. Haemolysin I belongs to the cholesterol-dependent cytolysin (CDC family whose best known members are listeriolysin O and perfringolysin O, produced by L. monocytogenes and C. perfringens respectively. HlyII and CytK are oligomeric ß-barrel pore-forming toxins related to the α-toxin of S. aureus or the ß-toxin of C. perfringens. The structure of haemolysin III, the least characterized haemolytic toxin from the B. cereus, group has not yet been determined.

  2. Setting risk-informed environmental standards for Bacillus anthracis spores.

    Science.gov (United States)

    Hong, Tao; Gurian, Patrick L; Ward, Nicholas F Dudley

    2010-10-01

    In many cases, human health risk from biological agents is associated with aerosol exposures. Because air concentrations decline rapidly after a release, it may be necessary to use concentrations found in other environmental media to infer future or past aerosol exposures. This article presents an approach for linking environmental concentrations of Bacillus. anthracis (B. anthracis) spores on walls, floors, ventilation system filters, and in human nasal passages with human health risk from exposure to B. anthracis spores. This approach is then used to calculate example values of risk-informed concentration standards for both retrospective risk mitigation (e.g., prophylactic antibiotics) and prospective risk mitigation (e.g., environmental clean up and reoccupancy). A large number of assumptions are required to calculate these values, and the resulting values have large uncertainties associated with them. The values calculated here suggest that documenting compliance with risks in the range of 10(-4) to 10(-6) would be challenging for small diameter (respirable) spore particles. For less stringent risk targets and for releases of larger diameter particles (which are less respirable and hence less hazardous), environmental sampling would be more promising.

  3. Quantum dot incorporated Bacillus spore as nanosensor for viral infection.

    Science.gov (United States)

    Zhang, Xinya; Zhou, Qian; Shen, Zhongfeng; Li, Zheng; Fei, Ruihua; Ji, Eoon Hye; Hu, Shen; Hu, Yonggang

    2015-12-15

    In this paper, we report a high-throughput biological method to prepare spore-based monodisperse microparticles (SMMs) and then form the nanocomposites of CdTe quantum dot (QD)-loaded SMMs by utilizing the endogenous functional groups from Bacillus spores. The SMMs and QD-incorporated spore microspheres (QDSMs) were characterized by using transmission electron microscopy, high-resolution transmission electron microscopy, fluorescence microscopy, fluorescence and UV-visible absorption spectroscopy, zeta potential analysis, Fourier-transform infrared spectroscopy, potentiometric titrations, X-ray photo-electron spectroscopy. The thermodynamics of QD/SMM interaction and antigen/QDSM interaction was also investigated by isothermal titration microcalorimetry (ITC). Fluorescent QDSMs coded either with a single luminescence color or with multiple colors of controlled emission intensity ratios were obtained. Green QDSMs were used as a model system to detect porcine parvovirus antibody in swine sera via flow cytometry, and the results demonstrated a great potential of QDSMs in high-throughput immunoassays. Due to the advantages such as simplicity, low cost, high throughput and eco-friendliness, our developed platform may find wide applications in disease detection, food safety evaluation and environmental assessment.

  4. Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance.

    Science.gov (United States)

    Badran, Ahmed H; Guzov, Victor M; Huai, Qing; Kemp, Melissa M; Vishwanath, Prashanth; Kain, Wendy; Nance, Autumn M; Evdokimov, Artem; Moshiri, Farhad; Turner, Keith H; Wang, Ping; Malvar, Thomas; Liu, David R

    2016-05-01

    The Bacillus thuringiensis δ-endotoxins (Bt toxins) are widely used insecticidal proteins in engineered crops that provide agricultural, economic, and environmental benefits. The development of insect resistance to Bt toxins endangers their long-term effectiveness. Here we have developed a phage-assisted continuous evolution selection that rapidly evolves high-affinity protein-protein interactions, and applied this system to evolve variants of the Bt toxin Cry1Ac that bind a cadherin-like receptor from the insect pest Trichoplusia ni (TnCAD) that is not natively bound by wild-type Cry1Ac. The resulting evolved Cry1Ac variants bind TnCAD with high affinity (dissociation constant Kd = 11-41 nM), kill TnCAD-expressing insect cells that are not susceptible to wild-type Cry1Ac, and kill Cry1Ac-resistant T. ni insects up to 335-fold more potently than wild-type Cry1Ac. Our findings establish that the evolution of Bt toxins with novel insect cell receptor affinity can overcome insect Bt toxin resistance and confer lethality approaching that of the wild-type Bt toxin against non-resistant insects. PMID:27120167

  5. Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance.

    Science.gov (United States)

    Badran, Ahmed H; Guzov, Victor M; Huai, Qing; Kemp, Melissa M; Vishwanath, Prashanth; Kain, Wendy; Nance, Autumn M; Evdokimov, Artem; Moshiri, Farhad; Turner, Keith H; Wang, Ping; Malvar, Thomas; Liu, David R

    2016-05-01

    The Bacillus thuringiensis δ-endotoxins (Bt toxins) are widely used insecticidal proteins in engineered crops that provide agricultural, economic, and environmental benefits. The development of insect resistance to Bt toxins endangers their long-term effectiveness. Here we have developed a phage-assisted continuous evolution selection that rapidly evolves high-affinity protein-protein interactions, and applied this system to evolve variants of the Bt toxin Cry1Ac that bind a cadherin-like receptor from the insect pest Trichoplusia ni (TnCAD) that is not natively bound by wild-type Cry1Ac. The resulting evolved Cry1Ac variants bind TnCAD with high affinity (dissociation constant Kd = 11-41 nM), kill TnCAD-expressing insect cells that are not susceptible to wild-type Cry1Ac, and kill Cry1Ac-resistant T. ni insects up to 335-fold more potently than wild-type Cry1Ac. Our findings establish that the evolution of Bt toxins with novel insect cell receptor affinity can overcome insect Bt toxin resistance and confer lethality approaching that of the wild-type Bt toxin against non-resistant insects.

  6. Adhesion of Spores of Bacillus thuringiensis on a Planar Surface

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Eunhyea [Georgia Institute of Technology; Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Lee, Ida [University of Tennessee, Knoxville (UTK); Joy, David Charles [ORNL; Palumbo, Anthony Vito [ORNL; Tsouris, Costas [ORNL

    2010-01-01

    Adhesion of spores of Bacillus thuringiensis (Bt) and spherical silica particles on surfaces was experimentally and theoretically investigated in this study. Topography analysis via atomic force microscopy (AFM) and electron microscopy indicates that Bt spores are rod shaped, {approx}1.3 {mu}m in length and {approx}0.8 {mu}m in diameter. The adhesion force of Bt spores and silica particles on gold-coated glass was measured at various relative humidity (RH) levels by AFM. It was expected that the adhesion force would vary with RH because the individual force components contributing to the adhesion force depend on RH. The adhesion force between a particle and a planar surface in atmospheric environments was modeled as the contribution of three major force components: capillary, van der Waals, and electrostatic interaction forces. Adhesion force measurements for Bt spore (silica particle) and the gold surface system were comparable with calculations. Modeling results show that there is a critical RH value, which depends on the hydrophobicity of the materials involved, below which the water meniscus does not form and the contribution of the capillary force is zero. As RH increases, the van der Waals force decreases while the capillary force increases to a maximum value.

  7. Degradation of ochratoxin A by Bacillus amyloliquefaciens ASAG1.

    Science.gov (United States)

    Chang, Xiaojiao; Wu, Zidan; Wu, Songling; Dai, Yanshi; Sun, Changpo

    2015-01-01

    Ochratoxin A (OTA) is widely found in food and feed products as a mycotoxin contaminant. It is produced by Penicillium species and several Aspergillus species. The identification OTA detoxification microorganisms is believed to be the best approach for decontamination. In this study, we isolated ASAG1, a bacterium with the ability to degrade OTA effectively, from grain depot-stored maize. A 16S rDNA sequencing approach was used to identify this strain as Bacillus amyloliquefaciens ASAG1. The degradation of OTA was detected in both medium and cell-free extracts after incubation with a culture of B. amyloliquefaciens ASAG1 cells. Subsequently, a hydrolysed enzyme (carboxypeptidase) related to the enzymatic conversion of OTA was cloned from the B. amyloliquefaciens ASAG1 genome. Using the Escherichia coli Expression System, we successfully expressed and purified this carboxypeptidase. When this enzyme was incubated with the engineered recombinant E. coli cells, the concentration of OTA was dramatically degraded. Our data therefore indicate that the carboxypeptidase produced by B. amyloliquefaciens ASAG1 is likely responsible for the biodegradation of OTA.

  8. Degradation of ochratoxin A by Bacillus amyloliquefaciens ASAG1.

    Science.gov (United States)

    Chang, Xiaojiao; Wu, Zidan; Wu, Songling; Dai, Yanshi; Sun, Changpo

    2015-01-01

    Ochratoxin A (OTA) is widely found in food and feed products as a mycotoxin contaminant. It is produced by Penicillium species and several Aspergillus species. The identification OTA detoxification microorganisms is believed to be the best approach for decontamination. In this study, we isolated ASAG1, a bacterium with the ability to degrade OTA effectively, from grain depot-stored maize. A 16S rDNA sequencing approach was used to identify this strain as Bacillus amyloliquefaciens ASAG1. The degradation of OTA was detected in both medium and cell-free extracts after incubation with a culture of B. amyloliquefaciens ASAG1 cells. Subsequently, a hydrolysed enzyme (carboxypeptidase) related to the enzymatic conversion of OTA was cloned from the B. amyloliquefaciens ASAG1 genome. Using the Escherichia coli Expression System, we successfully expressed and purified this carboxypeptidase. When this enzyme was incubated with the engineered recombinant E. coli cells, the concentration of OTA was dramatically degraded. Our data therefore indicate that the carboxypeptidase produced by B. amyloliquefaciens ASAG1 is likely responsible for the biodegradation of OTA. PMID:25517039

  9. Hepatitis in Disseminated Bacillus Calmette-Guérin Infection

    Directory of Open Access Journals (Sweden)

    Markus U Göttke

    2000-01-01

    Full Text Available Local immunotherapy with an attenuated live strain of Mycobacterium bovis, bacillus Calmette-Guérin (BCG, is an effective and frequently used treatment for in situ transitional cell carcinoma (TCC of the bladder. Success rates are high, and serious side effects are infrequent but can affect every organ system. A 79-year-old patient with recently diagnosed TCC who was treated with intravesical BCG for a recurrence after initial surgical treatment is reported. After unsuccessful attempts at bladder catheterization with the creation of a false passage for his third treatment, BCG was instilled via a suprapubic catheter the same day and again a week later. Two weeks after the third BCG instillation, the patient presented with profound lethargy and weakness to the point of not being able to get up out of a chair. He was febrile, anorexic, icteric and had hepatosplenomegaly. Disseminated BCG infection was suspected on the basis of history, clinical examination and a liver biopsy that showed noncaseating granulomatous hepatitis. Empirical treatment was started with antituberculous combination therapy. A short course of an oral corticosteroid was given. Clinical improvement was marked and sustained so that the patient could be discharged home for the full six-month course of his treatment. Disseminated BCG infection with granulomatous hepatitis can be severe and life-threatening in cases where a large intravascular inoculum of BCG may have been given inadvertently.

  10. Comparative sequence analyses on the 16S rRNA (rDNA) of Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus and proposal for creation of a new genus, Alicyclobacillus gen. nov

    Science.gov (United States)

    Wisotzkey, J. D.; Jurtshuk, P. Jr; Fox, G. E.; Deinhard, G.; Poralla, K.

    1992-01-01

    Comparative 16S rRNA (rDNA) sequence analyses performed on the thermophilic Bacillus species Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus revealed that these organisms are sufficiently different from the traditional Bacillus species to warrant reclassification in a new genus, Alicyclobacillus gen. nov. An analysis of 16S rRNA sequences established that these three thermoacidophiles cluster in a group that differs markedly from both the obligately thermophilic organisms Bacillus stearothermophilus and the facultatively thermophilic organism Bacillus coagulans, as well as many other common mesophilic and thermophilic Bacillus species. The thermoacidophilic Bacillus species B. acidocaldarius, B. acidoterrestris, and B. cycloheptanicus also are unique in that they possess omega-alicylic fatty acid as the major natural membranous lipid component, which is a rare phenotype that has not been found in any other Bacillus species characterized to date. This phenotype, along with the 16S rRNA sequence data, suggests that these thermoacidophiles are biochemically and genetically unique and supports the proposal that they should be reclassified in the new genus Alicyclobacillus.

  11. Multifactorial Resistance of Bacillus subtilis Spores to High-Energy Proton Radiation: Role of Spore Structural Components and the Homologous Recombination and Non-Homologous End Joining DNA Repair Pathways

    OpenAIRE

    Moeller, Ralf; Reitz, Günther; Li, Zuofeng; Klein, Stuart; Nicholson, Wayne L.

    2012-01-01

    The space environment contains high-energy charged particles (e.g., protons, neutrons, electrons, α-particles, heavy ions) emitted by the Sun and galactic sources or trapped in the radiation belts. Protons constitute the majority (87%) of high-energy charged particles. Spores of Bacillus species are one of the model systems used for astro- and radiobiological studies. In this study, spores of different Bacillus subtilis strains were used to study the effects of high energetic proton irradiati...

  12. Inactivation of Bacillus atrophaeus by OH radicals

    Science.gov (United States)

    Ono, Ryo; Yonetamari, Kenta; Tokumitsu, Yusuke; Yonemori, Seiya; Yasuda, Hachiro; Mizuno, Akira

    2016-08-01

    The inactivation of Bacillus atrophaeus by OH radicals is measured. This study aims to evaluate the bactericidal effects of OH radicals produced by atmospheric-pressure nonthermal plasma widely used for plasma medicine; however, in this study, OH radicals are produced by vacuum ultraviolet (VUV) photolysis of water vapor instead of plasma to allow the production of OH radicals with almost no other reactive species. A 172 nm VUV light from a Xe2 excimer lamp irradiates a He–H2O mixture flowing in a quartz tube to photodissociate H2O to produce OH, H, O, HO2, H2O2, and O3. The produced reactive oxygen species (ROS) flow out of the quartz tube nozzle to the bacteria on an agar plate and cause inactivation. The inactivation by OH radicals among the six ROS is observed by properly setting the experimental conditions with the help of simulations calculating the ROS densities. A 30 s treatment with approximately 0.1 ppm OH radicals causes visible inactivation.

  13. Bacillus anthracis diversity in Kruger National Park.

    Science.gov (United States)

    Smith, K L; DeVos, V; Bryden, H; Price, L B; Hugh-Jones, M E; Keim, P

    2000-10-01

    The Kruger National Park (KNP), South Africa, has a recorded history of periodic anthrax epidemics causing widespread disease among wild animals. Bacillus anthracis is the causative agent of anthrax, a disease primarily affecting ungulate herbivores. Worldwide there is little diversity among B. anthracis isolates, but examination of variable-number tandem repeat (VNTR) loci has identified six major clones, with the most dissimilar types split into the A and B branches. Both the A and B types are found in southern Africa, giving this region the greatest genetic diversity of B. anthracis worldwide. Consequently, southern Africa has been hypothesized to be the geographic origin of B. anthracis. In this study, we identify the genotypic types of 98 KNP B. anthracis isolates using multiple-locus VNTR analysis. Two major types are evident, the A branch and the B branch. The spatial and temporal distribution of the different genotypes indicates that anthrax epidemic foci are independent, though correlated through environmental cues. Kruger B isolates were found on significantly higher-calcium and higher-pH soils than were Kruger type A. This relationship between genotype and soil chemistry may be due to adaptive differences among divergent anthrax strains. While this association may be simply fortuitous, adaptation of A types to diverse environmental conditions is consistent with their greater geographic dispersal and genetic dissimilarity.

  14. A pangenomic study of Bacillus thuringiensis

    Institute of Scientific and Technical Information of China (English)

    Yongjun Fang; Songnian Hu; Jie Zhang; Ibrahim A1-Mssallem; Jun Yu; Zhaolong Li; Jiucheng Liu; Changlong Shu; Xumin Wang; Xiaowei Zhang; Xiaoguang Yu; Duojun Zhao; Guiming Liu

    2011-01-01

    Bacillus thuringiensis (B.thuringiensis) is a soil-dwelling Gram-positive bacterium and its plasmid-encoded toxins (Cry) are commonly used as biological alternatives to pesticides.In a pangenomic study,we sequenced seven B.thuringiensis isolates in both high coverage and base quality using the next-generation sequencing platform.The B.thuringiensis pangenome was extrapolated to have 4196 core genes and an asymptotic value of 558 unique genes when a new genome is added.Compared to the pangenomes of its closely related species of the same genus,B.thuringiensis pangenome shows an open characteristic,similar to B.cereus but not to B.anthracis; the latter has a closed pangenome.We also found extensive divergence among the seven B.thuringiensis genome assemblies,which harbor ample repeats and single nucleotide polymorphisms (SNPs).The identities among orthologous genes are greater than 84.5% and the hotspots for the genome variations were discovered in genomic regions of 2.3-2.8 Mb and 5.0-5.6 Mb.We concluded that high-coverage sequence assemblies from multiple strains,before all the gaps are closed,are very useful for pangenomic studies.

  15. Surface topography of the Bacillus stearothermophilus ribosome

    International Nuclear Information System (INIS)

    The surface topography of the intact 70S ribosome and free 30S and 50S subunits from Bacillus stearothermophilus strain 2,184 was investigated by lactoperoxidase-catalyzed iodination. Two-dimensional polyacrylamide gel electrophoresis was employed to separate ribosomal proteins for analysis of their reactivity. Free 50S subunits incorporated about 18% more 125I than did 50S subunits derived from 70S ribosomes, whereas free 30S subunits and 30S subunits derived from 70S ribosomes incorporated similar amounts of 125I. Iodinated 70S ribosomes and subunits retained 62-78% of the protein synthesis activity of untreated particles and sedimentation profiles showed no gross conformational changes due to iodination. The proteins most reactive to enzymatic iodination were S4, S7, S10 and Sa of the small subunit and L2, L4, L5/9, L6 and L36 of the large subunit. Proteins S2, S3, S7, S13, Sa, L5/9, L10, L11 and L24/25 were labeled substantially more in the free subunits than in the 70S ribosome. Other proteins, including S5, S9, S12, S15/16, S18 and L36 were more extensively iodinated in the 70S ribosome than in the free subunits. The locations of tyrosine residues in some homologus ribosomal proteins from B. stearothermophilus and E. coli are compared. (orig.)

  16. Extended genetic analysis of Brazilian isolates of Bacillus cereus and Bacillus thuringiensis

    Science.gov (United States)

    Zahner, Viviane; Silva, Ana Carolina Telles de Carvalho e; de Moraes, Gabriela Pinhel; McIntosh, Douglas; de Filippis, Ivano

    2013-01-01

    Multiple locus sequence typing (MLST) was undertaken to extend the genetic characterization of 29 isolates of Bacillus cereus and Bacillus thuringiensis previously characterized in terms of presence/absence of sequences encoding virulence factors and via variable number tandem repeat (VNTR). Additional analysis involved polymerase chain reaction for the presence of sequences (be, cytK, inA, pag, lef, cya and cap), encoding putative virulence factors, not investigated in the earlier study. MLST analysis ascribed novel and unique sequence types to each of the isolates. A phylogenetic tree was constructed from a single sequence of 2,838 bp of concatenated loci sequences. The strains were not monophyletic by analysis of any specific housekeeping gene or virulence characteristic. No clear association in relation to source of isolation or to genotypic profile based on the presence or absence of putative virulence genes could be identified. Comparison of VNTR profiling with MLST data suggested a correlation between these two methods of genetic analysis. In common with the majority of previous studies, MLST was unable to provide clarification of the basis for pathogenicity among members of the B. cereus complex. Nevertheless, our application of MLST served to reinforce the notion that B. cereus and B. thuringiensis should be considered as the same species. PMID:23440117

  17. Architecture and High-Resolution Structure of Bacillus thuringiensis and Bacillus cereus Spore Coat Surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Leighton, T; Wheeler, K; Malkin, A

    2005-02-18

    We have utilized atomic force microscopy (AFM) to visualize the native surface topology and ultrastructure of Bacillus thuringiensis and Bacillus cereus spores in water and in air. AFM was able to resolve the nanostructure of the exosporium and three distinctive classes of appendages. Removal of the exosporium exposed either a hexagonal honeycomb layer (B. thuringiensis) or a rodlet outer spore coat layer (B. cereus). Removal of the rodlet structure from B. cereus spores revealed an underlying honeycomb layer similar to that observed with B. thuringiensis spores. The periodicity of the rodlet structure on the outer spore coat of B. cereus was {approx}8 nm, and the length of the rodlets was limited to the cross-patched domain structure of this layer to {approx}200 nm. The lattice constant of the honeycomb structures was {approx}9 nm for both B. cereus and B. thuringiensis spores. Both honeycomb structures were composed of multiple, disoriented domains with distinct boundaries. Our results demonstrate that variations in storage and preparation procedures result in architectural changes in individual spore surfaces, which establish AFM as a useful tool for evaluation of preparation and processing ''fingerprints'' of bacterial spores. These results establish that high-resolution AFM has the capacity to reveal species-specific assembly and nanometer scale structure of spore surfaces. These species-specific spore surface structural variations are correlated with sequence divergences in a spore core structural protein SspE.

  18. Occurrence of Toxigenic Bacillus cereus and Bacillus thuringiensis in Doenjang, a Korean Fermented Soybean Paste.

    Science.gov (United States)

    Park, Kyung Min; Kim, Hyun Jung; Jeong, Moon Cheol; Koo, Minseon

    2016-04-01

    This study determined the prevalence and toxin profile of Bacillus cereus and Bacillus thuringiensis in doenjang, a fermented soybean food, made using both traditional and commercial methods. The 51 doenjang samples tested were broadly contaminated with B. cereus; in contrast, only one sample was positive for B. thuringiensis. All B. cereus isolates from doenjang were positive for diarrheal toxin genes. The frequencies of nheABC and hblACD in traditional samples were 22.7 and 0%, respectively, whereas 5.1 and 5.1% of B. cereus isolates from commercial samples possessed nheABC and hblACD, respectively. The detection rate of ces gene was 10.8%. The predominant toxin profile among isolates from enterotoxigenic B. cereus in doenjang was profile 4 (entFM-bceT-cytK). The major enterotoxin genes in emetic B. cereus were cytK, entFM, and nheA genes. The B. thuringiensis isolate was of the diarrheagenic type. These results provide a better understanding of the epidemiology of the enterotoxigenic and emetic B. cereus groups in Korean fermented soybean products.

  19. Extended genetic analysis of Brazilian isolates of Bacillus cereus and Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    Viviane Zahner

    2013-02-01

    Full Text Available Multiple locus sequence typing (MLST was undertaken to extend the genetic characterization of 29 isolates of Bacillus cereus and Bacillus thuringiensis previously characterized in terms of presence/absence of sequences encoding virulence factors and via variable number tandem repeat (VNTR. Additional analysis involved polymerase chain reaction for the presence of sequences (be, cytK, inA, pag, lef, cya and cap, encoding putative virulence factors, not investigated in the earlier study. MLST analysis ascribed novel and unique sequence types to each of the isolates. A phylogenetic tree was constructed from a single sequence of 2,838 bp of concatenated loci sequences. The strains were not monophyletic by analysis of any specific housekeeping gene or virulence characteristic. No clear association in relation to source of isolation or to genotypic profile based on the presence or absence of putative virulence genes could be identified. Comparison of VNTR profiling with MLST data suggested a correlation between these two methods of genetic analysis. In common with the majority of previous studies, MLST was unable to provide clarification of the basis for pathogenicity among members of the B. cereus complex. Nevertheless, our application of MLST served to reinforce the notion that B. cereus and B. thuringiensis should be considered as the same species.

  20. The Bacillus anthracis Exosporium: What's the Big "Hairy" Deal?

    Science.gov (United States)

    Bozue, Joel A; Welkos, Susan; Cote, Christopher K

    2015-10-01

    In some Bacillus species, including Bacillus subtilis, the coat is the outermost layer of the spore. In others, such as the Bacillus cereus family, there is an additional layer that envelops the coat, called the exosporium. In the case of Bacillus anthracis, a series of fine hair-like projections, also referred to as a "hairy" nap, extends from the exosporium basal layer. The exact role of the exosporium in B. anthracis, or for any of the Bacillus species possessing this structure, remains unclear. However, it has been assumed that the exosporium would play some role in infection for B. anthracis, because it is the outermost structure of the spore and would make initial contact with host and immune cells during infection. Therefore, the exosporium has been a topic of great interest, and over the past decade much progress has been made to understand its composition, biosynthesis, and potential roles. Several key aspects of this spore structure, however, are still debated and remain undetermined. Although insights have been gained on the interaction of exosporium with the host during infection, the exact role and significance of this complex structure remain to be determined. Furthermore, because the exosporium is a highly antigenic structure, future strategies for the next-generation anthrax vaccine should pursue its inclusion as a component to provide protection against the spore itself during the initial stages of anthrax. PMID:26542035

  1. Bacillus as a potential diagnostic marker for yellow tongue coating.

    Science.gov (United States)

    Ye, Juan; Cai, Xueting; Yang, Jie; Sun, Xiaoyan; Hu, Chunping; Xia, Junquan; Shen, Jianping; Su, Kelei; Yan, Huaijiang; Xu, Yuehua; Zhang, Yiyan; Zhang, Sujie; Yang, Lijun; Zhi, Hao; Gao, Sizhi Paul; Yu, Qiang; Hu, Jingqing; Cao, Peng

    2016-01-01

    Observation of tongue coating, a foundation for clinical diagnosis and treatment in traditional Chinese medicine (TCM), is a major indicator of the occurrence, development, and prognosis of disease. The biological basis of tongue diagnosis and relationship between the types and microorganisms of tongue coating remain elusive. Thirteen chronic erosive gastritis (CEG) patients with typical yellow tongue coating (YTC) and ten healthy volunteers with thin white tongue coating (WTC) were included in this study. Patients were provided a 2-course targeted treatment of a herbal medicine Ban Xia Xie Xin decoction, traditionally prescribed for CEG patients with YTC, to evaluate the relationship between tongue coating microbiota and diagnosis of CEG with typical YTC. The tongue coating segregation structure was determined using Illumina Miseq sequencing of the V4-V5 region of the 16S ribosomal RNA gene. Bacillus was significantly observed only in CEG patients with YTC, but not in patients who received the decoction. YTC (n = 22) and WTC (n = 29) samples were collected for bacterial culturing to illustrate the relationship between Bacillus and YTC. The Bacillus positivity rate of YTC samples was 72.7%; Bacillus was not observed in WTC samples. In conclusion, Bacillus was strongly associated with YTC. PMID:27578261

  2. Production of Alpha Amylase by Bacillus cereus in Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Helen H. Raplong

    2014-09-01

    Full Text Available Microorganisms have the ability to secrete enzymes when they are grown in the presence of certain substrates. Amylases are among the most important industrial enzymes and are of great significance in biotechnological studies. Bacteria belonging to the genus Bacillus were isolated using mannitol egg yolk polymyxin B (MYP agar a highly selective media for Bacillus cereus isolation. The isolates were tested for α-amylase production on nutrient agar supplemented with starch and in submerged fermentation. The bacteria isolated and identified (using the Microgen Bacillus identification kit were all Bacillus cereus and SB2 had the largest zone of hydrolysis of 12mm on nutrient agar supplemented with starch as well as the highest enzyme activity of 1.62U/ml. Amylase activity of 2.56U/ml was obtained after 24 hours incubation in submerged fermentation. When amylase enzyme production parameters where optimized, maximum amylase activity was obtained at a pH of 6.5, temperature of 350C, incubation time of 24 hours and 4% inoculums concentration. Bacillus cereus SB2 is a potential isolate for alpha-amylase production with soluble starch as the sole carbon source in submerged fermentation.

  3. The search and identification of the new immunodiagnostic targets of bacillus anthracis spore

    International Nuclear Information System (INIS)

    Spores of Bacillus anthracis have been used as bio warfare agent to bio terrorize purposes. As efficiency of anti-epidemic measures included urgent prevention and treatment is determined by terms within which the bio agent is identified. Direct and rapid spore detection by antibodies based detection system is very attractive alternative to current PCR-based assays or routine phenotyping which are the most accurate but are also complex, time-consumption and expensive. The main difficulty with respect to such kind of anthrax spores detection is a cross-reaction with spores of closely related bacteria. For development of species-specific antibodies to anthrax spores recombinant scFvs or hybridoma technique were used. In both case surface spore antigens contained species-specific epitopes are need. Among exosporium proteins only ExsF(BxpB), ExsK and SoaA are specific to B.cereus group. On the surface of B. anthracis spores, a unique tetrasaccharides containing an novel monosaccharide - anthrose, was discovered. It was shown that anthrose can be serving as species-specific target for B. anthracis spores detection. We have revealed that EA1 isolated from spore of Russians strain STI-1 contain carbohydrate which formed species-specific epitopes and determine immunogenicity of this antigen. Antibodies to this antigen specifically recognized the surface target of B. anthracis spores and do not reacted with others Bacillus spore. Based on these antibodies we developed the test-systems in different formats for rapid direct detection and identification of B. anthracis spores. The results of trial these test-systems with using more than 50 different Bacillus strains were indicated that carbohydrate of EA1 isolated from spore is effective immunodiagnostic target for anthrax spores bio detection.(author)

  4. UJI TOKSISITAS ISOLAT Bacillus thuringiensis dari Kabupaten Lahat, Palembang, Sumatera Selatan TERHADAP LARVA NYAMUK Culex sp.

    OpenAIRE

    Chandra, Welianto

    2016-01-01

    This study aims to determine the optimal concentration of isolates of Bacillus thuringiensis to control larvae of the mosquito Culex sp. The method used is the isolation of the bacterium Bacillus thuringiensis, then the inoculation of bacteria. Bacillus thuringiensis mud samples, as much as 25 grams, obtained in the area of Lahat, South Sumatra containing Bacillus thuringiensis which includes five districts, namely Sub Gumay Talang, Jaray, Kikim West, South Kikim, and Central Kikim. Gumay ...

  5. The influence of thermal shock on filamentous bacillus subtillis strain resistance to subsequent stress treatment

    International Nuclear Information System (INIS)

    When grown at 30 degC and heat shocked in a liquid medium at 45 degC, the filamentous cells of Bacillus subtilis 168 became more sensitive to subsequent killing with N-methyl-N'-nitro-N-nitrosoguanidine and UV light but not with γ-rays. Certain characteristics (for instance, the increased tolerance to damaging agents at 30 degC and the time-dependent changes in the sensitivity to MNNG induced by thermal shock) evidence against direct involvement of repair systems in this phenomenon. 29 refs., 3 figs., 1 tab

  6. Conversion of sub-megasized DNA to desired structures using a novel Bacillus subtilis genome vector

    OpenAIRE

    Kaneko, Shinya; Tsuge, Kenji; Takeuchi, Takashi; Itaya, Mitsuhiro

    2003-01-01

    A novel genome vector using the 4215 kb Bacillus subtilis genome provides for precise target cloning and processing of the cloned DNA to the desired structure. Each process highly dependent on homologous recombination in the host B.subtilis is distinguished from the other cloning systems. A 120 kb mouse jumonji (jmj) genomic gene was processed in the genome vector to give a series of truncated sub-megasized DNA. One of these truncated segments containing the first intron was copied in a plasm...

  7. Identification of a Bacillus subtilis secretion mutant using a beta-galactosidase screening procedure

    DEFF Research Database (Denmark)

    Jacobs, M F; Borchert, T V; Kontinen, V P;

    1995-01-01

    High-level synthesis of exportable beta-galactosidase (LacZ) fusion proteins in Bacillus subtilis results in a lethal phenotype, and has been suggested as a tool for the selection of secretion mutants. We tested a plasmid-based, inducible lacZ fusion gene system for this purpose, but frequent...... mutations in cis, which reduced expression of the fusion gene, forced abandonment of the induction-selection strategy. Instead, after modification of the indicator plasmid, a screening procedure for increased basal LacZ activity levels was adopted. This led to the identification of a conditional B. subtilis...

  8. Identification of a Bacillus subtilis secretion mutant using a ß-galactosidase screening procedure

    DEFF Research Database (Denmark)

    Jacobs, Myra F.; Andersen, Jens Bo; Borchert, Torben V.;

    1995-01-01

    High-level synthesis of exportable beta-galactosidase (LacZ) fusion proteins in Bacillus subtilis results in a lethal phenotype, and has been suggested as a tool for the selection of secretion mutants. We tested a plasmid-based, inducible lacZ fusion gene system for this purpose, but frequent...... mutations in cis, which reduced expression of the fusion gene, forced abandonment of the induction-selection strategy. Instead, after modification of the indicator plasmid, a screening procedure for increased basal LacZ activity levels was adopted. This led to the identification of a conditional B. subtilis...

  9. Production and characterization of biodiesel using nonedible castor oil by immobilized lipase from Bacillus aerius.

    Science.gov (United States)

    Narwal, Sunil Kumar; Saun, Nitin Kumar; Dogra, Priyanka; Chauhan, Ghanshyam; Gupta, Reena

    2015-01-01

    A novel thermotolerant lipase from Bacillus aerius was immobilized on inexpensive silica gel matrix. The immobilized lipase was used for the synthesis of biodiesel using castor oil as a substrate in a solvent free system at 55°C under shaking in a chemical reactor. Several crucial parameters affecting biodiesel yield such as incubation time, temperature, substrate molar ratio, and amount of lipase were optimized. Under the optimized conditions, the highest biodiesel yield was up to 78.13%. The characterization of synthesized biodiesel was done through FTIR spectroscopy, (1)H NMR spectra, and gas chromatography.

  10. Oral bacillus Calmette-Guérin vaccine against tuberculosis: why not?

    Directory of Open Access Journals (Sweden)

    Renata Monteiro-Maia

    2014-09-01

    Full Text Available The bacillus Calmette-Guérin (BCG vaccine is the only licensed vaccine for human use against tuberculosis (TB. Although controversy exists about its efficacy, the BCG vaccine is able to protect newborns and children against disseminated forms of TB, but fails to protect adults against active forms of TB. In the last few years, interest in the mucosal delivery route for the vaccine has been increasing owing to its increased capacity to induce protective immune responses both in the mucosal and the systemic immune compartments. Here, we show the importance of this route of vaccination in newly developed vaccines, especially for vaccines against TB.

  11. Oral bacillus Calmette-Guérin vaccine against tuberculosis: why not?

    Science.gov (United States)

    Monteiro-Maia, Renata; Pinho, Rosa Teixeira de

    2014-09-01

    The bacillus Calmette-Guérin (BCG) vaccine is the only licensed vaccine for human use against tuberculosis (TB). Although controversy exists about its efficacy, the BCG vaccine is able to protect newborns and children against disseminated forms of TB, but fails to protect adults against active forms of TB. In the last few years, interest in the mucosal delivery route for the vaccine has been increasing owing to its increased capacity to induce protective immune responses both in the mucosal and the systemic immune compartments. Here, we show the importance of this route of vaccination in newly developed vaccines, especially for vaccines against TB. PMID:25317714

  12. Cell wall mechanical properties as measured with bacterial thread made from Bacillus subtilis.

    OpenAIRE

    Mendelson, N H; Thwaites, J J

    1989-01-01

    Engineering approaches used in the study of textile fibers have been applied to the measurement of mechanical properties of bacterial cell walls by using the Bacillus subtilis bacterial thread system. Improved methods have been developed for the production of thread and for measuring its mechanical properties. The best specimens of thread produced from cultures of strain FJ7 grown in TB medium at 20 degrees C varied in diameter by a factor of 1.09 over a 30-mm thread length. The stress-strain...

  13. The Adverse Events, Signs, and Management of Neonatal Bacillus Calmette-Guérin Vaccine Overdose

    OpenAIRE

    Al-Maqbali, Ali Abdullah; Al-Maani, Amal Saif Suliman

    2016-01-01

    A two-day-old male neonate was inadvertently immunized with 20-times the recommended dose of Bacillus Calmette-Guérin (BCG) vaccine (1.0 ml instead of 0.05 ml). Examination of the neonate after one day revealed no signs of local reaction at the site of injection or systemic manifestation. However, nine days later, he developed a swelling (4 × 3 cm) at the site of injection without any enlargement of the lymph node, pallor, or jaundice. A needle aspiration of the swelling was done and sent for...

  14. 40 CFR 180.1128 - Bacillus subtilis MBI 600; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus subtilis MBI 600; exemption... FOOD Exemptions From Tolerances § 180.1128 Bacillus subtilis MBI 600; exemption from the requirement of... biofungicide Bacillus subtilis MBI 600 in or on all food commodities, including residues resulting from...

  15. Evaluation of in situ valine production by Bacillus subtilis in young pigs

    DEFF Research Database (Denmark)

    Nørgaard, Jan Værum; Canibe, Nuria; Assadi Soumeh, Elham;

    2016-01-01

    Mutants of Bacillus subtilis can be developed to overproduce Val in vitro. It was hypothesized that addition of Bacillus subtilis mutants to pig diets can be a strategy to supply the animal with Val. The objective was to investigate the effect of Bacillus subtilis mutants on growth performance an...

  16. Genetic Characterization of Bacillus anthracis 17 JB strain

    Directory of Open Access Journals (Sweden)

    Sakineh Seyed-Mohamadi

    2015-11-01

    Full Text Available Background and Objectives: Bacillus anthracis is one of the most homogenous bacteria ever described. Bacillus anthracis 17JB is a laboratory strain. It is broadly used as a challenge strain in guinea pigs for potency test of anthrax vaccine.Material and Methods: This work describes genetic characterization of B. anthracis 17 JB strain using the SNPs and MLVA genotyping.Results and Conclusion: In SNPs typing, the originally French 17JB strain represented the A. Br. 008/009 subgroup. In Levy's genotyping method, 843, 451 and 864 bp long fragments were identified at AA03, AJ03 and AA07 loci, respectively. In the vaccine manufacturer perspective these findings are much valuable on their own account, but similar research is required to extend molecular knowledge of B. anthracis epidemiology in Persia.Keywords: Bacillus anthracis 17JB, Genetic characterization, SNPs typing

  17. Bacillus cereus food poisoning: international and Indian perspective.

    Science.gov (United States)

    Tewari, Anita; Abdullah, Swaid

    2015-05-01

    Food borne illnesses result from eating food or drinking beverages that are contaminated with chemical matter, heavy metals, parasites, fungi, viruses and Bacteria. Bacillus cereus is one of the food-borne disease causing Bacteria. Species of Bacillus and related genera have long been troublesome to food producers on account of their resistant endospores. Their spores may be present on various types of raw and cooked foods, and their ability to survive high cooking temperatures requires that cooked foods be served hot or cooled rapidly to prevent the growth of this bacteria. Bacillus cereus is well known as a cause of food poisoning, and much more is now known about the toxins produced by various strains of this species, so that its significance in such episodes are clearer. However, it is still unclear why such cases are so rarely reported worldwide.

  18. Genotyping of Bacillus cereus strains by microarray-based resequencing.

    Directory of Open Access Journals (Sweden)

    Michael E Zwick

    Full Text Available The ability to distinguish microbial pathogens from closely related but nonpathogenic strains is key to understanding the population biology of these organisms. In this regard, Bacillus anthracis, the bacterium that causes inhalational anthrax, is of interest because it is closely related and often difficult to distinguish from other members of the B. cereus group that can cause diverse diseases. We employed custom-designed resequencing arrays (RAs based on the genome sequence of Bacillus anthracis to generate 422 kb of genomic sequence from a panel of 41 Bacillus cereus sensu lato strains. Here we show that RAs represent a "one reaction" genotyping technology with the ability to discriminate between highly similar B. anthracis isolates and more divergent strains of the B. cereus s.l. Clade 1. Our data show that RAs can be an efficient genotyping technology for pre-screening the genetic diversity of large strain collections to selected the best candidates for whole genome sequencing.

  19. BACILLUS THURINGIENSIS ELASTASES WITH INSECTICIDE ACTIVITY

    Directory of Open Access Journals (Sweden)

    E. V. Matseliukh

    2015-10-01

    Full Text Available The purpose of the research was a screening of proteases with elastase activity among Bacillus thuringiensis strains, their isolation, partially purification, study of physicochemical properties and insecticide activity in relation to the larvae of the Colorado beetle. The objects of the investigation were 18 strains of B. thuringiensis, isolated from different sources: sea water, dry biological product "Bitoksibatsillin" and also from natural populations of Colorado beetles of the Crimea, Kherson, Odesa, Mykolaiv and Zaporizhiia regions of Ukraine. Purification of enzymes with elastase activity isolated from above mentioned strains was performed by gel-chromatography and insecticide activity was studied on the 3–4 larvae instar of Colorado beetle. The ability of a number of B. thuringiensis strains to synthesize the proteases with elastase activity has been established. The most active were enzymes obtained from strains IMV B-7465, IMV B-7324 isolated from sea water, and strains 9, 902, Bt-H and 0-239 isolated from Colorado beetles. The study of the physicochemical properties of the partially purified proteases of these strains showed that they belonged to enzymes of the serine type. Peptidases of a number of B. thuringiensis strains (IMV B-7324, IMV B-7465, 902, 0-239, 9 are metal-dependent enzymes. Optimal conditions of action of all tested enzymes are the neutral and alkaline рН values and the temperatures of 30–40 °С. The studies of influence of the complex enzyme preparations and partially purified ones of B. thuringiensis strains on the larvae instar of Colorado beetles indicated that enzymes with elastase activity could be responsible for insecticide action of the tested strains.

  20. Production and Characterization of Bacillus firmus pectinase

    Directory of Open Access Journals (Sweden)

    Anna Roosdiana

    2013-03-01

    Full Text Available Pectinase is enzyme which functions to hydrolyze pectin become D-galacturonic acid unit. This enzyme is potential in various industries, especially in fruit juice industry.  Pectinase can be derived from various microorganisms resulting in different pectinase character. The aims of this research were to determine the optimum condition of pectinase production and to characterize the resulted pectinase including optimum condition of pectinase activity and the influence of metal ion.  The optimum condition of pectinase production was carried out by growing Bacillus firmus on basal media containing pectin as inducer at various  pH (5, 6, 7, 8, 9, 10, temperature (30, 35, 40, 45, 50 oC and fermentation time (6, 12, 18, 24, 30, 36 hours. while the optimum pectinase activity was done at various pH ( 4, 6, 7, 8, 10 , temperature (30, 35, 40, 45, 50 oC and reaction time (10, 20, 30, 40, 50 minutes. The influence of Zn2+, Mg2+, K+ at 2-10 mM to pectinase activity were also investigated. The result showed that optimum condition of pectinase production occurred at pH7-8, temperature 40-50 oC and fermentation time 18hours, while the optimum condition of pectinase activity was pH 7, temperature 50 oC and reaction time 30 minutes. The existence of Zn2+, Mg2+, K+ ions  affected significantly to pectinase activity.  Mg2+ acted as non competitive inhibitor; however K+ and Zn2+ acted as un competitive inhibitor.

  1. Effect of calcium ions on the evolution of biofouling by Bacillus subtilis in plate heat exchangers simulating the heat pump system used with treated sewage in the 2008 Olympic Village.

    Science.gov (United States)

    Tian, Lei; Chen, Xiao Dong; Yang, Qian Peng; Chen, Jin Chun; Shi, Lin; Li, Qiong

    2012-06-01

    Heat pump systems using treated sewage water as the heat source were used in the Beijing Olympic Village for domestic heating and cooling. However, considerable biofouling occurred in the plate heat exchangers used in the heat pump system, greatly limiting the system efficiency. This study investigates the biofouling characteristics using a plate heat exchanger in parallel with a flow cell system to focus on the effect of calcium ions on the biofilm development. The interactions between the microorganisms and Ca(2+) enhances both the extent and the rate of biofilm development with increasing Ca(2+) concentration, leading to increased heat transfer and flow resistances. Three stages of biofouling development were identified in the presence of Ca(2+) from different biofouling mass growth rates with an initial stage, a rapid growth stage and an extended growth stage. Each growth stage had different biofouling morphologies influenced by the Ca(2+) concentration. The effects of Ca(2+) on the biofouling heat transfer and flow resistances had a synergistic effect related to both the biofouling mass and the morphology. The effect of Ca(2+) on the biofouling development was most prominent during the rapid growth stage. PMID:22391321

  2. Effect of calcium ions on the evolution of biofouling by Bacillus subtilis in plate heat exchangers simulating the heat pump system used with treated sewage in the 2008 Olympic Village.

    Science.gov (United States)

    Tian, Lei; Chen, Xiao Dong; Yang, Qian Peng; Chen, Jin Chun; Shi, Lin; Li, Qiong

    2012-06-01

    Heat pump systems using treated sewage water as the heat source were used in the Beijing Olympic Village for domestic heating and cooling. However, considerable biofouling occurred in the plate heat exchangers used in the heat pump system, greatly limiting the system efficiency. This study investigates the biofouling characteristics using a plate heat exchanger in parallel with a flow cell system to focus on the effect of calcium ions on the biofilm development. The interactions between the microorganisms and Ca(2+) enhances both the extent and the rate of biofilm development with increasing Ca(2+) concentration, leading to increased heat transfer and flow resistances. Three stages of biofouling development were identified in the presence of Ca(2+) from different biofouling mass growth rates with an initial stage, a rapid growth stage and an extended growth stage. Each growth stage had different biofouling morphologies influenced by the Ca(2+) concentration. The effects of Ca(2+) on the biofouling heat transfer and flow resistances had a synergistic effect related to both the biofouling mass and the morphology. The effect of Ca(2+) on the biofouling development was most prominent during the rapid growth stage.

  3. Serological Correlate of Protection in Guinea Pigs for a Recombinant Protective Antigen Anthrax Vaccine Produced from Bacillus brevis

    OpenAIRE

    Chun, Jeong-Hoon; Choi, On-Jee; Cho, Min-Hee; Hong, Kee-Jong; Seong, Won Keun; Oh, Hee-Bok; Rhie, Gi-eun

    2012-01-01

    Objective Recombinant protective antigen (rPA) is the active pharmaceutical ingredient of a second generation anthrax vaccine undergoing clinical trials both in Korea and the USA. By using the rPA produced from Bacillus brevis pNU212 expression system, correlations of serological immune response to anthrax protection efficacy were analyzed in a guinea pig model. Methods Serological responses of rPA anthrax vaccine were investigated in guinea pigs that were given single or two injections (inte...

  4. Biodegradation of real petroleum wastewater by immobilized hyper phenol-tolerant strains of Bacillus cereus in a fluidized bed bioreactor

    OpenAIRE

    Banerjee, Aditi; Ghoshal, Aloke K.

    2016-01-01

    Microbial bioremediation of petroleum wastewater by phenol-degrading-bacteria holds promise in circumventing the issue of petroleum-spill related pollution. Herein, biodegradation of petroleum wastewater samples collected from oil refinery site was carried out in a fluidized bed bioreactor by Ca-alginate immobilized biomass of phenol-degrading strains of Bacillus cereus (AKG1 MTCC9817 and AKG2 MTCC9818). Degradation performance of the system was evaluated by measuring the changes in chemical ...

  5. Influence of Mycobacterium bovis bacillus Calmette-Guérin on antibody and cytokine responses to human neonatal vaccination

    OpenAIRE

    Martin O C Ota; Vekemans, Johan; Schlegel Heueter, Susanna; Fielding, Katherine; Sanneh, Mariama; Kidd, Michael; Newport, Melanie J.; Aaby, Peter; Whittle, Hilton; Lambert, Paul Henri; McAdam, Keith P. W. J.; Siegrist, Claire-Anne; Marchant, Arnaud

    2002-01-01

    The immaturity of the immune system increases the susceptibility of young infants to infectious diseases and prevents the induction of protective immune responses by vaccines. We previously reported that Mycobacterium bovis bacillus Calmette-Guérin (BCG) vaccination induces a potent Th1 response to mycobacterial Ags in newborns. In this study, we evaluated the influence of BCG on the response to unrelated vaccines given in early life. Newborns were randomly allocated to one of three study gro...

  6. Bacillus subtilis strain deficient for the protein-tyrosine kinase PtkA exhibits impaired DNA replication

    DEFF Research Database (Denmark)

    Petranovic, Dina; Michelsen, Ole; Zahradka, K;

    2007-01-01

    Bacillus subtilis has recently come into the focus of research on bacterial protein-tyrosine phosphorylation, with several proteins kinases, phosphatases and their substrates identified in this Gram-positive model organism. B. subtilis protein-tyrosine phosphorylation system PtkA/PtpZ was previou...... microscopy. B. subtilis cells lacking the kinase PtkA accumulated extra chromosome equivalents, exhibited aberrant initiation mass for DNA replication and an unusually long D period....

  7. The effects of orally administered Bacillus coagulans and inulin on prevention and progression of rheumatoid arthritis in rats

    OpenAIRE

    Khadijeh Abhari; Seyed Shahram Shekarforoush; Saeid Hosseinzadeh; Saeid Nazifi; Javad Sajedianfard; Mohammad Hadi Eskandari

    2016-01-01

    Background: Probiotics have been considered as an approach to addressing the consequences of different inflammatory disorders. The spore-forming probiotic strain Bacillus coagulans has demonstrated anti-inflammatory and immune-modulating effects in both animals and humans. The prebiotic inulin also potentially affects the immune system as a result of the change in the composition or fermentation profile of the gastrointestinal microbiota. Objective: In the present study, an in vivo model was ...

  8. The effects of orally administered Bacillus coagulans and inulin on prevention and progression of rheumatoid arthritis in rats

    OpenAIRE

    Seyed Shahram SHEKARFOROUSH; Abhari, Khadijeh; Hosseinzadeh, Saeid; Nazifi, Saeid; Sajedianfard, Javad; Eskandari, Mohammad Hadi

    2016-01-01

    Background: Probiotics have been considered as an approach to addressing the consequences of different inflammatory disorders. The spore-forming probiotic strain Bacillus coagulans has demonstrated anti-inflammatory and immune-modulating effects in both animals and humans. The prebiotic inulin also potentially affects the immune system as a result of the change in the composition or fermentation profile of the gastrointestinal microbiota.Objective: In the present study, an in vivo model was c...

  9. Developments in the use of Bacillus species for industrial production.

    Science.gov (United States)

    Schallmey, Marcus; Singh, Ajay; Ward, Owen P

    2004-01-01

    Bacillus species continue to be dominant bacterial workhorses in microbial fermentations. Bacillus subtilis (natto) is the key microbial participant in the ongoing production of the soya-based traditional natto fermentation, and some Bacillus species are on the Food and Drug Administration's GRAS (generally regarded as safe) list. The capacity of selected Bacillus strains to produce and secrete large quantities (20-25 g/L) of extracellular enzymes has placed them among the most important industrial enzyme producers. The ability of different species to ferment in the acid, neutral, and alkaline pH ranges, combined with the presence of thermophiles in the genus, has lead to the development of a variety of new commercial enzyme products with the desired temperature, pH activity, and stability properties to address a variety of specific applications. Classical mutation and (or) selection techniques, together with advanced cloning and protein engineering strategies, have been exploited to develop these products. Efforts to produce and secrete high yields of foreign recombinant proteins in Bacillus hosts initially appeared to be hampered by the degradation of the products by the host proteases. Recent studies have revealed that the slow folding of heterologous proteins at the membrane-cell wall interface of Gram-positive bacteria renders them vulnerable to attack by wall-associated proteases. In addition, the presence of thiol-disulphide oxidoreductases in B. subtilis may be beneficial in the secretion of disulphide-bond-containing proteins. Such developments from our understanding of the complex protein translocation machinery of Gram-positive bacteria should allow the resolution of current secretion challenges and make Bacillus species preeminent hosts for heterologous protein production. Bacillus strains have also been developed and engineered as industrial producers of nucleotides, the vitamin riboflavin, the flavor agent ribose, and the supplement poly

  10. Sigma A recognition sites in the Bacillus subtilis genome

    DEFF Research Database (Denmark)

    Jarmer, Hanne Østergaard; Larsen, Thomas Schou; Krogh, Anders Stærmose;

    2001-01-01

    A hidden Markov model of sigma (A) RNA polymerase cofactor recognition sites in Bacillus subtilis, containing either the common or the extended -10 motifs, has been constructed based on experimentally verified sigma (A) recognition sites. This work suggests that more information exists at the ini......A hidden Markov model of sigma (A) RNA polymerase cofactor recognition sites in Bacillus subtilis, containing either the common or the extended -10 motifs, has been constructed based on experimentally verified sigma (A) recognition sites. This work suggests that more information exists...

  11. Characterization of an L-arabinose isomerase from Bacillus subtilis

    DEFF Research Database (Denmark)

    Kim, Jin-Ha; Prabhu, Ponnandy; Jeya, Marimuthu;

    2010-01-01

    An isolated gene from Bacillus subtilis str. 168 encoding a putative isomerase was proposed as an L-arabinose isomerase (L-AI), cloned into Escherichia coli, and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1,491 bp, capable of encoding a polypep......An isolated gene from Bacillus subtilis str. 168 encoding a putative isomerase was proposed as an L-arabinose isomerase (L-AI), cloned into Escherichia coli, and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1,491 bp, capable of encoding...

  12. Analysis of myo-inositol hexakisphosphate hydrolysis by Bacillus phytase

    DEFF Research Database (Denmark)

    Kerovuo, J.; Rouvinen, J.; Hatzack, Frank-Andreas

    2000-01-01

    Phytic acid (myo-inositol hexakisphosphate, InsP(6)) hydrolysis by Bacillus phytase (PhyC) was studied. The enzyme hydrolyses only three phosphates from phytic acid. Moreover, the enzyme seems to prefer the hydrolysis of every second phosphate over that of adjacent ones. Furthermore, it is very...... a reaction mechanism different from that of other phytases. By combining the data presented in this study with (1) structural information obtained from the crystal structure of Bacillus amyloliquefaciens phytase [Ha, Oh, Shin, Kim, Oh, Kim, Choi and Oh (2000) Nat. Struct. Biol. 7, 147-153], and (2) computer...

  13. Aerobic granulation of pure bacterial strain Bacillus thuringiensis

    Institute of Scientific and Technical Information of China (English)

    Sunil S ADAV; Duu-Jong LEE

    2008-01-01

    The objective of this study is to cultivate aer-obic granules by pure bacterial strain, Bacillus thuringien-sis, in a sequencing batch reactor. Stable granules sized 2.0-2.2 mm were formed in the reactor after a five-week cultivation. These granules exhibited excellent settling attributes, and degraded phenol at rates of 1.49 and concentration, respectively. Confocal laser scanning microscopic test results show that Bacillus thuringiensis was distributed over the initial small aggregates, and the outer edge of the granule was away from the core regime in the following stage.

  14. Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model

    Directory of Open Access Journals (Sweden)

    Lopamudra Haldar

    2016-07-01

    Full Text Available Aim: To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. Materials and Methods: An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1 was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2 and (T3 groups received spore biomass of Bacillus coagulans B37 and Bacillus pumilus B9, respectively, suspended in sterilized skim milk at 8-9 log colony-forming units/ml plus basal diet for 28 days, while control group (T4 was supplied with clean water along with basal diet. There was a 14-day post-treatment period. A total of 288 fecal samples (8 fecal collections per rat were collected at every 7-day interval starting from 0 to 49 days and subjected to the enumeration of the counts of coliforms and lactobacilli and Bacillus spores using respective agar media. In vitro acid and bile tolerance tests on both the strains were performed. Results: The rats those (T2 and T3 received either B. coagulans B37 or B. pumilus B9 spore along with non-fermented skim milk showed decrease (p<0.01 in fecal coliform counts and increase (p<0.05 in both fecal lactobacilli and Bacillus spore counts as compared to the control group (T4 and the group fed only skim milk (T1. In vitro study indicated that both the strains were found to survive at pH 2.0 and 3.0 even up to 3 h and tolerate bile up to 2.0% concentration even after 12 h of exposure. Conclusions: This study revealed that oral administration of either B. coagulans B37 or B. pumilus B9 strains might be useful in reducing coliform counts accompanied by concurrent increase in lactobacilli counts in the intestinal flora in rats.

  15. Crossing of the epithelial barriers by Bacillus anthracis: the Known and the Unknown

    Directory of Open Access Journals (Sweden)

    Pierre L Goossens

    2015-10-01

    Full Text Available Anthrax, caused by Bacillus anthracis, a Gram-positive spore-forming bacterium, is initiated by the entry of spores into the host body. There are three types of human infection: cutaneous, inhalational, and gastrointestinal. For each form, B. anthracis spores need to cross the cutaneous, respiratory or digestive epithelial barriers, respectively, as a first obligate step to establish infection. Anthrax is a toxi-infection: an association of toxemia and rapidly spreading infection progressing to septicemia. The pathogenicity of Bacillus anthracis mainly depends on two toxins and a capsule. The capsule protects bacilli from the immune system, thus promoting systemic dissemination. The toxins alter host cell signalling, thereby paralysing the immune response of the host and perturbing the endocrine and endothelial systems.In this review, we will mainly focus on the events and mechanisms leading to crossing of the respiratory epithelial barrier, as the majority of studies have addressed inhalational infection. We will discuss the critical gaps of knowledge that need to be addressed to gain a comprehensive view of the initial steps of inhalational anthrax. We will then discuss the few data available on B. anthracis crossing the cutaneous and digestive epithelia.

  16. Ecological aspects of Bacillus thuringiensis in an Oxisol Ecologia do Bacillus thuringiensis num Latossolo

    Directory of Open Access Journals (Sweden)

    Lessandra Heck Paes Leme Ferreira

    2003-02-01

    Full Text Available Bacillus thuringiensis is a Gram positive, sporangial bacterium, known for its insecticidal habilities. Survival and conjugation ability of B. thuringiensis strains were investigated; vegetative cells were evaluated in non-sterile soil. Vegetative cells decreased rapidly in number, and after 48 hours the population was predominantly spores. No plasmid transfer was observed in non-sterile soil, probably because the cells died and the remaining cells sporulated quickly. Soil is not a favorable environment for B. thuringiensis multiplication and conjugation. The fate of purified B. thuringiensis toxin was analyzed by extractable toxin quantification using ELISA. The extractable toxin probably declined due to binding on surface-active particles in the soil.O comportamento de células vegetativas do Bacillus thuringiensis foi estudado em solo não esterilizado. Após o inóculo grande parte das células morrem e o restante esporula em 24 horas. Não foi observada conjugação provavelmente porque poucas células sobrevivem no solo e rapidamente esporulam, mostrando que este não é o ambiente propício para a multiplicação e conjugação desta bactéria. A toxina purificada, portanto livre de células, diminui rapidamente sua quantidade em solo não esterilizado. Provavelmente a ligação da toxina na fração argilosa do solo é a principal responsável por este fenômeno.

  17. CcpA-independent regulation of expression of the Mg2+-citrate transporter gene citM by arginine metabolism in Bacillus subtilis

    NARCIS (Netherlands)

    Warner, JB; Magni, C; Lolkema, JS; Warner, Jessica B.

    2003-01-01

    Transcriptional regulation of the Mg2+ -citrate transporter, CitM, the main citrate uptake system of Bacillus subtilis, was studied during growth in rich medium. Citrate in the growth medium was required for induction under all growth conditions. In Luria-Bertani medium containing citrate, citM expr

  18. Human monoclonal antibodies against anthrax lethal factor and protective antigen act independently to protect against Bacillus anthracis infection and enhance endogenous immunity to anthrax

    NARCIS (Netherlands)

    Albrecht, Mark T.; Li, Han; Williamson, E. Diane; LeButt, Chris S.; Flick-Smith, Helen C.; Quinn, Conrad P.; Westra, Hans; Galloway, Darrell; Mateczun, Alfred; Goldman, Stanley; Groen, Herman; Baillie, Les W. J.

    2007-01-01

    The unpredictable nature of bioterrorism and the absence of real-time detection systems have highlighted the need for an efficient postexposure therapy for Bacillus anthracis infection. One approach is passive immunization through the administration of antibodies that mitigate the biological action

  19. Conjugative plasmid pAW63 brings new insights into the genesis of the Bacillus anthracis virulence plasmid pXO2 and of the Bacillus thuringiensis plasmid pBT9727

    Directory of Open Access Journals (Sweden)

    Mahillon Jacques

    2005-07-01

    Full Text Available Abstract Background Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis belong to the genetically close-knit Bacillus cereus sensu lato group, a family of rod-shaped Gram-positive bacteria. pAW63 is the first conjugative plasmid from the B. cereus group to be completely sequenced. Results The 71,777 bp nucleotide sequence of pAW63 reveals a modular structure, including a 42 kb tra region encoding homologs of the Type IV secretion systems components VirB11, VirB4 and VirD4, as well as homologs of Gram-positive conjugation genes from Enterococcus, Lactococcus, Listeria, Streptococcus and Staphylococcus species. It also firmly establishes the existence of a common backbone between pAW63, pXO2 from Bacillus anthracis and pBT9727 from the pathogenic Bacillus thuringiensis serovar konkukian strain 97-27. The alignment of these three plasmids highlights the presence of well conserved segments, in contrast to distinct regions of high sequence plasticity. The study of their specific differences has provided a three-point reference framework that can be exploited to formulate solid hypotheses concerning the functionalities and the molecular evolution of these three closely related plasmids. This has provided insight into the chronology of their divergence, and led to the discovery of two Type II introns on pAW63, matching copies of the mobile element IS231L in different loci of pXO2 and pBT9727, and the identification on pXO2 of a 37 kb pathogenicity island (PAI containing the anthrax capsule genes. Conclusion The complete sequence determination of pAW63 has led to a functional map of the plasmid yielding insights into its conjugative apparatus, which includes T4SS-like components, as well as its resemblance to other large plasmids of Gram-positive bacteria. Of particular interest is the extensive homology shared between pAW63 and pXO2, the second virulence plasmid of B. anthracis, as well as pBT9727 from the pathogenic strain B. thuringiensis

  20. The isolation and identification of Bacillus subtilis from Suaeda salsa and its bacteriostatic activity detection%碱蓬内生枯草芽孢杆菌分离鉴定及抑菌活性检测

    Institute of Scientific and Technical Information of China (English)

    姜华; 修玉萍; 郑音; 任伟重; 侯婧; 张琳; 王有福

    2011-01-01

    Abstract:Seven endogenous strains were isolated from Liaoning Suaeda salsa using automatic microorganism identification system (VITEK) and molecular biology, including Bacillus stutilis, Bacillus circulans, Bacillus megatherium , Pseudomonas aeruginosa, Staphylococcus lentus, Staphylococcus sciuri and Streptococcus agalactiae. The results showed that Bacillus stutilis could inhibit the growth of 10 species of fungi in this experiment, especially Penicillium oxalicum, on which the inhibition rate reached 86. 4%, and the inhibition rate on Fusarium solani and Fusarium moniliforme were 52. 2% and 41. 4% respectively.%采用全自动微生物鉴定系统(VITEK)和分子生物学鉴定方法,从辽宁碱蓬中分离纯化出7种细菌:枯草芽孢杆菌(Bacillus stutilis)、环状芽孢杆菌(Bacillus circulans)、巨大芽孢杆菌(Bacillus megatherium)、铜绿假单胞菌(Pseudomonas aeruginosa)、缓慢葡萄球菌(Staphylococcus lentus)、松鼠葡萄球菌(Staphylococcus sciuri)和无乳链球菌(Streptococcus agalactiae).其中,枯草芽孢杆菌(Bacillus stutilis)对供试的10种真菌有抑菌作用,尤其对草酸青霉菌丝干物质抑制率可达86.4%,对茄病镰孢菌和串珠镰孢菌的抑制率分别为52.2%和41.4%.