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Sample records for bacillus licheniformis

  1. Biomineralization of Se Nanoshpere by Bacillus Licheniformis

    Institute of Scientific and Technical Information of China (English)

    Yongqiang Yuan; Jianming Zhu; Congqiang Liu; Shen Yu; Lei Lei

    2015-01-01

    Biological dissimilatory reduction of selenite (SeO32-) to elemental selenium (Se0) is com-mon, but the mineral formation and the biogenic process remain uncertain. In this study, we examined the Se0 formation during the selenite bioreduction by Bacillus licheniformis SeRB-1 through transmis-sion electron microscope (TEM), energy-dispersive spectrometry (EDS) and X-ray absorption fine structure (XAFS) techniques. Results showed that the reduction process occurred mostly during the exponential phase and early stationary phase, whilst the elemental selenium was produced in these pe-riods. From the TEM images and polyacrylamide gel electropheresis, it is known that the Se0 granule formation is a biologically-induced type, and the cell envelopes are the main biomineralization positions, and particles may go through a process from nucleation to crystallization, under the control of mi-crobes. In fact, the minerals are spherical nanoparticles, occurring as a microcrystal or amorphous form. It is vital to recognize which kinds of proteins and/or polysaccharides act as a template to direct nanoparticle nucleation and growth? This should focus for further studies. This study may shed light on the process of formation of Se(0) nanosphere.

  2. Diversity in the antibacterial potential of probiotic cultures Bacillus licheniformis MCC2514 and Bacillus licheniformis MCC2512.

    Science.gov (United States)

    Shobharani, Papanna; Padmaja, Radhakrishnan J; Halami, Prakash M

    2015-01-01

    The aim of the present study was to investigate the characteristic diversity and stability of antimicrobial compounds produced by two probiotic strains of Bacillus licheniformis (MCC2514 and MCC2512). Antimicrobial compounds from the two strains notably varied, related to stability and potency. The inhibitory spectrum of B. licheniformis MCC2512 was higher than MCC2514, but, related to the effect on Micrococcus luteus ATCC9341, MCC2514 (LD50 = 450 AU ml(-1)) was more potent than MCC2512 (LD50 = 750 AU ml(-1)). The compounds were thermo-resistant and stable at a wide range of pH and exhibited considerable resistance to digestive enzymes and bile salts (anionic biological detergents), contributing to their appropriate application in various food systems. The isolate B. licheniformis MCC2512 gave a positive response to Bacillus subtilis-based biosensors BSF2470 and BS168.BS2, confirming the mode of action on the cell wall and subtilin-type, respectively. For B. licheniformis MCC2514, the mode of action was characterized by constructing B. subtilis reporters that interfered in five major biosynthetic pathways, i.e., biosynthesis of DNA, RNA, protein, the cell wall and fatty acids. B. licheniformis MCC2514 responded to the yvgS reporter, indicating it as an RNA synthesis inhibitor. Overall, the investigation reveals variability of the antimicrobial compounds from B. licheniformis of different origins and for their possible application as biopreservative agents.

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

    Directory of Open Access Journals (Sweden)

    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.

  4. 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.

  5. Combined Bacillus licheniformis and Bacillus subtilis infection in a patient with oesophageal perforation.

    Science.gov (United States)

    Jeon, You La; Yang, John Jeongseok; Kim, Min Jin; Lim, Gayoung; Cho, Sun Young; Park, Tae Sung; Suh, Jin-Tae; Park, Yong Ho; Lee, Mi Suk; Kim, Soo Cheol; Lee, Hee Joo

    2012-12-01

    Species of the genus Bacillus are a common laboratory contaminant, therefore, isolation of these organisms from blood cultures does not always indicate infection. In fact, except for Bacillus anthracis and Bacillus cereus, most species of the genus Bacillus are not considered human pathogens, especially in immunocompetent individuals. Here, we report an unusual presentation of bacteraemia and mediastinitis due to co-infection with Bacillus subtilis and Bacillus licheniformis, which were identified by 16S RNA gene sequencing, in a patient with an oesophageal perforation.

  6. Chitinase production by Bacillus thuringiensis and Bacillus licheniformis: their potential in antifungal biocontrol.

    Science.gov (United States)

    Gomaa, Eman Zakaria

    2012-02-01

    Thirty bacterial strains were isolated from the rhizosphere of plants collected from Egypt and screened for production of chitinase enzymes. Bacillus thuringiensis NM101-19 and Bacillus licheniformis NM120-17 had the highest chitinolytic activities amongst those investigated. The production of chitinase by B. thuringiensis and B. licheniformis was optimized using colloidal chitin medium amended with 1.5% colloidal chitin, with casein as a nitrogen source, at 30°C after five days of incubation. An enhancement of chitinase production by the two species was observed by addition of sugar substances and dried fungal mats to the colloidal chitin media. The optimal conditions for chitinase activity by B. thuringiensis and B. licheniformis were at 40°C, pH 7.0 and pH 8.0, respectively. Na(+), Mg(2+), Cu(2+), and Ca(2+) caused enhancement of enzyme activities whereas they were markedly inhibited by Zn(2+), Hg(2+), and Ag(+). In vitro, B. thuringiensis and B. licheniformis chitinases had potential for cell wall lysis of many phytopathogenic fungi tested. The addition of B. thuringiensis chitinase was more effective than that of B. licheniformis in increasing the germination of soybean seeds infected with various phytopathogenic fungi.

  7. 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

    A retrospective study of bovine abortions associated with Bacillus licheniformis is described. The material consisted of 2445 bovine abortions submitted for diagnostics from 1986 through 1993. Initially, B, licheniformis had been isolated from 81 cases. Sections of these cases were reexamined mic...... 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....

  8. 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 inoculate...... intravenously with B. licheniformis bacteria at doses from...

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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.

  11. 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

  12. Production of Active Bacillus licheniformis Alpha-Amylase in Tobacco and its Application in Starch Liquefaction

    NARCIS (Netherlands)

    Pen, J; MOLENDIJK, L; Quax, Wim J.; SIJMONS, PC; VANOOYEN, AJJ; VANDENELZEN, PJM; RIETVELD, K; HOEKEMA, A

    1992-01-01

    As a first example of the feasibility of producing industrial bulk enzymes in plants, we have expressed Bacillus licheniformis alpha-amylase in transgenic tobacco, and applied the seeds directly in starch liquification. The enzyme was properly secreted into the intercellular space, and maximum expre

  13. 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

  14. Hydrolysis of Whey Protein Isolate with Bacillus licheniformis Protease: Aggregating Capacities of Peptide Fractions

    NARCIS (Netherlands)

    Creusot, N.P.; Gruppen, H.

    2008-01-01

    In a previous study, peptides aggregating at pH 7.0 derived from a whey protein hydrolysate made with Bacillus licheniformis protease were fractionated and identified. The objective of the present work was to investigate the solubility of the fractionated aggregating pepticles, as a function of conc

  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. Mode of action of Bacillus licheniformis pectin methylesterase on highly methylesterified and acetylated pectins

    NARCIS (Netherlands)

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

    2015-01-01

    A gene encoding a putative pectinesterase from Bacillus licheniformis DSM13 was cloned and expressed in Escherichia coli. The resulting recombinant enzyme (BliPME) was purified and characterized as a pectin methylesterase. The enzyme showed maximum activity at pH 8.0 and 50 °C. BliPME is able to rel

  17. 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.

  18. Global transcriptional analysis of Bacillus licheniformis reveals an overlap between heat shock and iron limitation stimulon.

    Science.gov (United States)

    Nielsen, Allan K; Breüner, Anne; Krzystanek, Marcin; Andersen, Jens T; Poulsen, Thomas A; Olsen, Peter B; Mijakovic, Ivan; Rasmussen, Michael D

    2010-01-01

    In this study, we characterized the heat shock stimulon of the important industrial microorganism Bacillus licheniformis using DNA microarrays. While sharing a high degree of homology with the closely related model organism Bacillus subtilis, the heat shock stimulon of B. licheniformis exhibited several novel and unexpected features. Most notably, heat shock in B. licheniformis resulted in decreased amounts of mRNA from the ytrABCEF operon, encoding a putative acetoin uptake system, and stimulated the transcription of purine biosynthesis and iron uptake genes. Unexpectedly, deletion of the ytrEF genes did not affect acetoin uptake, but increased heat sensitivity. To investigate the connection between heat stress and iron uptake further, we analyzed the iron limitation response of B. licheniformis by DNA microarrays and concluded that the response mostly involves the genes related to iron uptake and metabolism, while the only heat shock gene affected by iron limitation was clpE. We also attempted to delete the fur gene (encoding the ferric uptake repressor), but unexpectedly found it to be essential in B. licheniformis. Using the fluorescent protein-encoding reporter gene under control of the dhb promoter, which responded to both heat shock and iron-starvation, we confirmed the overlap between these responses.

  19. High yield recombinant thermostable α-amylase production using an improved Bacillus licheniformis system

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    Shi Gui-Yang

    2009-10-01

    Full Text Available Abstract Background Some strains of Bacillus licheniformis have been improved by target-directed screening as well as by classical genetic manipulation and used in commercial thermostable α-amylase and alkaline protease production for over 40 years. Further improvements in production of these enzymes are desirable. Results A new strain of B. licheniformis CBBD302 carrying a recombinant plasmid pHY-amyL for Bacillus licheniformis α-amylase (BLA production was constructed. The combination of target-directed screening and genetic recombination led to an approximately 26-fold improvement of BLA production and export in B. licheniformis. Furthermore, a low-cost fermentation medium containing soybean meal and cottonseed meal for BLA production in shake-flasks and in a 15 liter bioreactor was developed and a BLA concentration of up to 17.6 mg per ml growth medium was attained. Conclusion This production level of BLA by B. licheniformis CBBD302(pHY-amyL is amongst the highest levels in Gram-positive bacteria reported so far.

  20. Investigation of the Antimicrobial Activity of Bacillus licheniformis Strains Isolated from Retail Powdered Infant Milk Formulae.

    Science.gov (United States)

    Alvarez-Ordóñez, Avelino; Begley, Máire; Clifford, Tanya; Deasy, Thérèse; Considine, Kiera; O'Connor, Paula; Ross, R Paul; Hill, Colin

    2014-03-01

    This study investigated the potential antimicrobial activity of ten Bacillus licheniformis strains isolated from retail infant milk formulae against a range of indicator (Lactococcus lactis, Lactobacillus bulgaricus and Listeria innocua) and clinically relevant (Listeria monocytogenes, Staphylococcus aureus, Streptococcus agalactiae, Salmonella Typhimurium and Escherichia coli) microorganisms. Deferred antagonism assays confirmed that all B. licheniformis isolates show antimicrobial activity against the Gram-positive target organisms. PCR and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analyses indicated that four of the B. licheniformis isolates produce the bacteriocin lichenicidin. The remaining six isolates demonstrated a higher antimicrobial potency than lichenicidin-producing strains. Further analyses identified a peptide of ~1,422 Da as the most likely bioactive responsible for the antibacterial activity of these six isolates. N-terminal sequencing of the ~1,422 Da peptide from one strain identified it as ILPEITXIFHD. This peptide shows a high homology to the non-ribosomal peptides bacitracin and subpeptin, known to be produced by Bacillus spp. Subsequent PCR analyses demonstrated that the six B. licheniformis isolates may harbor the genetic machinery needed for the synthesis of a non-ribosomal peptide synthetase similar to those involved in production of subpeptin and bacitracin, which suggests that the ~1,422 Da peptide might be a variant of subpeptin and bacitracin.

  1. Modification of the activity of an a-amylase from Bacillus licheniformis by several surfactants

    OpenAIRE

    2006-01-01

    The influence of different commercial surfactants on the enzymatic activity of a commercial ??-amylase from Bacillus licheniformis (Termamyl 300 L) has been studied. As non-ionic surfactants, alkyl polyglycosides (Glucopon?? 215, Glucopon?? 600 and Glucopon?? 650) were studied, as were fatty alcohol ethoxylates (Findet 1214N/23 and Findet 10/15), and nonyl phenol ethoxylate (Findet 9Q/21.5NF). Also, an anionic surfactant, linear alkyl benzene sulfonate (LAS) was assayed. In general, none of t...

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

    Directory of Open Access Journals (Sweden)

    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.

  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

    Directory of Open Access Journals (Sweden)

    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. Bacillus licheniformis isolated from Korean traditional food sources enhances the resistance of Caenorhabditis elegans to infection by Staphylococcus aureus.

    Science.gov (United States)

    Yun, Hyun Sun; Heo, Ju Hee; Son, Seok Jun; Park, Mi Ri; Oh, Sangnam; Song, Min-Ho; Kim, Jong Nam; Go, Gwang-Woong; Cho, Ho-Seong; Choi, Nag-Jin; Jo, Seung-Wha; Jeong, Do-Youn; Kim, Younghoon

    2014-08-01

    We investigated whether Bacillus spp., newly isolated from Korean traditional food resources, influence the resistance of hosts to foodborne pathogens, by using Caenorhabditis elegans as a surrogate host model. Initially, we selected 20 Bacillus spp. that possess antimicrobial activity against various foodborne pathogens, including Staphylococcus aureus. Among the selected strains, six strains of Bacillus spp. used in preconditioning significantly prolonged the survival of nematodes exposed to S. aureus. Based on 16S rRNA gene sequencing, all six strains were identified as B. licheniformis. Our findings suggest that preconditioning with B. licheniformis may modulate the host defense response against S. aureus.

  6. 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.

  7. Expression of α-Amylase Gene of Bacillus licheniformis in Saccharomyces cerevisiae

    Institute of Scientific and Technical Information of China (English)

    罗进贤; 黎杨; 李文清; 张添元; 何鸣

    1994-01-01

    The secretive expression vector has been constructed using the promoter and signal se-quence of yeast MF-α1 factor,and the Bacillus licheniformis α-amylase gene without promoter and signal se-quence has been inserted into the downstream of the signal sequence on the vector.After the readjustment ofthe reading frame,the amylase gene was expressed in Saccharomyces cerevisiae and the product was secretedfrom it.The properties of enzymes secreted from yeast and B.subtilis are compared,and the mechanism ofthe gene expression and product secretion are discussed.

  8. Binding of Tris to Bacillus licheniformis alpha-amylase can affect its starch hydrolysis activity.

    Science.gov (United States)

    Ghalanbor, Zahra; Ghaemi, Nasser; Marashi, Sayed-Amir; Amanlou, Massoud; Habibi-Rezaei, Mehran; Khajeh, Khosro; Ranjbar, Bijan

    2008-01-01

    Bacillus licheniformis alpha-amylase (BLA) is routinely used as a model thermostable amylase in biochemical studies. Its starch hydrolysis activity has recently been studied in Tris buffer. Here, we address the question that whether the application of Tris buffer may influence the results of BLA activity analyses. Based on the inhibition studies and docking simulations, we suggest that Tris molecule is a competitive inhibitor of starch-hydrolyzing activity of BLA, and it has a high tendency to bind the enzyme active site. Hence, it is critically important to consider such effect when interpreting the results of activity studies of this enzyme in Tris buffer.

  9. Modification of the activity of an alpha-amylase from Bacillus licheniformis by several surfactants

    OpenAIRE

    2006-01-01

    The influence of different commercial surfactants on the enzymatic activity of a commercial alpha-amylase from Bacillus licheniformis (Termamyl 300 L) has been studied. As non-ionic surfactants, alkyl polyglycosides (Glucopon® 215, Glucopon® 600 and Glucopon® 650) were studied, as were fatty alcohol ethoxylates (Findet 1214N/23 and Findet 10/15), and nonyl phenol ethoxylate (Findet 9Q/21.5NF). Also, an anionic surfactant, linear alkyl benzene sulfonate (LAS) was assayed. In general, none of t...

  10. Purification and characterization of gamma poly glutamic acid from newly Bacillus licheniformis NRC20.

    Science.gov (United States)

    Tork, Sanaa E; Aly, Magda M; Alakilli, Saleha Y; Al-Seeni, Madeha N

    2015-03-01

    γ-poly glutamic acid (γ-PGA) has received considerable attention for pharmaceutical and biomedical applications. γ-PGA from the newly isolate Bacillus licheniformis NRC20 was purified and characterized using diffusion distance agar plate, mass spectrometry and thin layer chromatography. All analysis indicated that γ-PGA is a homopolymer composed of glutamic acid. Its molecular weight was determined to be 1266 kDa. It was composed of L- and D-glutamic acid residues. An amplicon of 3050 represents the γ-PGA-coding genes was obtained, sequenced and submitted in genbank database. Its amino acid sequence showed high similarity with that obtained from B. licheniformis strains. The bacterium NRC 20 was independent of L-glutamic acid but the polymer production enhanced when cultivated in medium containing L-glutamic acid as the sole nitrogen source. Finally we can conclude that γ-PGA production from B. licheniformis NRC20 has many promised applications in medicine, industry and nanotechnology.

  11. Metal-Binding Characteristics of the Gamma-Glutamyl Capsular Polymer of Bacillus licheniformis ATCC 9945.

    Science.gov (United States)

    McLean, R J; Beauchemin, D; Clapham, L; Beveridge, T J

    1990-12-01

    The metal-binding affinity of the anionic poly-gamma-d-glutamyl capsule of Bacillus licheniformis was investigated by using Na, Mg, Al, Ca, Cr, Mn, Fe, Ni, and Cu. Purified capsule was suspended in various concentrations of the chloride salts of the various metals, and after dialysis the bound metals were analyzed either by graphite furnace atomic absorption spectroscopy or by inductively coupled plasma-mass spectrometry. Exposure of purified capsule to excess concentrations of Na revealed it to contain 8.2 mumol of anionic sites per mg on the basis of Na binding. This was confirmed by titration of the capsule with HCl and NaOH. Other metal ions were then added in ionic concentrations equivalent to 25, 50, 75, 100, 200, and 400% of the available anionic sites. The binding characteristics varied with the metal being investigated. Addition of Cu, Al, Cr, or Fe induced flocculation. These metal ions showed the greatest affinity for B. licheniformis capsule in competitive-binding experiments. Flocculation was not seen with the addition of other metal ions. With the exception of Ni and Fe all capsule-metal-binding sites readily saturated. Ni had low affinity for the polymer, and its binding was increased at high metal concentrations. Fe binding resulted in the development of rust-colored ferrihydrite which itself could bind additional metal. Metal-binding characteristics of B. licheniformis capsule appear to be influenced by the chemical and physical properties of both the capsule and the metal ions.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    2005-01-01

    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. PMID:16192686

  14. Purification and characterization of a thermostable extracellular phytase from Bacillus licheniformis PFBL-03.

    Science.gov (United States)

    Fasimoye, Feyisola O; Olajuyigbe, Folasade M; Sanni, Morakinyo D

    2014-01-01

    Extracellular phytase from Bacillus licheniformis PFBL-03 was purified in three steps by using ammonium sulfate precipitation, ion-exchange chromatography on a DEAE Sephadex A-50 column, and gel filtration chromatography on Sephadex G-100. The single protein band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) suggested that the enzyme was homogeneous. The molecular mass determined from SDS-PAGE was 36 kD. The enzyme yield was 10% while the purification fold was 39. The purified phytase exhibited optimum activity at 55°C and was able to retain 55% of its original activity after 60 min of incubation at 80°C. The purified enzyme had optimum pH of 6.0 and was stable over a pH range of 4.0 to 7.5. The kinetic parameters K(m) and V(max) of the purified phytase for sodium phytate were 4.7 mM and 49.01 µmol/min. The activity of the enzyme was enhanced in the presence of Ca²⁺ but completely inhibited by Fe²⁺, Mn²⁺, and Cu²⁺. The exhibited characteristics of the purified phytase from Bacillus licheniformis PFBL-03 show that the enzyme has potential for applications in the animal feed industry.

  15. Construction and application of recombinant strain for the production of an alkaline protease from Bacillus licheniformis.

    Science.gov (United States)

    Lin, Songyi; Zhang, Meishuo; Liu, Jingbo; Jones, Gregory S

    2015-03-01

    The alkaline protease gene, Apr, from Bacillus licheniformis 2709 was cloned into an expression vector pET - 28b (+), to yield the recombinant plasmid pET-28b (+) - Apr. The pET-28b (+) - Apr was expressed in a high expression strain E. coli BL21. The amino acid sequence deduced from the DNA sequence analysis revealed a 98% identity to that of Bacillus licheniformis 2709. Sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE) was used to access the protein expression. SDS-PAGE analysis indicated a protein of Mr of 38.8 kDa. The medium components and condition of incubation were optimized for the growth state of a recombinant strain. The optimal composition of production medium was composed of glucose 8 g/L, peptone 8 g/L and salt solution 10 mL. The samples were incubated on a rotary shaker of 180 r/min at 37°C for 24 h.

  16. Proteomics study of extracellular fibrinolytic proteases from Bacillus licheniformis RO3 and Bacillus pumilus 2.g isolated from Indonesian fermented food

    Science.gov (United States)

    Nur Afifah, Diana; Rustanti, Ninik; Anjani, Gemala; Syah, Dahrul; Yanti; Suhartono, Maggy T.

    2017-02-01

    This paper presents the proteomics study which includes separation, identification and characterization of proteins. The experiment on Indonesian fermented food such as extracellular fibrinolytic protease from Bacillus licheniformis RO3 and Bacillus pumilus 2.g isolated from red oncom and tempeh gembus was conducted. The experimental works comprise the following steps: (1) a combination of one- and two-dimensional electrophoresis analysis, (2) mass spectrometry analysis using MALDI-TOF-MS and (3) investigation using protein database. The result suggested that there were new two protein fractions of B. licheniformis RO3 and three protein fractions of B. pumilus 2.g. These result has not been previously reported.

  17. Draft Genome Sequence of Bacillus licheniformis Strain YNP1-TSU Isolated from Whiterock Springs in Yellowstone National Park.

    Science.gov (United States)

    O'Hair, Joshua A; Li, Hui; Thapa, Santosh; Scholz, Matthew B; Zhou, Suping

    2017-03-02

    Novel cellulolytic microorganisms can potentially influence second-generation biofuel production. This paper reports the draft genome sequence of Bacillus licheniformis strain YNP1-TSU, isolated from hydrothermal-vegetative microbiomes inside Yellowstone National Park. The assembled sequence contigs predicted 4,230 coding genes, 66 tRNAs, and 10 rRNAs through automated annotation.

  18. Draft Genome Sequence of Bacillus licheniformis Strain GB2, a Hydrocarbon-Degrading and Plant Growth-Promoting Soil Bacterium.

    Science.gov (United States)

    Gkorezis, Panagiotis; Van Hamme, Jonathan; Bottos, Eric; Thijs, Sofie; Balseiro-Romero, Maria; Monterroso, Carmela; Kidd, Petra Suzan; Rineau, Francois; Weyens, Nele; Sillen, Wouter; Vangronsveld, Jaco

    2016-06-23

    We report the 4.39 Mb draft genome of Bacillus licheniformis GB2, a hydrocarbonoclastic Gram-positive bacterium of the family Bacillaceae, isolated from diesel-contaminated soil at the Ford Motor Company site in Genk, Belgium. Strain GB2 is an effective plant-growth promoter useful for diesel fuel remediation applications based on plant-bacterium associations.

  19. Draft Genome Sequence of Bacillus licheniformis Strain YNP1-TSU Isolated from Whiterock Springs in Yellowstone National Park

    Science.gov (United States)

    O'Hair, Joshua A.; Li, Hui; Thapa, Santosh; Scholz, Matthew B.

    2017-01-01

    ABSTRACT Novel cellulolytic microorganisms can potentially influence second-generation biofuel production. This paper reports the draft genome sequence of Bacillus licheniformis strain YNP1-TSU, isolated from hydrothermal-vegetative microbiomes inside Yellowstone National Park. The assembled sequence contigs predicted 4,230 coding genes, 66 tRNAs, and 10 rRNAs through automated annotation. PMID:28254968

  20. 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

    2015-12-02

    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.

  1. The fnr Gene of Bacillus licheniformis and the Cysteine Ligands of the C-Terminal FeS Cluster

    OpenAIRE

    Klinger, Anette; Schirawski, Jan; Glaser, Philippe; Unden, Gottfried

    1998-01-01

    In the facultatively anaerobic bacterium Bacillus licheniformis a gene encoding a protein of the fumarate nitrate reductase family of transcriptional regulators (Fnr) was isolated. Unlike Fnr proteins from gram-negative bacteria, but like Fnr from Bacillus subtilis, the protein contained a C-terminal cluster of cysteine residues. Unlike in Fnr from B. subtilis, this cluster (Cys226-X2-Cys229-X4-Cys234) is composed of only three Cys residues, which are supposed to serve together with an intern...

  2. Soybean fermentation with Bacillus licheniformis increases insulin sensitizing and insulinotropic activity.

    Science.gov (United States)

    Yang, Hye Jeong; Kwon, Dae Young; Moon, Na Rang; Kim, Min Jung; Kang, Hee Joo; Jung, Do Yeon; Park, Sunmin

    2013-11-01

    Traditionally fermented soybeans (chungkookjang; TFC) may have potent anti-diabetic activity, depending on the ambient microorganisms and conditions. We hypothesized that one of the major Bacillus species in TFC contributes to the anti-diabetic activity and could be used to standardize a highly functional TFC. We tested the hypothesis by using cell-based studies to evaluate insulin sensitizing and insulinotropic action of chungkookjangs fermented with various Bacillus spp. and fermentation periods. The 70% methanol and water extracts of chungkookjang fermented with Bacillus licheniformis (BL) for 48 h contained similar profiles of isoflavonoids and peptides to methanol and water extracts of TFC with potent anti-diabetic activity. Water extracts (mainly containing peptides) of TFC and BL fermented for 48 h and 72 h had a better insulin sensitizing action via activating peroxisome proliferator-activated receptor-γ (PPAR-γ) and increased the expression of PPAR-γ in 3T3-L1 adipocytes better than unfermented cooked soybeans (CSB). The 70% methanol extracts (predominantly isoflavone aglycones) of BL fermented for 48 h and 72 h improved glucose-stimulated insulin secretion and protected β-cell viability better than CSB in insulinoma cells, and the improvement by BL was similar to TFC. In conclusion, the BL water extract fermented for 48 h exhibited equal insulin sensitization as TFC and BL methanol extract exerted similar insulinotropic actions to those of TFC. B. licheniformis may be one of the major microorganisms responsible for anti-diabetic actions of chungkookjang. It is important to make chungkookjang that retains the anti-diabetic properties of the most efficacious traditional chungkookjang using a standardized method.

  3. 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...

  4. Isolation and characterization of different strains of Bacillus licheniformis for the production of commercially significant enzymes.

    Science.gov (United States)

    Ghani, Maria; Ansari, Asma; Aman, Afsheen; Zohra, Rashida Rahmat; Siddiqui, Nadir Naveed; Qader, Shah Ali Ul

    2013-07-01

    Utilization of highly specific enzymes for various industrial processes and applications has gained huge momentum in the field of white biotechnology. Selection of a strain by efficient plate-screening method for a specific purpose has also favored and boosted the isolation of several industrially feasible microorganisms and screening of a large number of microorganisms is an important step in selecting a potent culture for multipurpose usage. Five new bacterial isolates of Bacillus licheniformis were discovered from indigenous sources and characterized on the basis of phylogeny using 16S rDNA gene analysis. Studies on morphological and physiological characteristics showed that these isolates can easily be cultivated at different temperatures ranging from 30°C to 55°C with a wide pH values from 3.0 to 11.0 All these 05 isolates are salt tolerant and can grow even in the presences of high salt concentration ranging from 7.0 to 12.0%. All these predominant isolates of B. licheniformis strains showed significant capability of producing some of the major industrially important extracellular hydrolytic enzymes including α-amylase, glucoamylase, protease, pectinase and cellulase in varying titers. All these isolates hold great potential as commercial strains when provided with optimum fermentation conditions.

  5. Antimicrobial activity of a biosurfactant produced by Bacillus licheniformis strain M104 grown on whey

    Directory of Open Access Journals (Sweden)

    Eman Zakaria Gomaa

    2013-04-01

    Full Text Available The aim of the present study was to investigate the antimicrobial effect of the lipopeptide biosurfactants produced by Bacillus licheniformis strain M104 grown on whey. The biosurfactant was investigated for potential antimicrobial activity by using the disc-diffusion method against different Gram-positive bacteria {B subtilis, B. thuringiensis (two strains, B. cereus, Staphylococcus aureus (two strains and Listeria monocytogenes}, Gram-negative bacteria {(Pseudomonas aeruginosa, Escherichia coli (two strains, Salmonella typhimurium, Proteous vulgaris and Klebsiella pneumoniae and a yeast (Candida albicans}. The biosurfactant showed profoundly distinct antibacterial activity toward tested bacteria and displayed an antifungal activity against the tested yeast. Maximum antimicrobial activity of the biosurfactant was shown against S. aureus ATCC 25928. The biosurfactant had a broad inhibition effect on intracellular components of S. aureus ATCC 25928. The antimicrobial effect of lipopeptide biosurfactant produced by B. licheniformis strain M104 was time and concentration dependent. When biosurfactant was added to S. aureus medium in a concentration of (48 μg / ml, the maximum reduction of acid soluble phosphorous (53.06 %, total lipid (90.47 % total proteins (53.43%, RNA (83.29 % and DNA (48.50% were recorded after 12 h of incubation period. From the preliminary characterization results, it could be concluded that biosurfactants were a suitable alternative in potential applications of medical fields.

  6. Safety assessment of Bacillus licheniformis Me1 isolated from milk for probiotic application.

    Science.gov (United States)

    Nithya, Vadakedath; Muthukumar, Serva P; Halami, Prakash M

    2012-06-01

    In this study, an in vivo toxicological safety assessment of Bacillus licheniformis Me1, a native isolate from milk, was performed. An acute toxicity study in male albino Wistar rats demonstrated no treatment-related illness or mortality. A 90-day subchronic oral toxicity study using 2 doses (1.1 × 10(10) and 1.1 × 10(11) colony-forming unit [CFU]/kg body weight [BW], respectively) failed to show dose-dependent illness or mortality. Moreover, neither significant differences in serum biochemical and hematological analyses nor histopathological changes in organs or tissues were found when compared to the control groups. The no-observed-adverse-effect level (NOAEL) was found to be greater than 1.1 × 10(11) CFU/kg BW. The in vivo micronucleus assay in mice did not reveal any signs of genotoxic effect at any of the doses tested. Furthermore, dermal and acute eye irritation tests conducted in rabbits showed no edema or erythema and ocular lesions. These results suggest that B licheniformis Me1 can be considered safe for food industry applications.

  7. Microbial surfactant mediated degradation of anthracene in aqueous phase by marine Bacillus licheniformis MTCC 5514

    Directory of Open Access Journals (Sweden)

    Sreethar Swaathy

    2014-12-01

    Full Text Available The present study emphasizes the biosurfactant mediated anthracene degradation by a marine alkaliphile Bacillus licheniformis (MTCC 5514. The isolate, MTCC 5514 degraded >95% of 300 ppm anthracene in an aqueous medium within 22 days and the degradation percentage reduced significantly when the concentration of anthracene increased to above 500 ppm. Naphthalene, naphthalene 2-methyl, phthalic acid and benzene acetic acid are the products of degradation identified based on thin layer chromatography, high performance liquid chromatography, gas chromatography and mass analyses. It has been observed that the degradation is initiated by the biosurfactant of the isolate for solubilization through micellation and then the alkali pH and intra/extra cellular degradative enzymes accomplish the degradation process. Encoding of genes responsible for biosurfactant production (licA3 as well as catabolic reactions (C23O made with suitable primers designed. The study concludes in situ production of biosurfactant mediates the degradation of anthracene by B. licheniformis.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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.

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

    Science.gov (United States)

    Paraneeiswaran, Arunachalam; Shukla, Sudhir K; Prashanth, K; Rao, T Subba

    2015-01-01

    Naturally stressed habitats are known to be repositories for novel microorganisms with potential bioremediation applications. In this study, we isolated a [Co(III)-EDTA](-) 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](-) 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](2-) is less stable though it is toxic. This study showed that B. licheniformis SPB-2 reduced 1mM [Co(III)-EDTA](-) 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](-) 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](-) reduction. More detailed investigations showed that after [Co(III)-EDTA](-) reduction, i.e. [Co(II)-EDTA](2-) complex was removed by B. licheniformis SPB-2 from the bulk liquid by adsorption phenomenon. The bacterium showed a D10 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. Production of the novel two-peptide lantibiotic lichenicidin by Bacillus licheniformis DSM 13.

    Directory of Open Access Journals (Sweden)

    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

  13. Effect of Bacillus licheniformis and Bacillus subtilis supplementation of ewe's feed on sheep milk production and young lamb mortality.

    Science.gov (United States)

    Kritas, S K; Govaris, A; Christodoulopoulos, G; Burriel, A R

    2006-05-01

    The purpose of this pilot study was to evaluate under field conditions the effect of a probiotic containing Bacillus licheniformis and Bacillus subtilis on young lamb mortality and sheep milk production when administered in the late pregnancy and lactation feed of ewes. In a sheep farm, two groups of milking ewes with identical genetic material, management, nutrition, health status and similar production characteristics were formed. One group (46 ewes) served as control, while the other one (48 ewes) served as a probiotic-treated group. Both groups of ewes received a similar feeding regiment, but the ewes of the second group were additionally offered a probiotic product containing B. licheniformis and B. subtilis (BioPlus 2B, Chr. Hansen, Denmark) at the approximate dose of 2.56 x 10(9) viable spores per ewe per day. Lamb mortality during the 1.5 months suckling period, and milk yield during the 2 months of milk collection for commercial purposes have been recorded. In the non-treated control group, 13.1% mortality was observed versus 7.8% in the probiotic-treated group (P = 0.33), with mortality being mainly due to diarrhoea. Microbiological examination of diarrhoeic faeces from some of the dead lambs in both groups revealed the presence of Escherichia coli. The average daily milk yield per ewe was significantly lower in the control group (0.80 l) than that in the probiotic-treated group (0.93 l) (P milk in ewes that received probiotics was significantly (P milk yields, fat and protein content.

  14. Optimization of fermentation conditions for cellulases production by Bacillus licheniformis MVS1 and Bacillus sp. MVS3 isolated from Indian hot spring

    Directory of Open Access Journals (Sweden)

    Somen Acharya

    2012-08-01

    Full Text Available The aim of this work was to study the effect of some nutritional and environmental factors on the production of cellulases, in particular endoglucanase (CMCase and exoglucanases (FPase from Bacillus licheniformis MVS1 and Bacillus sp. MVS3 isolated from an Indian hot spring. The characterization study indicated that the optimum pH and temperature value was 6.5 to 7.0 and 50-55°C, respectively. Maximum cellulases production by both the isolates was detected after 60 h incubation period using wheat and rice straw. The combination of inorganic and organic nitrogen source was suitable for cellulases production. Overall, FPase production was much higher than CMCase production by both of the strains. Between the two thermophiles, the cellulolytic activity was more in B.licheniformis MVS1 than Bacillus sp. MVS3 in varying environmental and nutritional conditions.

  15. The fnr gene of Bacillus licheniformis and the cysteine ligands of the C-terminal FeS cluster.

    Science.gov (United States)

    Klinger, A; Schirawski, J; Glaser, P; Unden, G

    1998-07-01

    In the facultatively anaerobic bacterium Bacillus licheniformis a gene encoding a protein of the fumarate nitrate reductase family of transcriptional regulators (Fnr) was isolated. Unlike Fnr proteins from gram-negative bacteria, but like Fnr from Bacillus subtilis, the protein contained a C-terminal cluster of cysteine residues. Unlike in Fnr from B. subtilis, this cluster (Cys226-X2-Cys229-X4-Cys234) is composed of only three Cys residues, which are supposed to serve together with an internal residue (Cys71) as the ligands for an FeS center. Transfer of the B. licheniformis gene to an fnr mutant of B. subtilis complemented the ability for synthesis of nitrate reductase during anaerobic growth.

  16. 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.

  17. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  19. 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.

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

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    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.

  1. Concomitant production of protease and lipase by Bacillus licheniformis VSG1: production, purification and characterization

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    R. Sangeetha

    2010-03-01

    Full Text Available This study was aimed at producing protease and lipase simultaneously on a common medium by Bacillus licheniformis VSG1, which was isolated from a tannery effluent. The effect of media composition with respect to protein source, lipid source and emulsifier on the production of protease and lipase was analysed. Both those enzymes were produced under optimized conditions like pH, temperature and incubation time. The enzyme mixture comprising of both protease and lipase was purified by ammonium sulphate precipitation, dialysis and gel filtration chromatography to obtain 20-fold pure enzymes. The purified enzyme mixture was characterized to determine the optimum pH and temperature of protease and lipase, the response of the enzymes to inhibitors, additives and solvents. The molecular weight of both the enzymes was determined as 40 kDa on SDS-PAGE. The concomitant production of protease and lipase and the purification of both the enzymes in a single mixture have industrial significance, as many industrial processes use both protease and lipase together.

  2. 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.

  3. 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.

  4. Enhanced Production of Poly-γ-glutamic Acid by Bacillus licheniformis TISTR 1010 with Environmental Controls.

    Science.gov (United States)

    Kongklom, Nuttawut; Shi, Zhongping; Chisti, Yusuf; Sirisansaneeyakul, Sarote

    2016-12-24

    Bacillus licheniformis TISTR 1010 was used for glutamic acid-independent production of poly-γ-glutamic acid (γ-PGA). A fed-batch production strategy was developed involving feedings of glucose, citric acid, and ammonium chloride at specified stages of the fermentation. With the dissolved oxygen concentration controlled at ≥50% of air saturation and the pH controlled at ~7.4, the fed-batch operation at 37 °C afforded a peak γ-PGA concentration of 39.9 ± 0.3 g L(-1) with a productivity of 0.926 ± 0.006 g L(-1) h(-1). The observed productivity was nearly threefold greater than previously reported for glutamic acid-independent production using the strain TISTR 1010. The molecular weight of γ-PGA was in the approximate range of 60 to 135 kDa.

  5. 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 (... present. Solubility was studied in the pH range of 6 to 8. The lowest solubility without added salts was 60 mg.mL(-1) at pH 7. The addition of 0.1 mol.L-1 sodium salts of nitrate, sulfate, and thiocyanate had a small effect on solubility. However, solubility was lowered significantly by adding 0.5 mol.L-1...... 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...

  6. 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).

  7. Alcohol-induced structural transitions in the acid-denatured Bacillus licheniformis α-amylase

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    Adyani Azizah Abd Halim

    2017-01-01

    Full Text Available Alcohol-induced structural changes in the acid-denatured Bacillus licheniformis α-amylase (BLA at pH 2.0 were studied by far-ultra violet circular dichroism, intrinsic, three-dimensional and 8-anilino-1-naphthalene sulfonic acid (ANS fluorescence, acrylamide quenching and thermal denaturation. All the alcohols used in this study produced partial refolding in the acid-denatured BLA as evident from the increased mean residue ellipticity at 222 nm, increased intrinsic fluorescence and decreased ANS fluorescence. The order of effectiveness of these alcohols to induce a partially folded state of BLA was found to be: 2,2,2-trifluoroethanol/tert-butanol > 1-propanol/2-propanol > 2-chloroethanol > ethanol > methanol. Three-dimensional fluorescence and acrylamide quenching results obtained in the presence of 5.5 M tert-butanol also suggested formation of a partially folded state in the acid-denatured BLA. However, 5.5 M tert-butanol-induced state of BLA showed a non-cooperative thermal transition. All these results suggested formation of a partially folded state of the acid-denatured BLA in the presence of these alcohols. Furthermore, their effectiveness was found to be guided by their chain length, position of methyl groups and presence of the substituents.

  8. Hydrolysis of whey protein isolate with Bacillus licheniformis protease: aggregating capacities of peptide fractions.

    Science.gov (United States)

    Creusot, Nathalie; Gruppen, Harry

    2008-11-12

    In a previous study, peptides aggregating at pH 7.0 derived from a whey protein hydrolysate made with Bacillus licheniformis protease were fractionated and identified. The objective of the present work was to investigate the solubility of the fractionated aggregating peptides, as a function of concentration, and their aggregating capacities toward added intact proteins. The amount of aggregated material and the composition of the aggregates obtained were measured by nitrogen concentration and size exclusion chromatography, respectively. The results showed that of the four fractions obtained from the aggregating peptides, two were insoluble, while the other two consisted of 1:1 mixture of low and high solubility peptides. Therefore, insoluble peptides coaggregated, assumedly via hydrophobic interactions, other relatively more soluble peptides. It was also shown that aggregating peptides could aggregate intact protein nonspecifically since the same peptides were involved in the aggregation of whey proteins, beta-casein, and bovine serum albumin. Both insoluble and partly insoluble peptides were required for the aggregation of intact protein. These results are of interest for the applications of protein hydrolysates, as mixtures of intact protein and peptides are often present in these applications.

  9. Adsorption and reduction of palladium (Pd2+) by Bacillus licheniformis R08

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Preliminary study on the mechanism of Pd2+ biosorption by resting cells of Bacillus licheniformis R08 biomass has been carried out by means of chemical kinetics and AAS, TEM, XRD and FTIR methods. The results showed that at 30℃ and pH 3.5, when dry R08 biomass powder (800 mg/L) was mixed with Pd2+ (100 mg/L) for 45 min, the rate constant k of biosorption of Pd2+ attained a maximum of 5.97×10-2 min-1 and the half life period of the reaction reached 12 min. The part of Pd2+ adsorbed by R08 biomass was reduced to elemental, cell-bound Pd0 at the same condition. The cell wall of R08 biomass was the primary location for accumulating Pd2+, and aldoses, I. E. Hydrolysate of a part of polysaccharides on the peptidoglycan layer in the acidic medium, serving as the electron donor, in situ reduced the Pd2+ to Pd0.

  10. 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.

  11. 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.

  12. Cloning and characterization of a new laccase from Bacillus licheniformis catalyzing dimerization of phenolic acids.

    Science.gov (United States)

    Koschorreck, Katja; Richter, Sven M; Ene, Augusta B; Roduner, Emil; Schmid, Rolf D; Urlacher, Vlada B

    2008-05-01

    A new laccase gene (cotA) was cloned from Bacillus licheniformis and expressed in Escherichia coli. The recombinant protein CotA was purified and showed spectroscopic properties, typical for blue multi-copper oxidases. The enzyme has a molecular weight of approximately 65 kDa and demonstrates activity towards canonical laccase substrates 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), syringaldazine (SGZ) and 2,6-dimethoxyphenol (2,6-DMP). Kinetic constants KM and kcat for ABTS were of 6.5+/-0.2 microM and 83 s(-1), for SGZ of 4.3+/-0.2 microM and 100 s(-1), and for 2,6-DMP of 56.7+/-1.0 microM and 28 s(-1). Highest oxidizing activity towards ABTS was obtained at 85 degrees C. However, after 1 h incubation of CotA at 70 degrees C and 80 degrees C, a residual activity of 43% and 8%, respectively, was measured. Furthermore, oxidation of several phenolic acids and one non-phenolic acid by CotA was investigated. CotA failed to oxidize coumaric acid, cinnamic acid, and vanillic acid, while syringic acid was oxidized to 2,6-dimethoxy-1,4-benzoquinone. Additionally, dimerization of sinapic acid, caffeic acid, and ferulic acid by CotA was observed, and highest activity of CotA was found towards sinapic acid.

  13. Production, Characterization, and Application of Bacillus licheniformis W16 Biosurfactant in Enhancing Oil Recovery

    Science.gov (United States)

    Joshi, Sanket J.; Al-Wahaibi, Yahya M.; Al-Bahry, Saif N.; Elshafie, Abdulkadir E.; Al-Bemani, Ali S.; Al-Bahri, Asma; Al-Mandhari, Musallam S.

    2016-01-01

    The biosurfactant production by Bacillus licheniformis W16 and evaluation of biosurfactant based enhanced oil recovery (EOR) using core-flood under reservoir conditions were investigated. Previously reported nine different production media were screened for biosurfactant production, and two were further optimized with different carbon sources (glucose, sucrose, starch, cane molasses, or date molasses), as well as the strain was screened for biosurfactant production during the growth in different media. The biosurfactant reduced the surface tension and interfacial tension to 24.33 ± 0.57 mN m−1 and 2.47 ± 0.32 mN m−1 respectively within 72 h, at 40°C, and also altered the wettability of a hydrophobic surface by changing the contact angle from 55.67 ± 1.6 to 19.54°± 0.96°. The critical micelle dilution values of 4X were observed. The biosurfactants were characterized by different analytical techniques and identified as lipopeptide, similar to lichenysin-A. The biosurfactant was stable over wide range of extreme environmental conditions. The core flood experiments showed that the biosurfactant was able to enhance the oil recovery by 24–26% over residual oil saturation (Sor). The results highlight the potential application of lipopeptide biosurfactant in wettability alteration and microbial EOR processes. PMID:27933041

  14. Production, Characterization and Application of Bacillus licheniformis W16 Biosurfactant in Enhancing Oil Recovery

    Directory of Open Access Journals (Sweden)

    Sanket J. Joshi

    2016-11-01

    Full Text Available The biosurfactant production by Bacillus licheniformis W16 and evaluation of biosurfactant based enhanced oil recovery using core-flood under reservoir conditions were investigated. Previously reported nine different production media were screened for biosurfactant production, and two were further optimized with different carbon sources (glucose, sucrose, starch, cane molasses or date molasses, as well as the strain was screened for biosurfactant production during the growth in different media. The biosurfactant reduced the surface tension and interfacial tension to 24.33+0.57mN m-1 and 2.47+0.32mN m-1 respectively within 72h, at 40 C, and also altered the wettability of a hydrophobic surface by changing the contact angle from 55.67°+1.6° to 19.54°+0.96°. The critical micelle dilution values of 4X were observed. The biosurfactants were characterized by different analytical techniques and identified as lipopeptide, similar to lichenysin-A. The biosurfactant was stable over wide range of extreme environmental conditions. The core flood experiments showed that the biosurfactant was able to enhance the oil recovery by 24-26% over residual oil saturation (Sor. The results highlight the potential application of lipopeptide biosurfactant in wettability alteration and microbial enhanced oil recovery processes.

  15. 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.

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

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    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.

  17. Swapping of pro-sequences between keratinases of Bacillus licheniformis and Bacillus pumilus: altered substrate specificity and thermostability.

    Science.gov (United States)

    Rajput, Rinky; Tiwary, Ekta; Sharma, Richa; Gupta, Rani

    2012-08-10

    Pro-sequences were swapped in cis between keratinases from Bacillus licheniformis (Ker BL) and Bacillus pumilus (Ker BP) to construct Ker ProBP-BL and Ker ProBL-BP, respectively. Expression of these keratinases was carried out constitutively by E. coli HB101-pEZZ18 system. They were characterized with respect to their parent enzymes, Ker BL and Ker BP, respectively. Ker ProBP-BL became more thermostable with a t(1/2) of 45 min at 80°C contrary to Ker BL which was not stable beyond 60°C. Similarly, the activity of Ker ProBP-BL on keratin and casein substrate, i.e. K:C ratio increased to 1.2 in comparison to 0.1 for Ker BL. Hydrolysis of insulin B-chain revealed that the cleavage sites increased to six from four in case of Ker ProBP-BL in comparison to Ker BL. However, cleavage sites decreased from seven to four in case of Ker ProBL-BP in comparison to the parent keratinase, Ker BP. Likewise, Ker ProBL-BP revealed altered pH and temperature kinetics with optima at pH 10 and 60°C in comparison to Ker BP which had optima at pH 9 and 70°C. It also cleaved soluble substrates with better efficiency in comparison to Ker BP with K:C ratio of 1.6. Pro-sequence mediated conformational changes were also observed in trans and were almost similar to the features acquired by the chimeras constructed in cis by swapping the pro-sequence region.

  18. 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.

  19. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Inhibition of Bacillus licheniformis spore growth in milk by nisin, monolaurin, and pH combinations.

    Science.gov (United States)

    Mansour, M; Amri, D; Bouttefroy, A; Linder, M; Milliere, J B

    1999-02-01

    The effects of nisin and monolaurin, alone and in combination, were investigated on Bacillus licheniformis spores in milk at 37 degrees C. In the absence of inhibitors, germinated spores developed into growing vegetative cells and started sporulation at the end of the exponential phase. In the presence of nisin (25 IU ml-1), spore outgrowth was inhibited (4 log10 reduction at 10 h). Regrowth appeared between 10 and 24 h and reached a high population level (1.25 x 10(8) cfu ml-1) after 7 d. Monolaurin (250 micrograms ml-1) had a bacteriostatic effect during the first 10 h but thereafter, regrowth occurred slowly with a population level after 7 d (4 x 10(5) cfu ml-1) lower than that of nisin. Different combined effects of nisin (between 0 and 42 IU ml-1), monolaurin (ranging from 0 to 300 micrograms ml-1), pH values (between 5.0 and 7.0) and spore loads (10(3), 10(4), 10(5) spores ml-1) were investigated using a Doehlert matrix in order to study the main effects of these factors and the different interactions. Results were analysed using the Response Surface Methodology (RSM) and indicated that nisin and monolaurin had no action on spores before germination; only pH values had a significant effect (P monolaurin (100 micrograms ml-1) in combination acted synergistically on outgrown spores and vegetative cells, showing total inhibition at pH 6.0, without regrowth, within 7 d at 37 degrees C.

  2. 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.

  3. 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.

  4. Draft Genome Sequences of Three Cellulolytic Bacillus licheniformis Strains Isolated from Imperial Geyser, Amphitheater Springs, and Whiterock Springs inside Yellowstone National Park

    Science.gov (United States)

    O' Hair, Joshua A.; Li, Hui; Thapa, Santosh; Scholz, Matthew

    2017-01-01

    ABSTRACT Novel cellulolytic microorganisms are becoming more important for rapidly growing biofuel industries. This paper reports the draft genome sequences of Bacillus licheniformis strains YNP2-TSU, YNP3-TSU, and YNP5-TSU. These cellulolytic isolates were collected from several hydrothermal features inside Yellowstone National Park. PMID:28360181

  5. 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.

    OpenAIRE

    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.

  6. 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

    2016-05-12

    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.

  7. Crystal structure of thermostable alpha-amylase from Bacillus licheniformis refined at 1.7 A resolution.

    Science.gov (United States)

    Hwang, K Y; Song, H K; Chang, C; Lee, J; Lee, S Y; Kim, K K; Choe, S; Sweet, R M; Suh, S W

    1997-04-30

    alpha-Amylases (alpha-1,4-glucan-4-glucanohydrolase, E.C.3.2.1.1) catalyze the cleavage of alpha-1, 4-glucosidic linkages of starch components, glycogen, and various oligosaccharides. Thermostable alpha-amylases from Bacillus species are of great industrial importance in the production of corn syrup or dextrose. Thermostable alpha-amylase from Bacillus licheniformis, a monomeric enzyme with molecular mass of 55,200 Da (483 amino acid residues), shows a remarkable heat stability. This enzyme provides an attractive model for investigating the structural basis for thermostability of proteins. The three-dimensional structure of thermostable alpha-amylase from Bacillus licheniformis has been determined by the multiple isomorphous replacement method of X-ray crystallography. The structure has been refined to a crystallographic R-factor of 19.9% for 58,601 independent reflections with F0 > 2 sigma F0 between 8.0 and 1.7 A resolution, with root mean square deviations of 0.013 A from ideal bond lengths and 1.72 degrees from ideal bond angles. The final model consists of 469 amino acid residues and 294 water molecules. Missing from the model are the N- and C-termini and the segment between Trp182 and Asn192. Like other alpha-amylases, the polypeptide chain folds into three distinct domains. The first domain (domain A), consisting of 291 residues (from residue 3 to 103 and 207 to 396), forms a (beta/alpha)8-barrel structure. The second domain (domain B), consisting of residues 104 to 206, is inserted between the third beta-strand and the third alpha-helix of domain A. The third C-terminal domain (domain C), consisting of residues 397 to 482, folds into an eight-stranded antiparallel beta-barrel. Neither calcium ion nor chloride ion is located near the active site. This study reveals the architecture of the thermostable alpha-amylase from Bacillus licheniformis. By homology with other alpha-amylases, important active site residues can be identified as Asp231, Glu261, and Asp

  8. 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.

  9. Use of physiological information and process optimisation enhances production of extracellular nuclease by a marine strain of Bacillus licheniformis.

    Science.gov (United States)

    Rajarajan, Nithyalakshmy; Ward, Alan C; Burgess, J Grant; Glassey, Jarka

    2013-02-01

    The extracellular nuclease, NucB, from Bacillus licheniformis, can digest extracellular DNA in biofilms, causing biofilm dispersal, and may therefore be used commercially to remove biofilms. However, producing quantities of this secreted peptide is difficult and our aim was therefore to improve its laboratory scale production. This study builds on our understanding of B. licheniformis physiology to enhance NucB production. The addition of manganese, which triggers sporulation and enhances NucB expression, lead to a 5-fold increase in NucB production. Optimisation via Placket-Burman design of experiments identified 3 significant medium components and a subsequent Central Composite Design, to determine the optimum levels of these components, resulted in a 10-fold increase to 471U/ml. The optimal phosphate concentration was less than 0.3mM as this is known to inhibit nuclease production. The use of physiologically relevant information combined with optimisation represents a promising approach to increased enzyme production, which may also be widely applicable.

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

    Science.gov (United States)

    Baks, Tim; Bruins, Marieke E; Matser, Ariette M; Janssen, Anja E M; Boom, Remko M

    2008-01-23

    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 alpha-amylase from Bacillus licheniformis were compared. Suspensions of native starch or starch gelatinized at different conditions either with or without enzyme were hydrolyzed. During hydrolysis, the oligosaccharide concentration, the dextrose equivalent, and the enzyme activity were determined. We found that the hydrolyzate composition was affected by the type of starch pretreatment and the enzyme addition point but that it was just minimally affected by the pressure applied during hydrolysis, as long as gelatinization was complete. The differences between hydrolysis of thermally gelatinized, high-pressure gelatinized, and native starch were explained by considering the granule structure and the specific surface area of the granules. These results show that the hydrolyzate composition can be influenced by choosing different process sequences and conditions.

  11. Determination of the influence of substrate concentration on enzyme selectivity using whey protein Isolate and Bacillus licheniformis protease.

    Science.gov (United States)

    Butré, Claire I; Sforza, Stefano; Gruppen, Harry; Wierenga, Peter A

    2014-10-22

    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 cleavage site in the protein. It was determined from the identification and quantification of the peptides present in the hydrolysates. Solutions of 0.1-10% (w/v) whey protein isolate (WPI) were hydrolyzed by Bacillus licheniformis protease at constant enzyme-to-substrate ratio. The cleavage sites were divided into five groups, from very high (>10%) to very low selectivity (protein concentrations. This finding shows that both the rate of hydrolysis and the enzyme selectivity were influenced by the substrate concentration.

  12. 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.

  13. Ieodoglucomide C and Ieodoglycolipid, New Glycolipids from a Marine-Derived Bacterium Bacillus licheniformis 09IDYM23.

    Science.gov (United States)

    Tareq, Fakir Shahidullah; Lee, Hyi-Seung; Lee, Yeon-Ju; Lee, Jong Seok; Shin, Hee Jae

    2015-05-01

    Chemical examination of the ethyl acetate extract from the fermentation broth of the marine-derived bacterium Bacillus licheniformis resulted in the isolation of two new glycolipids, ieodoglucomide C (1) and ieodoglycolipid (2). The structural characterization of 1 and 2 was achieved by extensive spectroscopic evidence, including 2D NMR experiments. A combination of chemical derivatization techniques followed by NMR studies, LC-MS data analysis and a literature review was deployed for the establishment of the stereo-configurations of 1 and 2. Compounds 1 and 2 exhibited good antibiotic properties against Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Salmonella typhi, Escherichia coli and Pseudomonas aeruginosa with MICs ranging from 0.01 to 0.05 μM. Furthermore, the antifungal activity of 1 and 2 was evaluated against plant pathogenic fungi Aspergillus niger, Rhizoctonia solani, Botrytis cinerea and Colletotrichum acutatum as well as the human pathogen Candida albicans. Compounds 1 and 2 inhibited the mycelial growth of these pathogens with MIC values of 0.03-0.05 μM, revealing that these compounds are good candidates for the development of new fungicides.

  14. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  16. INMOVILIZACIÓN DE Bacillus licheniformis Y Saccharomyces cerevisiae PARA LA PRODUCCIÓN DE ETANOL A PARTIR DE ALMIDÓN DE PAPA

    Directory of Open Access Journals (Sweden)

    Diana Sossa-Urrego

    2008-09-01

    Full Text Available We evaluated a sequential discontinuous system composed by Bacillus licheniformis and Saccharomyces cerevisiae forethanol production. For the second phase of the process potato starch hydrolyzed were used, which was obtained from B.licheniformis cells. Both microorganisms were immobilized in a calcium alginate matrix of 3,2% and 2,5% (w/v, wherewas observed that these concentrations retained the majority of the cells (26x106 and 10x107 UFC/g and alloweddissemination of its products, gaining 3.3 g/L of reducing sugars and 642 AU/L (units Amylolytic for B. licheniformis and0,866% (v/v ethanol with S. cerevisiae. By means of a 22 factorial design were selected operating conditions at a reactorscale for production of hydrolyzed, finding that by cultivating B. licheniformis with 3 v.v.m. and 150 r.p.m. there were 3.7g/L of reducing sugars and 669 AU/L after 4 hours of the process. The hydrolyzed was characterized using HPLCchromatography, which determined that it is rich in oligomers and dextrin, and it has low concentration of glucose andmaltose. The use of hydrolyzed for ethanol production, generated low percentages (0,47% and 0,74% v/v in free andimmobilized cells respectively.

  17. Metabolome analysis reveals the effect of carbon catabolite control on the poly(γ-glutamic acid) biosynthesis of Bacillus licheniformis ATCC 9945.

    Science.gov (United States)

    Mitsunaga, Hitoshi; Meissner, Lena; Palmen, Thomas; Bamba, Takeshi; Büchs, Jochen; Fukusaki, Eiichiro

    2016-04-01

    Poly(γ-glutamic acid) (PGA) is a polymer composed of L- and/or D-glutamic acids that is produced by Bacillus sp. Because the polymer has various features as water soluble, edible, non-toxic and so on, it has attracted attention as a candidate for many applications such as foods, cosmetics and so on. However, although it is well known that the intracellular metabolism of Bacillus sp. is mainly regulated by catabolite control, the effect of the catabolite control on the PGA producing Bacillus sp. is largely unknown. This study is the first report of metabolome analysis on the PGA producing Bacillus sp. that reveals the effect of carbon catabolite control on the metabolism of PGA producing Bacillus licheniformis ATCC 9945. Results showed that the cells cultivated in glycerol-containing medium showed higher PGA production than the cells in glucose-containing medium. Furthermore, metabolome analysis revealed that the activators of CcpA and CodY, global regulatory proteins of the intracellular metabolism, accumulated in the cells cultivated in glycerol-containing and glucose-containing medium, respectively, with CodY apparently inhibiting PGA production. Moreover, the cells seemed to produce glutamate from citrate and ammonium using glutamine synthetase/glutamate synthase. Pulsed addition of di-ammonium hydrogen citrate, as suggested by the metabolome result, was able to achieve the highest value so far for PGA production in B. licheniformis.

  18. Presencia de Lactobacillus spp. y Bacillus licheniformis en margarina con yogur

    Directory of Open Access Journals (Sweden)

    González, Elisa

    1998-02-01

    Full Text Available The microbiological analysis of thirty samples of commercially produced margarine with incorporated yoghurt was carried out. After the initial control, the other tests were runned after 26, 56, 88,116 and 157 days of refrigerated storage. Constitutive biota of yoghurt was not detected. Occurrence (100% of samples of Lactobacillus spp. (Lactobacillus fermentum and Lactobacillus casei subsp. pseudoplantarum, being the first one slightly more numerous, and Bacillus licheniformis, which counts were mostly in a 103- 104ufc/g range, and only in 10% of the cases were < 103 ufc/g. Samples did not show signs of deterioration. Article calls upon about the convenience of developing a specific normative that clarifies the doubts about main microbiological hazards as well as the legal aspects regarding the product denomination.

    Se ha realizado el análisis de treinta muestras de margarina con yogur. Tras el control inicial, los restantes análisis se efectuaron a los 0,26, 56, 88,116 y 157 días de almacenamiento en refrigeración. No se detectó la biota constitutiva del yogur. Sí se demostró la presencia, en el 100% de las muestras, de Lactobacillus fermentum y Lactobacillus casei subsp. pseudoplantarum, siendo el primero ligeramente más numeroso, así como de Bacillus licheniformis, cuyos recuentos han sido en su mayoría comprendidos en el rango 103-104ufc/g, y sólo en el 10% de los casos fueron inferiores a 103ufc/g. Las muestras no mostraban signos de deterioro. El trabajo llama la atención sobre la conveniencia de desarrollar una normativa específica que aclare las dudas surgidas en torno a los principales riesgos microbiológicos, así como a aspectos legales relacionados con la denominación del producto.

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

    Directory of Open Access Journals (Sweden)

    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.

  20. 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.

    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...

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

    Directory of Open Access Journals (Sweden)

    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.

  2. Bacillus licheniformis BlaR1 L3 loop is a zinc metalloprotease activated by self-proteolysis.

    Directory of Open Access Journals (Sweden)

    Stéphanie Berzigotti

    Full Text Available In Bacillus licheniformis 749/I, BlaP β-lactamase is induced by the presence of a β-lactam antibiotic outside the cell. The first step in the induction mechanism is the detection of the antibiotic by the membrane-bound penicillin receptor BlaR1 that is composed of two functional domains: a carboxy-terminal domain exposed outside the cell, which acts as a penicillin sensor, and an amino-terminal domain anchored to the cytoplasmic membrane, which works as a transducer-transmitter. The acylation of BlaR1 sensor domain by the antibiotic generates an intramolecular signal that leads to the activation of the L3 cytoplasmic loop of the transmitter by a single-point cleavage. The exact mechanism of L3 activation and the nature of the secondary cytoplasmic signal launched by the activated transmitter remain unknown. However, these two events seem to be linked to the presence of a HEXXH zinc binding motif of neutral zinc metallopeptidases. By different experimental approaches, we demonstrated that the L3 loop binds zinc ion, belongs to Gluzincin metallopeptidase superfamily and is activated by self-proteolysis.

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

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    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.

  4. Purification and Characterization of Surfactant-Stable Protease from Bacillus Licheniformis: A Potential Additive for Laundry Detergent

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    Vivi Mardina

    2016-04-01

    Full Text Available This study purified and characterized the protease from Bacillus licheniformis that was cultured in skim latex serum fortified media. Ammonium sulphate precipitation and ion exchange chromatograph was employed in purification steps with the enzyme activity increase to 2.28 fold of purification compare to the crude enzyme. Assessment of the purified protein by SDS PAGE showed a single band with molecular mass of about 47 kDa. The enzyme was stable at temperature range of 35 oC to 65 oC and also at pH 6.0 and 7.0 for 60 min. The presence of Mn2+ and Ca2+ ions in the produced protease stimulated strongly the activity of the enzyme by 176.65% and 119.07% respectively, while inhibitory effects were found in the presence of Cu2+, Zn2+, Mg2+, and EDTA. The enzyme exhibited their stability toward surfactants (Triton X100, Tween 20, SDS, solvents (acetone, chloroform, hexane and toluene, oxidizing agent (H2O2 and Tesco Everyday Value® detergent with the residual activity around 80%. It also demonstrated the removal activity of blood stain completely with supplementation of the 7 mg/ml detergent solution. The established characteristics of the enzyme indicated their potentiality for detergent application.

  5. 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.

  6. Immobilization of pectin degrading enzyme from Bacillus licheniformis KIBGE IB-21 using agar-agar as a support.

    Science.gov (United States)

    Rehman, Haneef Ur; Aman, Afsheen; Zohra, Raheela Rahmat; Qader, Shah Ali Ul

    2014-02-15

    Pectinase from Bacillus licheniformis KIBGE IB-21 was immobilized in agar-agar matrix using entrapment technique. Effect of different concentrations of agar-agar on pectinase immobilization was investigated and it was found that maximum immobilization was achieved at 3.0% agar-agar with 80% enzyme activity. After immobilization, the optimum temperature of enzyme increased from 45 to 50 °C and reaction time from 5 to 10 minutes as compared to free enzyme. Due to the limited diffusion of high molecular weight substrate, K(m) of immobilized enzyme slightly increased from 1.017 to 1.055 mg ml(-1), while Vmax decreased from 23,800 to 19,392 μM min(-1) as compared to free enzyme. After 120 h entrapped pectinase retained their activity up to 82% and 71% at 30 °C and 40 °C, respectively. The entrapped pectinase showed activity until 10th cycle and maintain 69.21% activity even after third cycle.

  7. A comparative ecotoxicity analysis of α- and γ-phase aluminium oxide nanoparticles towards a freshwater bacterial isolate Bacillus licheniformis.

    Science.gov (United States)

    Pakrashi, Sunandan; Kumar, Deepak; Iswarya, V; Bhuvaneshwari, M; Chandrasekaran, N; Mukherjee, Amitava

    2014-12-01

    Crystalline structure of nanoparticles may influence their physicochemical behaviour as well as their toxicological impact on biota. The differences in orientation of the atoms result in the variations in chemical stability. Thus, toxicological impacts of different crystalline phases of aluminium oxide nanoparticles are expected to vary. The present study brings out a comparative toxicity analysis of γ-phase and α-phase aluminium oxide nanoparticles of comparable hydrodynamic size range towards a freshwater bacterial isolate Bacillus licheniformis at low exposure concentrations (5, 1, 0.5 and 0.05 µg/mL). Upon 2-h exposure, the α-aluminium oxide particles showed lower toxicity than the γ-phase aluminium oxide. The lower level of oxidative stress generation and cell membrane damage in case of the α-phase aluminium oxide nanoparticles substantiated the toxicity results. The involvement of protein, lipopolysaccharides in nanoparticle-cell surface interaction, was noted in both the cases. To conclude, the crystallinity of aluminium oxide nanoparticles played an important role in the interaction and the toxicity response.

  8. 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.

  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. Production of biosurfactant from Bacillus licheniformis for microbial enhanced oil recovery and inhibition the growth of sulfate reducing bacteria

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    H.S. El-Sheshtawy

    2015-06-01

    Full Text Available In this study, the bacterium Bacillus licheniformis has been isolated from oil reservoir; the ability of this bacterium to produce a biosurfactant was detected. Surface properties of the produced biosurfactant were confirmed by determining the emulsification power as well as surface and interfacial tension. The crude biosurfactant has been extracted from supernatant culture growth, and the yield of crude biosurfactant was about 1 g/l. Also, chemical structure of the produced biosurfactant was confirmed using FTIR analysis. Results revealed that, the emulsification power has been increased up to 96% and the surface tension decreased from 72 of distilled water to 36 mN/m after 72 h of incubation. The potential application of this bacterial species in microbial-enhanced oil recovery (MEOR was investigated. The percent of oil recovery was 16.6% upon application in a sand pack column designed to stimulate an oil recovery. It also showed antimicrobial activity against the growth of different strains of SRB (sulfate reducing bacteria. Results revealed that a complete inhibition of SRB growth using 1.0% crude biosurfactant is achieved after 3 h.

  11. Glutamate dehydrogenase (RocG) in Bacillus licheniformis WX-02: Enzymatic properties and specific functions in glutamic acid synthesis for poly-γ-glutamic acid production.

    Science.gov (United States)

    Tian, Guangming; Wang, Qin; Wei, Xuetuan; Ma, Xin; Chen, Shouwen

    2017-04-01

    Poly-γ-glutamic acid (γ-PGA), a natural biopolymer, is widely used in cosmetics, medicine, food, water treatment, and agriculture owing to its features of moisture sequestration, cation chelation, non-toxicity and biodegradability. Intracellular glutamic acid, the substrate of γ-PGA, is a limiting factor for high yield in γ-PGA production. Bacillus subtilis and Bacillus licheniformis are both important γ-PGA producing strains, and B. subtilis synthesizes glutamic acid in vivo using the unique GOGAT/GS pathway. However, little is known about the glutamate synthesis pathway in B. licheniformis. The aim of this work was to characterize the glutamate dehydrogenase (RocG) in glutamic acid synthesis from B. licheniformis with both in vivo and in vitro experiments. By re-directing the carbon flux distribution, the rocG gene deletion mutant WX-02ΔrocG produced intracellular glutamic acid with a concentration of 90ng/log(CFU), which was only 23.7% that of the wild-type WX-02 (380ng/log(CFU)). Furthermore, the γ-PGA yield of mutant WX-02ΔrocG was 5.37g/L, a decrease of 45.3% compared to the wild type (9.82g/L). In vitro enzymatic assays of RocG showed that RocG has higher affinity for 2-oxoglutarate than glutamate, and the glutamate synthesis rate was far above degradation. This is probably the first study to reveal the glutamic acid synthesis pathway and the specific functions of RocG in B. licheniformis. The results indicate that γ-PGA production can be enhanced through improving intracellular glutamic acid synthesis.

  12. 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.

  13. Effect of a Probiotic Containing Bacillus licheniformis and Bacillus subtilis and Ferroin Solution on Growth Performance, Body Composition and Haematological Parameters in Kutum (Rutilus frisii kutum) Fry.

    Science.gov (United States)

    Azarin, Hajar; Aramli, Mohammad Sadegh; Imanpour, Mohammad Reza; Rajabpour, Mina

    2015-03-01

    This study aimed to assess the efficacy of BioPlus 2B, a probiotic containing Bacillus licheniformis and B. subtilis and Ferroin solution on growth performance, body composition and haematological parameters in kutum, Rutilus frisii kutum, fry. The fish were fed dry pellets containing various ratios of probiotics and Ferroin for 60 days after absorption of the yolk sac. At the end of the trial, growth indices (final weight, weight gain, specific growth rate, daily growth rate, food conversion ratio and condition factor), body composition (crude protein, crude lipid, ash and moisture) and haematological parameters [haematocrit (Hct), haemoglobin (Hb), red blood cells (RBC), white blood cells (WBC), neutrophils (NEUTR), lymphocytes (LYM), mean cell volume (MCV), mean cell haemoglobin (MCH) and mean cell haemoglobin concentration (MCHC)] were assessed. Regarding body composition, total protein levels were higher, and ash, moisture and lipid levels were lower in fish receiving the probiotic and Ferroin treatments compared with the control group (P < 0.05). Fish receiving diets supplemented with probiotics and Ferroin solution showed significantly better growth than those fed the basal diet (control). RBC, Hct, Hb, MCV, MCH and LYM were all highest in fish fed probiotic (1.6 × 10(9) CFU/g dry pellet) + Ferroin solution (7 mg/kg dry pellet) + dry pellets. These results indicate that the combination of probiotic and Ferroin solution represents an effective dietary supplement for improving carcass quality, growth performance and haematological parameters in kutum fry.

  14. Valorización de residuos agroindustriales para la obtención de liquenisina por Bacillus licheniformis AL 1.1

    OpenAIRE

    Coronel León, Jonathan

    2015-01-01

    La cepa bacteriana AL 1.1 se aisló de una muestra de suelo de la isla Decepción del continente Antártico. Inicialmente se observó que dicha cepa tenía la capacidad de reducir la tensión superficial del medio, por lo que fue seleccionada para realizar la presente tesis doctoral. Después de confirmar la identificación del aislado como un Bacillus licheniformis se caracterizó el biotensioactivo (BT) producido como liquenisina (LchAL1.1). El estudio de las propiedades físico-químicas de la Lc...

  15. 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.).

  16. Purification and biochemical properties of a thermostable, haloalkaline cellulase from Bacillus licheniformis AMF-07 and its application for hydrolysis of different cellulosic substrates to bioethanol production

    Science.gov (United States)

    Azadian, Fatemeh; Badoei-dalfard, Arastoo; Namaki-Shoushtari, Abdolhamid; Hassanshahian, Mehdi

    2016-01-01

    A thermophilic strain AMF-07, hydrolyzing carboxymethylcellulose (CMC) was isolated from Kerman hot spring and was identified as Bacillus licheniformis based on 16S rRNA sequence homology. The carboxymethylcellulase (CMCase) enzyme produced by the B. licheniformis was purified by (NH4)2SO4 precipitation, ion exchange and gel filtration chromatography. The purified enzyme gave a single band on SDS- PAGE with a molecular weight of 37 kDa. The CMCase enzyme was highly active and stable over broad ranges of temperature (40-80ºC), pH (6.0-10.0) and NaCl concentration (10-25%) with an optimum at 70ºC, pH 9.0 and 20% NaCl, which showed excellent thermostable, alkali-stable and halostable properties. Moreover, it displayed high activity in the presence of cyclohexane (134%) and chloroform (120%). Saccharification of rice bran and wheat bran by the CMCase enzyme resulted in respective yields of 24 and 32 g L-1 reducing sugars. The enzymatic hydrolysates of rice bran were then used as the substrate for ethanol production by Saccharomyces cerevisiae. Fermentation of cellulosic hydrolysate using S. cerevisiae, reached maximum ethanol production about 0.125 g g-1 dry substrate (pretreated wheat bran). Thus, the purified cellulase from B. licheniformis AMF-07 utilizing lignocellulosic biomass could be greatly useful to develop industrial processes. PMID:28097168

  17. Purification and biochemical properties of a thermostable, haloalkaline cellulase from Bacillus licheniformis AMF-07 and its application for hydrolysis of different cellulosic substrates to bioethanol production

    Directory of Open Access Journals (Sweden)

    Fatemeh Azadian

    2016-09-01

    Full Text Available A thermophilic strain AMF-07, hydrolyzing carboxymethylcellulose (CMC was isolated from Kerman hot spring and was identified as Bacillus licheniformis based on 16S rRNA sequence homology. The carboxymethylcellulase (CMCase enzyme produced by the B. licheniformis was purified by (NH42SO4 precipitation, ion exchange and gel filtration chromatography. The purified enzyme gave a single band on SDS-PAGE with a molecular weight of 37 kDa. The CMCase enzyme was highly active and stable over broad ranges of temperature (40-80 ºC, pH (6.0-10.0 and NaCl concentration (10-25% with an optimum at 70 ºC, pH 9.0 and 20% NaCl, which showed excellent thermostable, alkali-stable and halostable properties. Moreover, it displayed high activity in the presence of cyclohexane (134% and chloroform (120%. Saccharification of rice bran and wheat bran by the CMCase enzyme resulted in respective yields of 24 and 32 g L-1 reducing sugars. The enzymatic hydrolysates of rice bran were then used as the substrate for ethanol production by Saccharomyces cerevisiae. Fermentation of cellulosic hydrolysate using S. cerevisiae, reached maximum ethanol production about 0.125 g g-1 dry substrate (pretreated wheat bran. Thus, the purified cellulase from B. licheniformis AMF-07 utilizing lignocellulosic biomass could be greatly useful to develop industrial processes.

  18. SHG10 keratinolytic alkaline protease from Bacillus licheniformis SHG10 DSM 28096: Robust stability and unusual non-cumbersome purification.

    Science.gov (United States)

    Embaby, Amira M; Saeed, Hesham; Hussein, Ahmed

    2016-12-01

    Present study underlines an unusual non-cumbersome-powerful strategy for purification of SHG10 keratinolytic alkaline protease from Bacillus licheniformis SHG10 DSM 28096 with robust stability properties. The enzyme was impressively purified to homogeneity with specific activity, purification fold, and yield of 613.82 U mg(-1) , 58.91 and 99%, respectively, via a sequential two-step purification strategy: precipitation with 65% (NH4 )2 SO4 and flow through fractions of DEAE-cellulose DE 53 column. SDS-PAGE conferred a monomeric enzyme with a molecular mass of 30.4 kDa. The enzyme demonstrated optimal activity at pH (10.0-11.0) and at 65 °C. It exhibited full stability at pH (6.0-11.0) over 38 h at 4 °C and at 65 °C for 15 min. Remarkable enhanced enzyme activity (130.15 and 126.37%) was retained in presence of commercial laundry detergents Oxi and Ariel after 1 h, respectively. Organic solvent stability of the enzyme was verified in butanol, ether, acetonitrile, isopropanol, and chloroform. Imposingly, full storage stability (100%) of the enzyme along 1 year in -20 °C was confirmed. Km -Vmax was 0.00174 mM-534.2 mM Sub · min(-1)  · mg protein(-1) and 1.266 mg-28.89 mg Sub · h(-1)  · mg protein(-1) on N-Suc-Ala-Ala-Pro-Phe-pNA and keratin azure, respectively. Robust stability properties of SHG10 keratinolytic alkaline protease along with rapid-efficient purification underpin its potential commercialization for industrial exploitation.

  19. 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.

  20. 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%.

  1. 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.

  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. Branched chain amino acids maintain the molecular weight of poly(γ-glutamic acid) of Bacillus licheniformis ATCC 9945 during the fermentation.

    Science.gov (United States)

    Mitsunaga, Hitoshi; Meissner, Lena; Büchs, Jochen; Fukusaki, Eiichiro

    2016-10-01

    Poly(γ-glutamic acid) mainly produced by Bacillus spp. is an industrially important compound due to several useful features. Among them, molecular weight is an important characteristic affecting on the physical properties such as viscosities and negative charge densities. However, it is difficult to control the molecular size of PGA since it decreases during fermentation. Previous study reported that PGA produced in the media containing different carbon sources such as glucose and glycerol showed differences in molecular weight. Therefore in this study, the effect of carbon source on the PGA molecular weight was examined; with the aim of developing a strategy to maintain the high molecular weight of PGA during fermentation. Our result showed that the weight average molecular weight (Mw) of PGA of Bacillus licheniformis ATCC 9945 cultivated in the media containing PTS-sugars were higher than the medium containing glycerol (non-PTS). The result of metabolome analysis indicated the possibility of CodY (a global regulator protein) activation in the cells cultivated in the media containing PTS-sugars. To mimic this effect, branched-chain amino acids (BCAAs), which are activators of CodY, were added to a medium containing glycerol. As the result, the Mw of PGA in the BCAAs-supplemented media were maintained and high during the early production phase compared to the non BCAAs-supplemented medium. These results indicate that BCAAs can repress the PGA molecular weight reduction during fermentation in B. licheniformis ATCC 9945.

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

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    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. Comparison of starch hydrolysis activity and thermal stability of two Bacillus licheniformis alpha-amylases and insights into engineering alpha-amylase variants active under acidic conditions.

    Science.gov (United States)

    Lee, Seunjae; Oneda, Hiroshi; Minoda, Masashi; Tanaka, Akiyoshi; Inouye, Kuniyo

    2006-06-01

    Bacillus licheniformis alpha-amylase (BLA) is widely used in various procedures of starch degradation in the food industry, and a BLA species with improved activity at higher temperature and under acidic conditions is desirable. Two BLA species, designated as PA and MA, have been isolated from the wild-type B. licheniformis strain and a mutant strain, respectively. In this study, their starch-hydrolysis activity and thermal stability were examined. MA showed higher activity than PA, especially at acidic pH (pH 5.0-5.5), and even after 1 h of treatment at 90 degrees C. MA was active in the range of pH 4.0-8.0, which is much wider than that (pH 4.5-7.5) of PA. It was shown that the proton dissociation constants on the acidic and alkaline sides (pKa1 and pKa2) were shifted to more acidic and basic values, respectively, by the mutation of PA to MA. The activation energy and thermodynamic parameters for their thermal inactivation indicate that MA is more thermally stable and catalytically active than PA, suggesting that MA could be useful for glucose-production process coupled with reactions catalyzed by beta-amylase.

  6. Oxygen and nitrate in utilization by Bacillus licheniformis of the arginase and arginine deiminase routes of arginine catabolism and other factors affecting their syntheses.

    Science.gov (United States)

    Broman, K; Lauwers, N; Stalon, V; Wiame, J M

    1978-09-01

    Bacillus licheniformis has two pathways of arginine catabolism. In well-aerated cultures, the arginase route is present, and levels of catabolic ornithine carbamoyltransferase were low. An arginase pathway-deficient mutant, BL196, failed to grow on arginine as a nitrogen source under these conditions. In anaerobiosis, the wild type contained very low levels of arginase and ornithine transaminase. BL196 grew normally on glucose plus arginine in anaerobiosis and, like the wild type, had appreciable levels of catabolic transferase. Nitrate, like oxygen, repressed ornithine carbamoyltransferase and stimulated arginase synthesis. In aerobic cultures, arginase was repressed by glutamine in the presence of glucose, but not when the carbon-energy source was poor. In anaerobic cultures, ammonia repressed catabolic ornithine carbamoyltransferase, but glutamate and glutamine stimulated its synthesis. A second mutant, derived from BL196, retained the low arginase and ornithine transaminase levels of BL196 but produced high levels of deiminase pathway enzymes in the presence of oxygen.

  7. Optimization of medium composition for keratinase production on feather by Bacillus licheniformis RG1 using statistical methods involving response surface methodology.

    Science.gov (United States)

    Ramnani, Priya; Gupta, Rani

    2004-10-01

    A 3.5-fold increase in keratinase production by Bacillus licheniformis RG1 was achieved by using statistical methods involving Plackett-Burman design and response surface methodology. Eight variables were screened using Plackett-Burman design. Of these, glucose, peptone and glutathione were found to affect the response signal positively, whereas CaCl(2) had a negative effect. Further interaction of these factors, along with phosphate and incubation time, was studied using response surface methodology. An optimum keratinase production of 1295 units/mg dry weight was obtained with the following medium composition: 1% glucose, 1% peptone, 1% phosphate, 0.05% glutathione, 0.5% feather and 2% inoculum under shaking at 250 rev./min with an incubation period of 72 h at 37 degrees C. Keratinase production was found to be a function of biomass and maximum production occurred during the stationary phase.

  8. An investigation into the preservation of microbial cell banks for α-amylase production during 5 l fed-batch Bacillus licheniformis fermentations.

    Science.gov (United States)

    Hancocks, Nichola H; Thomas, Colin R; Stocks, Stuart M; Hewitt, Christopher J

    2010-10-01

    Fluorescent staining techniques were used for a systematic examination of methods used to cryopreserve microbial cell banks. The aim of cryopreservation here is to ensure subsequent reproducible fermentation performance rather than just post thaw viability. Bacillus licheniformis cell physiology post-thaw is dependent on the cryopreservant (either Tween 80, glycerol or dimethyl sulphoxide) and whilst this had a profound effect on the length of the lag phase, during subsequent 5 l fed-batch fermentations, it had little effect on maximum specific growth rate, final biomass concentration or α-amylase activity. Tween 80 not only protected the cells during freezing but also helped them recover post-thaw resulting in shorter process times.

  9. 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.

  10. Influence of nitrogen source and pH value on undesired poly(γ-glutamic acid) formation of a protease producing Bacillus licheniformis strain.

    Science.gov (United States)

    Meissner, Lena; Kauffmann, Kira; Wengeler, Timo; Mitsunaga, Hitoshi; Fukusaki, Eiichiro; Büchs, Jochen

    2015-09-01

    Bacillus spp. are used for the production of industrial enzymes but are also known to be capable of producing biopolymers such as poly(γ-glutamic acid). Biopolymers increase the viscosity of the fermentation broth, thereby impairing mixing, gas/liquid mass and heat transfer in any bioreactor system. Undesired biopolymer formation has a significant impact on the fermentation and downstream processing performance. This study shows how undesirable poly(γ-glutamic acid) formation of an industrial protease producing Bacillus licheniformis strain was prevented by switching the nitrogen source from ammonium to nitrate. The viscosity was reduced from 32 to 2.5 mPa s. A constant or changing pH value did not influence the poly(γ-glutamic acid) production. Protease production was not affected: protease activities of 38 and 46 U mL(-1) were obtained for ammonium and nitrate, respectively. With the presented results, protease production with industrial Bacillus strains is now possible without the negative impact on fermentation and downstream processing by undesired poly(γ-glutamic acid) formation.

  11. 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%.

  12. 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.

  13. The Reduction of Selenite by Bacillus licheniformis%地衣芽孢杆菌对亚硒酸盐的还原

    Institute of Scientific and Technical Information of China (English)

    朱建明; 雷磊; 肖湘; 袁永强; 秦海波; 苏惠

    2011-01-01

    利用高硒碳质泥岩中筛选出的地衣芽孢杆菌(Bacillus licheni formi),研究了该菌对亚硒酸盐硒的耐受与还原行为.结果表明,液体培养基(YEG)中,它能耐受320 mM亚硒酸盐硒的浓度,酎受硒酸盐硒的浓度可高达1000mM.然而,高浓度的亚硒酸盐硒对它的生长有明显的抑制作用.在有氧和厌氧的环境中,地衣芽孢杆菌均能还原亚硒酸盐中的硒:将四价硒还原为纳米球状的元素硒颗粒,使其分布在菌体的周边和细胞内.在含5 mM亚硒酸钠的液体培养基中,还原亚硒酸钠硒成为元素硒的平均效率约为42%.地衣芽孢杆菌在生存环境无严格要求的条件下,其还原亚硒酸盐硒形成纳米元素硒颗粒的现象,是研究生物合成低毒的纳米活性元素硒和生物修复硒污染技术的基础,也为硒的微生物矿化过程提供了契机.%Bacillus Licheniformi, which can reduce the toxic selenite anion to red elemental selenium (Se°) , was isolated from a carbonaceous mudstone with high content of Se. The results showed that this strain can stand in 320 mM SeO2-3 and in more than 1000 mM SeO2-4. However,the high concentration of SeO2-3 can inhibit the growth of the strain in liquid medium (YEG). No matter in aerobic culture or anaerobic culture that the strain was inoculated , it can reduce selenite anion to nanospheric elemental selenium granules, distributed around or within the cells. In the liquid YEG medium contains 5mM of selenite,the transformed efficiency of Se4+ to Se° by Bacillus licheniformi was 42% approximately. Since no rigorous requirement for Bacillus licheniformi to live,it is suitable to be selected in microbial remediation techniques as the strains that cope with selenium pollutions, and to produce the nano-Se granules with bioavailability. At the same time,the phenomena of selenite anions reduced to elemental Se by the bacterial provides a chance to further understand the microbial mineralization of selenium

  14. Efficient recombinant expression and secretion of a thermostable GH26 mannan endo-1,4-β-mannosidase from Bacillus licheniformis in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Haltrich Dietmar

    2010-04-01

    Full Text Available Abstract Background Mannans are one of the key polymers in hemicellulose, a major component of lignocellulose. The Mannan endo-1,4-β-mannosidase or 1,4-β-D-mannanase (EC 3.2.1.78, commonly named β-mannanase, is an enzyme that can catalyze random hydrolysis of β-1,4-mannosidic linkages in the main chain of mannans, glucomannans and galactomannans. The enzyme has found a number of applications in different industries, including food, feed, pharmaceutical, pulp/paper industries, as well as gas well stimulation and pretreatment of lignocellulosic biomass for the production of second generation biofuel. Bacillus licheniformis is a Gram-positive endospore-forming microorganism that is generally non-pathogenic and has been used extensively for large-scale industrial production of various enzymes; however, there has been no previous report on the cloning and expression of mannan endo-1,4-β-mannosidase gene (manB from B. licheniformis. Results The mannan endo-1,4-β-mannosidase gene (manB, commonly known as β-mannanase, from Bacillus licheniformis strain DSM13 was cloned and overexpressed in Escherichia coli. The enzyme can be harvested from the cell lysate, periplasmic extract, or culture supernatant when using the pFLAG expression system. A total activity of approximately 50,000 units could be obtained from 1-l shake flask cultures. The recombinant enzyme was 6 × His-tagged at its C-terminus, and could be purified by one-step immobilized metal affinity chromatography (IMAC to apparent homogeneity. The specific activity of the purified enzyme when using locust bean gum as substrate was 1672 ± 96 units/mg. The optimal pH of the enzyme was between pH 6.0 - 7.0; whereas the optimal temperature was at 50 - 60°C. The recombinant β-mannanase was stable within pH 5 - 12 after incubation for 30 min at 50°C, and within pH 6 - 9 after incubation at 50°C for 24 h. The enzyme was stable at temperatures up to 50°C with a half-life time of activity (τ1

  15. A novel approach to improve poly-γ-glutamic acid production by NADPH Regeneration in Bacillus licheniformis WX-02

    Science.gov (United States)

    Cai, Dongbo; He, Penghui; Lu, Xingcheng; Zhu, Chengjun; Zhu, Jiang; Zhan, Yangyang; Wang, Qin; Wen, Zhiyou; Chen, Shouwen

    2017-01-01

    Poly-γ-glutamic acid (γ-PGA) is an important biochemical product with a variety of applications. This work reports a novel approach to improve γ-PGA through over expression of key enzymes in cofactor NADPH generating process for NADPH pool. Six genes encoding the key enzymes in NADPH generation were over-expressed in the γ-PGA producing strain B. licheniformis WX-02. Among various recombinants, the strain over-expressing zwf gene (coding for glucose-6-phosphate dehydrogenase), WX-zwf, produced the highest γ-PGA concentration (9.13 g/L), 35% improvement compared to the control strain WX-pHY300. However, the growth rates and glucose uptake rates of the mutant WX-zwf were decreased. The transcriptional levels of the genes pgsB and pgsC responsible for γ-PGA biosynthesis were increased by 8.21- and 5.26-fold, respectively. The Zwf activity of the zwf over expression strain increased by 9.28-fold, which led to the improvement of the NADPH generation, and decrease of accumulation of by-products acetoin and 2,3-butanediol. Collectively, these results demonstrated that NADPH generation via over-expression of Zwf is as an effective strategy to improve the γ-PGA production in B. licheniformis. PMID:28230096

  16. 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.

  17. 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.

    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...

  18. 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.

  19. 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.

  20. Biosynthesis of silver nanoparticles using a probiotic Bacillus licheniformis Dahb1 and their antibiofilm activity and toxicity effects in Ceriodaphnia cornuta.

    Science.gov (United States)

    Shanthi, Sathappan; Jayaseelan, Barbanas David; Velusamy, Palaniyandi; Vijayakumar, Sekar; Chih, Cheng Ta; Vaseeharan, Baskaralingam

    2016-04-01

    In the present study, we synthesized and characterized a probiotic Bacillus licheniformis cell free extract (BLCFE) coated silver nanoparticles (BLCFE-AgNPs). These BLCFE-AgNPs were characterized by UV-visible spectrophotometer, XRD, EDX, FTIR, TEM and AFM. A strong surface plasmon resonance centered at 422 nm in UV-visible spectrum indicates the formation of AgNPs. The XRD spectrum of silver nanoparticles exhibited 2θ values corresponding to the silver nanocrystal. TEM and AFM showed the AgNPs were spherical in shape within the range of 18.69-63.42 nm and the presence of silver was confirmed by EDX analysis. Light and Confocal Laser Scanning Microscope (CLSM) images showed a weak adherence and disintegrated biofilm formation of Vibrio parahaemolyticus Dav1 treated with BLCFE-AgNPs compared to control. This result suggests that BLCFE-AgNps may be used for the control of biofilm forming bacterial populations in the biomedical field. In addition, acute toxicity results concluded that BLCFE-AgNPs were less toxic to the fresh water crustacean Ceriodaphnia cornuta (50 μg/ml) when compared to AgNO3 (22 μg/ml). This study also reports a short term analysis (24 h) of uptake and depuration of BLCFE-AgNPs in C. cornuta.

  1. 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.

  2. Research Advances in the Application of Bioactive Substances Produced by Bacillus licheniformis%地衣芽胞杆菌生物活性物质应用研究进展

    Institute of Scientific and Technical Information of China (English)

    杨阳; 张付云; 苍桂璐; 王斌; 卢航

    2013-01-01

    Bacillus licheniformis is a common Gram-positive bacterium. It has good features of high heat-resistance,variouse enzyme production,good enzyme productivity,and high safety. It also can produce many kinds of bioactive substances such as polysaccharid,lipopeptide biosurfactant,bacitracin and small molecular substances. This paper summarizes the research progress of bioactive substances produced by Bacillus licheniformis and their biological activity and application,prospects its development direction of new strains construction by means of genetic engineering,so as to provide references for the future utilization of Bacillus licheniformis.%  地衣芽胞杆菌是一种常见革兰氏阳性菌,具有耐热、酶系丰富、产酶量高和安全等优良特性,同时在代谢过程中还可产生多种活性物质,例如多糖、脂肽类生物表面活性剂、杆菌肽及小分子等活性物质。本文对地衣芽胞杆菌的活性物质及其生物活性的应用研究进展进行概括,并且对其通过基因工程手段开发新型菌种的发展方向进行了展望,以期为地衣芽胞杆菌的利用提供参考。

  3. Preparation of β- Mannanase from Bacillus licheniformis%地衣芽孢杆菌β-甘露聚糖酶的制备

    Institute of Scientific and Technical Information of China (English)

    张峻; 何志敏; 胡鲲; 冯耀宇; 张志钢

    2001-01-01

    提出了一种地衣芽孢杆菌β-甘露聚糖酶制备的工艺。结果表明:在设定的操作条件下,6.6L自控罐发酵酶活可达260u/mL。发酵液用AlCl3与壳聚糖絮凝预处理后,絮凝体喷雾干燥可制成900u/g的酶粉,适合饲料和造纸工业使用;絮凝上清液先以平均截留分子质量5万的中空纤维聚砜膜分离,透过液再用平均截留分子质量1万的中空纤维聚砜膜浓缩后,喷雾干燥可制成11000u/g的酶粉,适合食品和医药工业使用。以上工艺酶活总收率88%,并易于工业放大。%A process has been developed for the preparation of β-mannanase from Bacillus licheniformis. The enzyme activity was 260 u/mL when incubated in a 6.6L fermentor under the designed conditions. After pretreatment of culture broth using flocculants of AlCl3 and chitosan,the floc was spray-dried (900 u/g) for use in feed or pulp industries, while the supernatant was fractionated by polysulfone membrane and then concentrated by polysulfone membrane. The final product was spray-dried (11 000u/g) for use in food or pharmaceutical industries. The proposed process resulted in the total yield of β-mannanase activity of 88 % and is easily scale-up.

  4. Molecular and biochemical characteristics of recombinant β-propeller phytase from Bacillus licheniformis strain PB-13 with potential application in aquafeed.

    Science.gov (United States)

    Kumar, Vinod; Sangwan, Punesh; Verma, A K; Agrawal, Sanjeev

    2014-05-01

    Phytic acid is the major storage form of organic phosphorus in nature- and plant-based animal feed. It forms insoluble complexes with nutritionally important metals and proteins that are unavailable for monogastric or agastric animals. Phytases initiate the stepwise hydrolysis of phytic acid and release inorganic orthophosphate. In the present investigation, the phytase gene from a phytase producing Bacillus licheniformis strain PB-13 was successfully expressed in Escherichia coli BL21. Recombinant phytase 'rPhyPB13' was found to be catalytically active, with an activity of 0.97 U/mL and specific activity of 0.77 U/mg. The rPhyPB13 was purified to 14.10-fold using affinity chromatography. Similar to other β-propeller phytases, purified rPhyPB13 exhibited maximal activity at pH 6.0-6.5 and 60 °C in the presence of 1 mM Ca(2+) and was highly active over a wider pH range (pH 4.0-8.0) and high temperature (80 °C). It has shown maximum activity towards Na-phytate as substrate. The observed K m , V max and k cat of purified rPhyPB13 were 1.064 mM, 1.32 μmol/min/mg and 27.46 s(-1), respectively. PhyPB13 was resistant to trypsin inactivation, activated in presence of Ca(2+) and inhibited in presence of EDTA. Crude rPhyPB13 has good digestion efficiency for commercial feed and soybean meal. These results indicate that PhyPB13 is a β-propeller phytase that has application potential in aquaculture feed.

  5. 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.

  6. Role of Bacillus licheniformis VS16-Derived Biosurfactant in Mediating Immune Responses in Carp Rohu and its Application to the Food Industry

    Science.gov (United States)

    Giri, Sib Sankar; Sen, Shib Sankar; Jun, Jin Woo; Sukumaran, V.; Park, Se Chang

    2017-01-01

    Multifarious applications of Bacillus licheniformis VS16-derived biosurfactant were explored. Labeo rohita fingerlings were injected intraperitoneally with 0.1 mL of phosphate-buffered saline (PBS) containing purified biosurfactant at 0 (control), 55 (S55), 110 (S110), 220 (S220), or 330 (S330) μg mL-1 concentrations. Various immunological parameters and the expression of immune-related genes were measured at 7, 14, and 21 days post-administration (dpa). At 21 dpa, fish were challenged with Aeromonas hydrophila and mortality was recorded for 14 days. Immune parameters such as lysozyme levels (39.29 ± 2.14 U mL-1), alternative complement pathway (61.21 ± 2.38 U mL-1), and phagocytic activities (33.37 ± 1.2%) were maximum (P < 0.05) in the S220 group at 14 dpa; but immunoglobulin levels (11.07 ± 0.83 mg mL-1) were highest in the S220 group at 7 dpa, compared to that in controls. Activities of digestive enzymes (amylase, protease, and lipase) were higher (P < 0.05) in the S220 and S330 groups than in the control group. Regarding cytokine gene expression, pro-inflammatory cytokines (TNF-α and IL-1β) were down-regulated (P < 0.05) in the S220 and S330 groups. Expression of IL-10, TGF-β, and IKB-α were up-regulated in the S220 and S330 groups at 14 dpa, with the highest levels in the S220 group. The expression of NF-κB p65 and IKK-β were down-regulated in treatment groups, and were lowest (P < 0.05) in the S220 group. The highest post-challenge survival rate (72.7%) was recorded in S220 group. Further, the potential of this substance to inhibit biofilm formation, and heavy metal removal from vegetables were also evaluated. Biosurfactant was effective in inhibiting biofilm formation up to 54.71 ± 1.27%. Moreover, it efficiently removed cadmium (Cd) from tested vegetables such as carrot, radish, ginger, and potato, with the highest removal efficiency (60.98 ± 1.29%) recorded in ginger contaminated with Cd. Collectively, these results suggest that isolated

  7. 地衣芽孢杆菌原生质体电转化方法的研究%Transformation of Undomesticated Strains of Bacillus licheniformis by Protoplast Electroporation

    Institute of Scientific and Technical Information of China (English)

    温赛; 杨建国

    2015-01-01

    为了解决地衣芽孢杆菌(Bacillus licheniformis)工业菌株难以转化的问题,将原生质体制备、电穿孔和原生质体再生技术相结合,建立了一种地衣芽孢杆菌原生质体电击转化方法.在对菌体生长状态、溶菌酶作用时间、电转电压、渗透压保护剂等条件进行优化后,试验了将不同类型的表达载体,即游离型质粒pGJ103 (3.3kb)和整合型质粒pAX01 (9.3kb),分别转入两株地衣芽孢杆菌工业生产菌株B.licheniformis CICC 10181和B.licheniformis CICC 20204中.实验结果显示,对数生长期后期的菌体酶解40min后制备的地衣芽孢杆菌原生质体得率为96%,再生率达25%以上.原生质体与质粒DNA在最适电压0.6kV/mm下电击转化,并以0.5mol/L山梨醇或甘露醇作为渗透压保护剂进行再生培养后,最终游离型质粒的转化率可达0.88×102~1.1×102 CFU/μgpGJ103,整合型质粒的转化率达到0.45×102~0.52×102 CFU/μg pAX01.该方法为地衣芽孢杆菌野生工业菌株的遗传改造提供了一种新的、高效的转化手段.

  8. 一株地衣芽孢杆菌的性质研究及发酵培养基优化%Research on the Character and Optimization of Fermentation Medium for Bacillus licheniformis, KL6

    Institute of Scientific and Technical Information of China (English)

    刘阳; 谭明; 潘宝平; 宋诙

    2012-01-01

    [目的]研究实验室自分离的地衣芽孢杆菌KL6作为微生物饲料添加剂的可行性,并对其发酵培养基进行优化,确定最佳培养条件,为工业化发酵提供依据.[方法]运用表型试验对其进行生理生化评价及产酶评价;运用4因素3水平正交试验和单因素试验对其培养基进行改进,并对其发酵条件进行优化.[结果]地衣芽孢杆菌KL6的D-葡萄糖产酸试验、硝酸盐还原试验、淀粉水解试验等均呈阳性;4种因素对活菌数影响的显著性次序为:MnSO4>NaCl>酵母粉>蛋白胨,对芽孢数影响的显著性次序为:MnSO4>蛋白胨>NaCl>酵母粉,培养基的最佳配比为:蛋白胨1.0%、酵母粉0.5%、NaCl 1.0%、25%浓度的硫酸锰0.2%;生长适宜温度为37℃,进行发酵罐放大培养,最佳放罐时间应在14~16 h,获得的菌体数量约101.63×109个/ml,芽孢率为97.11%.[结论]体外评价试验表明,地衣芽孢杆菌KL6具有作为微生物饲料添加剂的潜力;发酵培养基及条件优化试验表明,地衣芽孢杆菌KL6能够适于高密度液体发酵.该试验为以KL6为基础的微生物饲料添加剂的开发和大规模发酵培养提供了理论依据.%[Objective] The aim was to study the feasibility of separating Bacillus licheniformis, KL6, as microbial feed additives, optimized its fermentation medium, and determined the optimum culture condition so as to provide a basis of industrial fermentation. [Method] The physiological and biochemical and enzyme-producing of Bacillus licheniformis, KL6, were evaluated by phenotypic tests. Fermentation medium and conditions by and 4- factors and 3-levels orthogonal and single-factor expts improved the culture medium and optimized fermentation condition. [Result] The number of viable cell was influenced by four factors in order of significance as follows:MnSO4 > NaCl > Yeast extract > Peptone, the number of spores was MnSO4 > Peptone > NaCl > Yeast extract, and the

  9. 地衣芽孢杆菌抗菌肽的纯化及抗菌特性分析%Purification of Antimicrobial Peptide from Bacillus licheniformis LY12 and Analysis of Its Antibacterial Properties

    Institute of Scientific and Technical Information of China (English)

    樊陈; 高兆建; 张桂英; 鞠民友; 孙会刚; 杜永凯; 王东星

    2013-01-01

    以研究地衣芽孢杆菌(Bacilluslicheniformis)LY12所产抗菌肽特性,从发酵液中分离纯化该抗菌肽。通过超滤、离子交换层析、分子筛凝胶过滤层析、反向高效液相色谱等方法纯化该抗菌肽。结果显示:LC-ESI-MS质谱测得抗菌肽分子量为2750.785 Da,同Tricine SDS-PAGE分析结果基本一致。体外抗菌试验结果显示,抗菌肽LYF12能够抑制丝状真菌、细菌的生长。对枯草芽孢杆菌(Bacillussubtilis)、大肠杆菌(Escherichia coli)、铜绿假单胞菌(Pseudomonas aeruginosa)、金黄色葡萄球菌(Staphylococcus aureus)、藤黄微球菌(Micrococcus aureus)、蜡状芽孢杆菌(Bacillus cereus)、副溶血性弧菌(Vibrio parachaemolyticus)的最低抑菌浓度分别为9.8、19.8、20.7、7.2、9.6、10.2、15.7μg/mL。从地衣芽孢杆菌LY12分离的抗菌肽显示出对革兰氏阴性细菌、革兰氏阳性细菌及真菌具有较好的抗菌效果,故在食品防腐中具有潜在的应用价值。%To explore the characterization of a novel antimicrobial peptide from from Bacillus licheniformis LY12, the antimicrobial peptide was isolated. The novel antimicrobial peptide designated LYF12, was purified, with a procedure involving ultrafiltration, ion exchange chromatography (DEAE-Sepharose FF), gel filtration (Sephadex G-15), and reverse phase HPLCs on C8 column and C18 column. LC-ESI-MS indicated the molecular mass of the purified peptide was 2750.785 Da, which was in good agreement with the result of Tricine SDS-PAGE. .In vitro bioassays showed that LYF12 inhibits the growth of a variety of microbes, including filamentous fungi and bacteria. The minimum inhibitory concentrations (MIC) against Bacillus subtilis, Escherichiacoli, Pseudomonasaeruginosa, Staphylococcusaureus, Micrococcusaureus, Bacillus cereus, Vibrio parachaemolyticus were 9.8, 19.8, 20.7, 7.2, 9.6, 10.2, 15.7 μg/mL, respectively. Because the purified antimicrobial peptide exhibited antibacterial

  10. 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

  11. 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...

  12. 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.

  13. 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.

  14. 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.

  15. Genetic improvement of α-amylase producing Bacillus licheniformis by homolog-mediated α-amylase gene amplification%通过同源介导α-淀粉酶基因扩增改良地衣芽孢杆菌α-淀粉酶生产菌株

    Institute of Scientific and Technical Information of China (English)

    牛丹丹; 石贵阳; 王正祥

    2009-01-01

    地衣芽孢杆菌高温α-淀粉酶(BLA)是淀粉水解与生物加工过程中重要关键酶制剂之一.为了进一步提高地衣芽孢杆菌高温α-淀粉酶生产菌株的生产性能,本研究构建了一种含有地衣芽孢杆菌高温α-淀粉酶编码基因amyL的整合性重组质粒pBL-amyL.将重组质粒pBL-amyL转化入BLA工业生产菌株Bacillus licheniformis B0204,再在卡那霉素存在下介导其B.1icheniformis B0204染色体中的同源整合与高温α-淀粉酶编码基因amyL的扩增,由此获得了携带多个amyL拷贝的转化子.对转化子的amyL拷贝数及其BLA发酵水平分别用荧光实时定量PCR及摇瓶发酵试验进行评价与鉴定.与出发菌株B0204相比,含2~5倍amyL拷贝数的重组菌的BLA的合成水平显著提高.其中,重组菌REBL18生产BLA的水平提高了89.2%.%Bacillus licheniformis α-amylase (BLA) is one of the most important enzymes involved in starch hydrolysis and many biotechnological processes. To improve the BLA productivity, an integrative plasmid pBL-amyL carrying amyL gene encoding a thermophilic α-amylase of B. licheniformis was constructed and transformed into B. licheniformis B0204, an industrial α-amylase producer. The transformants harboring different copies of amyL were developed on kanamycin by using homolog-mediated chromosomal amplification of α-amylase gene. The recombinants with different multiple copies of amyL integrated in the chromosome were identified by real-time PCR and evaluated by shake-flask fermentation. Recombinants harboring 2-5 multiple copies of amyL produced more α-amylase comparison to the parental strain B0204.

  16. 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.

  17. 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

  18. 地衣芽胞杆菌与多烯磷脂酰胆碱联合治疗非酒精性脂肪性肝炎的疗效分析%The Effect Analysis of Bacillus Licheniformis and Polyene Phosphatidylcholine Combined Treatment of Nonalcoholic Steatohepatitis

    Institute of Scientific and Technical Information of China (English)

    曾云波

    2015-01-01

    Objective To analysis the effect of bacillus licheniformis and polyene phosphatidylcholine combined treatment of nonalcoholic steatohepatitis.Methods 92 patients with non -alcoholic steatohepatitis , selected from 2012 January to 2014 January in our hospital , were randomly divided into control group and study group , respectively after treatment, compared two groups of patients with clinical symptom score , BMI, WHR, blood routine and B ultrasound score.Results Study group patients after treatment of clinical symptoms and BMI , WHR score was significantly lower than that of the control group , the difference was statistically significant ( P<0.05 ) ,After the treatment of blood and B ultrasound groups were lower than the control group , the difference was statistically significant ( P<0.05 ).Conclusions Effect of Bacillus licheniformis and polyene phosphati-dylcholine combined treatment of nonalcoholic steatohepatitis than single polyene phosphatidylcholine in the treatment effect is good , no adverse reactions , it is worthy of popularization and application.%目的:分析地衣芽胞杆菌与多烯磷脂酰胆碱联合治疗非酒精性脂肪性肝炎的疗效。方法选取2012年1月~2014年1月收治的非酒精性脂肪性肝炎患者92例,随机分为对照组和研究组,分别治疗后对比两组患者临床症状评分、体重指数( BMI)、腰臀比( WHR)、血常规以及B超评分。结果治疗后,研究组的临床症状明显较好, BMI、WHR评分低于对照组,P<0.05;研究组的血常规与B超评分明显低于对照组,有统计学意义( P<0.05)。结论地衣芽胞杆菌与多烯磷脂酰胆碱联合治疗非酒精性脂肪性肝炎的疗效比单一多烯磷脂酰胆碱治疗效果好,无不良反应,值得推广应用。

  19. Co-production of surfactin and a novel bacteriocin by Bacillus subtilis subsp. subtilis H4 isolated from bikalga, an African alkaline Hibiscus sabdariffa seed fermented condiment

    DEFF Research Database (Denmark)

    Compaore, C. S.; Nielsen, Dennis S.; Ouoba, L. I. I.;

    2013-01-01

    Bikalga is a Hibiscus sabdariffa seed fermented condiment widely consumed in Burkina Faso and neighboring countries. The fermentation is dominated by Bacillus subtilis group species. Ten B. subtilis subsp. subtilis (six isolates) and Bacillus licheniformis (four isolates) isolated from traditiona...

  20. Isolation and Identification of Bacillus licheniformis and Optimization of the Culture Media%水产养殖用地衣芽孢杆菌的分离鉴定与其扩增培养基筛选

    Institute of Scientific and Technical Information of China (English)

    周冬仁; 罗毅志; 施伟达; 叶雪平; 沈振伟; 汤美锋

    2014-01-01

    为分离筛选适合水产养殖用的地衣芽孢杆菌,并探明不同培养基对其扩增效果,采用微生物学与分子生物学相结合的方法对养殖池塘淤泥中的细菌进行研究。结果表明:分离得到2株抑菌作用最强的菌株DSY002-2011和 DSY003-2011,其中,DSY002-2011菌株为地衣芽孢杆菌。由豆粕与红糖、玉米粉、大米粉和麸皮等分别构成的4种生产型培养基均能将DSY002-2011菌株的细菌数扩增至200亿个以上,其中,以红糖20 g/L+豆粕80 g/L的组合最优,其细菌扩增培养浓度可达39×109 cfu/mL。%The combined methods of microbiology and molecular biology were employed to isolate and screen the suitable B.licheniformis for aquaculture and explore the amplification effects of different culture media.Results:Two strains DSY002-2011 and DSY003-2011 with the strongest bacteriostatic action were isolated,in which DSY002-2011 was B.licheniformis,the bacterial count could be up to more than 20 billion in four production media composed of bean pulp,brown sugar,corn flour,rice meal and bran.The combination of browning sugar 20 g/L and bean pulp 80 g/L was of the best effects,the multiplication culture concentration of bacteria was up to 39 ×109 cfu/mL.

  1. 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

  2. Genetic Improvement of α-Amylase Producing Bacillus Licheniformis by Expression of Hemoglobin Gene from Vitreoscilla%透明颤菌vgb基因在地衣芽孢杆菌中表达用于改良α-淀粉酶生产菌株

    Institute of Scientific and Technical Information of China (English)

    王凤寰; 杨建国; 汪兵

    2012-01-01

    Bacillus licheniformis α-amylase is one of the most important enzymes involved in starch hydrolysis and food fermentation processes. To improve the α-amylase productivity and reduce the cost of industrial fermentation, an integrative vector pAX01-sacB-vgb carrying vgb gene encoding the Vitreoscilla hemoglobin was constructed and integrated into the chromosome of B. Licheniforms CICC 10181, an industrial α-amylase producer. The transformant showed remarkably improved cell growth and α-amylase production via vgb gene expression. In batch fermentations under same dissolved oxygen, the biomass and α-amylase of recombinant B. Licheniforms WY were 45.8% and 116.7% higher than those of the parent strain B. Licheniforms CICC 10181. This could be an effective way to enhance the industrial production of α-amylase.%地衣芽孢杆菌高温α-淀粉酶是淀粉加工与食品发酵工业上关键酶制剂之一.为了降低高温α-淀粉酶生产过程中所需要的高溶氧水平对设备的要求,进一步提高α-淀粉酶的生产性能,本研究构建同源整合载体pAX01-sacB-vgb,在地衣芽孢杆菌(B.licheniforms)CICC 10181染色体中表达增强摄氧的透明颤菌血红蛋白基因(Vitreoscilla hemoglobin gene,vgb).发酵实验结果表明,在同等溶氧条件下,重组菌比野生菌生物量提高了45.8%,高温α-淀粉酶酶活提高了116.7%,具有良好的工业应用前景.

  3. 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).

  4. Changing the thermostability of Bacillus licheniformis a-amylase

    OpenAIRE

    De Cordt, S.; Saraiva, J.; Hendrickx, M.; Maesmans, G.; Tobback, P

    1993-01-01

    We applied "solvent engineering" (i.e. variation of environmental conditions) to and/or irnrnobilized Baci//us licheniforrnis a-amylase covalently onto porous glass beads. In this way, important alterations in its thermostability characteristics (kinactrE Ainactw) ere achieved.

  5. Bacillus spp. among hospitalized patients with haematological malignancies: clinical features, epidemics and outcomes.

    Science.gov (United States)

    Ozkocaman, V; Ozcelik, T; Ali, R; Ozkalemkas, F; Ozkan, A; Ozakin, C; Akalin, H; Ursavas, A; Coskun, F; Ener, B; Tunali, A

    2006-10-01

    Between April 2000 and May 2005, 350 bacteraemic episodes occurred among patients treated in our haematology unit. Two hundred and twenty-eight of these episodes were caused by Gram-positive pathogens, most commonly coagulase-negative staphylococci and Staphylococcus aureus. One hundred and twenty-two episodes were due to Gram-negative pathogens, with a predominance of Escherichia coli, Acinetobacter baumannii and Pseudomonas aeruginosa. Bacillus bacteraemias constituted 12 of these episodes occurring in 12 patients, and accounted for 3.4% of all bacteraemic episodes. Of the 12 strains evaluated, seven were Bacillus licheniformis, three were Bacillus cereus and two were Bacillus pumilus. Seven episodes presented with bloodstream infection, three with pneumonia, one with severe abdominal pain and deterioration of liver function, and one with a catheter-related bloodstream infection. B. licheniformis was isolated from five patients who had been hospitalized at the same time. This outbreak was related to non-sterile cotton wool used during skin disinfection. B. cereus and B. licheniformis isolates were susceptible to cefepime, carbapenems, aminoglycosides and vancomycin, but B. pumilus isolates were resistant to all antibiotics except for quinolones and vancomycin. Two deaths were observed. In conclusion, Bacillus spp. may cause serious infections, diagnostic and therapeutic dilemmas, and high morbidity and mortality in patients with haematological malignancies. Both B. cereus and B. licheniformis may be among the 'new' Gram-positive pathogens to cause serious infection in patients with neutropenia.

  6. Analysis of a preferential action of α-amylase from B. licheniformis towards amorphous regions of waxy maize starch.

    Science.gov (United States)

    Foresti, María Laura; Williams, María del Pilar; Martínez-García, Ricardo; Vázquez, Analía

    2014-02-15

    Waxy maize starch was subjected to α-amylase (Bacillus licheniformis) hydrolysis in buffered medium to determine the evolution of reaction in quantitative terms and also in terms of the morphology and crystallinity of the partially hydrolyzed starch granules. Gathered data allowed studying the pattern of action of this α-amylase over waxy maize starch granules, with particular focus on a preferential hydrolysis of the amorphous regions of starch. Results showed that waxy maize starch hydrolysis followed a two-stage kinetic profile with an initial stage characterized by high reaction rate, followed by a slower second stage. The change of hydrolysis rate occurred at approximately 6h of reaction, a time for which X-ray diffraction data quantitatively analyzed by three different techniques showed a maximum of crystallinity in partially hydrolyzed granules. Scanning electron microscopy images illustrated the action of α-amylases which implied the exoerosion of the granules surface, the entry of α-amylases into the granules through radial channels, their endoerosion towards the granule exterior, and their fragmentation. Fragmentation of waxy maize starch granules revealed internal layered structures of starch which were interpreted as hydrolyzed/non-hydrolyzed growth rings. Under the conditions chosen, kinetic, electron microscopy and X-ray data all gave evidence of a preferential action of α-amylase from Bacillus licheniformis towards the less ordered regions of waxy maize starch. Results showed that, provided the proper hydrolysis time is chosen, starch granules with increased crystallinity can be obtained by a pure enzymatic treatment.

  7. Bacillus species isolated from tungrymbai and bekang, naturally fermented soybean foods of India.

    Science.gov (United States)

    Chettri, Rajen; Tamang, Jyoti Prakash

    2015-03-16

    Tungrymbai and bekang are naturally fermented soybean foods commonly consumed in Meghalaya and Mizoram states of India. A total of 39 samples of tungrymbai and 43 samples of bekang were collected from different villages and markets of Meghalaya and Mizoram, respectively and were analysed for microbial load. In both tungrymbai and bekang, the average population of Bacillus spp. was 8.2±0.1 log cfu/g. A total of 428 isolates of Bacillus were isolated from tungrymbai (211) and bekang (217) for detailed identification. On the basis of a combination of phenotypic and molecular characterisation using ARDRA, ITS-PCR and RAPD-PCR techniques, species of Bacillus isolated from tungrymbai were identified as Bacillus licheniformis (25.5%), Bacillus pumilus (19.5%) and Bacillus subtilis (55%), and species of Bacillus from bekang were Bacillus brevis (2%), Bacillus circulans (7.5%), Bacillus coagulans (6.5%), B. licheniformis (16.5%), B. pumilus (9.1%), Bacillus sphaericus (4.6%), B. subtilis (51.8%), and Lysinibacillus fusiformis (2%). The most dominant bacterium in both products was B. subtilis.

  8. 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

  9. Distribution and identification of proteolytic Bacillus spp. in paddy field soil under rice cultivation.

    Science.gov (United States)

    Watanabe, K; Hayano, K

    1993-07-01

    Proteolytic bacteria in paddy field soils under rice cultivation were characterized and enumerated using azocoll agar plates. Bacillus spp. were the proteolytic bacteria that were most frequently present, comprising 59% of the isolates. They were always the numerically dominant proteolytic bacteria isolated from three kinds of fertilizer treatments (yearly application of rice-straw compost and chemical fertilizer, yearly application of chemical fertilizer, and no fertilizer application) and at three different stages of rice development (vegetative growth stage, maximal tillering stage, and harvest stage). Of the 411 proteolytic bacteria isolated, 124 isolates had stronger proteolytic activity than others on the basis of gelatin liquefaction tests and most of them were Bacillus spp. (100% in 1989 and 92.4% in 1991). Bacillus subtilis and Bacillus cereus were the main bacteria of this group and Bacillus mycoides, Bacillus licheniformis, and Bacillus megaterium were also present. We conclude that these Bacillus spp. are the primary source of soil protease in these paddy fields.

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

    DEFF Research Database (Denmark)

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

    2008-01-01

    -hemolytic enterotoxin (NheA, NheB, NheC) and EM1 specific of emetic toxin producerswas 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......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 (BCETRPLA) and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (BDEVIA). Detection of genes encoding´cytotoxin K (CytK), haemolysin BL (Hbl A, Hbl C, Hbl D), non...

  11. Mechanisms of biofilm formation in paper machine by Bacillus species: the role of Deinococcus geothermalis.

    Science.gov (United States)

    Kolari, M; Nuutinen, J; Salkinoja-Salonen, M S

    2001-12-01

    Mechanisms for the undesired persistence of Bacillus species in paper machine slimes were investigated. Biofilm formation was measured for industrial Bacillus isolates under paper machine wet-end-simulating conditions (white water, pH 7, agitated at 45 degrees C for 1-2 days). None of the 40 tested strains of seven Bacillus species formed biofilm on polished stainless steel or on polystyrene surfaces as a monoculture. Under the same conditions, Deinococcus geothermalis E50051 covered all test surfaces as a patchy thick biofilm. The paper machine bacilli, however, formed mixed biofilms with D. geothermalis E50051 as revealed by confocal microscopy. Biofilm interactions between the bacilli and the deinococci varied from synergism to antagonism. Synergism in biofilm formation of D. geothermalis E50051 was strongest with Bacillus coagulans D50192, and with the type strains of B. coagulans, B. amyloliquefaciens or B. pumilus. Two B. licheniformis, one B. amyloliquefaciens, one B. pumilus and four B. cereus strains antagonized biofilm production by D. geothermalis. B. licheniformis D50141 and the type strain of B. licheniformis were the strongest antagonists. These bacteria inhibited deinococcal growth by emitting heat-stable, methanol-soluble metabolite(s). We conclude that the persistence of Bacillus species in paper machine slimes relates to their ability to conquer biofilms formed by primary colonizers, such as D. geothermalis.

  12. Identification of single nucleotide polymorphisms (SNPs) in the 16S rRNA gene of foodborne Bacillus spp.

    Science.gov (United States)

    Fernández-No, I C; Böhme, K; Caamaño-Antelo, S; Barros-Velázquez, J; Calo-Mata, P

    2015-04-01

    The main goal of this work was the identification of single nucleotide polymorphisms (SNPs) in the 16S rRNA gene of foodborne Bacillus spp. that may be useful for typing purposes. These species include, among others, Bacillus cereus, an important pathogenic species involved in food poisoning, and Bacillus licheniformis, Bacillus subtilis and Bacillus pumilus, which are causative agents of food spoilage described as responsible for foodborne disease outbreaks. With this purpose in mind, 52 Bacillus strains isolated from culture collections and fresh and processed food were considered. SNP type "Y" at sites 212 and 476 appeared in the majority of B. licheniformis studied strains. SNP type "R" at site 278 was detected in many strains of the B. subtilis/Bacillus amyloliquefaciens group, while polymorphism "Y" at site 173 was characteristic of the majority of strains of B. cereus/Bacillus thuringiensis group. The analysis of SNPs provided more intra-specific information than phylogenetic analysis in the cases of B. cereus and B. subtilis. Moreover, this study describes novel SNPs that should be considered when designing 16S rRNA-based primers and probes for multiplex-PCR, Real-Time PCR and microarray systems for foodborne Bacillus spp.

  13. 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.

  14. Bacillus coagulans

    Science.gov (United States)

    Bacillus coagulans is a type of bacteria. It is used similarly to lactobacillus and other probiotics as "beneficial" bacteria. People take Bacillus coagulans for diarrhea, including infectious types such as ...

  15. Construction of acetoin high-producing Bacillus subtilis strain

    Directory of Open Access Journals (Sweden)

    Yanjun Tian

    2016-07-01

    Full Text Available This paper describes the construction and selection of a high-producing mutant, Bacillus subtilis HB-32, with enhanced acetoin yield and productivity. The mutant was obtained by the protoplast fusion of a Bacillus subtilis mutant TH-49 (Val− producing acetoin and Bacillus licheniformis AD-30 producing α-acetolactate decarboxylase, with the fusogen polyethylene glycol and after the regeneration and selection, etc. of the fusant. The acetoin production reached 49.64 g/L, which is an increase of 61.8% compared to that of B. subtilis strain TH-49. Random amplified polymorphic DNA analysis was performed to determine the mutagenic and protoplast fusion effects and the genomic changes in the acetoin high-producing strain compared to the parent strains at the molecular level. The constructed strain was shown to be promising for large-scale acetoin production. Future studies should focus on the application of the mutant strain in practice.

  16. 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.

  17. 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.

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

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    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.

  19. Bacillus phytases: Current status and future prospects.

    Science.gov (United States)

    Borgi, Mohamed Ali; Boudebbouze, Samira; Mkaouar, Héla; Maguin, Emmanuelle; Rhimi, Moez

    2015-01-01

    Phytases catalyze the hydrolysis of phytic acid in a stepwise manner to lower inositol phosphates, myo-inositol (having important role in metabolism and signal transduction pathways), and inorganic phosphate. These enzymes have been widely used in animal feed in order to improve phosphorus nutrition and to decrease pollution in animal waste. Compared to previously described phytases, the phytase (PhyL) from Bacillus licheniformis ATCC 14580 has attractive biochemical properties which can increase the profitability of several biotechnological procedures (animal nutrition, humain health…etc). Due to its amino acid sequence with critical substitutions, the PhyL could be a model to enhance other phytases features, in terms of thermal stability and high activity. Otherwise, an engineered PhyL, with low pH optimum, will represent a challenge within the class of β- propeller phytases.

  20. Production of the Novel Two-Peptide Lantibiotic Lichenicidin by Bacillus licheniformis DSM 13

    OpenAIRE

    2009-01-01

    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 ...

  1. 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...

  2. THE KINETICS OF THE REACTIONS CATALYZED BY AN ENZYMATIC PREPARATION PRODUCED BY A BACILLUS LICHENIFORMIS STRAIN

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    DRAGOMIRESCU MONICA

    2007-01-01

    Full Text Available Robust immobilization techniques that preserve the activity of biomolecules havemany potential applications. In recent years, a number of new bioimobilisationmethods in sol-gel-derived materials were reported. The interactions between thebiomolecule and the inorganic material determine the degree to which thebiomolecule retains its native properties. The newer technological developments inthe field of immobilized biocatalysts can offer the possibility of a wider and moreeconomical exploitation of biocatalysts in biological applications, food and feedindustry, medicine, and in the development of bioprocess monitoring devices, like thebiosensors.The aim of this study was to obtain immobilized enzymatic preparations by methodswhich affect enzyme conformations and kinetic parameters as less as possible. Weimmobilized the enzymatic preparation with protease activity produced by a Bacilluslicheniformis B 40 local strain by physical bonding on ceramics and entrapment intosol-gel-derived glasses obtained from tetraethyl orthosilicate (TEOS, deposited inthin layer on a ceramic support (entrapment/deposition. Both physically adsorbedand entrapped/deposited enzymes follow Michaelis-Menten kinetics, similar with thesoluble enzyme. In the case of immobilized enzymes, the apparent Michaelisconstant, Km, was greater than that of the native one, as it was expected. The kineticparameters indicate that the enzymatic preparations adsorbed on ceramic supportand entrapped/deposited show less affinity for the substrate, Km being 1.3 and 2.1times higher than that of the native enzyme, respectively. The maximum velocityincreased also by 3.5 and 7.9 times respectively, compared with the free counterpart(according to Lineweaver-Burk linearization.

  3. 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.

  4. Prospecting Agro-waste Cocktail: Supplementation for Cellulase Production by a Newly Isolated Thermophilic B. licheniformis 2D55.

    Science.gov (United States)

    Kazeem, Muinat Olanike; Shah, Umi Kalsom Md; Baharuddin, Azhari Samsu; AbdulRahman, Nor' Aini

    2017-02-07

    Bacteria isolated from thermophilic environment that can produce cellulase as well as utilise agro-waste biomass have a high potential for developing thermostable cellulase required in the biofuel industry. The cost for cellulase represents a significant challenge in converting lignocellulose to fermentable sugars for biofuel production. Among three potential bacteria examined, Bacillus licheniformis 2D55 (accession no. KT799651) was found to produce the highest cellulolytic activity (CMCase 0.33 U/mL and FPase 0.09 U/mL) at 18-24 h fermentation when grown on microcrystalline cellulose (MCC) as a carbon source in shake flask at 50 °C. Cellulase production process was further conducted on the untreated and NaOH pretreated rice straw (RS), rice husk (RH), sugarcane bagasse (BAG) and empty fruit bunch (EFB). Untreated BAG produced the highest FPase (0.160 U/mL), while the highest CMCase (0.150 U/mL) was supported on the pretreated RH. The mixture of untreated BAG and pretreated RH as agro-waste cocktail has remarkably improved CMCase (3.7- and 1.4-fold) and FPase (2.5- and 11.5-fold) compared to the untreated BAG and pretreated RH, respectively. The mechanism of cellulase production explored through SEM analysis and the location of cellulase enzymes of the isolate was also presented. Agro-waste cocktail supplementation provides an alternative method for an efficient production of cellulase.

  5. Bacillus gobiensis sp. nov., isolated from a soil sample.

    Science.gov (United States)

    Liu, Bo; Liu, Guo-Hong; Cetin, Sengonca; Schumann, Peter; Pan, Zhi-Zhen; Chen, Qian-Qian

    2016-01-01

    A Gram-stain-positive, rod-shaped, endospore-forming, aerobic bacterium designated FJAT-4402T, was isolated from the weed rhizosphere soil of the Gobi desert in the Xinjiang Autonomous Region in the north-west of China. Isolate FJAT-4402T grew at 15-40 °C (optimum 30 °C), pH 5-10 (optimum pH 7) and in 0-3 % (w/v) NaCl (optimum 0 %). Phylogenetic analyses, based on 16S rRNA gene sequences, showed that isolate FJAT-4402T was a member of the genus Bacillus and was most closely related to Bacillus licheniformis DSM 13T (96.2 %). The isolate showed 33.3 % DNA-DNA relatedness to the closest reference isolate, B. licheniformis DSM 13T. The diagnostic diamino acid of the peptidoglycan of isolate FJAT-4402T was meso-diaminopimelic acid and the predominant isoprenoid quinone was MK-7. The major cellular fatty acids were anteiso-C15 : 0 (28.5 %), iso-C15 : 0 (20.1 %), anteiso-C17 : 0 (14.3 %), iso-C16 : 0 (9.6 %), C16 : 0 (8.4 %), iso-C17 : 0 (6.2 %) and iso-C14 : 0 (4.7 %) and the DNA G+C content was 42.0 mol%. The phenotypic, chemotaxonomic and genotypic properties indicated that strain FJAT-4402T represents a novel species within the genus Bacillus, for which the name Bacillus gobiensis sp. nov. is proposed. The type strain is FJAT-4402T ( = DSM 29500T = CGMCC 1.12902T).

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

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    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.

  7. 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.

  8. Identification and Molecular Characterization of Genes Coding Pharmaceutically Important Enzymes from Halo-Thermo Tolerant Bacillus

    Science.gov (United States)

    Safary, Azam; Moniri, Rezvan; Hamzeh-Mivehroud, Maryam; Dastmalchi, Siavoush

    2016-01-01

    Purpose: Robust pharmaceutical and industrial enzymes from extremophile microorganisms are main source of enzymes with tremendous stability under harsh conditions which make them potential tools for commercial and biotechnological applications. Methods: The genome of a Gram-positive halo-thermotolerant Bacillus sp. SL1, new isolate from Saline Lake, was investigated for the presence of genes coding for potentially pharmaceutical enzymes. We determined gene sequences for the enzymes laccase (CotA), l-asparaginase (ansA3, ansA1), glutamate-specific endopeptidase (blaSE), l-arabinose isomerase (araA2), endo-1,4-β mannosidase (gmuG), glutaminase (glsA), pectate lyase (pelA), cellulase (bglC1), aldehyde dehydrogenase (ycbD) and allantoinases (pucH) in the genome of Bacillus sp. SL1. Results: Based on the DNA sequence alignment results, six of the studied enzymes of Bacillus sp. SL-1 showed 100% similarity at the nucleotide level to the same genes of B. licheniformis 14580 demonstrating extensive organizational relationship between these two strains. Despite high similarities between the B. licheniformis and Bacillus sp. SL-1 genomes, there are minor differences in the sequences of some enzyme. Approximately 30% of the enzyme sequences revealed more than 99% identity with some variations in nucleotides leading to amino acid substitution in protein sequences. Conclusion: Molecular characterization of this new isolate provides useful information regarding evolutionary relationship between B. subtilis and B. licheniformis species. Since, the most industrial processes are often performed in harsh conditions, enzymes from such halo-thermotolerant bacteria may provide economically and industrially appealing biocatalysts to be used under specific physicochemical situations in medical, pharmaceutical, chemical and other industries. PMID:28101462

  9. A Thermostable α-Amylase Producing Natural Variant of Bacillus spp. Isolated From Soil in Iran

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    Iraj Rasooli

    2008-01-01

    Full Text Available Thermophilic processes appear more stable, rapid and less expensive and facilitate reactant activity and product recovery. Amylases have a quarter of the world enzyme market and thermostable α-amylases possess extensive commercial applications. Since little work has been done on strain isolation, growth and enzyme yield optimization, the level of thermophilic enzyme production remains relatively low. Therefore, large scale exploitation of thermophiles requires further intensive and integrated work. The present study describes isolation of an α-amylase producing bacillus from soil. The isolated bacillus was identified and named as Bacillus licheniformis Shahed-07. The strain was cultured in liquid media to produce α-amylase. The enzyme production conditions of the newly isolated bacillus revealed that the maximum enzyme production after 26 h of cultivation at pH 7.0 and 50°C. 0.5% tryptophan in production medium enhanced the enzyme productivity to two fold whereas peptone and lysin at 0.5% level showed a strong repression. Crude α-amylase characterization revealed that optimum activity was at pH 7.5 and 70°C. The crude enzyme was stable for 24 h at pH range of 6-7 at 70°C. Enzyme activity increased with temperature within the range of 40-70°C. The Bacillus licheniformis Shahed-07 strain produced thermostable α-amylase with characteristics suitable for application in starch processing and food industries.

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

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    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.

  11. Butyric acid released during milk lipolysis triggers biofilm formation of Bacillus species.

    Science.gov (United States)

    Pasvolsky, Ronit; Zakin, Varda; Ostrova, Ievgeniia; Shemesh, Moshe

    2014-07-02

    Bacillus species form biofilms within milking pipelines and on surfaces of equipment in the dairy industry which represent a continuous hygiene problem and can lead to serious economic losses due to food spoilage and equipment impairment. Although much is known about the mechanism by which the model organism Bacillus subtilis forms biofilms in laboratory mediums in vitro, little is known of how these biofilms are formed in natural environments such as milk. Besides, little is known of the signaling pathways leading to biofilm formation in other Bacillus species, such as Bacillus cereus and Bacillus licheniformis, both of which are known to contaminate milk. In this study, we report that milk triggers the formation of biofilm-related structures, termed bundles. We show this to be a conserved phenomenon among all Bacillus members tested. Moreover, we demonstrate that the tasA gene, which encodes a major portion of the matrix which holds the biofilm together, is vital for this process. Furthermore, we show that the free fatty acid (FFA) - butyric acid (BA), which is released during lipolysis of milk fat and demonstrates antimicrobial activity, is the potent trigger for biofilm bundle formation. We finally show that BA-triggered biofilm bundle formation is mediated by the histidine kinase, KinD. Taken together, these observations indicate that BA, which is a major FFA within milk triggers biofilm formation in a conserved mechanism among members of the Bacillus genus.

  12. In vitro susceptibility of Bacillus spp. to selected antimicrobial agents.

    Science.gov (United States)

    Weber, D J; Saviteer, S M; Rutala, W A; Thomann, C A

    1988-01-01

    Although often dismissed as contaminants when isolated from blood cultures, Bacillus spp. are increasingly recognized as capable of causing serious systemic infections. As part of a clinical-microbiological study, 89 strains of Bacillus spp. isolated from clinical blood cultures between 1981 and 1985 had their species determined and were tested for antimicrobial agent susceptibility to 18 antibiotics. Species of isolates were determined by the API 50CH and API 20E systems. Bacillus cereus (54 strains) was the most common species isolated, followed by B. megaterium (13 strains), B. polymyxa (5 strains), B. pumilus (4 strains), B. subtilis (4 strains), B. circulans (3 strains), B. amyloliquefaciens (2 strains), B. licheniformis (1 strain), and Bacillus spp. (3 strains). Microdilution MIC susceptibility tests revealed all B. cereus strains to be susceptible to imipenem, vancomycin, chloramphenicol, gentamicin, and ciprofloxacin. Non-B. cereus strains were most susceptible to imipenem, vancomycin, LY146032, and ciprofloxacin. Disk susceptibility testing suggested that B. cereus was rarely susceptible to penicillins, semisynthetic penicillins, or cephalosporins with the exception of mezlocillin. In contrast, many non-B. cereus strains were susceptible to penicillins, semisynthetic penicillins, and cephalosporins, but marked variability was noted among species. PMID:3395100

  13. Maintenance metabolism and carbon fluxes in Bacillus species

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    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

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

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    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.

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

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    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.

  16. 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.

  17. Inactivation of Bacillus spores inoculated in milk by Ultra High Pressure Homogenization.

    Science.gov (United States)

    Amador Espejo, Genaro Gustavo; Hernández-Herrero, M M; Juan, B; Trujillo, A J

    2014-12-01

    Ultra High-Pressure Homogenization treatments at 300 MPa with inlet temperatures (Ti) of 55, 65, 75 and 85 °C were applied to commercial Ultra High Temperature treated whole milk inoculated with Bacillus cereus, Bacillus licheniformis, Bacillus sporothermodurans, Bacillus coagulans, Geobacillus stearothermophilus and Bacillus subtilis spores in order to evaluate the inactivation level achieved. Ultra High-Pressure Homogenization conditions at 300 MPa with Ti = 75 and 85 °C were capable of a spore inactivation of ∼5 log CFU/mL. Furthermore, under these processing conditions, commercial sterility (evaluated as the complete inactivation of the inoculated spores) was obtained in milk, with the exception of G. stearothermophilus and B. subtilis treated at 300 MPa with Ti = 75 °C. The results showed that G. stearothermophilus and B. subtilis have higher resistance to the Ultra High-Pressure Homogenization treatments applied than the other microorganisms inoculated and that a treatment performed at 300 MPa with Ti = 85 °C was necessary to completely inactivate these microorganisms at the spore level inoculated (∼1 × 10(6) CFU/mL). Besides, a change in the resistance of B. licheniformis, B. sporothermodurans, G. stearothermophilus and B. subtilis spores was observed as the inactivation obtained increased remarkably in treatments performed with Ti between 65 and 75 °C. This study provides important evidence of the suitability of UHPH technology for the inactivation of spores in high numbers, leading to the possibility of obtaining commercially sterile milk.

  18. Bacillus probiotics.

    Science.gov (United States)

    Cutting, Simon M

    2011-04-01

    Bacterial spore formers are being used as probiotic supplements for use in animal feeds, for human dietary supplements as well as in registered medicines. Their heat stability and ability to survive the gastric barrier makes them attractive as food additives and this use is now being taken forward. While often considered soil organisms this conception is misplaced and Bacilli should be considered as gut commensals. This review summarises the current use of Bacillus species as probiotics, their safety, mode of action as well as their commercial applications.

  19. 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

  20. 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

  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. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. Use of Probiotic Bacillus spp. in Rotifer (Brachionus plicatilis) and Artemia (Artemia urmiana) Enrichment: Effects on Growth and Survival of Pacific White Shrimp, Litopenaeus vannamei, Larvae.

    Science.gov (United States)

    Jamali, Hadi; Imani, Ahmad; Abdollahi, Daruosh; Roozbehfar, Reza; Isari, Amin

    2015-06-01

    This study was to evaluate the effect of a preparation of Bacillus probiotic (Bacillus licheniformis and B. subtilis, 1:1) on growth and survival rate of Pacific white shrimp, Litopenaeus vannamei larvae. The larvae were fed on Artemia urmiana nauplii and Brachionus plicatilis enriched with the probiotic preparation at 1 × 10(6) CFU mL(-1) rate. The experimental setup was completely randomized design comprised of six treatments, namely solo Artemia nauplii (A) or rotifer (R), Artemia nauplii and rotifer without any enrichment (A + R), Artemia nauplii enrichment with probiotic bacilli (Bacillus licheniformis and B. subtilis) (A + B), rotifer enrichment with probiotic bacilli (R + B) and enriched Artemia nauplii and rotifer (A + R + B). All treatments were performed in triplicate. Chemical parameters of rearing water viz. pH, salinity and temperature were 7.5-8, 30-31 ppt and 31-32 °C, respectively. Photoperiod was 16L:8D. Shrimp larvae were fed Artemia nauplii and rotifers at 5-20 and 10-40 individuals per shrimp larvae four times a day, respectively. Growth and survival rate of larvae were determined at MII, MIII, PL1, PL4, PL7 and PL10 stages. Larvae in A + R + B treatment showed the highest total length (10.89 ± 0.51 mm), weight (674 ± 73 μg) and survival rate (65% ± 3.5). Lowest total length, weight and survival rate (7.96 ± 0.63 mm, 493 ± 52 μg and 24.5 ± 2.4%, respectively) were recorded in treatment B larvae. We concluded that Bacillus probiotic can improve growth and survival rate of Pacific white shrimp larvae without conceivably undesirable effects.

  10. 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.

  11. 76 FR 14289 - Bacillus thuringiensis

    Science.gov (United States)

    2011-03-16

    ... AGENCY 40 CFR Part 174 Bacillus thuringiensis eCry3.1Ab Protein in Corn; Temporary Exemption From the... regulation extends a temporary exemption from the requirement of a tolerance for residues of Bacillus... permissible level for residues of Bacillus thuringiensis eCry3.1Ab protein in corn. The temporary...

  12. 75 FR 34040 - Bacillus thuringiensis

    Science.gov (United States)

    2010-06-16

    ... AGENCY 40 CFR Part 174 Bacillus thuringiensis eCry3.1Ab Protein in Corn; Temporary Exemption from the... regulation establishes a temporary exemption from the requirement of a tolerance for residues of Bacillus... Bacillus thuringiensis eCry3.1Ab protein in corn under the FFDCA. The temporary tolerance exemption...

  13. 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.

  14. Halotolerant, biosurfactant-producing Bacillus species potentially useful for enhanced oil recovery

    Energy Technology Data Exchange (ETDEWEB)

    Jenneman, G.E.; McInerney, M.J.; Knapp, R.M.; Clark, J.B.; Feero, J.M.; Revus, D.E.; Menzie, D.E.

    1983-01-01

    A biosurfactant-producing Bacillus licheniformis was isolated from oil-field injection water with properties potentially useful for in situ enhanced oil recovery. Conventional miscible flooding procedures use expensive synthetic detergents such as petroleum sulfonates that precipitate in high NaCl brines and adsorb to rock surfaces. The Bacillus sp. produced a biosurfactant when grown at 40 C in a sucrose mineral salts medium containing 5% NaCl. The biosurfactant was produced during the log phase of growth in the presence or absence of either crude oil or hexadecane. The surface tension of a 5% NaCl solution decreased from 74.0 mN/m to 27 mN/m when the surfactant was added. Interfacial tension of a 5% NaCl brine/octane mixture was as low as 0.43 mN/m when measured by a spinning drop tensiometer. The surfactant was extracted by acid precipitation at a pH of 2.0. The extracted surfactant exhibited optimal surface tension-lowering ability in 4-5% NaCl solutions between pH's of 6.0 to 10.0. The addition of calcium up to 340 mg/liter and incubation temperatures up to 100 C did not alter appreciably the surfactant activity. Mobilization of crude oil and oil bank formation occurred in a sandpack column after addition of the biosurfactant. 16 references, 1 figure, 2 tables.

  15. Recovery of Bacillus and Pseudomonas spp. from the 'fired plots' under shifting cultivation in northeast India.

    Science.gov (United States)

    Pandey, Anita; Chaudhry, Shivaji; Sharma, Avinash; Choudhary, Vipin Singh; Malviya, Mukesh Kumar; Chamoli, Swati; Rinu, K; Trivedi, Pankaj; Palni, Lok Man S

    2011-01-01

    Soil samples, collected after the fire operations at agricultural sites under shifting cultivation in northeast India, were subjected to physico-chemical and microbial analysis. The fire affected various physico-chemical properties of the soil. Significant differences in pH and electrical conductivity were recorded in soil of fired and fallow plots. Significantly higher amounts of total organic carbon and nitrogen were estimated in fallow plots as compared to the fired. Difference in total phosphates was not significant. The fire operations resulted in stimulation of microbial communities. The bacteria were the most affected group followed by actinomycetes and fungi, respectively. The bacterial and actinomycetes counts were significantly higher in fired plots as compared to the fallow plots. The representative bacterial species recovered from the 'fired plots' belonged to the genus Bacillus and Pseudomonas. 16S rRNA analysis revealed their maximum similarity with B. clausii, B. licheniformis, B. megaterium, B. subtilis, B. thuringiensis, P. aeruginosa and P. stutzeri. Most of these species were found to be positive for phosphate solubilization and antagonism in plate based assays. In view of the importance of Bacillus and Pseudomonas species in plant growth promotion and biocontrol, recovery of these species after fire operations is indicative of the microbiological merit of shifting cultivation.

  16. Bacillus velezensis is not a later heterotypic synonym of Bacillus amyloliquefaciens; Bacillus methylotrophicus, Bacillus amyloliquefaciens subsp plantarum and ‘Bacillus oryzicola’ are later heterotypic synonyms of Bacillus

    Science.gov (United States)

    The rhizosphere isolated bacteria belonging to the Bacillus amyloliquefaciens subsp. plantarum and Bacillus methylotrophicus clades are an important group of strains that are used as plant growth promoters and antagonists of plant pathogens. These properties have made these strains the focus of comm...

  17. 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 were...

  18. 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

  19. 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

  20. 地衣芽孢杆菌基因文库的构建及分析%Construction and Analysis of Genomic Library of Bacillus licheni formis

    Institute of Scientific and Technical Information of China (English)

    熊裕焱; 彭雅娟; 卢瑞; 于怡; 王娜; 李清彪; 何宁

    2011-01-01

    作为一种高效无毒、无二次污染以及能够生物降解的水处理剂,生物絮凝剂受到越来越多的关注.以一株具有强絮凝活性地衣芽孢杆菌(Bacillus licheni formis)为实验菌,通过构建基因组文库,尝试筛选絮凝基因阳性克隆子.结果表明,该菌株基因文库构建成功,共得到2.1×103克隆子.随机挑取17个阳性克隆子进行DNA测序分析,发现一些相对保守蛋白及2个不具有明确功能的基因.该结果为进一步研究地衣芽孢杆菌全基因组结构功能及细菌絮凝基因奠定了基础.%As an efficient,innoxious, and biodegradable wastewater treatment agent with no secondary pollution, bioflocculant is receiving more and more concerns nowadays. In this paper,Bacillus licheniformis with excellent flocculating activity was used to construct the genomic library and tried to screen out the positive clones with flocculation gene. The results showcd that the genomic library was successfully consrructed,2. 1× 103 clones were screened out. 17 clones were randomly selected for DNA sequencing and analysis. Some relative conserved proteins and 2 novel genes with unknown functions were found. The results lay a foundation for the further study of gene encoding bacteria flocculant and the whole genomic structure and function of Bacillus licheniformis.

  1. Heterologous expression and enzymatic characterization of γ-glutamyltranspeptidase from Bacillus amyloliquefaciens.

    Science.gov (United States)

    Lee, Jung-Min; Lee, Jaejung; Nam, Gyeong-Hwa; Son, Byung-Sam; Jang, Myoung-Uoon; Lee, So-Won; Hurh, Byung-Serk; Kim, Tae-Jip

    2017-02-01

    γ-Glutamyltranspeptidase (GGT) catalyzes the cleavage of γ-glutamyl compounds and the transfer of γ-glutamyl moiety to water or to amino acid/peptide acceptors. GGT can be utilized for the generation of γ-glutamyl peptides or glutamic acid, which are used as food taste enhancers. In the present study, Bacillus amyloliquefaciens SMB469 with high GGT activity was isolated from Doenjang, a traditional fermented soy food of Korea. The gene encoding GGT from B. amyloliquefaciens SMB469 (BaGGT469) was cloned from the isolate, and heterologously expressed in E. coli and B. subtilis. For comparison, three additional GGT genes were cloned from B. subtilis 168, B. licheniformis DSM 13, and B. amyloliquefaciens FZB42. The BaGGT469 protein was composed of 591 amino acids. The final protein comprises two separate polypeptide chains of 45.7 and 19.7 kDa, generated via autocatalytic cleavage. The specific activity of BaGGT469 was determined to be 17.8 U/mg with γ-L-glutamyl-p-nitroanilide as the substrate and diglycine as the acceptor. GGTs from B. amyloliquefaciens showed 1.4- and 1.7-fold higher transpeptidase activities than those from B. subtilis and B. licheniformis, respectively. Especially, recombinant B. subtilis expressing BaGGT469 demonstrated 11- and 23-fold higher GGT activity than recombinant E. coli and the native B. amyloliquefaciens, respectively, did. These results suggest that BaGGT469 can be utilized for the enzymatic production of various γ-glutamyl compounds.

  2. Draft Genome Sequences of Three Alkaliphilic Bacillus Strains, Bacillus wakoensis JCM 9140T, Bacillus akibai JCM 9157T, and Bacillus hemicellulosilyticus JCM 9152T

    OpenAIRE

    Yuki, Masahiro; Oshima, Kenshiro; Suda, Wataru; OSHIDA, Yumi; Kitamura, Keiko; Iida, Toshiya; Hattori, Masahira; Ohkuma, Moriya

    2014-01-01

    Here, we report the draft genome sequences of the type strains of three cellulolytic or hemicellulolytic alkaliphilic Bacillus species: Bacillus wakoensis, Bacillus akibai, and Bacillus hemicellulosilyticus. The genome information for these three strains will be useful for studies of alkaliphilic Bacillus species, their evolution, and biotechnological applications for their enzymes.

  3. 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)

    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-41617**T and Bacillus malacitensis NRRL B-41618**T. Compara...

  4. Nano-Mechanical Properties of Heat Inactivated Bacillus anthracis and Bacillus thuringiensis Spores

    Science.gov (United States)

    2008-03-01

    NANO-MECHANICAL PROPERTIES OF HEAT INACTIVATED BACILLUS ANTHRACIS AND BACILLUS THURINGIENSIS ...GAP/ENP/08-M07 NANO-MECHANICAL PROPERTIES OF HEAT INACTIVATED BACILLUS ANTHRACIS AND BACILLUS THURINGIENSIS SPORES THESIS...AFIT/GAP/ENP/08-M07 NANO-MECHANICAL PROPERTIES OF HEAT INACTIVATED BACILLUS ANTHRACIS AND BACILLUS THURINGIENSIS SPORES Jessica

  5. Effects of two probiotic additives containing Bacillus spores on carcass characteristics, blood lipids and cecal volatile fatty acids in meat type chickens.

    Science.gov (United States)

    Novak, R; Bogovič Matijašić, B; Terčič, D; Cervek, M; Gorjanc, G; Holcman, A; Levart, A; Rogelj, I

    2011-08-01

    The objective of this study was to evaluate effects of two commercially available probiotic additives, containing Bacillus spores, on carcass and meat characteristics, serum lipids and concentration of cecal volatile fatty acids of meat type chickens. Birds were fed regular corn-soy meal based feed (control), supplemented with additive A, containing 1.6 × 10(6) spores per gram of feed of Bacillus subtilis and Bacillus licheniformis (group A) or additive B, containing the same concentration of Bacillus cereus var. toyoi spores (group B). One hundred and twenty birds (20 per replicate) were slaughtered at the age of 55 days. Results showed that birds in group B had higher (p blood serum cholesterol profile. Both probiotics influenced the cecal fermentation, which was observed as decrease in cecal concentrations of propionic, butyric, n-butyric and n-valeric acids, but the differences compared to control group were statistically significant for group A only. It was established that probiotic additive B was more effective regarding carcass and meat part weights than additive A, however the animals from group B also had more abdominal fat and their meat had significantly higher conductivity than control group, which is not considered as beneficial.

  6. 芽孢杆菌生物防治植物病害研究进展%Research Progress of Biological Control in Plant Diseases by Bacillus spp

    Institute of Scientific and Technical Information of China (English)

    朱明妍; 刘姣; 杜春梅

    2012-01-01

    The study mainly introduced and expounded the research and application development of biological control agent made by Bacillus suhtilis , B. laterosporus, B. cereus, B. licheniformis, B. thuringiensis and Paenibacillus polymyxa et al. against plant diseases. The main bio-control mechanism of the Bacillus spp. against on plant diseases was reviewed, including antagonism, competition and induced plant systemic resistance. Moreover, the genetic improvement of Bacillus spp. was mentioned and described. At last, the prospects for the development of the Bacillus spp. in the control of plant diseases was analyzed in briefly.%综述了枯草芽孢杆菌、侧孢芽孢杆菌、蜡样芽孢杆菌、地衣芽孢杆菌、苏云金芽孢杆菌和多粘芽类芽孢杆菌等芽孢杆菌防治植物病害的研究和应用现状;分析了芽孢杆菌防治植物病害的主要作用机制,包括拮抗作用、竞争作用、诱导植物系统抗性等方面;总结了生防芽孢杆菌的遗传改良研究进展,并对芽孢杆菌及其制剂在生物防治领域的发展前景进行了展望.

  7. 一株土壤源高产纤维素酶芽孢杆菌的分离与鉴定%Isolation and Identification of a High Yield Cellulase Bacillus Strain from Soil

    Institute of Scientific and Technical Information of China (English)

    董鹤娟; 丁轲; 恒子钤; 贾艳艳; 彭春平; 罗伟光; 李旺

    2013-01-01

    In order to obtain the high yield cellulase bacillus,58 soil samples from different areas in Henan province were collected,the bacillus producing cellulase was isolated by Congo red medium,and the cellulase activity was detected by 3,5-dinitro salicylic acid method.The high cellulase producing bacillus were screened and identified by the colony morphology,microscopic morphology in combination with the physiological and biochemical feature and 16S rDNA sequence.The results showed that the strain B.LY02 with the highest cellulase activity of 0.5351 U · mL-1 was obtained,and it was short rod,forming spores,gram strain positive,and had the ability of fermenting many sugars.The 16S rDNA sequence homology comparative analysis showed that it belonged to the genus Bacillus and most closery related to Bacillus licheniformis strain CICC10095 with 96.5% sequence simmarity.So the strain Bacillus LY02 was identified to be Bacillus licheniformis.%为了获得产纤维素酶的芽孢杆菌,从河南省不同地方采集玉米地、秸杆垛的土壤样品58份,利用刚果红平板法分离产纤维素酶的芽孢杆菌,采用3,5—二硝基水杨酸法测定纤维素酶活,结合茵落特征、显微形态、生理生化试验和16S rDNA序列进行鉴定.结果表明,从样品中分离出的42株产纤维素酶菌株中筛选出一株产纤维素酶较高的菌株B.LY02,纤维素酶活力可达0.5351 U/mL.该菌株呈短杆状,革兰氏染色阳性,能形成芽孢.基于16S rDNA序列同源性比较分析表明该菌株与Bacillus licheni ormis strain CICC10095的亲缘关系最近,基因序列的同源性为96.5%,因此鉴定该菌株为地衣芽孢杆菌.

  8. 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 ...

  9. Silver nanoparticles synthesis mediated by newly isolates of Bacillus spp., nanoparticles characterization and their activity against Bean Yellow Mosaic Virus and human pathogens

    Directory of Open Access Journals (Sweden)

    Essam K.F. Elbeshehy

    2015-05-01

    Full Text Available Extracellular agents produced by newly isolated bacterial strains were able to catalyze the synthesis of silver nanoparticles (AgNPs. The most effective isolates were identified as Bacillus pumilus, B. persicus and B. licheniformis using molecular identification. DLS analysis revealed that the AgNPs synthesized by the above strains were in the size range of 77-92 nm. TEM observations shown that the nanoparticles were coated with a capping agent, which was probably involved in nanoparticles stabilization allowing their perfect dispersion in aqueous solutions. FTIR analyses indicated the presence of proteins in the capping agent of the nanoparticles and suggested that the oxidation of hydroxyl groups of peptide hydrolysates (originated from the growth medium is coupled to the reduction of silver ions. Energy Dispersive X-ray Spectroscopy confirmed the above results. The nanoparticles, especially those synthesized by B. licheniformis, were stable (zeta potential ranged from -16.6 to -21.3 mV and showed an excellent in vitro antimicrobial activity against important human pathogens and a considerable antiviral activity against the Bean Yellow Mosaic Virus. The significance of the particular antiviral activity is highlighted, given the significant yield reduction in fava bean crops resulting from Bean Yellow Mosaic Virus infections, in many African countries.

  10. 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

  11. 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.

  12. Bacillus piscis sp. nov., a novel bacterium isolated from the muscle of the antarctic fish Dissostichus mawsoni.

    Science.gov (United States)

    Lee, Jae-Bong; Jeon, Seon Hwa; Choi, Seok-Gwan; Jung, Hee-Young; Kim, Myung Kyum; Srinivasan, Sathiyaraj

    2016-12-01

    In this paper, a new bacterial strain designated as 16MFT21(T) is isolated from the muscle of a fish caught in the Antarctic Ocean. Strain 16MFT21(T) is a Gram-staining-positive, catalase-oxidase-positive, rod-shaped facultative-aerobic bacterium. The phylogenetic analysis that is based on the 16S-rRNA gene sequence of strain 16MFT21(T) revealed that it belongs to the genus Bacillus in the family Bacillaceae in the class Bacilli. The highest degrees of the sequence similarity of the strain 16MFT21(T) is with Bacillus licheniformis ATCC 14580(T) (96.6%) and Bacillus sonorensis NBRC 101234(T) (96.6%). The isolate formed a pale-yellow pigment, and it grew in the presence of 0% to 10% (w/v) NaCl (optimum at 2% NaCl), a pH of 6.0 to 10.0 (optimum pH from 7.0 to 8.0), and from 4°C to 30°C (optimum at 30°C). The major polar lipids consist of diphosphatidylglycerol (DPG) and phosphatidylglycerol (PG). The predominant fatty acids are iso-C15:0, anteiso-C15:0, iso-C17:0, and anteiso-C17:0. The main respiratory quinone is menaquinone-7 (MK-7), and based on the use of the meso-diaminopimelic acid as the diagnostic diamino acid, the peptidoglycan cell-wall type is A1γ. Based on the phylogenetic, phenotypic, and chemotaxonomic data, strain 16MFT21(T) (=KCTC 18866(T) =JCM 31664(T)) for which the name Bacillus piscis sp. nov. is proposed should be classified as a new species.

  13. 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

  14. Thermostability enhancement and change in starch hydrolysis profile of the maltohexaose-forming amylase of Bacillus stearothermophilus US100 strain.

    Science.gov (United States)

    Ben Ali, Mamdouh; Khemakhem, Bassem; Robert, Xavier; Haser, Richard; Bejar, Samir

    2006-02-15

    The implications of Asn315 and Val450 in the atypical starch hydrolysis profile of Bacillus stearothermophilus Amy (a-amylase) US100 have been suggested previously [Ben Ali, Mhiri, Mezghani and Bejar (2001) Enzyme Microb. Tech. 28, 537-542]. In order to confirm this hypothesis, three mutants were generated. Of these two have a single mutation, N315D or V450G, whereas the third contains both mutations. Analysis of the starch breakdown-profile of these three mutants, as well as of the wild-type, allowed us to conclude that each single mutation induces a small variation in the hydrolysis product. However, the major end product produced by the double mutant shifts from maltopentaose/maltohexaose to maltose/maltotriose, confirming the involvement of these two residues in starch hydrolysis. The superimposition of AmyUS100 model with that of Bacillus licheniformis shows in AmyUS100 an additional loop containing residues Ile214 and Gly215. Remarkably, the deletion of these two residues increases the half-life at 100 degrees C from 15 min to approx. 70 min. Moreover, this engineered amylase requires less calcium, 25 p.p.m. instead of 100 p.p.m., to reach maximal thermostability.

  15. Keratinase Production by Three Bacillus spp. Using Feather Meal and Whole Feather as Substrate in a Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Ana Maria Mazotto

    2011-01-01

    Full Text Available Three Bacillus species (B. subtilis LFB-FIOCRUZ 1270, B. subtilis LFB-FIOCRUZ 1273, and B. licheniformis LFB-FIOCRUZ 1274, isolated from the poultry industry, were evaluated for keratinase production using feathers or feather meal as the sole carbon and nitrogen sources in a submerged fermentation. The three Bacillus spp. produced extracellular keratinases and peptidases after 7 days. Feather meal was the best substrate for keratinase and peptidase production in B. subtilis 1273, with 412 U/mL and 463 U/ml. The three strains were able to degrade feather meal (62–75% and feather (40–95% producing 3.9–4.4 mg/ml of soluble protein in feather meal medium and 1.9–3.3 mg/ml when feather medium was used. The three strains produced serine peptidases with keratinase and gelatinase activity. B. subtilis 1273 was the strain which exhibited the highest enzymatic activity.

  16. Anti-adhesion activity of two biosurfactants produced by Bacillus spp. prevents biofilm formation of human bacterial pathogens.

    Science.gov (United States)

    Rivardo, F; Turner, R J; Allegrone, G; Ceri, H; Martinotti, M G

    2009-06-01

    In this work, two biosurfactant-producing strains, Bacillus subtilis and Bacillus licheniformis, have been characterized. Both strains were able to grow at high salinity conditions and produce biosurfactants up to 10% NaCl. Both extracted-enriched biosurfactants showed good surface tension reduction of water, from 72 to 26-30 mN/m, low critical micelle concentration, and high resistance to pH and salinity. The potential of the two lipopeptide biosurfactants at inhibiting biofilm adhesion of pathogenic bacteria was demonstrated by using the MBEC device. The two biosurfactants showed interesting specific anti-adhesion activity being able to inhibit selectively biofilm formation of two pathogenic strains. In particular, Escherichia coli CFT073 and Staphylococcus aureus ATCC 29213 biofilm formation was decreased of 97% and 90%, respectively. The V9T14 biosurfactant active on the Gram-negative strain was ineffective against the Gram-positive and the opposite for the V19T21. This activity was observed either by coating the polystyrene surface or by adding the biosurfactant to the inoculum. Two fractions from each purified biosurfactant, obtained by flash chromatography, fractions (I) and (II), showed that fraction (II), belonging to fengycin-like family, was responsible for the anti-adhesion activity against biofilm of both strains.

  17. Comparative genome analysis of Bacillus cereus group genomes with Bacillus subtilis

    OpenAIRE

    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

    2005-01-01

    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-...

  18. 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.

  19. 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.

  20. Smallpox and pan-Orthodox Virus Detection by Real-Time 3’-Minor Groove Binder TaqMan Assays Oil the Roche LightCycler and the Cepheid Smart Cycler Platforms

    Science.gov (United States)

    2007-11-02

    Bacillus anthracis BA0068 Ames Sterne SPS 97.13.213 Bacillus cereus Bacillus coagulans Bacillus licheniformis Bacillus macerans Bacillus ...megaterium Bacillus polymyxa Bacillus sphaericus Bacillus stearothermophilus Bacillus subtilis subsp. niger Bacillus thuringiensis Bacillus popilliae...HA-MGB assay presented here has been used to monitor the viral load in monkey blood and tissues after infection with

  1. Bacillus novalis sp. nov., Bacillus vireti sp. nov., Bacillus soli sp. nov., Bacillus bataviensis sp. nov. and Bacillus drentensis sp. nov., from the Drentse A grasslands.

    Science.gov (United States)

    Heyrman, Jeroen; Vanparys, Bram; Logan, Niall A; Balcaen, An; Rodríguez-Díaz, Marina; Felske, Andreas; De Vos, Paul

    2004-01-01

    A group of 42 isolates were isolated from the soil of several disused hay fields, in the Drentse A agricultural research area (The Netherlands), that were taken out of production at different times. The group represents hitherto-uncultured Bacillus lineages that have previously been found, by a non-cultural method, to be predominant in soil. The strains were subjected to a polyphasic taxonomic study, including (GTG)5-PCR, 16S rDNA sequence analysis, DNA-DNA hybridizations, DNA base-ratio determination, fatty acid analysis and morphological and biochemical characterization. By comparing the groupings obtained by (GTG)5-PCR and 16S rDNA sequence analysis, six clusters of similar strains could be recognized. A DNA-DNA relatedness study showed that these clusters represented five novel genospecies. Further analysis supported the proposal of five novel species in the genus Bacillus, namely Bacillus novalis sp. nov. (type strain IDA3307T=R-15439T=LMG 21837T=DSM 15603T), Bacillus vireti sp. nov. (type strain IDA3632T=R-15447T=LMG 21834T=DSM 15602T), Bacillus soli sp. nov. (type strain IDA0086T=R-16300T=LMG 21838T=DSM 15604T), Bacillus bataviensis sp. nov. (type strain IDA1115T=R-16315T=LMG 21833T=DSM 15601T) and Bacillus drentensis sp. nov. (type strain IDA1967T=R-16337T=LMG 21831T=DSM 15600T).

  2. Use of Nonspecific, Glutamic Acid-Free, Media and High Glycerol or High Amylase as Inducing Parameters for Screening Bacillus Isolates Having High Yield of Polyglutamic Acid.

    Science.gov (United States)

    Baxi, Nandita N

    2014-01-01

    Out of fifty-five Bacillus isolates obtained from ten different regional locations and sources, seven showed the ability to consistently produce specific extracellular polymeric substance (EPS) on rich as well as synthetic but nonspecific media which did not contain glutamic acid. The isolates were identified as either Bacillus licheniformis or Bacillus subtilis. The EPS from all isolates was resistant to alpha protease, proteinase K, and was thus of high molecular weight. Further it was detected after SDS-PAGE by methylene blue but not by coomassie blue R staining as in case of proteins with high proportion of acidic amino acids. Cell-free EPS, after acid hydrolysis, showed absence of carbohydrates and presence of only glutamic acid. Thus the native the EPS from all seven isolates was confirmed to be gamma polyglutamic acid (PGA) and not exopolysaccharide. The Bacillus isolate T which produced maximum polymer on all media tested had higher amylase: protease activity as compared to other strains. If inoculum was developed in rich medium as compared to synthetic medium, the PGA produced increased by twofold in the subsequent synthetic production medium. Similarly, use of inoculum consisting of young and vegetative cells also increased the PGA production by twofold though amount of inoculum did not affect yield of PGA. Though PGA was produced in even in the absence of glutamic acid supplementation in the production medium by all isolates, the yield of PGA increased by fourfold in the presence glutamic acid and the maximum yield was 30 g/l for isolate K. The supplementation of glutamine instead of glutamic acid into the medium caused an increase in the viscosity of the non-Newtonian solution of PGA.

  3. COMPARISON BETWEEN THE PERCENTAGE OF INCIDENCE OF MASTITIS CAUSED BY Bacillus spp. AND Staphylococcus spp. IN WINTER SEASON IN KHARTOUM STATE, SUDAN

    Directory of Open Access Journals (Sweden)

    Reem Rabie MOHAMMED SALIH

    2016-07-01

    Full Text Available This study was conducted in certain area at Khartoum State (Eltebna, Falasteen, Shambat, Hilat Kuku, Elhalfaia, Elsamrab and The University of Khartoum farms in winter season to determine the type of mastitis and to compare between the incidence of mastitis caused by Stapylococcus spps and Bacillus spp. The total number of dairy cows, which were examined in 34 investigated farms, amounted to 500 animals, but the number of positive cows infected with mastitis were 100. The milk samples were collected from cows due to complain of owners from clinical cases of mastitis. Hundred milk samples were collected from apparent cases of mastitis. All mastitic cases were examined by visual examination and palpation of the udder: 55% acute mastitis, 44% chronic mastitis and 1% gangrenous mastitis were diagnosed. Milk samples were cultured in Blood agar and MacConkey´s agar for 24 hours at 37º C. The isolation of Bacillus spp. amounted 74% , these constituted 31% Bacillus coagulans, 11% B. cereus, 9% B. subtilis, 9% B. licheniformis, 4% B. circulans, 2% B. lentus, 3% B. mycoides, 3% B. amyloliquefaciens and 2% B. megaterium. The percentages of acute mastitis caused by B. coagulans was 14%, B. subtilis 8%, B. lichneformes 7% and 2% for every followed Bacillus spp. (B. cereus, B. circulans B. lentus, B. mycoides, B. amyloliquefaciens and B. megaterium. The percentage of chronic mastitis caused by Bacillus spp. were as follows: B. coagulans was 17%, B. cereus 9%, 2% for every Bacillus spp. (B. lichneformes, B. circulans and B. lentus and 1% for every followed Bacillus spp. (B. subtilis, B. mycoides, B. amyloliquefaciens and B. megaterium. Staph aureus and Staph hyicus amounted to 24% and the percentage of chronic mastitis caused by Staph aureus was 44% and that caused by Staph hyicus was 8%. The percentage of acute mastitis caused by each species of Staph was the same 24%. Other bacteria were isolated from mastitic cows Corynebaccterium spp. 1% and Klebsiella

  4. NCBI nr-aa BLAST: CBRC-DDIS-04-0058 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DDIS-04-0058 ref|YP_080398.1| Metallophosphoesterase [Bacillus licheniformis A...TCC 14580] ref|YP_092816.1| YkoQ [Bacillus licheniformis ATCC 14580] gb|AAU24760.1| Metallophosphoesterase [Bacillus lichen...iformis ATCC 14580] gb|AAU42123.1| YkoQ [Bacillus licheniformis DSM 13] YP_080398.1 2e-10 31% ...

  5. 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% ...

  6. 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% ...

  7. 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.

  8. 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% ...

  9. Photothermal spectroscopy of Bacillus anthracis and Bacillus cereus with microcantilevers

    Energy Technology Data Exchange (ETDEWEB)

    Wig, Andrew G [ORNL; Arakawa, Edward T [ORNL; Passian, Ali [ORNL; Ferrell, Thomas L [ORNL; Thundat, Thomas George [ORNL

    2006-03-01

    Microcalorimetric optical and infrared spectroscopy is a method of determining the spectral absorption of small quantities of materials over a wide range of incident wavelengths. In this paper, the first spectroscopic results for microcantilevers coated with Bacillus anthracis (BA) are presented. These results, for B. anthracis from 2.5 to 14.5 {micro}m, are compared with results from microcantilevers coated with Bacillus cereus (BC) and standard spectroscopic absorption data. The results demonstrate strong correlation between the deflection measurements and the reference spectroscopic absorption peaks. An advantage of this microcantilever-based method over traditional spectroscopy is that much smaller amounts of material (nanogram quantities) can be detected in comparison with the milligram amounts needed for standard methods. Another advantage is that the complete system can be relatively small without sacrificing spectral resolution.

  10. 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...

  11. 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.

  12. Isolation and identification of Bacillus spp. from compost material, compost and mushroom casing soil active against Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Stanojević Olja

    2016-01-01

    Full Text Available The isolation of bacteria was carried out from samples of straw and chicken manure, compost at various stages of the composting process and casing soil used for growing button mushrooms. A preliminary screening of 108 bacterial isolates for antagonistic activity against Trichoderma aggressivum f. europaeum showed that 23 tested isolates inhibited mycelial growth of the pathogenic fungus. Further screening with four indicator isolates of fungi revealed that all 23 bacterial isolates inhibited the growth of T. aggressivum f. europaeum, T. harzianum and T. koningii, while only 13 isolates inhibited the growth of T. atroviride. T. aggressivum f. europaeum proved to be the most sensitive, with many bacterial isolates generating a high percentage of growth inhibition. Only two bacterial isolates (B-129 and B-268 were successful in inhibiting the growth of all 4 tested pathogens. All 23 bacterial isolates were characterized as Gram-positive and catalase-positive and were subjected to molecular identification based on the partial sequence, the hypervariant region of the 16S rDNA. It was shown that the obtained bacterial strains belong to Bacillus subtilis, B. amyloliquefaciens, B. licheniformis and B. pumilus species. [Projekat Ministarstva nauke Republike Srbije, br. 31043 i br. 173026

  13. 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

  14. beta-Amylase production by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [correction of polymaxa] strains.

    Science.gov (United States)

    Niziołek, S

    1997-01-01

    The production of extracellular beta-amylase by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [corrected] strains was investigated, and the maximal yields of the enzyme were 3.6; 9.3 and 20.4 U/mL of the culture fluid, respectively (U, 1 mumol of maltose equivalent per min at 30 degrees C). Several cultivation media were used for beta-amylase production. Bacillus cereus and some strains of Bacillus megaterium gave good yields of beta-amylase only in medium with the addition of nutrient broth. However, beta-amylase produced during growth in protein rich medium (nutrient broth) was highly unstable, probably due to inactivation by proteolytic enzymes co-existing in the culture fluid. Bacillus polymyxa [corrected] strains can produce good yields of beta-amylase on a semi-synthetic medium consisting of inorganic salts, potato starch and inexpensive soybean extract instead of costly peptone and meat extract. The most potential beta-amylase producer was the strain Bacillus polymyxa [corrected] NCIB 8524. The tested Bacillus megaterium and Bacillus polymyxa [corrected] strains were apparently differentiated by temperature cultivation (30 and 37 degrees C) suitable for beta-amylase amylase yield.

  15. Antimicrobials of Bacillus species: mining and engineering

    OpenAIRE

    Zhao, Xin

    2016-01-01

    Bacillus sp. have been successfully used to suppress various bacterial and fungal pathogens. Due to the wide availability of whole genome sequence data and the development of genome mining tools, novel antimicrobials are being discovered and updated,;not only bacteriocins, but also NRPs and PKs. A new classification system of known and putative antimicrobial compounds of Bacillus by genome mining is presented in Chapter 2. Importantly, predicting, isolating and screening of Bacillus strains w...

  16. Microarray-based Resequencing of Multiple Bacillus anthracis Isolates

    Science.gov (United States)

    2004-12-17

    al.: Iden- tification of anthrax toxin genes in a Bacillus cereus associ- ated with an illness resembling inhalation anthrax. Proc Natl Acad Sci USA...Norwegian Bacillus cereus and Bacillus thuringiensis soil isolates. Appl Environ Microbiol 2001, 67:4863-4873. 26. Radnedge L, Agron PG, Hill KK, Jackson PJ...Ticknor LO, Keim P, Andersen GL: Genome differences that distinguish Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis . Appl

  17. 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.

  18. A selective chromogenic agar that distinguishes Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis.

    Science.gov (United States)

    Juergensmeyer, Margaret A; Gingras, Bruce A; Restaino, Lawrence; Frampton, Elon W

    2006-08-01

    A selective and differential plating medium, R & F anthracis chromogenic agar (ACA), has been developed for isolating and identifying presumptive colonies of Bacillus anthracis. ACA contains the chromogenic substrate 5-bromo-4-chloro-3-indoxyl-choline phosphate that upon hydrolysis yields teal (blue green) colonies indicating the presence of phosphatidylcholine-specific phospholipase C (PC-PLC) activity. Among seven Bacillus species tested on ACA, only members of the Bacillus cereus group (B. anthracis, B. cereus, and B. thuringiensis) produced teal colonies (PC-PLC positive) having cream rings. Examination of colony morphology in 18 pure culture strains of B. anthracis (15 ATCC strains plus AMES-1-RIID, ANR-1, and AMED-RIID), with one exception, required 48 h at 35 to 37 degrees C for significant color production, whereas only 24 h was required for B. cereus and B. thuringiensis. This differential rate of PC-PLC synthesis in B. anthracis (due to the truncated plcR gene and PlcR regulator in B. anthracis) allowed for the rapid differentiation on ACA of presumptive colonies of B. anthracis from B. cereus and B. thuringiensis in both pure and mixed cultures. Effective recovery of B. anthracis from a variety of matrices having both high (soil and sewage) and low microbial backgrounds (cloth, paper, and blood) spiked with B. anthracis ANR-1 spores suggests the probable utility of ACA plating for B. anthracis recovery in a diversity of applications.

  19. Pacemaker-associated Bacillus cereus endocarditis.

    Science.gov (United States)

    Barraud, Olivier; Hidri, Nadia; Ly, Kim; Pichon, Nicolas; Manea, Petrus; Ploy, Marie-Cécile; Garnier, Fabien

    2012-11-01

    We report the case of a pacemaker-associated Bacillus cereus endocarditis in a nonimmunocompromised patient. Antibiotic treatment was ineffective, and the pacemaker had to be removed. B. cereus was cultured from several blood samples and from the pacemaker electrodes. This case underlines the contribution of the rpoB gene for Bacillus species determination.

  20. 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.

  1. Bacillus cereus endocarditis in native aortic valve.

    Science.gov (United States)

    Ngow, H A; Wan Khairina, W M N

    2013-02-01

    Bacillus cereus endocarditis is rare. It has been implicated in immunocompromised individuals, especially in intravenous drug users as well as in those with a cardiac prosthesis. The patient was a 31-year-old ex-intravenous drug addict with a past history of staphylococcal pulmonary valve endocarditis, who presented with symptoms of decompensated cardiac failure. Echocardiography showed severe aortic regurgitation with an oscillating vegetation seen on the right coronary cusp of the aortic valve. The blood cultures grew Bacillus cereus. We report this as a rare case of Bacillus cereus endocarditis affecting a native aortic valve.

  2. Microbial Transformation of Quercetin by Bacillus cereus

    OpenAIRE

    Rao, Koppaka V.; Weisner, Nghe T.

    1981-01-01

    Biotransformation of quercetin was examined with a number of bacterial cultures. In the presence of a bacterial culture (Bacillus cereus), quercetin was transformed into two crystalline products, identified as protocatechuic acid and quercetin-3-glucoside (isoquercitrin).

  3. 75 FR 862 - Bacillus subtilis; Registration Review Proposed Decision; Notice of Availability

    Science.gov (United States)

    2010-01-06

    ... AGENCY Bacillus subtilis; Registration Review Proposed Decision; Notice of Availability AGENCY... proposed registration review decision for the pesticide Bacillus subtilis (case 6012) and opens a public... EPA's proposed registration review decision Bacillus subtilis (case 6012). The Bacillus subtilis...

  4. Bacillus cereus Bacteremia in a Preterm Neonate

    OpenAIRE

    Hilliard, Nicholaus J.; Schelonka, Robert L.; Waites, Ken B.

    2003-01-01

    Bacillus cereus is an uncommon but potentially serious bacterial pathogen causing infections of the bloodstream, lungs, and central nervous system of preterm neonates. A case of bacteremia caused by B. cereus in a 19-day-old preterm neonate who was successfully treated with vancomycin, tobramycin, meropenem, and clindamycin is described. Implications for the diagnostic laboratory and clinicians when Bacillus species are detected in normally sterile sites are discussed, and the small numbers o...

  5. 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.

  6. Role of the salt bridge between Arg176 and Glu126 in the thermal stability of the Bacillus amyloliquefaciens α-amylase (BAA).

    Science.gov (United States)

    Zonouzi, Roseata; Khajeh, Khosro; Monajjemi, Majid; Ghaemi, Naser

    2013-01-01

    In the Bacillus amyloliquefaciens α-amylase (BAA), the loop (residues 176-185; region I) that is the part of the calcium-binding site (CaI, II) has two more amino acid residues than the α-amylase from Bacillus licheniformis (BLA). Arg176 in this region makes an ionic interaction with Glu126 from region II (residues 118-130), but this interaction is lost in BLA owing to substitution of R176Q and E126V. The goal of the present work was to quantitatively estimate the effect of ionic interaction on the overall stability of the enzyme. To clarify the functional and structural significance of the corresponding salt bridge, Glu126 was deleted (ΔE126) and converted to Val (E126V), Asp (E126D), and Lys (E126K) by site-directed mutagenesis. Kinetic constants, thermodynamic parameters, and structural changes were examined for the wild-type and mutated forms using UV-visible, atomic absoption, and fluorescence emission spectroscopies. Wild type exhibited higher k(cat) and K(m) but lower catalytic efficiency than the mutant enzymes. A decreased thermostability and an increased flexibility were also found in all of the mutant enzymes when compared with the wild type. Additionally, the calcium content of the wild type was more than ΔE126. Thus, it may be suggested that ionic interaction could decrease the mobility of the discussed region, prevent the diffusion of cations, and improve the thermostability of the whole enzyme. Based on these observations, the contribution of loop destabilization may be compensated by the formation of a salt bridge that has been used as an evolutionary mechanism or structural adaptation by the mesophilic enzyme.

  7. Comparative Study on Biological Characteristics of Six Bacillus Strains%6株芽孢杆菌的生物学特性比较研究

    Institute of Scientific and Technical Information of China (English)

    李雅丽; 秦艳; 周绪霞; 李卫芬

    2011-01-01

    The main biological characteristics of six Bacillus strains isolated were studied,including the activity of enzyme-producing,the endurance to NaCl,bile salt and artificial gastric juice as well as artificial intestinal fluid and the sensitivity to antibiotics. The results showed that,Bacillus subtilis strain 1, B. coagulans and B. cereus had the capability of producing high yield of protease and amylase. Two strains of B. subtilis and B. coagulans showed well resistance to high concentration of NaCl.B. subtilis strain 2,B. coagulans and B. natto as well as B. cereus had relatively strong tolerance for bovine bile salt. Except B. licheniformis,the other five strains of Bacillus could resist the artificial gastric and intestinal fluid well. Six strains of Bacillus were sensitive to common forage antibiotics. These results showedthat B. subtilis 1, B. coagulans and B. cereus were excellent probiotics.%以实验室分离保存的6株芽孢杆菌为材料,对其主要生物学特性进行了研究,包括产酶活力,对NaCl、牛胆盐、人工胃液和人工肠液的耐受性及对抗生素的敏感性.结果显示.枯草芽孢杆菌1、凝结芽孢杆菌和蜡样芽孢杆菌均能产生较高活性的中性蛋白酶和淀粉酶;2株枯草芽孢杆菌和凝结芽孢杆菌能耐较高浓度的NaCl;枯草芽孢杆菌2、凝结、纳豆和蜡样芽孢杆菌对牛胆盐的耐受性较强;除地衣芽孢杆菌外,其他5株芽孢杆菌对胃液和肠液的耐受性较好;6株芽孢杆菌均对常用饲用抗生素敏感.结果表明,枯草1、凝结、蜡样芽孢杆菌是优良的候选益生菌菌株.

  8. Heterologous Expression of Enzymes for Trehalose Synthesis from Sulfolobus acidocaldarius in Bacillus subtilis%1株古细菌海藻糖合成酶系在枯草芽孢杆菌中的诱导表达

    Institute of Scientific and Technical Information of China (English)

    王珊瑛; 李由然; 顾正华; 张梁; 丁重阳; 石贵阳

    2016-01-01

    来源于古细菌嗜酸硫化叶菌(Sulfolobus acidocaldarius)的麦芽寡糖基海藻糖合成酶(maltooligosylterhalose synthase,MTSase)和麦芽寡糖基海藻糖水解酶(maltooligosyltrehalose trehalohydrolase,MTHase)联合作用,可以利用淀粉为底物生成海藻糖.本研究构建了6种枯草芽孢杆菌-大肠杆菌穿梭表达载体,将密码子优化后的MTSase和MTHase编码基因在木糖异构酶基因的启动子及其阻遏蛋白介导下实现了在枯草芽孢杆菌中的功能表达,木糖启动子分别来源于枯草芽孢杆菌(Bacillus subtilis)、巨大芽孢杆菌(Bacillus megaterium)、地衣芽孢杆菌(Bacillus licheniformis).在以5 g/L甘油为碳源,24 g/L蛋白胨为氮源时,菌体培养8h后加入终质量浓度为6g/L的木糖,37℃诱导16h后重组MTSase、MTHase产生联合作用,海藻糖转化率达到33.57%.实现了MTSase、MTHase在枯草芽孢杆菌中的功能表达.

  9. Fatal Bacillus cereus bacteremia in a patient with diabetes.

    Science.gov (United States)

    Orrett, F A

    2000-04-01

    This report describes a fatal case of Bacillus cereus septicemia in a patient with uncontrolled diabetes and re-emphasizes the potential seriousness of Bacillus infections in patients with compromised immune function.

  10. Human Neutrophils Kill Bacillus anthracis.

    Directory of Open Access Journals (Sweden)

    2005-11-01

    Full Text Available Bacillus anthracis spores cause natural infections and are used as biological weapons. Inhalation infection with B. anthracis, the etiological agent of anthrax, is almost always lethal, yet cutaneous infections usually remain localized and resolve spontaneously. Neutrophils are typically recruited to cutaneous but seldom to other forms of anthrax infections, raising the possibility that neutrophils kill B. anthracis. In this study we infected human neutrophils with either spores or vegetative bacteria of a wild-type strain, or strains, expressing only one of the two major virulence factors. The human neutrophils engulfed B. anthracis spores, which germinated intracellularly and were then efficiently killed. Interestingly, neutrophil killing was independent of reactive oxygen species production. We fractionated a human neutrophil granule extract by high-performance liquid chromatography and identified alpha-defensins as the component responsible for B. anthracis killing. These data suggest that the timely recruitment of neutrophils can control cutaneous infections and possibly other forms of B. anthracis infections, and that alpha-defensins play an important role in the potent anti-B. anthracis activity of neutrophils.

  11. Human neutrophils kill Bacillus anthracis.

    Directory of Open Access Journals (Sweden)

    Anne Mayer-Scholl

    2005-11-01

    Full Text Available Bacillus anthracis spores cause natural infections and are used as biological weapons. Inhalation infection with B. anthracis, the etiological agent of anthrax, is almost always lethal, yet cutaneous infections usually remain localized and resolve spontaneously. Neutrophils are typically recruited to cutaneous but seldom to other forms of anthrax infections, raising the possibility that neutrophils kill B. anthracis. In this study we infected human neutrophils with either spores or vegetative bacteria of a wild-type strain, or strains, expressing only one of the two major virulence factors. The human neutrophils engulfed B. anthracis spores, which germinated intracellularly and were then efficiently killed. Interestingly, neutrophil killing was independent of reactive oxygen species production. We fractionated a human neutrophil granule extract by high-performance liquid chromatography and identified alpha-defensins as the component responsible for B. anthracis killing. These data suggest that the timely recruitment of neutrophils can control cutaneous infections and possibly other forms of B. anthracis infections, and that alpha-defensins play an important role in the potent anti-B. anthracis activity of neutrophils.

  12. Human neutrophils kill Bacillus anthracis.

    Science.gov (United States)

    Mayer-Scholl, Anne; Hurwitz, Robert; Brinkmann, Volker; Schmid, Monika; Jungblut, Peter; Weinrauch, Yvette; Zychlinsky, Arturo

    2005-11-01

    Bacillus anthracis spores cause natural infections and are used as biological weapons. Inhalation infection with B. anthracis, the etiological agent of anthrax, is almost always lethal, yet cutaneous infections usually remain localized and resolve spontaneously. Neutrophils are typically recruited to cutaneous but seldom to other forms of anthrax infections, raising the possibility that neutrophils kill B. anthracis. In this study we infected human neutrophils with either spores or vegetative bacteria of a wild-type strain, or strains, expressing only one of the two major virulence factors. The human neutrophils engulfed B. anthracis spores, which germinated intracellularly and were then efficiently killed. Interestingly, neutrophil killing was independent of reactive oxygen species production. We fractionated a human neutrophil granule extract by high-performance liquid chromatography and identified alpha-defensins as the component responsible for B. anthracis killing. These data suggest that the timely recruitment of neutrophils can control cutaneous infections and possibly other forms of B. anthracis infections, and that alpha-defensins play an important role in the potent anti-B. anthracis activity of neutrophils.

  13. 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.

  14. Measurement of Metabolic Activity in Dormant Spores of Bacillus Species

    Science.gov (United States)

    2015-01-14

    SECURITY CLASSIFICATION OF: Spores of Bacillus megaterium and Bacillus subtilis were harvested shortly after release from sporangia, incubated under...Dec-2014 Approved for Public Release; Distribution Unlimited Final Report: Measurement of Metabolic Activity in Dormant Spores of Bacillus Species...Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 spores, Bacillus , spore dormancy, 3-phosphoglycerate REPORT DOCUMENTATION PAGE 11

  15. Diversity and applications of Bacillus bacteriocins.

    Science.gov (United States)

    Abriouel, Hikmate; Franz, Charles M A P; Ben Omar, Nabil; Gálvez, Antonio

    2011-01-01

    Members of the genus Bacillus are known to produce a wide arsenal of antimicrobial substances, including peptide and lipopeptide antibiotics, and bacteriocins. Many of the Bacillus bacteriocins belong to the lantibiotics, a category of post-translationally modified peptides widely disseminated among different bacterial clades. Lantibiotics are among the best-characterized antimicrobial peptides at the levels of peptide structure, genetic determinants and biosynthesis mechanisms. Members of the genus Bacillus also produce many other nonmodified bacteriocins, some of which resemble the pediocin-like bacteriocins of the lactic acid bacteria (LAB), while others show completely novel peptide sequences. Bacillus bacteriocins are increasingly becoming more important due to their sometimes broader spectra of inhibition (as compared with most LAB bacteriocins), which may include Gram-negative bacteria, yeasts or fungi, in addition to Gram-positive species, some of which are known to be pathogenic to humans and/or animals. The present review provides a general overview of Bacillus bacteriocins, including primary structure, biochemical and genetic characterization, classification and potential applications in food preservation as natural preservatives and in human and animal health as alternatives to conventional antibiotics. Furthermore, it addresses their environmental applications, such as bioprotection against the pre- and post-harvest decay of vegetables, or as plant growth promoters.

  16. 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...

  17. The supercoiling of Bacillus subtilis

    Science.gov (United States)

    Mendelson, Neil H.

    2003-03-01

    Cylindrical shaped cells of Bacillus subtilis (0.7 X 4 mm) grow with twist and when prevented from separating at cell division form long filaments that writhe and supercoil to produce plectonemic fibers. By repetition macrofibers arise consisting of structures mm in length with loops at both ends of a twisted shaft. The entire structure is topologically a single filament. All the cells in a macrofiber also grow with twist consequently as a fiber elongates its loop ends rotate about the axis of the fiber shaft in opposite directions relative to one another. This holds for both right and left-handed structures, with any degree of twist. Although the individual cells grow with constant twist, the rate of loop rotation increases as a function of fiber length. Theory suggests that there is a gradient of rotation rates along the length of a fiber ranging from maxima at the loop ends to zero at the center of its length. In fibers prevented from rotating at one end the rotation rate gradient ranges from zero at the blocked end to maximum at the free end as shown here. When loop rotation at both ends is blocked fibers supercoil and their loop ends move toward one another. Newly designed force gauges were used to measure the tension engendered by supercoiling of such fibers. The findings illustrate a micromachine -like behavior of macrofibers, powered by cell growth, twisting and supercoiling. Biological functions of the micromachine such as self-assembly, translational motions over solid surfaces, and the dragging objects over surfaces appear to utilize only a small fraction of the total power available from the macrofiber micromachine. Collaborators: J.J. Thwaites, P. Shipman, D. Roy, and L. Cheng.

  18. 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.

  19. Bacillus cereus bacteremia in a preterm neonate.

    Science.gov (United States)

    Hilliard, Nicholaus J; Schelonka, Robert L; Waites, Ken B

    2003-07-01

    Bacillus cereus is an uncommon but potentially serious bacterial pathogen causing infections of the bloodstream, lungs, and central nervous system of preterm neonates. A case of bacteremia caused by B. cereus in a 19-day-old preterm neonate who was successfully treated with vancomycin, tobramycin, meropenem, and clindamycin is described. Implications for the diagnostic laboratory and clinicians when Bacillus species are detected in normally sterile sites are discussed, and the small numbers of infant infections proven to be due to this organism that have been described previously are reviewed.

  20. 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

  1. Enterotoxin Production in Natural Isolates of Bacillaceae outside the Bacillus cereus Group

    OpenAIRE

    Phelps, Rebecca J.; McKillip, John L.

    2002-01-01

    Thirty-nine Bacillus strains obtained from a variety of environmental and food sources were screened by PCR for the presence of five gene targets (hblC, hblD, hblA, nheA, and nheB) in two enterotoxin operons (HBL and NHE) traditionally harbored by Bacillus cereus. Seven isolates exhibited a positive signal for at least three of the five possible targets, including Bacillus amyloliquefaciens, B. cereus, Bacillus circulans, Bacillus lentimorbis, Bacillus pasteurii, and Bacillus thuringiensis su...

  2. 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

  3. 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...

  4. 地衣芽孢杆菌对非酒精性脂肪性肝病的治疗作用%The effect of Bnfillus licheniformis on nonalcoholic fatty liver disease

    Institute of Scientific and Technical Information of China (English)

    李海涛; 王雯; 张志坚

    2012-01-01

    Objective To evaluate the effect of Bnfillus licheniformis on nonalcoholic fatty liver disease (NAFLD). Methods 40 healthy volunteers were as normal control group. 82 patients with nonalcoholic fatty liver disease were randomly divided into two groups: 41 patients received the treatment with polyene phosphatdyl choline (456 mg po, tid) (polyene phosphatdyl choline group); 41 patients received polyene phosphatdyl choline (456 mg po, tid) and Bnfillus licheniformis (0.5 g po, tid) (polyene phosphatdyl choline and Bnfillus licheniformis group). The both groups were given low-calorie diet, exercise and other treatment for 6 weeks. The parameters were measured, which including fasting plasma resistin, insulin sensitive index (ISI) by fasting insulin (FINS), alanine aminotransferase (ALT) before and after treatment. Results Before treatment, FINS, ALT, ISI and RETN were higher in polyene phosphatdyl choline group and polyene phosphatdyl choline and Bnfillus licheniformis group than those in normal control group (P < 0.05). FINS, ALT, RETN and ISI significantly decreased (P < 0.05) after treatment. Polyene phosphatdyl choline and Bnfillus licheniformis group's changes were more significant than those of polyene phosphatdyl choline group (P < 0.05). Conclusion Bnfillus licheniformis can decrease IR and the levels of serum endotoxin, improve liver function of NAFLD. It is a choice for NAFLD patients.%目的 探讨地衣芽孢杆菌制剂对非酒精性脂肪性肝病的治疗作用.方法 选择正常健康人40例作为正常对照组.选择临床诊断非酒精性脂肪性肝病患者82例,随机将其分为易善复治疗组和易善复加整肠生治疗组,易善复治疗组41例,口服易善复456 mg,3次/d;易善复加整肠生治疗组41例,口服易善复456 mg及整肠生0.5 g,3次/d,两组均结合低热量饮食、运动等基础治疗,共6周.治疗前、后分别测定肝功能丙氨酸转移酶(ALT)、空腹胰岛素水平(FINS)、胰岛素敏感指

  5. 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...

  6. High Production of Thermostable β-Galactosidase of Bacillus stearothermophilus in Bacillus subtilis

    OpenAIRE

    1985-01-01

    By cloning the β-galactosidase gene of Bacillus stearothermophilus IAM11001 (ATCC 8005) into Bacillus subtilis, enzyme production was enhanced 50 times. β-Galactosidase could be purified to 80% homogeneity by incubating the cell extract of B. subtilis at 70°C for 15 min, followed by centrifugation to remove the denatured proteins. Because of its heat stability and ease of production, β-galactosidase is suitable for application in industrial processes.

  7. 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.

  8. Bacillus cereus panophthalmitis: source of the organism.

    Science.gov (United States)

    Shamsuddin, D; Tuazon, C U; Levy, C; Curtin, J

    1982-01-01

    Serious infections with the "nonpathogenic" Bacillus species are increasingly being recognized, especially in drug abusers. Cases of panophthalmitis secondary to infection with Bacillus cereus, with and without associated bacteremia, have been reported. Three drug abusers with panophthalmitis seen in our hospitals during a three-year period are described, and the similar cases reported in the literature are reviewed. The syndrome is characterized by an acute onset with a rapid fulminating course that eventually leads to enucleation or evisceration of the eye. The pathogenic mechanism is unknown, but is probably related to the production of toxin (lecithinase) by B. cereus. Clindamycin appears to be the antibiotic of choice in the treatment of this infection. In order to identify a possible source of the organism, 59 samples of heroin and injection paraphernalia were cultured. Twenty cultures yielded organisms; Bacillus species were the predominant isolates. Thirty-eight percent of the isolates were identified as B. cereus. Thus, infections caused by Bacillus species in drug abusers can probably be associated with intravenous heroin abuse because heroin mixtures and injection paraphernalia are frequently contaminated with this organism.

  9. 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...

  10. 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...

  11. Complete Genome of Bacillus subtilis Myophage Grass

    OpenAIRE

    Miller, Stanton Y.; Colquhoun, Jennifer M.; Perl, Abbey L.; Chamakura, Karthik R.; Kuty Everett, Gabriel F.

    2013-01-01

    Bacillus subtilis is a ubiquitous Gram-positive model organism. Here, we describe the complete genome of B. subtilus myophage Grass. Aside from genes encoding core proteins pertinent to the life cycle of the phage, Grass has several interesting features, including an FtsK/SpoIIIE protein.

  12. 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.

  13. Complete Genome of Bacillus megaterium Podophage Pookie

    OpenAIRE

    Ladzekpo, Tsonyake N.; DeCrescenzo, Andrew J.; Hernandez, Adriana C.; Kuty Everett, Gabriel F.

    2015-01-01

    Bacteriophage Pookie is a novel podophage, isolated from soil, which infects Bacillus megaterium. B. megaterium is an important host for large-scale recombinant protein production. Here, we present the complete genome of phage Pookie and describe its core features.

  14. Methyl Iodide Fumigation of Bacillus anthracis Spores.

    Science.gov (United States)

    Sutton, Mark; Kane, Staci R; Wollard, Jessica R

    2015-09-01

    Fumigation techniques such as chlorine dioxide, vaporous hydrogen peroxide, and paraformaldehyde previously used to decontaminate items, rooms, and buildings following contamination with Bacillus anthracis spores are often incompatible with materials (e.g., porous surfaces, organics, and metals), causing damage or residue. Alternative fumigation with methyl bromide is subject to U.S. and international restrictions due to its ozone-depleting properties. Methyl iodide, however, does not pose a risk to the ozone layer and has previously been demonstrated as a fumigant for fungi, insects, and nematodes. Until now, methyl iodide has not been evaluated against Bacillus anthracis. Sterne strain Bacillus anthracis spores were subjected to methyl iodide fumigation at room temperature and at 550C. Efficacy was measured on a log-scale with a 6-log reduction in CFUs being considered successful compared to the U.S. Environmental Protection Agency biocide standard. Such efficacies were obtained after just one hour at 55 °C and after 12 hours at room temperature. No detrimental effects were observed on glassware, PTFE O-rings, or stainless steel. This is the first reported efficacy of methyl iodide in the reduction of Bacillus anthracis spore contamination at ambient and elevated temperatures.

  15. Antimicrobials of Bacillus species: mining and engineering

    NARCIS (Netherlands)

    Zhao, Xin

    2016-01-01

    Bacillus sp. have been successfully used to suppress various bacterial and fungal pathogens. Due to the wide availability of whole genome sequence data and the development of genome mining tools, novel antimicrobials are being discovered and updated,;not only bacteriocins, but also NRPs and PKs. A n

  16. 地衣芽孢杆菌1801的分离鉴定及其活性研究%Studies on Is01ation,Classification and Active Substance of Bacillus licheniformis 1801

    Institute of Scientific and Technical Information of China (English)

    王燕; 王开梅; 杨自文

    2007-01-01

    从泡菜样品中筛选到一株对多种食品腐败指示茵都有抑菌活性的菌株,经初步鉴定为地衣芽孢杆菌1801.用TYG培养基进行发酵培养,其抑菌活性在发酵18 h达到高峰,该茵发酵培养物的抑茵活性对蛋白酶K、热、酸碱较稳定.采用pH3沉淀法可将该茵的活性物质进行初步提取.

  17. Bacillus endolithicus sp. nov., isolated from pebbles.

    Science.gov (United States)

    Parag, B; Sasikala, Ch; Ramana, Ch V

    2015-12-01

    Strain JC267T was isolated from pebbles collected from Pingleshwar beach, Gujarat, India. Cells are Gram-stain-positive, facultatively anaerobic, non-motile rods forming sub-terminal endospores in swollen ellipsoidal to oval sporangia. Strain JC267T contains anteiso-C15 : 0, iso-C15 : 0, iso-C14 : 0, iso-C16 : 0, C16 : 0 and anteiso-C17 : 0 as major (>5 %) cellular fatty acids. Polar lipids include phosphatidylglycerol, phospholipids (PL1-3), glycolipids (GL1-2) and an unidentified lipid. Cell-wall amino acids are composed of diagnostic meso-diaminopimelic acid, dl-alanine and a small amount of d-glutamic acid. The genomic DNA G+C content of strain JC267T is 45.5 mol%. The 16S rRNA gene sequence of strain JC267T showed highest sequence similarities of Bacillus when subjected to EzTaxon-e blast analysis. The reassociation values based on DNA-DNA hybridization of strain JC267T with Bacillus halosaccharovorans IBRC-M 10095T and Bacillus niabensis JCM 16399T were 26 ± 1 % and 34 ± 3 %, respectively. Based on taxonomic data obtained using a polyphasic approach, strain JC267T represents a novel species of the genus Bacillus, for which the name Bacillus endolithicus sp. nov. is proposed. The type strain is JC267T ( = IBRC-M 10914T = KCTC 33579T).

  18. 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.

  19. Genetic diversity among Bacillus anthracis, Bacillus cereus and Bacillus thuringiensis strains using repetitive element polymorphism-PCR.

    Science.gov (United States)

    Brumlik, Michael J; Bielawska-Drózd, Agata; Zakowska, Dorota; Liang, Xudong; Spalletta, Ronald A; Patra, Guy; Delvecchio, Vito G

    2004-01-01

    Repetitive element polymorphism-PCR (REP-PCR) is one of the tools that has been used to elucidate genetic diversity of related microorganisms. Using the MB1 primer, REP-PCR fingerprints from 110 Bacillus strains within the "B. cereus group" have identified eighteen distinct categories, while other more distantly related bacterial species fell within six additional categories. All Bacillus anthracis strains tested were found to be monomorphic by fluorophore-enhanced REP-PCR (FERP) fingerprinting using the MB1 primer. In contrast, other non- B. anthracis isolates displayed a high degree of polymorphism. Dendrogramic analysis revealed that the non- B. anthracis strains possessing the Ba813 chromosomal marker were divided into two clusters. One of the clusters shared identity with the B. cereus strains examined.

  20. 75 FR 16113 - Bacillus subtilis; Registration Review Final Decision; Notice of Availability

    Science.gov (United States)

    2010-03-31

    ... AGENCY Bacillus subtilis; Registration Review Final Decision; Notice of Availability AGENCY... final registration review decision for the pesticide Bacillus subtilis, case 6012. Registration review... availability of EPA's final registration review decision for Bacillus subtilis, case 6012. The...

  1. 1

    NARCIS (Netherlands)

    Yan, Zheng; Zheng, Xiaowei; Han, Bei Zhong; Yan, Yin Zhuo; Zhang, Xin; Chen, Jing Yu

    2015-01-01

    Bacillus licheniformis has been found to be one of the persistent dominant microorganisms in Daqu, which is a traditional fermentation starter, and it has been used to intensify certain strains. To understand the impact of B. licheniformis on Daqu, the fermentation behaviour of B. licheniformis w

  2. 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.

  3. Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes

    Science.gov (United States)

    2014-10-30

    2010 31-May-2014 Approved for Public Release; Distribution Unlimited Final Report: (Life Science Division/Biochemistry) Inactivation of Bacillus ...S) AND ADDRESS (ES) U.S. Army Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 Bacillus Anthracis, Spores, Biofilm, Inhibition...Biochemistry) Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes Report Title The Specific Aims of the project were to investigate: 1) the

  4. Transport of Bacillus thuringiensis var. Kurstaki Via Fomites

    Science.gov (United States)

    2011-01-01

    Special Feature: Remediation Transport of Bacillus Thuringiensis var. Kurstaki Via Fomites Sheila Van Cuyk, Lee Ann B. Veal, Beverley Simpson, and...evaluate biodefense concepts of operations using routine spraying of Bacillus thuringiensis var. kurstaki (Btk). Btk is dispersed in large quantities as...used is a water-based slurry containing Bacillus thuringiensis var. kurstaki (Btk). This bacterium produces a toxin that is lethal to gypsy moth

  5. In vitro susceptibility of Bacillus spp. to selected antimicrobial agents.

    OpenAIRE

    1988-01-01

    Although often dismissed as contaminants when isolated from blood cultures, Bacillus spp. are increasingly recognized as capable of causing serious systemic infections. As part of a clinical-microbiological study, 89 strains of Bacillus spp. isolated from clinical blood cultures between 1981 and 1985 had their species determined and were tested for antimicrobial agent susceptibility to 18 antibiotics. Species of isolates were determined by the API 50CH and API 20E systems. Bacillus cereus (54...

  6. 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...

  7. Mutations determining mitomycin resistance in Bacillus subtilis.

    Science.gov (United States)

    Iyer, V N

    1966-12-01

    Iyer, V. N. (Microbiology Research Institute, Canada Department of Agriculture, Ottawa, Canada). Mutations determining mitomycin resistance in Bacillus subtilis. J. Bacteriol. 92:1663-1669. 1966.-The pattern of development of genetic resistance in Bacillus subtilis to mitomycin C was studied, and spontaneous single and multistep mutants were obtained. The transmission and expression of these mutations in sensitive strains proved possible by means of genetic transformation. The mutations were genetically studied in relation to a chromosomal mutation, mac-1, which confers resistance to the macrolide antibiotic erythromycin and which has been previously localized in the early-replicating segment of the B. subtilis chromosome. The results indicate that all of three primary mutations studied in this manner, as well as a secondary and tertiary mutation derived from one of the primary mutations, are clustered in this early-replicating segment. It appears that the secondary and tertiary mutations enhance the resistance conferred by the primary mutation, apparently without themselves conferring any resistance.

  8. Bacillus thuringiensis crystal proteins that target nematodes

    OpenAIRE

    Wei, Jun-Zhi; Hale, Kristina; Carta, Lynn; Platzer, Edward; Wong, Cynthie; Fang, Su-Chiung; Aroian, Raffi V.

    2003-01-01

    Bacillus thuringiensis (Bt) crystal proteins are pore-forming toxins used as insecticides around the world. Previously, the extent to which these proteins might also target the invertebrate phylum Nematoda has been mostly ignored. We have expressed seven different crystal toxin proteins from two largely unstudied Bt crystal protein subfamilies. By assaying their toxicity on diverse free-living nematode species, we demonstrate that four of these crystal proteins are active against multiple nem...

  9. Bacillus thuringiensis and Its Pesticidal Crystal Proteins

    OpenAIRE

    Schnepf, E.; Crickmore, N; Van Rie, J.; Lereclus, D.; Baum, J; Feitelson, J.; Zeigler, D. R.; Dean, D H

    1998-01-01

    During the past decade the pesticidal bacterium Bacillus thuringiensis has been the subject of intensive research. These efforts have yielded considerable data about the complex relationships between the structure, mechanism of action, and genetics of the organism’s pesticidal crystal proteins, and a coherent picture of these relationships is beginning to emerge. Other studies have focused on the ecological role of the B. thuringiensis crystal proteins, their performance in agricultural and o...

  10. BACILLUS THURINGIENSIS ELASTASES WITH INSECTICIDE ACTIVITY

    OpenAIRE

    E. V. Matseliukh; N. A. Nidialkova; V. V. Krout'; L. D. Varbanets; A. V. Kalinichenko; V. F. Patyka

    2015-01-01

    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, Kh...

  11. Pseudomembranous tracheobronchitis due to Bacillus cereus.

    Science.gov (United States)

    Strauss, R; Mueller, A; Wehler, M; Neureiter, D; Fischer, E; Gramatzki, M; Hahn, E G

    2001-09-01

    We present a case of a rapidly progressive pseudomembranous tracheobronchitis and pneumonia in a 52-year-old woman with severe aplastic anemia. Bacillus cereus was isolated from bronchoalveolar lavage fluids, blood cultures, and pseudomembrane biopsy specimens; despite intensive antibiotic treatment, the patient's condition deteriorated rapidly. To our knowledge, this is the first report of a B. cereus infection that has caused pseudomembranous tracheobronchitis, possibly because of the production of bacterial toxins.

  12. 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

  13. Disinfection of Vegetative Cells of Bacillus anthracis

    Science.gov (United States)

    2016-03-01

    and the fate of vegetative cells resulting from augmented germination . In this study, data were generated on the inactivation of vegetative B...all the dilutions. First, a solution of 1000 mg chlorine solution was prepared in two steps . Sodium hypochlorite solution was diluted 1:5, and then 1... Germinant -Enhanced Decontamination of Bacillus Spores Adhered to Iron and Cement-Mortar Drinking Water Infrastructures. Appl. Environ. Microbiol. 2012, 78

  14. Complete Genomes of Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, Two Phylogenetically Distinct Probiotics.

    Directory of Open Access Journals (Sweden)

    Indu Khatri

    Full Text Available Several spore-forming strains of Bacillus are marketed as probiotics due to their ability to survive harsh gastrointestinal conditions and confer health benefits to the host. We report the complete genomes of two commercially available probiotics, Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, and compare them with the genomes of other Bacillus and Lactobacillus. The taxonomic position of both organisms was established with a maximum-likelihood tree based on twenty six housekeeping proteins. Analysis of all probiotic strains of Bacillus and Lactobacillus reveal that the essential sporulation proteins are conserved in all Bacillus probiotic strains while they are absent in Lactobacillus spp. We identified various antibiotic resistance, stress-related, and adhesion-related domains in these organisms, which likely provide support in exerting probiotic action by enabling adhesion to host epithelial cells and survival during antibiotic treatment and harsh conditions.

  15. Complete Genomes of Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, Two Phylogenetically Distinct Probiotics.

    Science.gov (United States)

    Khatri, Indu; Sharma, Shailza; Ramya, T N C; Subramanian, Srikrishna

    2016-01-01

    Several spore-forming strains of Bacillus are marketed as probiotics due to their ability to survive harsh gastrointestinal conditions and confer health benefits to the host. We report the complete genomes of two commercially available probiotics, Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, and compare them with the genomes of other Bacillus and Lactobacillus. The taxonomic position of both organisms was established with a maximum-likelihood tree based on twenty six housekeeping proteins. Analysis of all probiotic strains of Bacillus and Lactobacillus reveal that the essential sporulation proteins are conserved in all Bacillus probiotic strains while they are absent in Lactobacillus spp. We identified various antibiotic resistance, stress-related, and adhesion-related domains in these organisms, which likely provide support in exerting probiotic action by enabling adhesion to host epithelial cells and survival during antibiotic treatment and harsh conditions.

  16. Meningitis and bacteremia due to Bacillus cereus. A case report and a review of Bacillus infections.

    Science.gov (United States)

    Siegman-Igra, Y; Lavochkin, J; Schwartz, D; Konforti, N

    1983-06-01

    A patient with meningitis and bacteremia due to Bacillus cereus is described. The patient had transsphenoidal hypophysectomy for chromophobe adenoma, complicated by rhinorrhea, which was corrected by subarachnoid drainage. Three weeks after removal of the drain, the patient presented with meningitis and died the following day. The causative organism was identified as B. cereus. The literature on Bacillus infections is reviewed with special attention to severe infections. A modified classification is proposed, dividing infections into superficial, closed-space and systemic ones. Sixty-one previously reported cases of systemic Bacillus infections are reviewed according to type of infection (endocarditis, meningitis or pulmonary infection), and the underlying conditions, ways of acquiring the infection, clinical picture and mortality are discussed.

  17. 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

  18. 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.

  19. 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

    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...... 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...

  20. Bacillus luteus sp. nov., isolated from soil.

    Science.gov (United States)

    Subhash, Y; Sasikala, Ch; Ramana, Ch V

    2014-05-01

    Two bacterial strains (JC167T and JC168) were isolated from a soil sample collected from Mandpam, Tamilnadu, India. Colonies of both strains were orange and cells Gram-stain-positive. Cells were small rods, and formed terminal endospores of ellipsoidal to oval shape. Both strains were positive for catalase, oxidase and hydrolysis of starch/gelatin, and negative for chitin hydrolysis, H2S production, indole production and nitrate reduction activity. Major fatty acids of both strains (>5%) were anteiso-C15:0, iso-C16:0, iso-C15:0, anteiso-C17:0, iso-C14:0 and C16:0 with minor (1%) amounts of iso-C17:0, anteiso-C17:0 B/iso-C17:0 I and C16:1ω11c. Diphosphatydilglycerol, phosphatidylethanolamine and phosphatidylglycerol were the major polar lipids of both strains. Cell wall amino acids were L-alanine, D-alanine, D-glutamic acid and meso-diaminopimelic acid. β-Carotene and five unidentified carotenoids were present in both strains. Mean genomic DNA G+C content was 53.4±1 mol% and the two strains were closely related (mean DNA-DNA hybridization>90%). 16S rRNA gene sequence comparisons of both strains indicated that they represent species of the genus Bacillus within the family Bacillaceae of the phylum Firmicutes. Both strains had a sequence similarity of 97.6% with Bacillus saliphilus 6AGT and Bacillus. Sequence similarity between strain JC167T and 168 was 100%. Strain JC167T showed 25.8±1% reassociation (based on DNA-DNA hybridization) with B. saliphilus DSM 15402T (=6AGT). Distinct morphological, physiological and genotypic differences from previously described taxa support the classification of strain JC167T as a representative of a novel species of the genus Bacillus, for which the name Bacillus luteus sp. nov. is proposed. The type strain is JC167T (=KCTC 33100T=LMG 27257T).

  1. Role of fatty acids in Bacillus environmental adaptation

    Directory of Open Access Journals (Sweden)

    Sara Esther Diomande

    2015-08-01

    Full Text Available The large bacterial genus genus Bacillus is widely distributed in the environment and is able to colonize highly diverse niches. Some Bacillus species harbour pathogenic characteristics. The fatty acid (FA composition is among the essential criteria used to define Bacillus species. Some elements of the FA pattern composition are common to Bacillus species, whereas others are specific and can be categorized in relation to the ecological niches of the species. Bacillus species are able to modify their FA patterns to adapt to a wide range of environmental changes, including changes in the growth medium, temperature, food processing conditions, and pH. Like many other Gram-positive bacteria, Bacillus strains display a well-defined FA synthesis II system that is equilibrated with a FA degradation pathway and regulated to efficiently respond to the needs of the cell. Like endogenous FAs, exogenous FAs may positively or negatively affect the survival of Bacillus vegetative cells and the spore germination ability in a given environment. Some of these exogenous FAs may provide a powerful strategy for preserving food against contamination by the Bacillus pathogenic strains responsible for foodborne illness.

  2. Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation.

    Science.gov (United States)

    Romero, Diego; Pérez-García, Alejandro; Veening, Jan-Willem; de Vicente, Antonio; Kuipers, Oscar P

    2006-09-01

    A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool for molecular genetic analysis of interesting Bacillus strains, which are hard to transform by conventional methods.

  3. Intractable Bacillus cereus bacteremia in a preterm neonate.

    Science.gov (United States)

    John, Anna B; Razak, Eissa A S A; Razak, Emad E M H; Al-Naqeeb, Niran; Dhar, Rita

    2007-04-01

    Although often regarded as a contaminant, Bacillus spp. have been implicated in serious systemic infections. The incidence of such infections is low with only a few cases reported in the literature. We describe the clinical course of early-onset Bacillus cereus bacteremia in a preterm neonate who was successfully treated with vancomycin.

  4. Dendritic Cells Endocytose Bacillus Anthracis Spores: Implications for Anthrax Pathogenesis

    Science.gov (United States)

    2007-11-02

    Dendritic Cells Endocytose Bacillus anthracis Spores: Implications for Anthrax Pathogenesis1 Katherine C. Brittingham,* Gordon Ruthel,* Rekha G...germination and dissemination of spores. Found in high frequency throughout the respiratory track, dendritic cells (DCs) routinely take up foreign...COVERED - 4. TITLE AND SUBTITLE Dendritic cells endocytose Bacillus anthracis spores: implications for anthrax pathogenesis, The Journal of

  5. 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.

  6. Complete Genome Sequence of Bacillus megaterium Siphophage Silence

    OpenAIRE

    Solis, Jonathan A.; Farmer, Nicholas G.; Cahill, Jesse L.; Rasche, Eric S.; Kuty Everett, Gabriel F.

    2015-01-01

    Silence is a newly isolated siphophage that infects Bacillus megaterium, a soil bacterium that is used readily in research and commercial applications. A study of B. megaterium phage Silence will enhance our knowledge of the diversity of Bacillus phages. Here, we describe the complete genome sequence and annotated features of Silence.

  7. Draft Genome Sequence of Bacillus tequilensis Strain FJAT-14262a

    OpenAIRE

    Chen, Qian-Qian; Liu, Bo; Liu, Guo-hong; Wang, Jie-ping; Che, Jian-Mei

    2015-01-01

    Bacillus tequilensis FJAT-14262a is a Gram-positive rod-shaped bacterium. Here, we report the 4,038,551-bp genome sequence of B. tequilensis FJAT-14262a, which will provide useful information for genomic taxonomy and phylogenomics of Bacillus.

  8. Draft Genome Sequence of Bacillus tequilensis Strain FJAT-14262a.

    Science.gov (United States)

    Chen, Qian-Qian; Liu, Bo; Liu, Guo-Hong; Wang, Jie-Ping; Che, Jian-Mei

    2015-11-12

    Bacillus tequilensis FJAT-14262a is a Gram-positive rod-shaped bacterium. Here, we report the 4,038,551-bp genome sequence of B. tequilensis FJAT-14262a, which will provide useful information for genomic taxonomy and phylogenomics of Bacillus.

  9. 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.

  10. Quantitative immunofluorescence studies of the serology of Bacillus anthracis spores.

    OpenAIRE

    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.

  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. The Poly-γ-d-Glutamic Acid Capsule Surrogate of the Bacillus anthracis Capsule Is a Novel Toll-Like Receptor 2 Agonist.

    Science.gov (United States)

    Jeon, Jun Ho; Lee, Hae-Ri; Cho, Min-Hee; Park, Ok-Kyu; Park, Jungchan; Rhie, Gi-eun

    2015-10-01

    Bacillus anthracis is a pathogenic Gram-positive bacterium that causes a highly lethal infectious disease, anthrax. The poly-γ-d-glutamic acid (PGA) capsule is one of the major virulence factors of B. anthracis, along with exotoxins. PGA enables B. anthracis to escape phagocytosis and immune surveillance. Our previous study showed that PGA activates the human macrophage cell line THP-1 and human dendritic cells, resulting in the production of the proinflammatory cytokine interleukin-1β (IL-1β) (M. H. Cho et al., Infect Immun 78:387-392, 2010, http://dx.doi.org/10.1128/IAI.00956-09). Here, we investigated PGA-induced cytokine responses and related signaling pathways in mouse bone marrow-derived macrophages (BMDMs) using Bacillus licheniformis PGA as a surrogate for B. anthracis PGA. Upon exposure to PGA, BMDMs produced proinflammatory mediators, including tumor necrosis factor alpha (TNF-α), IL-6, IL-12p40, and monocyte chemoattractant protein 1 (MCP-1), in a concentration-dependent manner. PGA stimulated Toll-like receptor 2 (TLR2) but not TLR4 in Chinese hamster ovary cells expressing either TLR2 or TLR4. The ability of PGA to induce TNF-α and IL-6 was retained in TLR4(-/-) but not TLR2(-/-) BMDMs. Blocking experiments with specific neutralizing antibodies for TLR1, TLR6, and CD14 showed that TLR6 and CD14 also were necessary for PGA-induced inflammatory responses. Furthermore, PGA enhanced activation of mitogen-activated protein (MAP) kinases and nuclear factor-kappa B (NF-κB), which are responsible for expression of proinflammatory cytokines. Additionally, PGA-induced TNF-α production was abrogated not only in MyD88(-/-) BMDMs but also in BMDMs pretreated with inhibitors of MAP kinases and NF-κB. These results suggest that immune responses induced by PGA occur via TLR2, TLR6, CD14, and MyD88 through activation of MAP kinase and NF-κB pathways.

  13. Bacillus oryzisoli sp. nov., isolated from rice rhizosphere.

    Science.gov (United States)

    Zhang, Xiao-Xia; Gao, Ju-Sheng; Zhang, Lei; Zhang, Cai-Wen; Ma, Xiao-Tong; Zhang, Jun

    2016-09-01

    The taxonomy of strain 1DS3-10T, a Gram-staining-positive, endospore-forming bacterium isolated from rice rhizosphere, was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences demonstrated that the novel strain was grouped with established members of the genus Bacillus and appeared to be closely related to the type strains Bacillus benzoevorans DSM 5391T (97.9 %), Bacillus circulans DSM 11T (97.7 %), Bacillus novalis JCM 21709T (97.3 %), Bacillus soli JCM 21710T (97.3 %), Bacillus oceanisediminis CGMCC 1.10115T (97.3 %) and BacillusnealsoniiFO-92T (97.1 %). The fatty acid profile of strain 1DS3-10T, which showed a predominance of iso-C15 : 0 and anteiso-C15 : 0, supported the allocation of the strain to the genus Bacillus. The predominant menaquinone was MK-7 (100 %). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and unknown aminolipids. Cell-wall peptidoglycan contained meso-diaminopimelic acid. DNA-DNA hybridization values between strain 1DS3-10T and the type strains of closely related species were 25-33 %, which supported that 1DS3-10T represented a novel species in the genus Bacillus. The results of some physiological and biochemical tests also allowed the phenotypic differentiation of strain 1DS3-10T from the most closely related recognized species. On the basis of the phylogenetic and phenotypic evidence, strain 1DS3-10T represents a novel species of the genus Bacillus, for which the name Bacillus oryzisoli sp. nov. is proposed. The type strain of the novel species is 1DS3-10T (=ACCC 19781T=DSM 29761T).

  14. Occurrence of natural Bacillus thuringiensis contaminants and residues of Bacillus thuringiensis-based insecticides on fresh fruits and vegetables

    DEFF Research Database (Denmark)

    Frederiksen, Kristine; Rosenquist, Hanne; Jørgensen, Kirsten

    2006-01-01

    A total of 128 Bacillus cereus-like strains isolated from fresh fruits and vegetables for sale in retail shops in Denmark were characterized. Of these strains, 39% (50/128) were classified as Bacillus thuringiensis on the basis of their content of cry genes determined by PCR or crystal proteins...

  15. 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

  16. Novel cloning vectors for Bacillus thuringiensis.

    OpenAIRE

    Baum, J A; Coyle, D M; Gilbert, M P; Jany, C S; Gawron-Burke, C

    1990-01-01

    Seven replication origins from resident plasmids of Bacillus thuringienis subsp. kurstaki HD263 and HD73 were cloned in Escherichia coli. Three of these replication origins, originating from plasmids of 43, 44, and 60 MDa, were used to construct a set of compatible shuttle vectors that exhibit structural and segregational stability in the Cry- strain B. thuringiensis HD73-26. These shuttle vectors, pEG597, pEG853, and pEG854, were designed with rare restriction sites that permit various adapt...

  17. Regulation of protoxin synthesis in Bacillus thuringiensis.

    OpenAIRE

    Minnich, S A; Aronson, A I

    1984-01-01

    A derivative of Bacillus thuringiensis subsp. kurstaki (HD-1) formed parasporal inclusions at 25 degrees C, but not at 32 degrees C. This strain differed from the parent only in the loss of a 110-megadalton (Md) plasmid, but plasmid and chromosomal copies of protoxin genes were present in both strains. On the basis of temperature shift experiments, the sensitive period appeared to be during midexponential growth, long before the time of protoxin synthesis at 3 to 4 h after the end of exponent...

  18. Features of Bacillus cereus swarm cells.

    Science.gov (United States)

    Senesi, Sonia; Salvetti, Sara; Celandroni, Francesco; Ghelardi, Emilia

    2010-11-01

    When propagated on solid surfaces, Bacillus cereus can produce differentiated swarm cells under a wide range of growth conditions. This behavioural versatility is ecologically relevant, since it allows this bacterium to adapt swarming to environmental changes. Swarming by B. cereus is medically important: swarm cells are more virulent and particularly prone to invade host tissues. Characterisation of swarming-deficient mutants highlights that flagellar genes as well as genes governing different metabolic pathways are involved in swarm-cell differentiation. In this review, the environmental and genetic requirements for swarming and the role played by swarm cells in the virulence this pathogen exerts will be outlined.

  19. Bacillus anthracis factors for phagosomal escape.

    Science.gov (United States)

    Tonello, Fiorella; Zornetta, Irene

    2012-07-01

    The mechanism of phagosome escape by intracellular pathogens is an important step in the infectious cycle. During the establishment of anthrax, Bacillus anthracis undergoes a transient intracellular phase in which spores are engulfed by local phagocytes. Spores germinate inside phagosomes and grow to vegetative bacilli, which emerge from their resident intracellular compartments, replicate and eventually exit from the plasma membrane. During germination, B. anthracis secretes multiple factors that can help its resistance to the phagocytes. Here the possible role of B. anthracis toxins, phospholipases, antioxidant enzymes and capsules in the phagosomal escape and survival, is analyzed and compared with that of factors of other microbial pathogens involved in the same type of process.

  20. Bacillus crescens sp. nov., isolated from soil.

    Science.gov (United States)

    Shivani, Y; Subhash, Y; Dave Bharti, P; Sasikala, Ch; Ramana, Ch V

    2015-08-01

    Two bacterial strains (JC247T and JC248) were isolated from soil samples collected from Rann of Kutch, Gujarat, India. Colonies of both strains were creamy white. Cells were Gram-stain-positive, rods-to-curved rods (crescent-shaped), and produced centrally located oval-shaped endospores. Major (>5 %) fatty acids of both strains were iso-C16  :  0, iso-C14  :  0, iso-C15  :  0, C16  :  1ω11c and C16  :  0, with minor ( 1 %) amounts of anteiso-C15  :  0, anteiso-C17  :  0, iso-C16  :  1 H, iso-C17  :  0, iso-C18  :  0, C14  :  0, C17  :  0, C18  :  0, C18  :  1ω9c, iso-C17  :  1ω10c and anteiso-C17  :  0B/isoI. Diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol were the major polar lipids of both strains. Cell-wall amino acids were l-alanine, d-alanine, d-glutamic acid and meso-diaminopimelic acid. The genomic DNA G+C content of strains JC247T and JC248 was 48.2 and 48.1 mol%, respectively. Both strains were closely related with mean DNA-DNA hybridization >90 %. 16S rRNA gene sequence analysis of both strains indicated that they are members of the genus Bacillus within the family Bacillaceae of the phylum Firmicutes. Both strains had a 16S rRNA gene sequence similarity of 96.93 % with Bacillus firmus NCIMB 9366T and Bacillus. Sequence similarity between strain JC247T and JC248 was 100 %. Distinct morphological, physiological and genotypic differences from previously described taxa support the classification of strains JC247T and JC248 as representatives of a novel species of the genus Bacillus, for which the name Bacilluscrescens sp. nov. is proposed. The type strain is JC247T ( = KCTC 33627T = LMG 28608T).

  1. Synthesis of lipoteichoic acids in Bacillus anthracis.

    Science.gov (United States)

    Garufi, Gabriella; Hendrickx, Antoni P; Beeri, Karen; Kern, Justin W; Sharma, Anshika; Richter, Stefan G; Schneewind, Olaf; Missiakas, Dominique

    2012-08-01

    Lipoteichoic acid (LTA), a glycerol phosphate polymer, is a component of the envelope of Gram-positive bacteria that has hitherto not been identified in Bacillus anthracis, the causative agent of anthrax. LTA synthesis in Staphylococcus aureus and other microbes is catalyzed by the product of the ltaS gene, a membrane protein that polymerizes polyglycerol phosphate from phosphatidyl glycerol. Here we identified four ltaS homologues, designated ltaS1 to -4, in the genome of Bacillus anthracis. Polyglycerol phosphate-specific monoclonal antibodies were used to detect LTA in the envelope of B. anthracis strain Sterne (pXO1(+) pXO2(-)) vegetative forms. B. anthracis mutants lacking ltaS1, ltaS2, ltaS3, or ltaS4 did not display defects in growth or LTA synthesis. In contrast, B. anthracis strains lacking both ltaS1 and ltaS2 were unable to synthesize LTA and exhibited reduced viability, altered envelope morphology, aberrant separation of vegetative forms, and decreased sporulation efficiency. Expression of ltaS1 or ltaS2 alone in B. anthracis as well as in other microbes was sufficient for polyglycerol phosphate synthesis. Thus, similar to S. aureus, B. anthracis employs LtaS enzymes to synthesize LTA, an envelope component that promotes bacterial growth and cell division.

  2. Antimicrobial Effects of Honey on Bacillus Cereus

    Directory of Open Access Journals (Sweden)

    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

    2014-05-01

    Full Text Available 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 Razavi Province and A3 of South Khorasan province (were prepared and studied in terms of chemical parameters .The antibacterial effect of honey was surveyed throughTurbidimeter using spectrometer with incubator time of 2, 4, 6, and 8hrs. the level of turbidity caused by bacterium growth was measured at different times with a wavelength of 600nm. Results: According to the study, the samples containing higher concentration of polyphenol has more antimicrobial activity. The samples of A2, A3, and A1 had the highest concentration of polyphenol, respectively. Conclusion: The results indicate the prebiotic effect of honey that can be justified by the presence of fructo-oligosacharids and vitamin B. Keywords: Honey, Bacillus Cereus, Antibacterial, Turbidimetry.

  3. 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.

  4. Real-Time PCR Identification of Unique Bacillus anthracis Sequences.

    Science.gov (United States)

    Cieślik, P; Knap, J; Kolodziej, M; Mirski, T; Joniec, J; Graniak, G; Zakowska, D; Winnicka, I; Bielawska-Drózd, A

    2015-01-01

    Bacillus anthracis is a spore-forming, Gram-positive microorganism. It is a causative agent of anthrax, a highly infectious disease. It belongs to the "Bacillus cereus group", which contains other closely related species, including Bacillus cereus, Bacillus thuringiensis, Bacillus mycoides, Bacillus weihenstephanensis, and Bacillus pseudomycoides. B. anthracis naturally occurs in soil environments. The BA5345 genetic marker was used for highly specific detection of B. anthracis with TaqMan probes. The detection limit of a real-time PCR assay was estimated at the level of 16.9 copies (CI95% - 37.4 to 37.86, SD = 0.2; SE = 0.118). Oligonucleotides designed for the targeted sequences (within the tested locus) revealed 100 % homology to B. anthracis strain reference sequences deposited in the database (NCBI) and high specificity to all tested B. anthracis strains. Additional in silico analysis of plasmid markers pag and cap genes with B. anthracis strains included in the database was carried out. Our study clearly indicates that the BA5345 marker can be used with success as a chromosomal marker in routine identification of B. anthracis; moreover, detection of plasmid markers indicates virulence of the examined strains.

  5. Identification of Bacillus strains for biological control of catfish pathogens.

    Science.gov (United States)

    Ran, Chao; Carrias, Abel; Williams, Malachi A; Capps, Nancy; Dan, Bui C T; Newton, Joseph C; Kloepper, Joseph W; Ooi, Ei L; Browdy, Craig L; Terhune, Jeffery S; Liles, Mark R

    2012-01-01

    Bacillus strains isolated from soil or channel catfish intestine were screened for their antagonism against Edwardsiella ictaluri and Aeromonas hydrophila, the causative agents of enteric septicemia of catfish (ESC) and motile aeromonad septicaemia (MAS), respectively. Twenty one strains were selected and their antagonistic activity against other aquatic pathogens was also tested. Each of the top 21 strains expressed antagonistic activity against multiple aquatic bacterial pathogens including Edwardsiella tarda, Streptococcus iniae, Yersinia ruckeri, Flavobacterium columnare, and/or the oomycete Saprolegnia ferax. Survival of the 21 Bacillus strains in the intestine of catfish was determined as Bacillus CFU/g of intestinal tissue of catfish after feeding Bacillus spore-supplemented feed for seven days followed by normal feed for three days. Five Bacillus strains that showed good antimicrobial activity and intestinal survival were incorporated into feed in spore form at a dose of 8×10(7) CFU/g and fed to channel catfish for 14 days before they were challenged by E. ictaluri in replicate. Two Bacillus subtilis strains conferred significant benefit in reducing catfish mortality (Pbiological control in vivo was also investigated in terms of whether the strains contain plasmids or express resistance to clinically important antibiotics. The Bacillus strains identified from this study have good potential to mediate disease control as probiotic feed additives for catfish aquaculture.

  6. Screening of Bacillus Species with Potentials of Antibiotics Production

    OpenAIRE

    Faruk Adamu KUTA; Lohya NIMZING; Priscilla Yahemba ORKA’A

    2009-01-01

    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 cer...

  7. 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.

  8. 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.''

  9. 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 ℃ .

  10. 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℃.

  11. Multilocus sequence analysis of Bacillus thuringiensis serovars navarrensis, bolivia and vazensis and Bacillus weihenstephanensis reveals a common phylogeny.

    Science.gov (United States)

    Soufiane, Brahim; Baizet, Mathilde; Côté, Jean-Charles

    2013-01-01

    The Bacillus cereus group sensu lato includes six closely-related bacterial species: Bacillus cereus, Bacillus anthracis, Bacillus thuringiensis, Bacillus mycoides, Bacillus pseudomycoides and Bacillus weihenstephanensis. B. thuringiensis is distinguished from the other species mainly by the appearance of an inclusion body upon sporulation. B. weihenstephanensis is distinguished based on its psychrotolerance and the presence of specific signature sequences in the 16S rRNA gene and cspA genes. A total of seven housekeeping genes (glpF, gmK, ilvD, pta, purH, pycA and tpi) from different B. thuringiensis serovars and B. weihenstephanensis strains were amplified and their nucleotide sequences determined. A maximum likelihood phylogenetic tree was inferred from comparisons of the concatenated sequences. B. thuringiensis serovars navarrensis, bolivia and vazensis clustered not with the other B. thuringiensis serovars but rather with the B. weihenstephanensis strains, indicative of a common phylogeny. In addition, specific signature sequences and single nucleotide polymorphisms common to B. thuringiensis serovars navarrensis, bolivia and vazensis and the B. weihenstephanensis strains, and absent in the other B. thuringiensis serovars, were identified.

  12. 77 FR 73934 - Bacillus subtilis Strain QST 713 Variant Soil; Amendment to an Exemption From the Requirement of...

    Science.gov (United States)

    2012-12-12

    ... AGENCY 40 CFR Part 180 Bacillus subtilis Strain QST 713 Variant Soil; Amendment to an Exemption From the Requirement of a Tolerance for Bacillus subtilis Strain QST 713 To Include Residues of Bacillus subtilis... Bacillus subtilis strain QST 713 in or on all food commodities by including residues of Bacillus...

  13. Bacillus thuringiensis as a surrogate for Bacillus anthracis in aerosol research.

    Science.gov (United States)

    Tufts, Jenia A M; Calfee, M Worth; Lee, Sang Don; Ryan, Shawn P

    2014-05-01

    Characterization of candidate surrogate spores prior to experimental use is critical to confirm that the surrogate characteristics are as closely similar as possible to those of the pathogenic agent of interest. This review compares the physical properties inherent to spores of Bacillus anthracis (Ba) and Bacillus thuringiensis (Bt) that impact their movement in air and interaction with surfaces, including size, shape, density, surface morphology, structure and hydrophobicity. Also evaluated is the impact of irradiation on the physical properties of both Bacillus species. Many physical features of Bt and Ba have been found to be similar and, while Bt is considered typically non-pathogenic, it is in the B. cereus group, as is Ba. When cultured and sporulated under similar conditions, both microorganisms share a similar cylindrical pellet shape, an aerodynamic diameter of approximately 1 μm (in the respirable size range), have an exosporium with a hairy nap, and have higher relative hydrophobicities than other Bacillus species. While spore size, morphology, and other physical properties can vary among strains of the same species, the variations can be due to growth/sporulation conditions and may, therefore, be controlled. Growth and sporulation conditions are likely among the most important factors that influence the representativeness of one species, or preparation, to another. All Bt spores may, therefore, not be representative of all Ba spores. Irradiated spores do not appear to be a good surrogate to predict the behavior of non-irradiated spores due to structural damage caused by the irradiation. While the use of Bt as a surrogate for Ba in aerosol testing appears to be well supported, this review does not attempt to narrow selection between Bt strains. Comparative studies should be performed to test the hypothesis that viable Ba and Bt spores will behave similarly when suspended in the air (as an aerosol) and to compare the known microscale characteristics

  14. Bacillus thuringiensis and Bacillus weihenstephanensis Inhibit the Growth of Phytopathogenic Verticillium Species.

    Science.gov (United States)

    Hollensteiner, Jacqueline; Wemheuer, Franziska; Harting, Rebekka; Kolarzyk, Anna M; Diaz Valerio, Stefani M; Poehlein, Anja; Brzuszkiewicz, Elzbieta B; Nesemann, Kai; Braus-Stromeyer, Susanna A; Braus, Gerhard H; Daniel, Rolf; Liesegang, Heiko

    2016-01-01

    Verticillium wilt causes severe yield losses in a broad range of economically important crops worldwide. As many soil fumigants have a severe environmental impact, new biocontrol strategies are needed. Members of the genus Bacillus are known as plant growth-promoting bacteria (PGPB) as well as biocontrol agents of pests and diseases. In this study, we isolated 267 Bacillus strains from root-associated soil of field-grown tomato plants. We evaluated the antifungal potential of 20 phenotypically diverse strains according to their antagonistic activity against the two phytopathogenic fungi Verticillium dahliae and Verticillium longisporum. In addition, the 20 strains were sequenced and phylogenetically characterized by multi-locus sequence typing (MLST) resulting in 7 different Bacillus thuringiensis and 13 Bacillus weihenstephanensis strains. All B. thuringiensis isolates inhibited in vitro the tomato pathogen V. dahliae JR2, but had only low efficacy against the tomato-foreign pathogen V. longisporum 43. All B. weihenstephanensis isolates exhibited no fungicidal activity whereas three B. weihenstephanensis isolates showed antagonistic effects on both phytopathogens. These strains had a rhizoid colony morphology, which has not been described for B. weihenstephanensis strains previously. Genome analysis of all isolates revealed putative genes encoding fungicidal substances and resulted in identification of 304 secondary metabolite gene clusters including 101 non-ribosomal polypeptide synthetases and 203 ribosomal-synthesized and post-translationally modified peptides. All genomes encoded genes for the synthesis of the antifungal siderophore bacillibactin. In the genome of one B. thuringiensis strain, a gene cluster for zwittermicin A was detected. Isolates which either exhibited an inhibitory or an interfering effect on the growth of the phytopathogens carried one or two genes encoding putative mycolitic chitinases, which might contribute to antifungal activities

  15. Bacillus thuringiensis and Bacillus weihenstephanensis Inhibit the Growth of Phytopathogenic Verticillium Species

    Science.gov (United States)

    Hollensteiner, Jacqueline; Wemheuer, Franziska; Harting, Rebekka; Kolarzyk, Anna M.; Diaz Valerio, Stefani M.; Poehlein, Anja; Brzuszkiewicz, Elzbieta B.; Nesemann, Kai; Braus-Stromeyer, Susanna A.; Braus, Gerhard H.; Daniel, Rolf; Liesegang, Heiko

    2017-01-01

    Verticillium wilt causes severe yield losses in a broad range of economically important crops worldwide. As many soil fumigants have a severe environmental impact, new biocontrol strategies are needed. Members of the genus Bacillus are known as plant growth-promoting bacteria (PGPB) as well as biocontrol agents of pests and diseases. In this study, we isolated 267 Bacillus strains from root-associated soil of field-grown tomato plants. We evaluated the antifungal potential of 20 phenotypically diverse strains according to their antagonistic activity against the two phytopathogenic fungi Verticillium dahliae and Verticillium longisporum. In addition, the 20 strains were sequenced and phylogenetically characterized by multi-locus sequence typing (MLST) resulting in 7 different Bacillus thuringiensis and 13 Bacillus weihenstephanensis strains. All B. thuringiensis isolates inhibited in vitro the tomato pathogen V. dahliae JR2, but had only low efficacy against the tomato-foreign pathogen V. longisporum 43. All B. weihenstephanensis isolates exhibited no fungicidal activity whereas three B. weihenstephanensis isolates showed antagonistic effects on both phytopathogens. These strains had a rhizoid colony morphology, which has not been described for B. weihenstephanensis strains previously. Genome analysis of all isolates revealed putative genes encoding fungicidal substances and resulted in identification of 304 secondary metabolite gene clusters including 101 non-ribosomal polypeptide synthetases and 203 ribosomal-synthesized and post-translationally modified peptides. All genomes encoded genes for the synthesis of the antifungal siderophore bacillibactin. In the genome of one B. thuringiensis strain, a gene cluster for zwittermicin A was detected. Isolates which either exhibited an inhibitory or an interfering effect on the growth of the phytopathogens carried one or two genes encoding putative mycolitic chitinases, which might contribute to antifungal activities

  16. Synergy between toxins of Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus.

    Science.gov (United States)

    Wirth, Margaret C; Jiannino, Joshua A; Federici, Brian A; Walton, William E

    2004-09-01

    Synergistic interactions among the multiple endotoxins of Bacillus thuringiensis subsp. israelensis de Barjac play an important role in its high toxicity to mosquito larvae and the absence of insecticide resistance in populations treated with this bacterium. A lack of toxin complexity and synergism are the apparent causes of resistance to Bacillus sphaericus Neide in particular Culex field populations. To identify endotoxin combinations of the two Bacillus species that might improve insecticidal activity and manage mosquito resistance to B. sphaericus, we tested their toxins alone and in combination. Most combinations of B. sphaericus and B. t. subsp. israelensis toxins were synergistic and enhanced toxicity relative to B. sphaericus, particularly against Culex quinquefasciatus Say larvae resistant to B. sphaericus and Aedes aegypti (L.), a species poorly susceptible to B. sphaericus. Toxicity also improved against susceptible Cx. quinquefasciatus. For example, when the CytlAa toxin from B. t. subsp. israelensis was added to Bin and Cry toxins, or when native B. t. subsp. israelensis was combined with B. sphaericus, synergism values as high as 883-fold were observed and combinations were 4-59,000-fold more active than B. sphaericus. These data, and previous studies using cytolytic toxins, validate proposed strategies for improving bacterial larvicides by combining B. sphaericus with B. t. subsp. israelensis or by engineering recombinant bacteria that express endotoxins from both strains. These combinations increase both endotoxin complexity and synergistic interactions and thereby enhance activity and help avoid insecticide resistance.

  17. Partial purification and characterization of protease enzyme from Bacillus subtilis and Bacillus cereus.

    Science.gov (United States)

    Orhan, Elif; Omay, Didem; Güvenilir, Yüksel

    2005-01-01

    The aim of this experimental study was to isolate and partially purify protease enzyme from Bacillus cereus and Bacillus subtilis. Protease enzyme is obtained by inducing spore genesis of bacteria from Bacillus species in suitable nutrient plates. The partial purification was realized by applying, respectively, ammonium sulfate precipitation, dialysis, and DEAE-cellulose ion-exchange chromatography to the supernatant that was produced later. Optimum pH, optimum temperature, pH stability, and temperature stability were determined, as well as the effects of pH, temperature, substrate concentration, reaction time, and inhibitors and activators on enzyme activity. In addition, the molecular mass of the obtained enzyme was investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The specific activity of partially purified enzyme from B. subtilis was determined to be 84 U/mg. The final enzyme preparation was eight-fold more pure than the crude homogenate. The molecular mass of the partially purified enzyme was found to be 45 kDa by using SDS-PAGE. The protease enzyme that was partially purified from B. cereus was purified 1.2-fold after ammonium sulfate precipitation. The molecular mass of the partially purified enzyme was determined to be 37 kDa by using SDS-PAGE.

  18. 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.

  19. Bacillus cereus panophthalmitis after intravenous heroin.

    Science.gov (United States)

    Hatem, G; Merritt, J C; Cowan, C L

    1979-03-01

    Two healthy young black men developed panophthalmitis after intravenous heroin injections. Bacillus cereus, considered to be a relatively noncommon pathogen for man, was found to be the causative agent as it was recovered from the anterior chamber and viterous cavity of both cases. The ocular findings were unilateral in each case, and neither patient had any sistemic involvement from the bacteremia. The onset of visual symptoms varied from 24 to 36 hours after the last intravenous injection with the eye becoming rapidly blind. Photographs of the early fundus lesions included preretinal hypopyon-like lesions and peculiar changes in the blood vasculature. Intracameral gentamicin and steroids did not alter the cause, and treatment was enucleation.

  20. Bacillus anthracis Bioterrorism Incident, Kameido, Tokyo, 1993

    Science.gov (United States)

    Keim, Paul; Kaufmann, Arnold F.; Keys, Christine; Smith, Kimothy L.; Taniguchi, Kiyosu; Inouye, Sakae; Kurata, Takeshi

    2004-01-01

    In July 1993, a liquid suspension of Bacillus anthracis was aerosolized from the roof of an eight-story building in Kameido, Tokyo, Japan, by the religious group Aum Shinrikyo. During 1999 to 2001, microbiologic tests were conducted on a liquid environmental sample originally collected during the 1993 incident. Nonencapsulated isolates of B. anthracis were cultured from the liquid. Multiple-locus, variable-number tandem repeat analysis found all isolates to be identical to a strain used in Japan to vaccinate animals against anthrax, which was consistent with the Aum Shinrikyo members’ testimony about the strain source. In 1999, a retrospective case-detection survey was conducted to identify potential human anthrax cases associated with the incident, but none were found. The use of an attenuated B. anthracis strain, low spore concentrations, ineffective dispersal, a clogged spray device, and inactivation of the spores by sunlight are all likely contributing factors to the lack of human cases. PMID:15112666

  1. Sludge based Bacillus thuringiensis biopesticides: viscosity impacts.

    Science.gov (United States)

    Brar, S K; Verma, M; Tyagi, R D; Valéro, J R; Surampalli, R Y

    2005-08-01

    Viscosity studies were performed on raw, pre-treated (sterilised and thermal alkaline hydrolysed or both types of treatment) and Bacillus thuringiensis (Bt) fermented sludges at different solids concentration (10-40 g/L) for production of biopesticides. Correlations were established among rheological parameter (viscosity), solids (total and dissolved) concentration and entomotoxicity (Tx) of Bt fermented sludges. Exponential and power laws were preferentially followed by hydrolysed fermented compared to raw fermented sludge. Soluble chemical oxygen demand variation corroborated with increase in dissolved solids concentration on pre-treatments, contributing to changes in viscosity. Moreover, Tx was higher for hydrolysed fermented sludge in comparison to raw fermented sludge owing to increased availability of nutrients and lower viscosity that improved oxygen transfer. The shake flask results were reproducible in fermenter. This study will have major impact on selecting fermentation, harvesting and formulation techniques of Bt fermented sludges for biopesticide production.

  2. Antagonistic competition moderates virulence in Bacillus thuringiensis.

    Science.gov (United States)

    Garbutt, Jennie; Bonsall, Michael B; Wright, Denis J; Raymond, Ben

    2011-08-01

    Classical models of the evolution of virulence predict that multiple infections should select for elevated virulence, if increased competitiveness arises from faster growth. However, diverse modes of parasite competition (resource-based, antagonism, immunity manipulation) can lead to adaptations with different implications for virulence. Using an experimental evolution approach we investigated the hypothesis that selection in mixed-strain infections will lead to increased antagonism that trades off against investment in virulence. Selection in mixed infections led to improved suppression of competitors in the bacterial insect pathogen Bacillus thuringiensis. Increased antagonism was associated with decreased virulence in three out of four selected lines. Moreover, mixed infections were less virulent than single-strain infections, and between-strain competition tended to decrease pathogen growth in vivo and in vitro. Spiteful interactions among these bacteria may be favoured because of the high metabolic costs of virulence factors and the high risk of mixed infections.

  3. 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).

  4. Global network reorganization during dynamic adaptations of Bacillus subtilis metabolism

    DEFF Research Database (Denmark)

    Buescher, Joerg Martin; Liebermeister, Wolfram; Jules, Matthieu

    2012-01-01

    Adaptation of cells to environmental changes requires dynamic interactions between metabolic and regulatory networks, but studies typically address only one or a few layers of regulation. For nutritional shifts between two preferred carbon sources of Bacillus subtilis, we combined statistical...

  5. 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...

  6. 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.

  7. 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...

  8. AFB (Acid-Fast Bacillus) Smear and Culture

    Science.gov (United States)

    ... Smear; Mycobacteria Culture; TB NAAT Formal name: Acid-Fast Bacillus Smear and Culture and Sensitivity; Mycobacteria tuberculosis ... used to detect several different types of acid-fast bacilli, but it is most commonly used to ...

  9. 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.

  10. Lantibiotics, class I bacteriocins from the genus Bacillus.

    Science.gov (United States)

    Lee, Hyungjae; Kim, Hae-Yeong

    2011-03-01

    Antimicrobial peptides exhibit high levels of antimicrobial activity against a broad range of spoilage and pathogenic microorganisms. Compared with bacteriocins produced by lactic acid bacteria, antimicrobial peptides from the genus Bacillus have been relatively less recognized despite their broad antimicrobial spectra. These peptides can be classified into two different groups based on whether they are ribosomally (bacteriocins) or nonribosomally (polymyxins and iturins) synthesized. Because of their broad spectra and high activity, antimicrobial peptides from Bacillus spp. may have great potential for applications in the food, agricultural, and pharmaceutical industries to prevent or control spoilage and pathogenic microorganisms. In this review, we introduce ribosomally synthesized antimicrobial peptides, the lantibiotic bacteriocins produced by members of Bacillus. In addition, the biosynthesis, genetic organization, mode of action, and regulation of subtilin, a well-investigated lantibiotic from Bacillus subtilis, are discussed.

  11. 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.

  12. Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation

    OpenAIRE

    Romero, Diego; Perez-Garcia, Alejandro; Veening, Jan-Willem; Vicente, Antonio; Oscar P. Kuipers; Vicente A.

    2006-01-01

    A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool for molecular genetic analysis of interesting Bacillus strains, which are hard to transform by conventional methods. (c) 2006 Elsevier B.V. All rights reserved.

  13. Plant growth regulation of Bt-cotton through Bacillus species

    OpenAIRE

    Pindi, Pavan Kumar; Sultana, Tasleem; Vootla, Praveen Kumar

    2013-01-01

    Deccan plateau in India periodically experiences droughts due to irregular rain fall and the soil in many parts of the region is considered to be poor for farming. Plant growth promoting rhizobacteria are originally defined as root-colonizing bacteria, i.e., Bacillus that cause either plant growth promotion or biological control of plant diseases. The study aims at the isolation of novel Bacillus species and to assess the biotechnological potential of the novel species as a biofertilizer, wit...

  14. 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...

  15. 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...

  16. Regulation of cry Gene Expression in Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    Chao Deng

    2014-07-01

    Full Text Available 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-transcriptional, metabolic and post-translational levels.

  17. Identification of Bacillus strains for biological control of catfish pathogens.

    Directory of Open Access Journals (Sweden)

    Chao Ran

    Full Text Available Bacillus strains isolated from soil or channel catfish intestine were screened for their antagonism against Edwardsiella ictaluri and Aeromonas hydrophila, the causative agents of enteric septicemia of catfish (ESC and motile aeromonad septicaemia (MAS, respectively. Twenty one strains were selected and their antagonistic activity against other aquatic pathogens was also tested. Each of the top 21 strains expressed antagonistic activity against multiple aquatic bacterial pathogens including Edwardsiella tarda, Streptococcus iniae, Yersinia ruckeri, Flavobacterium columnare, and/or the oomycete Saprolegnia ferax. Survival of the 21 Bacillus strains in the intestine of catfish was determined as Bacillus CFU/g of intestinal tissue of catfish after feeding Bacillus spore-supplemented feed for seven days followed by normal feed for three days. Five Bacillus strains that showed good antimicrobial activity and intestinal survival were incorporated into feed in spore form at a dose of 8×10(7 CFU/g and fed to channel catfish for 14 days before they were challenged by E. ictaluri in replicate. Two Bacillus subtilis strains conferred significant benefit in reducing catfish mortality (P<0.05. A similar challenge experiment conducted in Vietnam with four of the five Bacillus strains also showed protective effects against E. ictaluri in striped catfish. Safety of the four strains exhibiting the strongest biological control in vivo was also investigated in terms of whether the strains contain plasmids or express resistance to clinically important antibiotics. The Bacillus strains identified from this study have good potential to mediate disease control as probiotic feed additives for catfish aquaculture.

  18. Antagonism of Bacillus spp. against Xanthomonas campestris pv. campestris

    OpenAIRE

    Leila Monteiro; Rosa de Lima Ramos Mariano; Ana Maria Souto-Maior

    2005-01-01

    The antagonism of eight Bacillus isolates was investigated against nine strains of Xanthomonas campestris pv. campestris (causal agent of crucifers black rot) to assess the role of lipopeptides in this process. Antimicrobial and hemolytic (surfactant) activity tests were performed in vitro using agar diffusion methods. Antibiosis and hemolysis were positive for four Bacillus isolates against all X. campestris pv. campestris strains. The correlation observed between antimicrobial and hemolytic...

  19. Plant growth regulation of Bt-cotton through Bacillus species.

    Science.gov (United States)

    Pindi, Pavan Kumar; Sultana, Tasleem; Vootla, Praveen Kumar

    2014-06-01

    Deccan plateau in India periodically experiences droughts due to irregular rain fall and the soil in many parts of the region is considered to be poor for farming. Plant growth promoting rhizobacteria are originally defined as root-colonizing bacteria, i.e., Bacillus that cause either plant growth promotion or biological control of plant diseases. The study aims at the isolation of novel Bacillus species and to assess the biotechnological potential of the novel species as a biofertilizer, with respect to their plant growth promoting properties as efficient phosphate-solubilizing bacteria. Seven different strains of Bacillus were isolated from cotton rhizosphere soil near boys' hostel of Palamuru University which belongs to Deccan plateau. Among seven isolated strains, Bacillus strain-7 has shown maximum support for good growth of eight cotton cultivars. This bacterial species is named Bacillus sp. PU-7 based on the phenotypic and phylogenetic analysis. Among eight cotton cultivars, Mahyco has shown high levels of IAA, proteins, chlorophyll, sugars and low level of proline. Efficacy of novel Bacillus sp. PU-7 with Mahyco cultivar has been checked experimentally at field level in four different cotton grown agricultural soils. The strains supported plant growth in almost all the cases, especially in the deep black soil, with a clear evidence of maximum plant growth by increased levels of phytohormone production and biochemical analysis, followed by shallow black soil. Hence, it is inferred that the novel isolate can be used as bioinoculant in the cotton fields.

  20. Clinical significance of Bacillus species isolated from blood cultures.

    Science.gov (United States)

    Weber, D J; Saviteer, S M; Rutala, W A; Thomann, C A

    1989-06-01

    To determine the clinical significance of blood isolates of Bacillus, we reviewed all blood cultures obtained at North Carolina Memorial Hospital between 1981 and 1985. Over the five-year study period the number of patients (incidence per 10,000 hospital admissions) from whom Bacillus was isolated increased from 4.97 in 1981 to 12.5 in 1985. The incidence per 1,000 blood cultures also increased from 1.12 in 1981 to 2.33 in 1985. Review of the medical records of 78 of the 95 patients (82%) with positive cultures allowed retrospective classification of five isolates (6.4%) as clinically significant, 33 isolates (42.3%) as possibly significant, and 40 isolates (51.3%) as nonsignificant. Underlying diseases in patients with clinically significant Bacillus bacteremia included burn trauma in two, leukemia in one, carcinoma in one, and gastrointestinal hemorrhage in one. All isolates judged to be clinically significant and the majority of possibly significant isolates were B cereus. We conclude that the isolation of Bacillus species from blood cultures is clinically significant in 5% to 10% of cases, that the incidence of Bacillus bacteremia is increasing, and that burn trauma should be added to the list of conditions known to predispose to clinically significant Bacillus bacteremia.

  1. Mortality of adult Stomoxys calcitrans fed isolates of Bacillus thuringiensis.

    Science.gov (United States)

    Lysyk, T J; Kalischuk-Tymensen, L D; Selinger, L B

    2012-10-01

    We examined the ability of five isolates of Bacillus thuringiensis Berliner to cause mortality in adult stable flies, Stomoxys calcitrans (L.). Isolates Bacillus thuringiensis tolworthi 4L3 (serotype 9), Bacillus thuringiensis darmstadiensis 4M1 (serotype 10a10b), Bacillus thuringiensis thompsoni 401 (serotype 12), Bacillus thuringiensis thuringiensis HD2 (serotype 1), and Bacillus thuringiensis kurstaki HD945 (serotype 3a3b3c) were administered to adult flies in diets containing blood only, sugar only, and both sugar and blood combined. B. t. tolworthi 4L3 had no effect on adult mortality regardless of the feeding substrate. The remaining isolates tended to cause the greatest mortality when administered in blood alone. B. t. thompsoni 401 was the only isolate that consistently caused adult mortality when fed in blood at concentrations ranging from 0.21 to 50.0 microg of protein per ml of blood. This isolate also caused mortality when applied topically. The time to 50% mortality declined with dose and reached a lower asymptote at approximately equal to 1.3 d at an oral dose of 8.75 microg/ml and at a topical dose of 0.14 microg per fly.

  2. 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.

  3. Draft Genome Sequences of Type Strains Bacillus drentensis DSM 15600T and Bacillus novalis DSM 15603T.

    Science.gov (United States)

    Liu, Bo; Liu, Guo-Hong; Zhu, Yu-Jing; Wang, Jie-Ping; Che, Jian-Mei; Chen, Qian-Qian; Chen, Zheng

    2016-12-15

    Here, we report the draft genome sequences of Bacillus drentensis DSM 15600(T) and Bacillus novalis DSM 15603(T) with 5,305,306 bp and 5,667,584 bp, respectively, which will provide useful information for the functional gene mining and application of these two species. The average DNA G+C contents were 38.91% and 40.01%, respectively.

  4. Transfer of the toxin protein genes of Bacillus sphaericus into Bacillus thuringiensis subsp. israelensis and their expression.

    OpenAIRE

    Bourgouin, C.; Delécluse, A; de 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...

  5. Label-free, non-invasive light scattering sensor for rapid screening of Bacillus colonies.

    Science.gov (United States)

    Singh, Atul K; Sun, Xiulan; Bai, Xingjian; Kim, Huisung; Abdalhaseib, Maha Usama; Bae, Euiwon; Bhunia, Arun K

    2015-02-01

    Bacillus species are widely distributed in nature and have great significance both as industrially beneficial microbes and as public health burdens. We employed a novel light-scattering sensor, BARDOT (bacterial rapid detection using optical scattering technology) for instant screening of colonies of Bacillus species on agar plates. A total of 265 Bacillus and non-Bacillus isolates from our collection were used to develop and verify scatter image libraries including isolates from food, environmental and clinical samples. All Bacillus species (n=118) were detected with a high positive predictive value, PPV (≥90%) while non-Bacillus spp. had very low PPV (Bacillus colonies on phenol red mannitol (PRM) generated the highest differential scatter patterns and were used in subsequent studies. Surface plot analysis of scatter patterns confirmed differences for Bacillus and non-Bacillus isolates. BARDOT successfully detected Bacillus from inoculated baby formula, cheese, and naturally contaminated bovine unpasteurized milk in 7-16h. Ten of 129 colonies (isolates) from seven milk samples were Bacillus and remainders were non-Bacillus spp. BARDOT results were confirmed by PCR and 16S rDNA sequencing. This study demonstrates that BARDOT could be used as a screening tool to identify relevant Bacillus colonies from a community prior to genome sequencing.

  6. Biochemical and molecular characterizaion of Bacillus pumilus isolated from coastal environment in Cochin, India

    Digital Repository Service at National Institute of Oceanography (India)

    Parvathi, A.; Krishna, K.; Jose, J.; Joseph, N.; Nair, S.

    Bacillus species constitute a diverse group of bacteria widely distributed in soil and the aquatic environment. In this study, Bacillus strains isolated from the coastal environment of Cochin, India were identified by detailed conventional...

  7. Novel cloning vectors for Bacillus thuringiensis.

    Science.gov (United States)

    Baum, J A; Coyle, D M; Gilbert, M P; Jany, C S; Gawron-Burke, C

    1990-11-01

    Seven replication origins from resident plasmids of Bacillus thuringienis subsp. kurstaki HD263 and HD73 were cloned in Escherichia coli. Three of these replication origins, originating from plasmids of 43, 44, and 60 MDa, were used to construct a set of compatible shuttle vectors that exhibit structural and segregational stability in the Cry- strain B. thuringiensis HD73-26. These shuttle vectors, pEG597, pEG853, and pEG854, were designed with rare restriction sites that permit various adaptations, including the construction of small recombinant plasmids lacking antibiotic resistance genes. The cryIA(c) and cryIIA insecticidal crystal protein genes were inserted into these vectors to demonstrate crystal protein production in B. thuringiensis. Introduction of a cloned cryIA(c) gene from strain HD263 into a B. thuringiensis subsp. aizawai strain exhibiting good insecticidal activity against Spodoptera exigua resulted in a recombinant strain with an improved spectrum of insecticidal activity. Shuttle vectors of this sort should be valuable in future genetic studies of B. thuringiensis as well as in the development of B. thuringiensis strains for use as microbial pesticides.

  8. Bacillus cereus, a volatile human pathogen.

    Science.gov (United States)

    Bottone, Edward J

    2010-04-01

    Bacillus cereus is a Gram-positive aerobic or facultatively anaerobic, motile, spore-forming, rod-shaped bacterium that is widely distributed environmentally. While B. cereus is associated mainly with food poisoning, it is being increasingly reported to be a cause of serious and potentially fatal non-gastrointestinal-tract infections. The pathogenicity of B. cereus, whether intestinal or nonintestinal, is intimately associated with the production of tissue-destructive exoenzymes. Among these secreted toxins are four hemolysins, three distinct phospholipases, an emesis-inducing toxin, and proteases. The major hurdle in evaluating B. cereus when isolated from a clinical specimen is overcoming its stigma as an insignificant contaminant. Outside its notoriety in association with food poisoning and severe eye infections, this bacterium has been incriminated in a multitude of other clinical conditions such as anthrax-like progressive pneumonia, fulminant sepsis, and devastating central nervous system infections, particularly in immunosuppressed individuals, intravenous drug abusers, and neonates. Its role in nosocomial acquired bacteremia and wound infections in postsurgical patients has also been well defined, especially when intravascular devices such as catheters are inserted. Primary cutaneous infections mimicking clostridial gas gangrene induced subsequent to trauma have also been well documented. B. cereus produces a potent beta-lactamase conferring marked resistance to beta-lactam antibiotics. Antimicrobials noted to be effective in the empirical management of a B. cereus infection while awaiting antimicrobial susceptibility results for the isolate include ciprofloxacin and vancomycin.

  9. 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.

  10. 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.

  11. A pangenomic study of Bacillus thuringiensis.

    Science.gov (United States)

    Fang, Yongjun; Li, Zhaolong; Liu, Jiucheng; Shu, Changlong; Wang, Xumin; Zhang, Xiaowei; Yu, Xiaoguang; Zhao, Duojun; Liu, Guiming; Hu, Songnian; Zhang, Jie; Al-Mssallem, Ibrahim; Yu, Jun

    2011-12-20

    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 2.3-2.8Mb and 5.0-5.6Mb. We concluded that high-coverage sequence assemblies from multiple strains, before all the gaps are closed, are very useful for pangenomic studies.

  12. 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.

  13. Bacillus cereus cellulitis from contaminated heroin.

    Science.gov (United States)

    Dancer, S J; McNair, D; Finn, P; Kolsto, A B

    2002-03-01

    Concern exists over recent unexplained deaths among intravenous drug users. This report describes a patient with crepitant cellulitis who was admitted complaining of severe pain in the right forearm. Ultrasonography demonstrated gas in the tissues and he was referred for early surgical debridement of the arm. He was treated with intravenous benzyl penicillin, gentamicin and metronidazole and made a full recovery. Aspirate samples grew Bacillus cereus, morphologically similar to the isolate obtained from a sample of the patient's own heroin. Antibiogram and API 50CHB profiles were also similar. Further typing included 'H' flagellar serotyping, which found both blood and heroin strains to be non-typable, and amplified fragment polymorphism analysis, which showed that the strains were indistinguishable. Genotyping of two selected genes from B. cereus confirmed almost certain identity between the two strains. This case illustrates the potential virulence of B. cereus when inoculated into tissues, and to our knowledge, is the first report to demonstrate a conclusive microbiological link between contaminated heroin and serious sepsis in a drug user due to B. cereus.

  14. Bacillus cereus infection outbreak in captive psittacines.

    Science.gov (United States)

    Godoy, S N; Matushima, E R; Chaves, J Q; Cavados, C F G; Rabinovitch, L; Teixeira, R H F; Nunes, A L V; Melville, P; Gattamorta, M A; Vivoni, A M

    2012-12-28

    This study reports an uncommon epizootic outbreak of Bacillus cereus that caused the sudden death of 12 psittacines belonging to the species Anodorhynchus hyacinthinus (1 individual), Diopsittaca nobilis (1 individual), Ara severa (1 individual) and Ara ararauna (9 individuals) in a Brazilian zoo. Post-mortem examination of the animals reveled extensive areas of lung hemorrhage, hepatic congestion, hemorrhagic enteritis and cardiac congestion. Histopathological examination of the organs showed the presence of multiple foci of vegetative cells of Gram-positive bacilli associated with discrete and moderate mononuclear inflammatory cell infiltrate. Seventeen B. cereus strains isolated from blood and sterile organs of nine A. ararauna were analyzed in order to investigate the genetic diversity (assessed by Rep-PCR) and toxigenic profiles (presence of hblA, hblC and hblD; nheA, nheB and nheC as well as cytK, ces and entFM genes) of such strains. Amplification of genomic DNA by Rep-PCR of B. cereus strains generated two closely related profiles (Rep-PCR types A and B) with three bands of difference. All strains were classified as belonging to the toxigenic profile I which contained HBL and NHE gene complexes, entFM and cytK genes. Altogether, microbiological and histopathological findings and the evidence provided by the success of the antibiotic prophylaxis, corroborate that B. cereus was the causative agent of the infection that killed the birds.

  15. Specific identification of Bacillus anthracis strains

    Science.gov (United States)

    Krishnamurthy, Thaiya; Deshpande, Samir; Hewel, Johannes; Liu, Hongbin; Wick, Charles H.; Yates, John R., III

    2007-01-01

    Accurate identification of human pathogens is the initial vital step in treating the civilian terrorism victims and military personnel afflicted in biological threat situations. We have applied a powerful multi-dimensional protein identification technology (MudPIT) along with newly generated software termed Profiler to identify the sequences of specific proteins observed for few strains of Bacillus anthracis, a human pathogen. Software termed Profiler was created to initially screen the MudPIT data of B. anthracis strains and establish the observed proteins specific for its strains. A database was also generated using Profiler containing marker proteins of B. anthracis and its strains, which in turn could be used for detecting the organism and its corresponding strains in samples. Analysis of the unknowns by our methodology, combining MudPIT and Profiler, led to the accurate identification of the anthracis strains present in samples. Thus, a new approach for the identification of B. anthracis strains in unknown samples, based on the molecular mass and sequences of marker proteins, has been ascertained.

  16. 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.

  17. TOXINS AND ANTITOXINS OF BACILLUS DYSENTERIAE SHIGA.

    Science.gov (United States)

    Olitsky, P K; Kligler, I J

    1920-01-01

    With the methods which have been described we have separated an exotoxin and an endotoxin from cultures of the Shiga dysenteric bacillus. The study of the nature and effect of the poison of this microorganism is thus simplified. The two toxins are physically and biologically distinct. The exotoxin is relatively heat-labile, arises in the early period of growth, and yields an antiexotoxic immune serum. The endotoxin, on the other hand, is heat-stable, is formed in the later period of growth, and is not neutralized by the antiexotoxic serum. The exotoxin exhibits a specific affinity for the central nervous organs in the rabbit, giving rise to a characteristic lesion-mainly, hemorrhages, necroses, and possibly a perivascular infiltration in the gray matter of the upper spinal cord and medulla. The endotoxin exerts a typical action on the intestinal tract, producing edema, hemorrhages, necroses, and ulcerations, especially in the large intestine. In dysentery in man the intestinal lesions predominate, but in severe epidemics paralysis and neuritis have been observed (Osler(17)). These facts become specially significant from the standpoint of the serum therapy of bacillary dysentery. A potent antidysenteric serum should contain antibodies against the exotoxin as well as the endotoxin. That such a serum can be produced in horses has been experimentally demonstrated.

  18. Transferrin Impacts Bacillus thuringiensis Biofilm Levels

    Directory of Open Access Journals (Sweden)

    Bianca Garner

    2016-01-01

    Full Text Available The present study examined the impact of transferrin on Bacillus thuringiensis biofilms. Three commercial strains, an environmental strain (33679, the type strain (10792, and an isolate from a diseased insect (700872, were cultured in iron restricted minimal medium. All strains produced biofilm when grown in vinyl plates at 30°C. B. thuringiensis 33679 had a biofilm biomass more than twice the concentration exhibited by the other strains. The addition of transferrin resulted in slightly increased growth yields for 2 of the 3 strains tested, including 33679. In contrast, the addition of 50 μg/mL of transferrin resulted in an 80% decrease in biofilm levels for strain 33679. When the growth temperature was increased to 37°C, the addition of 50 μg/mL of transferrin increased culture turbidity for only strain 33679. Biofilm levels were again decreased in strain 33679 at 37°C. Growth of B. thuringiensis cultures in polystyrene resulted in a decrease in overall growth yields at 30°C, with biofilm levels significantly decreased for 33679 in the presence of transferrin. These findings demonstrate that transferrin impacts biofilm formation in select strains of B. thuringiensis. Identification of these differences in biofilm regulation may be beneficial in elucidating potential virulence mechanisms among the differing strains.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. Requirements for the Development of Bacillus Anthracis Spore Reference Materials Used to Test Detection Systems

    Science.gov (United States)

    2006-01-01

    in some strains of Bacillus cereus and Bacillus thuringiensis [55, 56]. The Ba813 marker has been used for a real time PCR assay using Taqman-type...pXO1, the large Bacillus anthracis plasmid harboring the anthrax toxin genes, J. Bacteriol. 181, 6509-6515 (1999). [36] L.B. Price, M. Hugh-Jones, P. J...useful results. The spores of Bacillus anthracis (BA) are particular- ly dangerous because they persist in the environment, and relatively small numbers

  6. Simple detection of Bacillus anthracis spores by precipitation method with goat antibody anti anthrosa

    OpenAIRE

    2016-01-01

    Background: Bacillus anthracis has a potential for biological weapon or bioterorism. Attack of Bacillus anthracis is very fatal, and the distribution is very easy and cheap through the spores. The aim of this was study to detect the spores of Bacillus anthracis. Methods: Bacillus anthracis isolates were grown on serum agar and then sheep blood medium, to stimulate capsule formation. Spores which formed painted using the method of Schaefer and Fultton. The methods of precipitation and immun...

  7. 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.

  8. Studies on the characterisation of Biosealant properties of Bacillus sphaericus

    Directory of Open Access Journals (Sweden)

    Kantha D.Arunachalam

    2010-03-01

    Full Text Available In previous works Bacillus pasteurii was the only well known species used to precipitate the calcium carbonate. Bacillus spharecius was yet another partially characterized species with similar entity, having the capability of precipitating calcium carbonate. Earlier researchers have shown very less implementation of the organism inremediation aspect. Bacillus spharecius was sub cultured and temperature, pH were optimized at 7.4 and 37°C. Growth curve for Bacillus spharecius showed that the log phase was between 4-11 hours and after 21 hours the bacterial growth was inhibited. EDTA titration was performed to find out the amount of CaCO3 precipitate and itwas highest at pH 8. The broth culture was subjected to Atomic Force Microscope studies. The analysis confirmed the presence of calcite in both the bacterial solution and dry scrapes. Optimum nickel ion concentration for calcium carbonate precipitation was found to be 80μm. The cubes were treated for 5 days in laboratory scale and to pilot scale in the second phase for 25 days. At the end of the study, the potential of Bacillus pasteurii in Bio-concrete was well established.

  9. Bacillus anthracis lethal toxin reduces human alveolar epithelial barrier function.

    Science.gov (United States)

    Langer, Marybeth; Duggan, Elizabeth Stewart; Booth, John Leland; Patel, Vineet Indrajit; Zander, Ryan A; Silasi-Mansat, Robert; Ramani, Vijay; Veres, Tibor Zoltan; Prenzler, Frauke; Sewald, Katherina; Williams, Daniel M; Coggeshall, Kenneth Mark; Awasthi, Shanjana; Lupu, Florea; Burian, Dennis; Ballard, Jimmy Dale; Braun, Armin; Metcalf, Jordan Patrick

    2012-12-01

    The lung is the site of entry for Bacillus anthracis in inhalation anthrax, the deadliest form of the disease. Bacillus anthracis produces virulence toxins required for disease. Alveolar macrophages were considered the primary target of the Bacillus anthracis virulence factor lethal toxin because lethal toxin inhibits mouse macrophages through cleavage of MEK signaling pathway components, but we have reported that human alveolar macrophages are not a target of lethal toxin. Our current results suggest that, unlike human alveolar macrophages, the cells lining the respiratory units of the lung, alveolar epithelial cells, are a target of lethal toxin in humans. Alveolar epithelial cells expressed lethal toxin receptor protein, bound the protective antigen component of lethal toxin, and were subject to lethal-toxin-induced cleavage of multiple MEKs. These findings suggest that human alveolar epithelial cells are a target of Bacillus anthracis lethal toxin. Further, no reduction in alveolar epithelial cell viability was observed, but lethal toxin caused actin rearrangement and impaired desmosome formation, consistent with impaired barrier function as well as reduced surfactant production. Therefore, by compromising epithelial barrier function, lethal toxin may play a role in the pathogenesis of inhalation anthrax by facilitating the dissemination of Bacillus anthracis from the lung in early disease and promoting edema in late stages of the illness.

  10. 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.

  11. Bacillus filamentosus sp. nov., isolated from sediment sample.

    Science.gov (United States)

    Sonalkar, Vidya V; Mawlankar, Rahul; Venkata Ramana, V; Joseph, Neetha; Shouche, Yogesh S; Dastager, Syed G

    2015-02-01

    A novel Gram-stain positive, endospore-forming bacterium, designated SGD-14(T), was isolated from a marine sediment sample in Goa Province, India. Cells of the isolate were found to be strictly aerobic. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SGD-14(T) showed a similarity of 99.5 % with Bacillus endophyticus and similarities to other Bacillus type strains were below 96 %. The whole-cell sugar pattern was found to consist of ribose, xylose and glucose. The predominant menaquinone was identified as MK-7 and the major fatty acids as anteiso-C15:0, iso-C15:0, iso-C16:0, anteiso-C17:0, C16:0 and iso-C14:0. The strain was found to grow optimally at 30 °C and pH 7.0-7.5. DNA G + C content was determined to be 39.6 mol%. The phospholipid pattern was found to consist of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. DNA-DNA hybridization studies between strain SGD-14(T) and B. endophyticus CIP106778(T) showed that strain SGD-14(T) exhibited Bacillus, for which the name Bacillus filamentosus sp. nov. is proposed. The type strain of Bacillus filamentosus is SGD-14(T) = (=NCIM 5491(T) = DSM 27955(T)).

  12. 77 FR 19109 - Bacillus Pumilus Strain GHA 180; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2012-03-30

    ... AGENCY 40 CFR Part 180 Bacillus Pumilus Strain GHA 180; Exemption From the Requirement of a Tolerance... an exemption from the requirement of a tolerance for residues of Bacillus pumilus strain GHA 180 in... permissible level for residues of Bacillus pumilus strain GHA 180. DATES: This regulation is effective...

  13. 40 CFR 180.1107 - Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Delta endotoxin of Bacillus... From Tolerances § 180.1107 Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into... Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens is exempt from...

  14. 40 CFR 180.1269 - Bacillus mycoides Isolate J: 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 mycoides Isolate J: exemption... FOOD Exemptions From Tolerances § 180.1269 Bacillus mycoides Isolate J: exemption from the requirement of a tolerance. Bacillus mycoides isolate J is temporarily exempt from the requirement of a...

  15. 77 FR 2910 - Bacillus Amyloliquefaciens Strain D747; Exemption From the Requirement of a Tolerance; Technical...

    Science.gov (United States)

    2012-01-20

    ... AGENCY 40 CFR Part 180 Bacillus Amyloliquefaciens Strain D747; Exemption From the Requirement of a... establishment of an exemption from the requirement of a tolerance for residues of Bacillus amyloliquefaciens strain D747 (formerly known as Bacillus subtilis variant amyloliquefaciens strain D747). This document...

  16. 77 FR 745 - Bacillus Amyloliquefaciens Strain D747; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2012-01-06

    ... AGENCY 40 CFR Part 180 Bacillus Amyloliquefaciens Strain D747; Exemption From the Requirement of a... establishes an exemption from the requirement of a tolerance for residues of the Bacillus amyloliquefaciens strain D747 (formerly known as Bacillus subtilis variant amyloliquefaciens strain D747) in or on all...

  17. 40 CFR 180.1282 - Bacillus firmus I-1582; 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 firmus I-1582; exemption from... FOOD Exemptions From Tolerances § 180.1282 Bacillus firmus I-1582; exemption from the requirement of a..., for residues of Bacillus firmus I-1582 when used as a soil application or seed treatment....

  18. 78 FR 35147 - Bacillus pumilus Strain BU F-33; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2013-06-12

    ... AGENCY 40 CFR Part 180 Bacillus pumilus Strain BU F-33; Exemption From the Requirement of a Tolerance... an exemption from the requirement of a tolerance for residues of Bacillus pumilus strain BU F-33 in... residues of Bacillus pumilus strain BU F-33 under the FFDCA. DATES: This regulation is effective June...

  19. 76 FR 28689 - Microbiology Devices; Classification of In Vitro Diagnostic Device for Bacillus Species Detection

    Science.gov (United States)

    2011-05-18

    ... Vitro Diagnostic Device for Bacillus Species Detection AGENCY: Food and Drug Administration, HHS. ACTION... devices for Bacillus species (spp). detection into ] class II (special controls), in accordance with the.... Regulatory History of In Vitro Diagnostic Devices for Bacillus Spp. Detection After the enactment of...

  20. 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...

  1. 40 CFR 180.1111 - Bacillus subtilis GB03; 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 GB03; exemption from... FOOD Exemptions From Tolerances § 180.1111 Bacillus subtilis GB03; exemption from the requirement of a tolerance. The biofungicide Bacillus subtilis GB03 is exempted from the requirement of a tolerance in or...

  2. 40 CFR 180.1202 - Bacillus sphaericus; 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 sphaericus; exemption from... FOOD Exemptions From Tolerances § 180.1202 Bacillus sphaericus; exemption from the requirement of a... pesticides, Bacillus sphaericus when used in or on all food crops....

  3. 21 CFR 184.1012 - α-Amylase enzyme preparation from Bacillus stearothermophilus.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true α-Amylase enzyme preparation from Bacillus... preparation from Bacillus stearothermophilus. (a) α-Amylase enzyme preparation is obtained from the culture... Bacillus stearothermophilus. Its characterizing enzyme activity is α-amylase (1,4 α-D...

  4. 77 FR 1633 - Bacillus Subtilis Strain CX-9060; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2012-01-11

    ... AGENCY 40 CFR Part 180 Bacillus Subtilis Strain CX-9060; Exemption From the Requirement of a Tolerance... an exemption from the requirement of a tolerance for residues of the microbial pesticide Bacillus... eliminates the need to establish a maximum permissible level for residues of Bacillus subtilis strain...

  5. 40 CFR 180.1224 - Bacillus pumilus GB34; 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 pumilus GB34; exemption from... FOOD Exemptions From Tolerances § 180.1224 Bacillus pumilus GB34; exemption from the requirement of a... pesticide Bacillus pumilus GB34 when used as a seed treatment in or on all food commodities. An exemption...

  6. 40 CFR 180.1011 - Viable spores of the microorganism Bacillus thuringiensis Berliner; exemption from the...

    Science.gov (United States)

    2010-07-01

    ... Bacillus thuringiensis Berliner; exemption from the requirement of a tolerance. 180.1011 Section 180.1011... microorganism Bacillus thuringiensis Berliner; exemption from the requirement of a tolerance. (a) For the... authentic strain of Bacillus thuringiensis Berliner conforming to the morphological and...

  7. Malate dehydrogenases from actinomycetes: structural comparison of Thermoactinomyces enzyme with other actinomycete and Bacillus enzymes.

    OpenAIRE

    1984-01-01

    Malate dehydrogenases from bacteria belonging to the genus Thermoactinomyces are tetrameric, like those from Bacillus spp., and exhibit a high degree of structural homology to Bacillus malate dehydrogenase as judged by immunological cross-reactivity. Malate dehydrogenases from other actinomycetes are dimers and do not cross-react with antibodies to Bacillus malate dehydrogenase.

  8. Wide Area Recovery and Resiliency Program (WARRP) Interim Clearance Strategy for Environments Contaminated with Bacillus anthracis

    Science.gov (United States)

    2012-07-01

    Interim Clearance Strategy for Environments Contaminated with Bacillus anthracis July 2012...WARRP) Interim Clearance Strategy for Environments Contaminated with Bacillus anthracis 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT...contains color images. 14. ABSTRACT If a Bacillus anthracis incident occurs in the United States or within its territories, the public health and

  9. 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...

  10. Genomic characterization of six novel Bacillus pumilus bacteriophages.

    Science.gov (United States)

    Lorenz, Laura; Lins, Bridget; Barrett, Jonathan; Montgomery, Andrew; Trapani, Stephanie; Schindler, Anne; Christie, Gail E; Cresawn, Steven G; Temple, Louise

    2013-09-01

    Twenty-eight bacteriophages infecting the local host Bacillus pumilus BL-8 were isolated, purified, and characterized. Nine genomes were sequenced, of which six were annotated and are the first of this host submitted to the public record. The 28 phages were divided into two groups by sequence and morphological similarity, yielding 27 cluster BpA phages and 1 cluster BpB phage, which is a BL-8 prophage. Most of the BpA phages have a host range restricted to distantly related strains, B. pumilus and B. simplex, reflecting the complexities of Bacillus taxonomy. Despite isolation over wide geographic and temporal space, the six cluster BpA phages share most of their 23 functionally annotated protein features and show a high degree of sequence similarity, which is unique among phages of the Bacillus genera. This is the first report of B. pumilus phages since 1981.

  11. A novel secreted metzincin metalloproteinase from Bacillus intermedius.

    Science.gov (United States)

    Sabirova, Albina R; Rudakova, Natalya L; Balaban, Nelly P; Ilyinskaya, Olga N; Demidyuk, Ilya V; Kostrov, Sergey V; Rudenskaya, Galina N; Sharipova, Margarita R

    2010-11-05

    The mprBi gene from Bacillus intermedius 3-19 encoding a novel secreted metalloproteinase was identified. The mpriBi gene was expressed in an extracellular proteinase-deficient Bacillus subtilis BG 2036 strain and the corresponding protein was characterized biochemically. The 19 kDa MprBi protein was purified to homogeneity and sequenced by mass spectroscopy and Edman degradation methods. Amino acid sequence analysis of MprBi identified an active site motif HEYGHNFGLPHD and a conserved structural component Met-turn, both of which are unique features of the metzincin clan. Furthermore, MprBi harbors a number of distinct sequence elements characteristic of proteinase domains in eukaryotic adamalysins. We conclude that MprBi and similar proteins from other Bacillus species form a novel group of metzincin metalloproteinases in prokaryotes.

  12. 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

  13. 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.

  14. 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.

  15. Genotyping of Bacillus cereus strains by microarray-based resequencing.

    Science.gov (United States)

    Zwick, Michael E; Kiley, Maureen P; Stewart, Andrew C; Mateczun, Alfred; Read, Timothy D

    2008-07-02

    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.

  16. 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.

  17. 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.

  18. 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...

  19. 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...

  20. Antagonism of Bacillus spp. against Xanthomonas campestris pv. campestris

    Directory of Open Access Journals (Sweden)

    Leila Monteiro

    2005-01-01

    Full Text Available The antagonism of eight Bacillus isolates was investigated against nine strains of Xanthomonas campestris pv. campestris (causal agent of crucifers black rot to assess the role of lipopeptides in this process. Antimicrobial and hemolytic (surfactant activity tests were performed in vitro using agar diffusion methods. Antibiosis and hemolysis were positive for four Bacillus isolates against all X. campestris pv. campestris strains. The correlation observed between antimicrobial and hemolytic activities indicated that lipopeptides were involved in the antibiosis mechanism of the studied antagonists. Fermentation studies were carried out with the isolates that showed highest antimicrobial and hemolytic activities, to follow up growth and production of bioactive and surfactant compounds. Production of bioactive and surfactant compounds was observed during the late growth phase of the Bacillus isolates.Investigação sobre o antagonismo de oito isolados de Bacillus: B. subtilis R14, B. megaterium pv. cerealis RAB7, B. megaterium pv. cerealis C211, B. megaterium C116, Bacillus sp. RAB9, B. cereus C240, Bacillus sp. C11 e B. cereus C210, contra nove linhagens de X. campestris pv. campestris (bactéria responsável pela podridão negra das crucíferas foi realizada para se verificar a participação de lipopeptídeos neste mecanismo. Testes de atividades antimicrobiana e hemolítica (surfactante foram realizados, utilizando-se o método de difusão em ágar. Antibiose e hemólise foram positivas para quatro isolados de Bacillus: R14, RAB7, C116 e C210. A correlação observada entre as atividades antimicrobiana e a hemolítica indica que lipopeptídeos estão envolvidos no mecanismo de antibiose dos isolados investigados. As fermentações foram realizadas com os isolados que demonstraram melhores resultados nos testes de atividades antimicrobiana e hemolítica: R14, RAB7 e C116, para acompanhar o crescimento e a produção de compostos bioativos e

  1. 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.

  2. Cloning of the Protective Antigen Gene of Bacillus anthracis

    Science.gov (United States)

    1983-09-01

    of the complicated precedents of duplicate toxin genes in chro- muumm mosomall and plasmid DNA of B. thuringiensis (Schnepf and Whitely, 1981; Klier...OiL V4. 34. S-W7. SW 1v 99 CwI 0193 by MT 0 009-7483/06O-002.00/0 mU"- - 1*;)-0Cloning of the Protective Antigen Gene OCT 19 MI L Sof Bacillus ...Sumnler uncertain, it is probably caused by other Bacillus antigens, 4 t which may include LF and EF. PA produced from recom- A The - "w t of a

  3. Osteomyelitis due to Bacillus cereus in an adolescent: case report and review.

    Science.gov (United States)

    Schricker, M E; Thompson, G H; Schreiber, J R

    1994-06-01

    Non-anthracis Bacillus species associated with clinical infections are usually dismissed as contaminants or nonpathogens. As opportunists, however, Bacillus organisms can cause significant systemic infections including bacteremia, endophthalmitis, and pneumonia. Osteomyelitis with non-anthracis Bacillus organisms has been described in adults, although to our knowledge it has been described only once in a child. We report a case of chronic osteomyelitis due to Staphylococcus aureus and superinfection with Bacillus cereus in a 13-year-old adolescent. A Bacillus isolate should be considered a true pathogen in children with chronic osteomyelitis who have a poor clinical response to antistaphylococcal therapy.

  4. 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.

  5. Identification and Classification of bcl Genes and Proteins of Bacillus cereus Group Organisms and Their Application in Bacillus anthracis Detection and Fingerprinting▿ †

    OpenAIRE

    Leski, Tomasz A.; Caswell, Clayton C.; Pawlowski, Marcin; Klinke, David J.; Bujnicki, Janusz M.; Hart, Sean J.; Lukomski, Slawomir

    2009-01-01

    The Bacillus cereus group includes three closely related species, B. anthracis, B. cereus, and B. thuringiensis, which form a highly homogeneous subdivision of the genus Bacillus. One of these species, B. anthracis, has been identified as one of the most probable bacterial biowarfare agents. Here, we evaluate the sequence and length polymorphisms of the Bacillus collagen-like protein bcl genes as a basis for B. anthracis detection and fingerprinting. Five genes, designated bclA to bclE, are p...

  6. 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.

  7. PCR screening for the surfactin (sfp) gene in marine Bacillus strains and its molecular characterization from Bacillus tequilensis NIOS11

    OpenAIRE

    POROB, Seema; NAYAK, Sagar; FERNANDES, Areena; PADMANABHAN, Priyanka

    2013-01-01

    The sfp gene responsible for surfactin production was screened from the DNA extracts of 37 Bacillus spp. whose identity was confirmed by 16S rRNA gene sequence analyses. PCR screening revealed amplification of sfp gene fragments in a total of 25 isolates. Several isolates belonging to Bacillus tequilensis were found to be positive for this gene. A gene fragment coding for the sfp gene was amplified and cloned from genomic DNA of the isolate B. tequilensis NIOS11. The cloned gene has an open r...

  8. The Genome Sequence of Bacillus cereus ATCC 10987 Reveals Metabolic Adaptations and a Large Plasmid Related to Bacillus anthracis pXO1

    Science.gov (United States)

    2004-01-01

    R.L. and Waites,K.B. (2003) Bacillus cereus bacteremia in a preterm neonate. J. Clin. Microbiol., 41, 3441±3444. 9. Ginsburg,A.S., Salazar,L.G., True... bacteremia and pneumonia due to Bacillus cereus . J. Clin. Microbiol., 35, 504±507. 12. Okinaka,R., Cloud,K., Hampton,O., Hoffmaster,A., Hill,K., Keim,P...The genome sequence of Bacillus cereus ATCC 10987 reveals metabolic adaptations and a large plasmid related to Bacillus anthracis pXO1 David A. Rasko

  9. Virulence of Bacillus cereus: a multivariate analysis.

    Science.gov (United States)

    Minnaard, J; Delfederico, L; Vasseur, V; Hollmann, A; Rolny, I; Semorile, L; Pérez, P F

    2007-05-10

    Biological activity and presence of DNA sequences related to virulence genes were studied in 21 strains of the Bacillus cereus group. The activity of spent culture supernatants and the effect of infection by vegetative bacterial cells were assessed on cultured human enterocytes (Caco-2 cells). The effect of extracellular factors on the detachment, necrosis and mitochondrial dehydrogenase activity of cultured human enterocytes was studied. Hemolytic activity on rabbit red blood cells was also evaluated and the effect of direct procaryotic-eucaryotic interactions was assessed in infection assays with vegetative bacterial cells. Concerning virulence genes, presence of the DNA sequences corresponding to the genes entS, entFM, nhe (A, B and C), sph, hbl (A, B, C and D), piplC and bceT was assessed by PCR. Ribopatterns were determined by an automated riboprinting analysis after digestion of the DNA with EcoRI. Principal component analysis and biplots were used to address the relationship between variables. Results showed a wide range of biological activities: decrease in mitochondrial dehydrogenase activity, necrosis, cell detachment and hemolytic activity. These effects were strain-dependent. Concerning the occurrence of the DNA sequences tested, different patterns were found. In addition, ribotyping showed that strains under study grouped into two main clusters. One of these clusters includes all the strains that were positive for all the DNA sequences tested. Positive and negative correlations between variables under study were evidenced. Interestingly, high detaching strains were positively correlated with the presence of the sequences entS, nheC and sph. Within gene complexes, high correlation was found between sequences of the hbl complex. In contrast, sequences of the nhe complex were not correlated. Some strains clustered together in the biplots. These strains were positive for all the DNA sequences tested and they were able to detach enterocytes upon infection

  10. 14C Analysis of protein extracts from Bacillus spores.

    Science.gov (United States)

    Cappuccio, Jenny A; Falso, Miranda J Sarachine; Kashgarian, Michaele; Buchholz, Bruce A

    2014-07-01

    Investigators of bioagent incidents or interdicted materials need validated, independent analytical methods that will allow them to distinguish between recently made bioagent samples versus material drawn from the archives of a historical program. Heterotrophic bacteria convert the carbon in their food sources, growth substrate or culture media, into the biomolecules they need. The F(14)C (fraction modern radiocarbon) of a variety of media, Bacillus spores, and separated proteins from Bacillus spores was measured by accelerator mass spectrometry (AMS). AMS precisely measures F(14)C values of biological materials and has been used to date the synthesis of biomaterials over the bomb pulse era (1955 to present). The F(14)C of Bacillus spores reflects the radiocarbon content of the media in which they were grown. In a survey of commercial media we found that the F(14)C value indicated that carbon sources for the media were alive within about a year of the date of manufacture and generally of terrestrial origin. Hence, bacteria and their products can be dated using their (14)C signature. Bacillus spore samples were generated onsite with defined media and carbon free purification and also obtained from archived material. Using mechanical lysis and a variety of washes with carbon free acids and bases, contaminant carbon was removed from soluble proteins to enable accurate (14)C bomb-pulse dating. Since media is contemporary, (14)C bomb-pulse dating of isolated soluble proteins can be used to distinguish between historical archives of bioagents and those produced from recent media.

  11. Global Network Reorganization During Dynamic Adaptations of Bacillus subtilis Metabolism

    NARCIS (Netherlands)

    Buescher, Joerg Martin; Liebermeister, Wolfram; Jules, Matthieu; Uhr, Markus; Muntel, Jan; Botella, Eric; Hessling, Bernd; Kleijn, Roelco Jacobus; Le Chat, Ludovic; Lecointe, Francois; Maeder, Ulrike; Nicolas, Pierre; Piersma, Sjouke; Ruegheimer, Frank; Becher, Doerte; Bessieres, Philippe; Bidnenko, Elena; Denham, Emma L.; Dervyn, Etienne; Devine, Kevin M.; Doherty, Geoff; Drulhe, Samuel; Felicori, Liza; Fogg, Mark J.; Goelzer, Anne; Hansen, Annette; Harwood, Colin R.; Hecker, Michael; Hubner, Sebastian; Hultschig, Claus; Jarmer, Hanne; Klipp, Edda; Leduc, Aurelie; Lewis, Peter; Molina, Frank; Noirot, Philippe; Peres, Sabine; Pigeonneau, Nathalie; Pohl, Susanne; Rasmussen, Simon; Rinn, Bernd; Schaffer, Marc; Schnidder, Julian; Schwikowski, Benno; Van Dijl, Jan Maarten; Veiga, Patrick; Walsh, Sean; Wilkinson, Anthony J.; Stelling, Joerg; Aymerich, Stephane; Sauer, Uwe

    2012-01-01

    Adaptation of cells to environmental changes requires dynamic interactions between metabolic and regulatory networks, but studies typically address only one or a few layers of regulation. For nutritional shifts between two preferred carbon sources of Bacillus subtilis, we combined statistical and mo

  12. Novel giant siphovirus from Bacillus anthracis features unusual genome characteristics.

    Directory of Open Access Journals (Sweden)

    Holly H Ganz

    Full Text Available Here we present vB_BanS-Tsamsa, a novel temperate phage isolated from Bacillus anthracis, the agent responsible for anthrax infections in wildlife, livestock and humans. Tsamsa phage is a giant siphovirus (order Caudovirales, featuring a long, flexible and non-contractile tail of 440 nm (not including baseplate structure and an isometric head of 82 nm in diameter. We induced Tsamsa phage in samples from two different carcass sites in Etosha National Park, Namibia. The Tsamsa phage genome is the largest sequenced Bacillus siphovirus, containing 168,876 bp and 272 ORFs. The genome features an integrase/recombinase enzyme, indicative of a temperate lifestyle. Among bacterial strains tested, the phage infected only certain members of the Bacillus cereus sensu lato group (B. anthracis, B. cereus and B. thuringiensis and exhibited moderate specificity for B. anthracis. Tsamsa lysed seven out of 25 B. cereus strains, two out of five B. thuringiensis strains and six out of seven B. anthracis strains tested. It did not lyse B. anthracis PAK-1, an atypical strain that is also resistant to both gamma phage and cherry phage. The Tsamsa endolysin features a broader lytic spectrum than the phage host range, indicating possible use of the enzyme in Bacillus biocontrol.

  13. Complete Genome Sequence of Bacillus thuringiensis Strain 407 Cry-

    OpenAIRE

    Poehlein, Anja; Liesegang, Heiko

    2013-01-01

    Bacillus thuringiensis is an insect pathogen that has been used widely as a biopesticide. Here, we report the genome sequence of strain 407 Cry-, which is used to study the genetic determinants of pathogenicity. The genome consists of a 5.5-Mb chromosome and nine plasmids, including a novel 502-kb megaplasmid.

  14. Bacillus nakamurai sp. nov., a black pigment producing strain

    Science.gov (United States)

    Two isolates of a Gram-positive, strictly aerobic, motile, rod-shaped, endospore-forming bacterium were identified during a survey of the Bacillus diversity of the Agriculture Research Service Culture Collection. These strains were originally isolated from soil and have a phenotype of producing a da...

  15. Bacillus Calmette-Guérin vaccination at birth

    DEFF Research Database (Denmark)

    Kjærgaard, Jesper; Stensballe, Lone Graff; Birk, Nina Marie

    2016-01-01

    BACKGROUND: Bacillus Calmette-Guérin vaccine (BCG) induces a complex, pro-inflammatory immune response. Obesity is associated with low-grade inflammation. AIMS: The purpose of the study was to test whether BCG at birth has effects on infant growth and body composition. STUDY DESIGN, SUBJECTS...

  16. Development of Bacillus subtilis mutants to produce tryptophan in pigs

    DEFF Research Database (Denmark)

    Bjerre, Karin; Cantor, Mette D.; Nørgaard, Jan Værum

    2017-01-01

    Objectives To generate tryptophan-overproducing Bacillus subtilis strains for in situ use in pigs, to reduce the feed cost for farmers and nitrogen pollution. Results A novel concept has been investigated—to generate B. subtilis strains able to produce tryptophan (Trp) in situ in pigs. Mutagenesis...

  17. Bacillusin A, an antibacterial macrodiolide from Bacillus amyloliquefaciens

    Science.gov (United States)

    Bioassay-guided fractionation of the organic extracts of a Bacillus amyloliquefaciens strain (AP183) led to the discovery of a new macrocyclic polyene antibiotic, bacillusin A (1). Its structure was assigned by interpretation of NMR and MS spectroscopic data as a novel macrodiolide composed of dimer...

  18. Biodegradation of 2-hydroxyquinoxaline (2-HQ) by Bacillus sp.

    Science.gov (United States)

    Reddy, G V Subba; Reddy, B R; Tlou, M G

    2014-08-15

    An aerobic Gram +ve bacterial strain capable of utilizing 2-Hydroxyquinoxaline (2-HQ) as sole source of carbon and energy was isolated from Chrysanthemum indicum Indian agricultural soil and named as HQ2. On the basis of morphology, physico-biochemical characteristics and 16S rRNA sequence analysis, strain HQ2 was identified as Bacillus sp. The generation time of Bacillus sp. in log phase during growth on 2-HQ is 0.79 h or 47.4 min. The optimal conditions for 2-HQ degradation by Bacillus sp. were inoculum density of 1.0 OD, pH of 6-8, temperature of 37-45 °C and 2-HQ concentration of 500 ppm. Among the additional carbon and nitrogen sources, carbon sources did not influence the degradation rate of 2-HQ, but nitrogen sources-yeast extract marginally enhanced the rate of degradation of 2-HQ. GC-MS analysis of the culture Bacillus sp. grown on 2-HQ indicated the formation of dimers from 2 molecules of 2-hydroxyquinoxaline. The formation of dimer for degradation of 2-HQ by the culture appears to be the first report to our scientific knowledge.

  19. Bacillus Calmette-Guérin vaccination at birth

    DEFF Research Database (Denmark)

    Kjærgaard, Jesper

    2016-01-01

    The Bacillus Calmette-Guérin vaccine (BCG), which is used to protect against tuberculosis, has been associated with a variety of other effects since it was developed almost 100 years ago. Most notably, observational studies and randomized clinical trials from low-income countries indicate...

  20. Factors affecting the solubility of Bacillus halmapalus alpha-amylase

    DEFF Research Database (Denmark)

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

    2008-01-01

    A detailed study of the solubility of recombinant Bacillus halmapalus alpha-amylase has been conducted. A semi-purified preparation from a bulk crystallisation was chos en that contained six isoforms with pI-values of between 5.5 and 6.1. The solubility was strongly affected by pH and could...

  1. The transcriptionally active regions in the genome of Bacillus subtilis

    DEFF Research Database (Denmark)

    Rasmussen, Simon; Nielsen, Henrik Bjørn; Jarmer, Hanne Østergaard

    2009-01-01

    The majority of all genes have so far been identified and annotated systematically through in silico gene finding. Here we report the finding of 3662 strand-specific transcriptionally active regions (TARs) in the genome of Bacillus subtilis by the use of tiling arrays. We have measured the genome...

  2. Engineering of Bacillus subtilis 168 for increased nisin resistance

    DEFF Research Database (Denmark)

    Hansen, Mette; Wangari, Romilda; Hansen, Egon Bech

    2009-01-01

    . Bacillus subtilis had been suggested as a potential host for the biosynthesis of nisin but was discarded due to its sensitivity to the lethal action of nisin. In this study, we have reevaluated the potential of B. subtilis as a host organism for the heterologous production of nisin. We applied...

  3. A New Saponin Transformed from Ginsenoside Rhl by Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    Guo Hong LI; Yue Mao SHEN; Ke Qin ZHANG

    2005-01-01

    A novel saponin was isolated from the transformed products of ginsenoside Rh1 by Bacillus subtilis. It's structure was determined to be 3-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosyl-20 (S)-protopanaxatriol on the basis of the spectral data.

  4. Thermostable, Raw-Starch-Digesting Amylase from Bacillus stearothermophilus

    OpenAIRE

    Kim, Jaeyoung; Nanmori, Takashi; Shinke, Ryu

    1989-01-01

    An endospore-forming thermophilic bacterium, which produced amylase and was identified as Bacillus stearothermophilus, was isolated from soil. The amylase had an optimum temperature of 70°C and strongly degraded wheat starch granules (93%) and potato starch granules (80%) at 60°C.

  5. Characterization of germination receptors of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

    Hornstra, L.M.; Vries, de Y.P.; Wells-Bennik, M.H.J.; Vos, de W.M.; Abee, T.

    2006-01-01

    Specific amino acids, purine ribonucleosides, or a combination of the two is required for efficient germination of endospores of Bacillus cereus ATCC 14579. A survey including 20 different amino acids showed that L-alanine, L-cysteine, L-threonine, and L-glutamine are capable of initiating the germi

  6. Complete Genome Sequence of Bacillus megaterium Myophage Mater

    OpenAIRE

    Lancaster, Jacob C.; Hodde, Mary K.; Hernandez, Adriana C.; Kuty Everett, Gabriel F.

    2015-01-01

    Bacillus megaterium is a ubiquitous, soil inhabiting Gram-positive bacterium that is a common model organism and is used in industrial applications for protein production. The following reports the complete sequencing and annotation of the genome of B. megaterium myophage Mater and describes the major features identified.

  7. Novel giant siphovirus from Bacillus anthracis features unusual genome characteristics.

    Science.gov (United States)

    Ganz, Holly H; Law, Christina; Schmuki, Martina; Eichenseher, Fritz; Calendar, Richard; Loessner, Martin J; Getz, Wayne M; Korlach, Jonas; Beyer, Wolfgang; Klumpp, Jochen

    2014-01-01

    Here we present vB_BanS-Tsamsa, a novel temperate phage isolated from Bacillus anthracis, the agent responsible for anthrax infections in wildlife, livestock and humans. Tsamsa phage is a giant siphovirus (order Caudovirales), featuring a long, flexible and non-contractile tail of 440 nm (not including baseplate structure) and an isometric head of 82 nm in diameter. We induced Tsamsa phage in samples from two different carcass sites in Etosha National Park, Namibia. The Tsamsa phage genome is the largest sequenced Bacillus siphovirus, containing 168,876 bp and 272 ORFs. The genome features an integrase/recombinase enzyme, indicative of a temperate lifestyle. Among bacterial strains tested, the phage infected only certain members of the Bacillus cereus sensu lato group (B. anthracis, B. cereus and B. thuringiensis) and exhibited moderate specificity for B. anthracis. Tsamsa lysed seven out of 25 B. cereus strains, two out of five B. thuringiensis strains and six out of seven B. anthracis strains tested. It did not lyse B. anthracis PAK-1, an atypical strain that is also resistant to both gamma phage and cherry phage. The Tsamsa endolysin features a broader lytic spectrum than the phage host range, indicating possible use of the enzyme in Bacillus biocontrol.

  8. Complete Genome Sequence of Bacillus megaterium Myophage Moonbeam

    OpenAIRE

    Cadungog, Joshua N.; Khatemi, Brontee E.; Hernandez, Adriana C.; Kuty Everett, Gabriel F.

    2015-01-01

    Moonbeam is a newly isolated myophage of Bacillus megaterium, a common Gram-positive bacterium that is routinely used for large-scale protein production. Bacteriophages have potential to be useful tools for industrial applications. Here, we describe the complete genome of Moonbeam and describe its features.

  9. Bilirubin Oxidase Activity of Bacillus subtilis CotA

    OpenAIRE

    Sakasegawa, S; Ishikawa, H.; Imamura, S.; Sakuraba, H.; Goda, S.; Ohshima, T.

    2006-01-01

    The spore coat protein CotA from Bacillus subtilis was previously identified as a laccase. We have now found that CotA also shows strong bilirubin oxidase activity and markedly higher affinity for bilirubin than conventional bilirubin oxidase. This is the first characterization of bilirubin oxidase activity in a bacterial protein.

  10. Live-imaging of Bacillus subtilis spore germination and outgrowth

    NARCIS (Netherlands)

    Pandey, R.

    2014-01-01

    Spores of Gram-positive bacteria such as Bacillus and Clostridium cause huge economic losses to the food industry. In food products, spores survive under food preservation conditions and subsequent germination and outgrowth eventually causes food spoilage. Therefore efforts are being made to elimina

  11. The impact of manganese on biofilm development of Bacillus subtilis

    NARCIS (Netherlands)

    Mhatre, Eisha; Troszok, Agnieszka; Gallegos-Monterrosa, Ramses; Lindstädt, Stefanie; Hölscher, Theresa; Kuipers, Oscar P.; Kovács, Ákos T.

    2016-01-01

    Bacterial biofilms are dynamic and structurally complex communities, involving cell-to-cell interactions. In recent years, various environmental signals were identified that induce the complex biofilm development of the Gram-positive bacterium Bacillus subtilis. These signaling molecules are often m

  12. Bacillus subtilis Biosensor Engineered To Assess Meat Spoilage

    NARCIS (Netherlands)

    Daszczuk, Alicja; Dessalegne, Yonathan; Drenth, Ismael; Hendriks, Elbrich; Jo, Emeraldo; van Lente, Tom; Oldebesten, Arjan; Parrish, Jonathon; Poljakova, Wlada; Purwanto, Annisa A.; van Raaphorst, Renske; Boonstra, Mirjam; van Heel, Auke; Herber, Martijn; van der Meulen, Sjoerd; Siebring, Jeroen; Sorg, Robin A.; Heinemann, Matthias; Kuipers, Oscar P.; Veening, Jan-Willem

    2014-01-01

    Here, we developed a cell-based biosensor that can assess meat freshness using the Gram-positive model bacterium Bacillus subtilis as a chassis. Using transcriptome analysis, we identified promoters that are specifically activated by volatiles released from spoiled meat. The most strongly activated

  13. Bacillus thuringiensis-based Products for Insect Pest Control

    NARCIS (Netherlands)

    Maagd, de R.A.

    2015-01-01

    Bacillus thuringiensis (or Bt, as it has become generally known) is one of the oldest and widely used biological control agents and has a long history of use. Bt and a number of related bacteria produce a variety of toxins, mostly—but not exclusively- localized in the parasporal crystals, which are,

  14. Environmental regulation of alcohol metabolism in thermotolerant methylotrophic Bacillus strains

    NARCIS (Netherlands)

    Arfman, N.; Moezelaar, H.R.; Attwood, M.M.; Robinson, G.K.; Geel, M. van; Dijkhuizen, L.

    1992-01-01

    The thermotolerant methylotroph Bacillus sp. C1 possesses a novel NAD-dependent methanol dehydrogenase (MDH), with distinct structural and mechanistic properties. During growth on methanol and ethanol, MDH was responsible for the oxidation of both these substrates. MDH activity in cells grown on met

  15. Binding Affinity of Glycoconjugates to BACILLUS Spores and Toxins

    Science.gov (United States)

    Rasol, Aveen; Eassa, Souzan; Tarasenko, Olga

    2010-04-01

    Early recognition of Bacillus cereus group species is important since they can cause food-borne illnesses and deadly diseases in humans. Glycoconjugates (GCs) are carbohydrates covalently linked to non-sugar moieties including lipids, proteins or other entities. GCs are involved in recognition and signaling processes intrinsic to biochemical functions in cells. They also stimulate cell-cell adhesion and subsequent recognition and activation of receptors. We have demonstrated that GCs are involved in Bacillus cereus spore recognition. In the present study, we have investigated whether GCs possess the ability to bind and recognize B. cereus spores and Bacillus anthracis recombinant single toxins (sTX) and complex toxins (cTX). The affinity of GCs to spores + sTX and spores + cTX toxins was studied in the binding essay. Our results demonstrated that GC9 and GC10 were able to selectively bind to B. cereus spores and B. anthracis toxins. Different binding affinities for GCs were found toward Bacillus cereus spores + sTX and spores + cTX. Dilution of GCs does not impede the recognition and binding. Developed method provides a tool for simultaneous recognition and targeting of spores, bacteria toxins, and/or other entities.

  16. A new Bacillus pasteurii urease inhibitor from Euphorbia decipiens.

    Science.gov (United States)

    Lodhi, Muhammad Arif; Hussain, Javid; Abbasi, Muhammad Athar; Jassbi, Amir Reza; Choudhary, Muhammad Iqbal; Ahmad, Viqar Uddin

    2006-10-01

    Inhibition of Bacillus pasteurii urease enzyme by 3,7,15-tri-O-acetyl-5-O-nicotinoyl-13,14-dihydroxymyrsinol (1), a diterpene ester with a myrsinol-type skeleton, isolated from Euphorbia decipiens Boiss. and Buhse, was un-competitive consistent with the molecular docking results. The Ki value was 117.40 +/- 0.7 microM.

  17. Successful treatment of Bacillus cereus infection with ciprofloxacin.

    OpenAIRE

    Gascoigne, A.D.; Richards, J.; Gould, K.; Gibson, G. J.

    1991-01-01

    Bacillus cereus is rarely a pulmonary pathogen but may cause pneumonia in immunocompromised patients. A patient with bronchiectasis and no recognisable immunodeficiency had this organism isolated during two infective exacerbations, once from respiratory secretions and once by blood culture. Ciprofloxacin treatment was effective on both occasions.

  18. Purification and characterization of two polyhydroxyalcanoates from Bacillus cereus.

    Science.gov (United States)

    Zribi-Maaloul, Emna; Trabelsi, Imen; Elleuch, Lobna; Chouayekh, Hichem; Ben Salah, Riadh

    2013-10-01

    This work aimed to study the potential of 155 strains of Bacillus sp., isolated from a collection of Tunisian microorganisms, for polyhydroxyalcanoates production. The strains were submitted to a battery of standard tests commonly used for determining bioplastic properties. The findings revealed that two of the isolates, namely Bacillus US 163 and US 177, provided red excitations at a wavelength of approximately 543 nm. The polyhydroxyalcanoates produced by the two strains were purified. Gas chromatography-mass spectroscopy (GC-MS), Fourier transformed infrared spectroscopy (FTIR), and gel permeation chromatography (GPC) were used to characterize the two biopolymers. Bacillus US 163 was noted to produce a poly methyl-3-hydroxy tetradecanoic acid (P-3HTD) with an average molecular weight of 455 kDa, a completely amorphous homopolymer without crystallinity. The US 177 strain produced a homopolymer of methyl-3-hydroxy octadecanoic acid (P3-HOD) with an average molecular weight of 555 kDa. Exhibiting the highest performance, US 163 and US 177 were submitted to 16S rRNA gene sequencing, and the results revealed that they belonged to the Bacillus cereus species. Overall, the findings indicated that the Bacilli from petroleum soil have a number of promising properties that make them promising candidates for bioplastic production.

  19. 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

  20. Bacillus subtilis Vegetative Catalase Is an Extracellular Enzyme

    OpenAIRE

    Naclerio, G; Baccigalupi, L; Caruso, C; De Felice, M; Ricca, E

    1995-01-01

    Strong catalase activity was secreted by Bacillus subtilis cells during stationary growth phase in rich medium but not in sporulation-inducing medium. N-terminal sequencing indicated that the secreted activity was due to the vegetative catalase KatA, previously considered an endocellular enzyme. Extracellular catalase protected B. subtilis cells from oxidative assault.

  1. Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation

    NARCIS (Netherlands)

    Romero, Diego; Perez-Garcia, Alejandro; Veening, Jan-Willem; de Vicente, Antonio; Kuipers, Oscar P.; de, Vicente A.

    2006-01-01

    A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool

  2. Bacillus cereus: emetic toxin production and gamma hypothesis for growth

    NARCIS (Netherlands)

    Biesta-Peters, E.G.

    2011-01-01

    Bacillus cereus is a food spoilage microorganism and a pathogen. Growth of B. cereus can be prevented or delayed by adding growth limiting compounds to the food product or by altered storage conditions. Combinations of growth limiting factors

  3. Nucleotide Sequence of the Protective Antigen Gene of Bacillus Anthracis

    Science.gov (United States)

    1988-02-02

    Montie, S. Kadis, and S. I. Ajl (ed.), Microbial toxins, vol. 3. Academic Press, Inc., New York. 23. Little, S. F., and G. B. Knudaon. 1986...Takkinen, and L. Kaariainen. 1981. Nucleotide sequence of the promoter and NHa-terminal signal peptide region of the a- amylase gene from Bacillus

  4. Linking Bacillus cereus genotypes and carbohydrate utilization capacity

    NARCIS (Netherlands)

    Warda, Alicja K.; Siezen, Roland J.; Boekhorst, Jos; Wells-Bennik, Marjon H.J.; Jong, de 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 wi

  5. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity

    NARCIS (Netherlands)

    Warda, Alicja K.; Siezen, Roland J.; Boekhorst, Jos; Wells-Bennik, Marjon H.J.; Jong, de 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

  6. Induction of natural competence in Bacillus cereus ATCC14579

    NARCIS (Netherlands)

    Mironczuk, Aleksandra M.; Kovács, Ákos T.; Kuipers, O.P.

    2008-01-01

    Natural competence is the ability of certain microbes to take up exogenous DNA from the environment and integrate it in their genome. Competence development has been described for a variety of bacteria, but has so far not been shown to occur in Bacillus cereus. However, orthologues of most proteins

  7. Progress in food-related research focussing on Bacillus cereus

    NARCIS (Netherlands)

    Vries, de Y.P.; Voort, van der M.; Schaik, van W.; Hornstra, L.M.; Vos, de W.M.; Abee, T.

    2004-01-01

    Bacillus cereus is a gram-positive, rod-shaped, endospore-forming bacterium that occurs ubiquitously and is frequently isolated from soil and food products. When B. cereus is present in foods, it can cause spoilage and poisoning. The work of our group is focussed on several properties of B. cereus t

  8. Domains of Bacillus thuringiensis crystal proteins involved in insecticidal activity

    NARCIS (Netherlands)

    Bosch, H.J.; Schipper, B.; Kleij, van der H.; Maagd, de R.A.; Stiekema, W.J.

    1994-01-01

    The expected increase in application of Bacillus thuringiensis (Bt) in crop protection makes it necessary to anticipate the development of Bt-resistant insects. To safeguard the long-term use of Bt-based insecticides, we studied the mode of action of Bt crystal proteins. CryIA(b), CryIC and CryIE ar

  9. Bacillus vini sp. nov. isolated from alcohol fermentation pit mud.

    Science.gov (United States)

    Ma, Kedong; Chen, Xiaorong; Guo, Xiang; Wang, Yanwei; Wang, Huimin; Zhou, Shan; Song, Jinlong; Kong, Delong; Zhu, Jie; Dong, Weiwei; He, Mingxiong; Hu, Guoquan; Zhao, Bingqiang; Ruan, Zhiyong

    2016-08-01

    A novel aerobic, Gram-stain-positive, sporogenous, rod-shaped bacterium, designated LAM0415(T), was isolated from an alcohol fermentation pit mud sample collected from Sichuan Luzhou-flavour liquor enterprise in China. The isolate was found to be able to grow at NaCl concentrations of 0-10 % (w/v) (optimum: 1.0 %), 10-50 °C (optimum: 30-35 °C) and pH 3.0-10.0 (optimum: 7.0-8.0). Phylogenetic analysis of 16S rRNA gene sequences indicated that the new isolate belonged to the genus Bacillus and was closely related to Bacillus sporothermodurans DSM 10599(T) and Bacillus oleronius DSM 9356(T), with 98.4 and 97.2 % sequence similarity, respectively. The DNA-DNA hybridization values between strain LAM0415(T) and the two reference strains were 33.3 ± 1.2 and 42.8 ± 0.8 %, respectively. The genomic DNA G+C content was 35.2 mol% as determined by the T m method. The major fatty acids were determined to be iso-C15:0, anteiso-C15:0 and anteiso-C17:0. The predominant menaquinones were identified as MK7 and MK8. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, one unidentified phospholipid and four unidentified glycolipids. The diagnostic amino acid of the cell wall peptidoglycan was determined to be meso-diaminopimelic acid. On the basis of its phenotypic, phylogenetic and chemotaxonomic characteristics, strain LAM0415(T) (=ACCC 06413(T) = JCM 19841(T)) represents the type strain of a novel species of the genus Bacillus, for which the name Bacillus vini sp. nov. is proposed.

  10. PCR screening for the surfactin (sfp) gene in marine Bacillus strains and its molecular characterization from Bacillus tequilensis NIOS11

    Digital Repository Service at National Institute of Oceanography (India)

    Porob, S.; Nayak, S.; Fernandes, Areena; Padmanabhan, P.; Patil, B.A.; Meena, R.M.; Ramaiah, N.

    The sfp gene responsible for surfactin production was screened from the DNA extracts of 37 Bacillus spp. whose identity was confirmed by 16S rRNA gene sequence analyses. PCR screening revealed amplification of sfp gene fragments in a total of 25...

  11. 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.

  12. Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific.

    Science.gov (United States)

    Maes, Emmanuel; Krzewinski, Frederic; Garenaux, Estelle; Lequette, Yannick; Coddeville, Bernadette; Trivelli, Xavier; Ronse, Annette; Faille, Christine; Guerardel, Yann

    2016-04-29

    The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets.

  13. Efficacy and efficiency of new Bacillus thuringiensis var. israelensis and Bacillus sphaericus formulations against Afrotropical anophelines in Western Kenya

    NARCIS (Netherlands)

    Fillinger, U.; Knols, B.G.J.; Becker, N.

    2002-01-01

    We evaluated the efficacy of new water-dispersible granular (WDG) formulations of Bacillus thuringienis var. israelensis (Bti; VectoBac?) and B. sphaericus (Bs; VectoLex?, Valent BioScience Corp., Illinois, USA) for the control of larval Anopheles gambiae sensu lato Giles mosquitoes in a malaria-end

  14. Isolation and characterization of protease from Bacillus subtilis 1012M15

    Directory of Open Access Journals (Sweden)

    ELFI SUSANTI

    2003-01-01

    Full Text Available A local strain of Bacillus sp. BAC4, is known to produce penicillin G acylase (PGA enzyme with relatively high activity. This strain secretes the PGA into the culture medium. However, it has been reported that PGA activity fall and rise during culture, and the activity plummets during storege at –200C, which probably due to usage protease activity of Bacillus sp. BAC4. To study the possible use of Bacillus subtilis 1012M15 as a host cell for cloning the pga gene from Bacillus sp. BAC4, the protease activity of Bacillus subtilis 1012M15 were studied. Protease activity was determined by Horikoshi method. In this experiment, maximum protease activity in Bacillus subtilis 1012M15 culture was obsereved after 8 hours. At this optimum condition, protease activity of Bacillus sp. BAC4 is five time higher than that of Bacillus subtilis 1012M15. This situation promised the possible usage of Bacillus subtilis 1012M15 as a host cell for pga expression. For protease characterization, the bacterial culture had been separated from the cell debris by centrifugation. The filtrate was concentrated by freeze drying, fractionated by ammonium sulphate, dialyzed in selovan tube, and then fractionated by ion exchance chromatography employing DEAE-cellulose. The five peaks resulted indicated the presence of five protease. Based on inhibitor and activator influence analysis, it could be concluded that proteases from Bacillus subtilis 1012M15 contained of serin protease as well as metalloprotease and serin protease mixture.

  15. Bacillus thuringiensis: legado para el siglo XXI Bacillus thuringiensis: the legacy to the XXI century

    Directory of Open Access Journals (Sweden)

    Orduz S.

    1998-06-01

    Full Text Available

    Los insecticidas basados en la bacteria Bacillus thuringiensis son el principal renglón productivo del mercado mundial de biopesticidas. La investigación dedicada a esta área, promovida por la urgente necesidad de resolver problemas agrícolas y de salud pública, ha dado lugar a un conocimiento exhaustivo de su biología. La diversidad de cepas diferentes de B. thuringiensis ha permitido desarrollar productos principalmente, pero no exclusivamente, para el control de insectos. Con los nuevos desarrollos de la biología molecular, se ha logrado comprender su mecanismo de acción a nivel molecular y también se ha logrado extender sus capacidades entomopatógenas. Como producto de su amplio uso en muchos países, se han presentado casos de resistencia en poblaciones de insectos susceptibles. Con esta revisión se pretende elaborar un contexto teórico del estado actual de la investigación sobre B. thuringiensis, describiendo brevemente el conocimiento sobre esta bacteria, haciendo hincapié en los fenómenos biológicos que subyacen su actividad tóxica y la problemática que se avecina en el próximo siglo con los fenómenos de resistencia cada vez más comunes, todo esto analizado desde una perspectiva biotecnológica.

    Bacillus thuringiensis-based insecticides are the main production line of the biopesticides world market. The research devoted to this area, promoted by the necessity to solve problems in agriculture and public health has resulted in an exhaustive knowledge of its biology. The diversity of the B. thuringiensis strains has permitted to develop several products mainly, but not exclusively, for insect control. With the new developments in the field of molecular biology, it has been possible to understand the molecular basis of the mode of action and to increase the range of activity as well. As a result

  16. Genome Sequencing of Bacillus subtilis SC-8, Antagonistic to the Bacillus cereus Group, Isolated from Traditional Korean Fermented-Soybean Food

    Science.gov (United States)

    Yeo, In-Cheol; Lee, Nam Keun

    2012-01-01

    Bacillus subtilis SC-8 is a Gram-positive bacterium displaying narrow antagonistic activity for the Bacillus cereus group. B. subtilis SC-8 was isolated from Korean traditional fermented-soybean food. Here we report the draft genome sequence of B. subtilis SC-8, including biosynthetic genes for antibiotics that may have beneficial effects for control of food-borne pathogens. PMID:22207744

  17. Expression of the neutral protease gene from a thermophilic Bacillus sp BT1 strain in Bacillus subtilis and its natural host : Identification of a functional promoter

    NARCIS (Netherlands)

    Vecerek, B; Venema, G

    2000-01-01

    The expression of the neutral protease gene (npr) from the thermophilic Bacillus sp. BT1 strain was studied in its natural host and in mesophilic Bacillus subtilis. In the thermophilic BT1 strain, the transcription of the protease gene is initiated from its own promoter, just 5' to the gene. In cont

  18. Sporulation of Bacillus spp. within biofilms: a potential source of contamination in food processing environments.

    Science.gov (United States)

    Faille, C; Bénézech, T; Midelet-Bourdin, G; Lequette, Y; Clarisse, M; Ronse, G; Ronse, A; Slomianny, C

    2014-06-01

    Bacillus strains are often isolated from biofilms in the food industries. Previous works have demonstrated that sporulation could occur in biofilms, suggesting that biofilms would be a significant source of food contamination with spores. In this study, we investigated the properties of mono-species and mixed Bacillus biofilms and the ability of Bacillus strains to sporulate inside biofilms. Bacillus strains were able to form mono-species biofilms on stainless steel coupons, with up to 90% spores after a 48 h-incubation. These spores were highly resistant to cleaning but were easily transferred to agar, mimicking the cross-contamination of food, thereby suggesting that biofilms would be of particular concern due to a potential for Bacillus spore food contamination. This hypothesis was strengthened by the fact that Bacillus strains were able to form mixed biofilms with resident strains and that sporulation still occurred easily in these complex structures.

  19. Bacillus composti sp. nov. and Bacillus thermophilus sp. nov., two thermophilic, Fe(III)-reducing bacteria isolated from compost.

    Science.gov (United States)

    Yang, Guiqin; Chen, Ming; Yu, Zhen; Lu, Qin; Zhou, Shungui

    2013-08-01

    Two novel thermophilic bacteria, designated SgZ-9(T) and SgZ-10(T), were isolated from compost. Cells of the two strains were catalase-positive, endospore-forming and Gram-staining-positive rods. Strain SgZ-9(T) was oxidase-positive and non-motile, and strain SgZ-10(T) was oxidase-negative and motile. The highest 16S rRNA gene sequence similarity for both strains SgZ-9(T) and SgZ-10(T) was observed with Bacillus fortis (97.5 % and 96.9 %, respectively). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SgZ-9(T) formed a cluster with B. fortis R-6514(T) and Bacillus fordii R-7190(T), and SgZ-10(T) formed a cluster with Bacillus farraginis R-6540(T). The DNA-DNA pairing studies showed that SgZ-9(T) displayed 41.6 % and 30.7 % relatedness to the type strains of B. fortis and B. fordii, respectively. The 16S rRNA gene sequence similarity between strains SgZ-9(T) and SgZ-10(T) was 97.2 %, and the level of DNA-DNA relatedness between them was 39.2 %. The DNA G+C content of SgZ-9(T) and SgZ-10(T) was 45.3 and 47.9 mol%, respectively. Chemotaxonomic analysis revealed that both strains contained the menaquinone 7 (MK-7) as the predominant respiratory quinone. The major cellular fatty acids (>5 %) were iso-C15 : 0, anteiso-C15 : 0, anteiso-C17 : 0, iso-C16 : 0 and iso-C17 : 0 in SgZ-9(T) and iso-C15 : 0, anteiso-C15 : 0, iso-C17 : 0, anteiso-C17 : 0 and iso-C16 : 0 in SgZ-10(T). Based on the phenotypic characteristics, chemotaxonomic features, DNA-DNA hybridization with the nearest phylogenetic neighbours and phylogenetic analysis based on the 16S rRNA gene sequences, the two strains were determined to be two distinct novel species in the genus Bacillus, and the names proposed are Bacillus composti sp. nov. SgZ-9(T) ( = CCTCC AB2012109(T) = KACC 16872(T)) and Bacillus thermophilus sp. nov. SgZ-10(T) (CCTCC AB2012110(T) = KACC 16873(T)).

  20. Impedance Measurements Could Accelerate Phage-Based Identification of Bacillus anthracis and Other Bacteria

    Science.gov (United States)

    2016-09-01

    Impedance Measurements Could Accelerate Phage-Based Identification of Bacillus anthracis And Other Bacteria Thomas Brown, Salwa Shan, Teresa...This is particularly true in the field of biodefense where phage have a long history of being used to identify Bacillus anthracis and Yersinia pestis...based diagnostic assays for this pathogen. After exposing small quantities of Bacillus cultures to ɣ phage, we tracked the cultures for up to 90