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Sample records for bacillus subtilis spore

  1. Structural Analysis of Bacillus subtilis Spore Peptidoglycan During Sporulation

    OpenAIRE

    2000-01-01

    Structural analysis of Bacillus subtilis spore peptidoglycan during sporulation:Jennifer L. Meador-Parton:David L. Popham, Chairman:Department of Biology:(ABSTRACT):Bacterial spore peptidoglycan (PG) is very loosely cross-linked relative to vegetative PG. Theories suggest that loosely cross-linked spore PG may have a flexibility which contributes to the attainment of spore core dehydration. The structure of the PG found in fully dormant spores has previously been examined in wild type and m...

  2. 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...

  3. INCORPORATION OF BACTERIOPHAGE GENOME BY SPORES OF BACILLUS SUBTILIS.

    Science.gov (United States)

    TAKAHASHI, I

    1964-06-01

    Takahashi, I. (Microbiology Research Institute, Ottawa, Ontario, Canada). Incorporation of bacteriophage genome by spores of Bacillus subtilis. J. Bacteriol. 87:1499-1502. 1964-The buoyant density in a CsCl gradient of deoxyribonucleic acid (DNA) extracted from spores of Bacillus subtilis was found to be identical to that of DNA from vegetative cells. Density-gradient centrifugation of DNA of spores derived from cultures infected with phage PBS 1 revealed the presence of a minor band whose density corresponded to that of the phage DNA in addition to the spore DNA. No intermediate bands were present. The relative amount of the phage DNA present in the spores was estimated to be 11%, suggesting that spores of this organism may incorporate several copies of the phage genome. Although the possibility that true lysogeny may occur cannot be entirely eliminated, the results seem to indicate that the phage genomes incorporated into spores are not attached to the host chromosome in this system.

  4. 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...

  5. Sporicidal characteristics of heated dolomite powder against Bacillus subtilis spores.

    Science.gov (United States)

    Yasue, Syogo; Sawai, Jun; Kikuchi, Mikio; Nakakuki, Takahito; Sano, Kazuo; Kikuchi, Takahide

    2014-01-01

    Dolomite is a double salt composed of calcium carbonate (CaCO3) and magnesium carbonate (MgCO3). The heat treatment of CaCO3 and MgCO3 respectively generates calcium oxide (CaO) and magnesium oxide (MgO), which have antimicrobial activity. In this study, heated dolomite powder (HDP) slurry was investigated for its sporicidal activity against Bacillus subtilis ATCC 6633 spores. The B. subtilis spores used in this study were not affected by acidic (pH 1) or alkaline (pH 13) conditions, indicating that they were highly resistant. However, dolomite powder heated to 1000℃ for 1 h could kill B. subtilis spores, even at pH 12.7. Sporicidal activity was only apparent when the dolomite powder was heated to 800℃ or higher, and sporicidal activity increased with increases in the heating temperature. This temperature corresponded to that of the generation of CaO. We determined that MgO did not contribute to the sporicidal activity of HDP. To elucidate the sporicidal mechanism of the HDP against B. subtilis spores, the generation of active oxygen from HDP slurry was examined by chemiluminescence analysis. The generation of active oxygen increased when the HDP slurry concentration rose. The results suggested that, in addition to its alkalinity, the active oxygen species generated from HDP were associated with sporicidal activity.

  6. Vacuum-induced Mutations In Bacillus Subtilis Spores

    Science.gov (United States)

    Munakata, N.; Maeda, M.; Hieda, K.

    During irradiation experiments with vacuum-UV radiation using synchrotron sources, we made unexpected observation that Bacillus subtilis spores of several recombination-deficient strains lost colony-forming ability by the exposure to high vacuum alone. Since this suggested the possible injury in spore DNA, we looked for mutation induction using the spores of strains HA101 (wild-type repair capability) and TKJ6312 (excision and spore repair deficient) that did not lose survivability. It was found that the frequency of nalidixic-acid resistant mutation increased several times in both of these strains by the exposure to high vacuum (10e-4 Pa after 24 hours). The analysis of sequence changes in gyrA gene showed that the majority of mutations carried a unique allele (gyrA12) of tandem double-base substitutions from CA to TT. The observation has been extended to rifampicin resistant mutations, the majority of that carried substitutions from CA to TT or AT in rpoB gene. On the other hand, when the spores of strains PS578 and PS2319 (obtained from P. Setlow) that are defective in a group of small acidic proteins (alpha/beta-type SASP) were similarly treated, none of the mutants analyzed carried such changes. This suggests that the unique mutations might be induced by the interaction of small acidic proteins with spore DNA under forced dehydration. The results indicate that extreme vacuum causes severe damage in spore DNA, and provide additional constraint to the long-term survival of bacterial spores in the space environment.

  7. Architecture and Assembly of the Bacillus subtilis Spore Coat

    Science.gov (United States)

    2014-09-26

    icandy contaminated with germinated spores and these germinat ed spores were removed by centrifugation in a one step HistodenzTM (Sigma, St. Louis...spore resistance but also because some coat proteins play significant roles in spore germination . However, much recent work on the spore coat has... germinating spores of various Bacillus [14,21 30] and Clostridium [3 1] species. H owever, this analysis has generally been conducted on wild type

  8. Mutagenesis of Bacillus subtilis spores exposed to simulated space environment

    Science.gov (United States)

    Munakata, N.; Natsume, T.; Takahashi, K.; Hieda, K.; Panitz, C.; Horneck, G.

    Bacterial spores can endure in a variety of extreme earthly environments. However, some conditions encountered during the space flight could be detrimental to DNA in the spore, delimiting the possibility of transpermia. We investigate the genetic consequences of the exposure to space environments in a series of preflight simulation project of EXPOSE. Using Bacillus subtilis spores of repair-proficient HA101 and repair-deficient TKJ6312 strains, the mutations conferring resistance to rifampicin were detected, isolated and sequenced. Most of the mutations were located in a N-terminal region of the rpoB gene encoding RNA polymerase beta-subunit. Among several potentially mutagenic factors, high vacuum, UV radiation, heat, and accelerated heavy ions induced mutations with varying efficiencies. A majority of mutations induced by vacuum exposure carried a tandem double-base change (CA to TT) at a unique sequence context of TCAGC. Results indicate that the vacuum and high temperature may act synergistically for the induction of mutations.

  9. High Pressure Germination of Bacillus subtilis Spores with Alterations in Levels and Types of Germination Proteins

    Science.gov (United States)

    2014-01-01

    1ITLE AND SUBTITLE 5a CONTRACTNUMBER High pressure germination of Bacillus subtilis spores with W911NF-09-l-0286 alterations in levels and types of...A moderate high pressure (mHP) of 150 megaPascals (MPa) triggers germination of Bacillus subtilis spores via germinant receptors (GRs), while...germination by a very high pressure (vHP) of550 MPa is GR-independent. The mHP and vHP germination of Bacillus subtilis spores with different levels ofGRs

  10. Mechanisms of Induction of Germination of Bacillus subtilis Spores by High Pressure

    OpenAIRE

    Paidhungat, Madan; Setlow, Barbara; Daniels, William B.; Hoover, Dallas; Papafragkou, Efstathia; Setlow, Peter

    2002-01-01

    Spores of Bacillus subtilis lacking all germinant receptors germinate >500-fold slower than wild-type spores in nutrients and were not induced to germinate by a pressure of 100 MPa. However, a pressure of 550 MPa induced germination of spores lacking all germinant receptors as well as of receptorless spores lacking either of the two lytic enzymes essential for cortex hydrolysis during germination. Complete germination of spores either lacking both cortex-lytic enzymes or with a cortex not att...

  11. UV resistance of Bacillus anthracis spores revisited: validation of Bacillus subtilis spores as UV surrogates for spores of B. anthracis Sterne.

    Science.gov (United States)

    Nicholson, Wayne L; Galeano, Belinda

    2003-02-01

    Recent bioterrorism concerns have prompted renewed efforts towards understanding the biology of bacterial spore resistance to radiation with a special emphasis on the spores of Bacillus anthracis. A review of the literature revealed that B. anthracis Sterne spores may be three to four times more resistant to 254-nm-wavelength UV than are spores of commonly used indicator strains of Bacillus subtilis. To test this notion, B. anthracis Sterne spores were purified and their UV inactivation kinetics were determined in parallel with those of the spores of two indicator strains of B. subtilis, strains WN624 and ATCC 6633. When prepared and assayed under identical conditions, the spores of all three strains exhibited essentially identical UV inactivation kinetics. The data indicate that standard UV treatments that are effective against B. subtilis spores are likely also sufficient to inactivate B. anthracis spores and that the spores of standard B. subtilis strains could reliably be used as a biodosimetry model for the UV inactivation of B. anthracis spores.

  12. 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

  13. Dynamic localization of penicillin-binding proteins during spore development in Bacillus subtilis

    NARCIS (Netherlands)

    Scheffers, Dirk-Jan

    2005-01-01

    During Bacillus subtilis spore formation, many membrane proteins that function in spore development localize to the prespore septum and, subsequently, to the outer prespore membrane. Recently, it was shown that the cell-division-specific penicillin-binding proteins (PBPs) 1 and 2b localize to the as

  14. Bacillus subtilis spore protein SpoVAC functions as a mechanosensitive channel

    NARCIS (Netherlands)

    Velasquez Guzman, Jeanette; Schuurman-Wolters, Geesina; Birkner, Jan Peter; Abee, Tjakko; Poolman, Bert

    2014-01-01

    A critical event during spore germination is the release of Ca-DPA (calcium in complex with dipicolinic acid). The mechanism of release of Ca-DPA through the inner membrane of the spore is not clear, but proteins encoded by the Bacillus subtilis spoVA operon are involved in the process. We cloned an

  15. Effects of Electrolyzed Oxidizing Water on Inactivation of Bacillus subtilis and Bacillus cereus Spores in Suspension and on Carriers.

    Science.gov (United States)

    Zhang, Chunling; Li, Baoming; Jadeja, Ravirajsinh; Hung, Yen-Con

    2016-01-01

    Spores of some Bacillus species are responsible for food spoilage and foodborne disease. These spores are highly resistant to various interventions and cooking processes. In this study, the sporicidal efficacy of acidic electrolyzed oxidizing (EO) water (AEW) and slightly acidic EO water (SAEW) with available chlorine concentration (ACC) of 40, 60, 80, 100, and 120 mg/L and treatment time for 1, 2, 3, 4, 5, and 6 min were tested on Bacillus subtilis and Bacillus cereus spores in suspension and on carrier with or without organics. The reduction of spore significantly increased with increasing ACC and treatment time (P waters containing 120 mg/L ACC, while only SAEW at 120 mg/L and 2 min treatment achieved >6 log reductions of B. subtilis spore. Both types of EO water with ACC of 60 mg/L and 6 min treatment achieved a reduction of B. subtilis and B. cereus spores to nondetectable level. EO water with ACC of 80 mg/L and treatment time of 3 min on carrier test without organics addition resulted in reductions of B. subtilis spore to nondetectable level. But, addition of 0.3% organics on carrier decreased the inactivation effect of EO water. This study indicated that EO water was highly effective in inactivation of B. subtilis and B. cereus spores in suspension or on carrier, and therefore, rendered it as a promising disinfectant to be applied in food industry.

  16. Function of the SpoVAEa and SpoVAF Proteins of Bacillus subtilis Spores

    Science.gov (United States)

    2014-06-01

    1ITLE AND SUBTITLE 5a CONTRACTNUMBER Function of the SpoVAEa and SpoVAF proteins of Bacillus W911NF-09-1-0286 subtilis spores 5b. GRANT NUMBER 5c...ABSTRACT The Bacillus subtilis spoVAEa and spoVAF genes are expressed in developng spores as members of the spoVA operon that encodes proteins essential...8217\\ ;~ 1~~~4-~,.1. A\\ C’~~1T 1\\ D~ ~~,.1 C’~~1T 1\\ T’\\ ~-~ ,.1;~~1. •• 4-~,.1 ~:-:1~-1 •• ;~ ~~~~~~~ ~f:’ 15. SUBJECT TERMS Bacillus , spores SpoVA

  17. Evaluation of germination, distribution, and persistence of Bacillus subtilis spores through the gastrointestinal tract of chickens.

    Science.gov (United States)

    Latorre, J D; Hernandez-Velasco, X; Kallapura, G; Menconi, A; Pumford, N R; Morgan, M J; Layton, S L; Bielke, L R; Hargis, B M; Téllez, G

    2014-07-01

    Spores are popular as direct-fed microbials, though little is known about their mode of action. Hence, the first objective of the present study was to evaluate the in vitro germination and growth rate of Bacillus subtilis spores. Approximately 90% of B. subtilis spores germinate within 60 min in the presence of feed in vitro. The second objective was to determine the distribution of these spores throughout different anatomical segments of the gastrointestinal tract (GIT) in a chicken model. For in vivo evaluation of persistence and dissemination, spores were administered to day-of-hatch broiler chicks either as a single gavage dose or constantly in the feed. During 2 independent experiments, chicks were housed in isolation chambers and fed sterile corn-soy-based diets. In these experiments one group of chickens was supplemented with 10(6) spores/g of feed, whereas a second group was gavaged with a single dose of 10(6) spores per chick on day of hatch. In both experiments, crop, ileum, and cecae were sampled from 5 chicks at 24, 48, 72, 96, and 120 h. Viable B. subtilis spores were determined by plate count method after heat treatment (75°C for 10 min). The number of recovered spores was constant through 120 h in each of the enteric regions from chickens receiving spores supplemented in the feed. However, the number of recovered B. subtilis spores was consistently about 10(5) spores per gram of digesta, which is about a 1-log10 reduction of the feed inclusion rate, suggesting approximately a 90% germination rate in the GIT when fed. On the other hand, recovered B. subtilis spores from chicks that received a single gavage dose decreased with time, with only approximately 10(2) spores per gram of sample by 120 h. This confirms that B. subtilis spores are transiently present in the GIT of chickens, but the persistence of vegetative cells is presently unknown. For persistent benefit, continuous administration of effective B. subtilis direct-fed microbials as vegetative

  18. Responses of Bacillus subtilis spores to space environment: results from experiments in space.

    Science.gov (United States)

    Horneck, G

    1993-02-01

    Onboard of several spacecrafts (Apollo 16, Spacelab 1, LDEF), spores of Bacillus subtilis were exposed to selected parameters of space, such as space vacuum, different spectral ranges of solar UV-radiation and cosmic rays, applied separately or in combination, and we have studied their survival and genetic changes after retrieval. The spores survive extended periods of time in space--up to several years--, if protected against the high influx of solar UV-radiation. Water desorption caused by the space vacuum leads to structural changes of the DNA; the consequences are an increased mutation frequency and altered photobiological properties of the spores. UV-effects, such as killing and mutagenesis, are augmented, if the spores are in space vacuum during irradiation. Vacuum-specific photoproducts which are different from the 'spore photoproduct' may cause the synergistic response of spores to the simultaneous action of UV and vacuum. The experiments provide an experimental test of certain steps of the panspermia hypothesis.

  19. Role of dipicolinic acid in the germination, stability, and viability of spores of Bacillus subtilis.

    Science.gov (United States)

    Magge, Anil; Granger, Amanda C; Wahome, Paul G; Setlow, Barbara; Vepachedu, Venkata R; Loshon, Charles A; Peng, Lixin; Chen, De; Li, Yong-Qing; Setlow, Peter

    2008-07-01

    Spores of Bacillus subtilis spoVF strains that cannot synthesize dipicolinic acid (DPA) but take it up during sporulation were prepared in medium with various DPA concentrations, and the germination and viability of these spores as well as the DPA content in individual spores were measured. Levels of some other small molecules in DPA-less spores were also measured. These studies have allowed the following conclusions. (i) Spores with no DPA or low DPA levels that lack either the cortex-lytic enzyme (CLE) SleB or the receptors that respond to nutrient germinants could be isolated but were unstable and spontaneously initiated early steps in spore germination. (ii) Spores that lacked SleB and nutrient germinant receptors and also had low DPA levels were more stable. (iii) Spontaneous germination of spores with no DPA or low DPA levels was at least in part via activation of SleB. (iv) The other redundant CLE, CwlJ, was activated only by the release of high levels of DPA from spores. (v) Low levels of DPA were sufficient for the viability of spores that lacked most alpha/beta-type small, acid-soluble spore proteins. (vi) DPA levels accumulated in spores prepared in low-DPA-containing media varied greatly between individual spores, in contrast to the presence of more homogeneous DPA levels in individual spores made in media with high DPA concentrations. (vii) At least the great majority of spores of several spoVF strains that contained no DPA also lacked other major spore small molecules and had gone through some of the early reactions in spore germination.

  20. Structural Analysis of Bacillus subtilis Spore Peptidoglycan during Sporulation

    OpenAIRE

    2000-01-01

    A major structural element of bacterial endospores is a peptidoglycan (PG) wall. This wall is produced between the two opposed membranes surrounding the developing forespore and is composed of two layers. The inner layer is the germ cell wall, which appears to have a structure similar to that of the vegetative cell wall and which serves as the initial cell wall following spore germination. The outer layer, the cortex, has a modified structure, is required for maintenance of spore dehydration,...

  1. Decrease in spermidine content during logarithmic phase of cell growth delays spore formation of Bacillus subtilis.

    Science.gov (United States)

    Ishii, I; Takada, H; Terao, K; Kakegawa, T; Igarashi, K; Hirose, S

    1994-11-01

    Bacillus subtilis 168M contained a large amount of spermidine during the logarithmic phase of growth, but the amount decreased drastically during the stationary phase. The extracts, prepared from B. subtilis cells harvested in the logarithmic phase, contained activity of arginine decarboxylase (ADC) rather than the activity of ornithine decarboxylase. In the presence of alpha-difluoromethylarginine (DFMA), a specific and irreversible inhibitor of ADC, the amount of spermidine in B. subtilis during the logarithmic phase decreased to about 25% of the control cells. Under these conditions, spore formation of B. subtilis 168M delayed greatly without significant inhibition of cell growth. The decrease in spermidine content in the logarithmic phase rather than in the stationary phase was involved in the delay of sporulation. Electron microscopy of cells at 24 hrs. of culture confirmed the delay of spore formation by the decrease of spermidine content. Furthermore, the delay of sporulation was negated by the addition of spermidine. These data suggest that a large amount of spermidine existing during the logarithmic phase plays an important role in the sporulation of B. subtilis.

  2. Investigating the thermodynamic stability of Bacillus subtilis spore-uranium(VI) adsorption though surface complexation modeling

    Science.gov (United States)

    Harrold, Z.; Hertel, M.; Gorman-Lewis, D.

    2012-12-01

    Dissolved uranium speciation, mobility, and remediation are increasingly important topics given continued and potential uranium (U) release from mining operations and nuclear waste. Vegetative bacterial cell surfaces are known to adsorb uranium and may influence uranium speciation in the environment. Previous investigations regarding U(VI) adsorption to bacterial spores, a differentiated and dormant cell type with a tough proteinaceous coat, include U adsorption affinity and XAFS data. We investigated the thermodynamic stability of aerobic, pH dependent uranium adsorption to bacterial spore surfaces using purified Bacillus subtilis spores in solution with 5ppm uranium. Adsorption reversibility and kinetic experiments indicate that uranium does not precipitate over the duration of the experiments and equilibrium is reached within 20 minutes. Uranium-spore adsorption edges exhibited adsorption at all pH measured between 2 and 10. Maximum adsorption was achieved around pH 7 and decreased as pH increased above 7. We used surface complexation modeling (SCM) to quantify uranium adsorption based on balanced chemical equations and derive thermodynamic stability constants for discrete uranium-spore adsorption reactions. Site specific thermodynamic stability constants provide insight on interactions occurring between aqueous uranium species and spore surface ligands. The uranium adsorption data and SCM parameters described herein, also provide a basis for predicting the influence of bacterial spores on uranium speciation in natural systems and investigating their potential as biosorption agents in engineered systems.

  3. Disinfection and regrowth potential of bacillus subtilis spores by ozone, ultraviolet rays and gamma irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hae Yeon; Lee, O Mi; Kim, Tae Hun; Lee, Myun Joo; Yu, Seung Ho [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of)

    2009-06-15

    Chlorination has been the most commonly adopted disinfection process for the treatment of drinking water. However, Cryptosporidium parvum oocysts and Giardia lamblia cysts were not treated effectively by the common chlorine-based disinfectants. Additionally the regrowth of pathogenic microorganisms is associated with hygienic and aesthetic problems for the consumers of drinking water. Study on alternative disinfection processes such as ozone, UV-C, VUV and gamma irradiation were conducted. Bacillus subtilis spores have been used as a surrogate microorganism for Cryptosporidium parvum oocysts and Giardia lamblia cyst. Inactivation efficiency by ozone was from 30% to 96% within the range of 5 min to 120 min exposures. Inactivation efficiencies by UV-C and VUV were 95.18%, 95.07% at 30 sec, respectively. Inactivation efficiency at gamma irradiation dose of 2 kGy was 99.4%. Microbial regrowths after ozone, UV-C, VUV and gamma irradiation disinfections were also evaluated for 4 days. Bacillus subtilis spores after ozone treatment for 120 min exposure at the rate of 1.68 mg {center_dot} min{sup -1} showed 96.02% disinfection efficiency and significant microbial regrowth. Bacillus subtilis spores after UV-C (99.25% disinfection efficiency) and VUV (99.67% disinfection efficiency) treatments for 5 min showed gradual regrowth. However, inactivation efficiency of gamma irradiation at dose of 1 kGy was 98.8% and the disinfected sample showed no microbial regrowth for 4 days. Therefore, gamma irradiation is the most effective process for the disinfection of pathogenic microorganisms such as oocysts of protozoan parasites among four disinfection process.

  4. Involvement of Coat Proteins in Bacillus subtilis Spore Germination in High-Salinity Environments.

    Science.gov (United States)

    Nagler, Katja; Setlow, Peter; Reineke, Kai; Driks, Adam; Moeller, Ralf

    2015-10-01

    The germination of spore-forming bacteria in high-salinity environments is of applied interest for food microbiology and soil ecology. It has previously been shown that high salt concentrations detrimentally affect Bacillus subtilis spore germination, rendering this process slower and less efficient. The mechanistic details of these salt effects, however, remained obscure. Since initiation of nutrient germination first requires germinant passage through the spores' protective integuments, the aim of this study was to elucidate the role of the proteinaceous spore coat in germination in high-salinity environments. Spores lacking major layers of the coat due to chemical decoating or mutation germinated much worse in the presence of NaCl than untreated wild-type spores at comparable salinities. However, the absence of the crust, the absence of some individual nonmorphogenetic proteins, and the absence of either CwlJ or SleB had no or little effect on germination in high-salinity environments. Although the germination of spores lacking GerP (which is assumed to facilitate germinant flow through the coat) was generally less efficient than the germination of wild-type spores, the presence of up to 2.4 M NaCl enhanced the germination of these mutant spores. Interestingly, nutrient-independent germination by high pressure was also inhibited by NaCl. Taken together, these results suggest that (i) the coat has a protective function during germination in high-salinity environments; (ii) germination inhibition by NaCl is probably not exerted at the level of cortex hydrolysis, germinant accessibility, or germinant-receptor binding; and (iii) the most likely germination processes to be inhibited by NaCl are ion, Ca(2+)-dipicolinic acid, and water fluxes.

  5. Bacillus subtilis spores on artificial meteorites survive hypervelocity atmospheric entry: implications for Lithopanspermia.

    Science.gov (United States)

    Fajardo-Cavazos, Patricia; Link, Lindsey; Melosh, H Jay; Nicholson, Wayne L

    2005-12-01

    An important but untested aspect of the lithopanspermia hypothesis is that microbes situated on or within meteorites could survive hypervelocity entry from space through Earth's atmosphere. The use of high-altitude sounding rockets to test this notion was explored. Granite samples permeated with spores of Bacillus subtilis strain WN511 were attached to the exterior telemetry module of a sounding rocket and launched from White Sands Missile Range, New Mexico into space, reaching maximum atmospheric entry velocity of 1.2 km/s. Maximum recorded temperature during the flight was measured at 145 degrees C. The surfaces of the post-flight granite samples were swabbed and tested for recovery and survival of WN511 spores, using genetic markers and the unique DNA fingerprint of WN511 as recovery criteria. Spore survivors were isolated at high frequency, ranging from 1.2% to 4.4% compared with ground controls, from all surfaces except the forward-facing surface. Sporulation-defective mutants were noted among the spaceflight survivors at high frequency (4%). These experiments constitute the first report of spore survival to hypervelocity atmospheric transit, and indicate that sounding rocket flights can be used to model the high-speed atmospheric entry of bacteria-laden artificial meteorites.

  6. Investigating synergism during sequential inactivation of Bacillus subtilis spores with several disinfectants.

    Science.gov (United States)

    Cho, Min; Kim, Jae-Hong; Yoon, Jeyong

    2006-08-01

    The sequential application of ozone, chlorine dioxide, or UV followed by free chlorine was performed to investigate the synergistic inactivation of Bacillus subtilis spores. The greatest synergism was observed when chlorine dioxide was used as a primary disinfectant followed by secondary disinfection with free chlorine. A lesser synergistic effect was observed when ozone was used as the primary disinfectant, but no synergism was observed when UV was used as the primary disinfectant. When free chlorine was used as the primary disinfectant (i.e., sequential application in the reverse order), the synergistic effect was shown only when chlorine dioxide was applied as the secondary disinfectant. The synergistic effect observed could be related to damage to the spore coat during primary disinfection, suggested by the loss of proteins from spores during disinfectant treatment. The greatest synergism observed by the chlorine dioxide/free chlorine pair suggested that common reaction sites might exist for these disinfectants. The concept of percent synergistic effect was introduced to quantitatively compare the extent of synergistic effects in the sequential disinfection processes.

  7. Role of Spore Coat Proteins in the Resistance of Bacillus subtilis Spores to Caenorhabditis elegans Predation▿

    OpenAIRE

    2008-01-01

    Bacterial spores are resistant to a wide range of chemical and physical insults that are normally lethal for the vegetative form of the bacterium. While the integrity of the protein coat of the spore is crucial for spore survival in vitro, far less is known about how the coat provides protection in vivo against predation by ecologically relevant hosts. In particular, assays had characterized the in vitro resistance of spores to peptidoglycan-hydrolyzing enzymes like lysozyme that are also imp...

  8. HtrC is involved in proteolysis of YpeB during germination of Bacillus anthracis and Bacillus subtilis spores.

    Science.gov (United States)

    Bernhards, Casey B; Chen, Yan; Toutkoushian, Hannah; Popham, David L

    2015-01-01

    Bacterial endospores can remain dormant for decades yet can respond to nutrients, germinate, and resume growth within minutes. An essential step in the germination process is degradation of the spore cortex peptidoglycan wall, and the SleB protein in Bacillus species plays a key role in this process. Stable incorporation of SleB into the spore requires the YpeB protein, and some evidence suggests that the two proteins interact within the dormant spore. Early during germination, YpeB is proteolytically processed to a stable fragment. In this work, the primary sites of YpeB cleavage were identified in Bacillus anthracis, and it was shown that the stable products are comprised of the C-terminal domain of YpeB. Modification of the predominant YpeB cleavage sites reduced proteolysis, but cleavage at other sites still resulted in loss of full-length YpeB. A B. anthracis strain lacking the HtrC protease did not generate the same stable YpeB products. In B. anthracis and Bacillus subtilis htrC mutants, YpeB was partially stabilized during germination but was still degraded at a reduced rate by other, unidentified proteases. Purified HtrC cleaved YpeB to a fragment similar to that observed in vivo, and this cleavage was stimulated by Mn(2+) or Ca(2+) ions. A lack of HtrC did not stabilize YpeB or SleB during spore formation in the absence of the partner protein, indicating other proteases are involved in their degradation during sporulation.

  9. Decontamination of Bacillus subtilis var. niger spores on selected surfaces by chlorine dioxide gas*

    Science.gov (United States)

    Li, Yan-ju; Zhu, Neng; Jia, Hai-quan; Wu, Jin-hui; Yi, Ying; Qi, Jian-cheng

    2012-01-01

    Objective: Chlorine dioxide (CD) gas has been used as a fumigant in the disinfection of biosafety laboratories. In this study, some experiments were conducted to assess the inactivation of spores inoculated on six materials [stainless steel (SS), painted steel (PS), polyvinyl chlorid (PVC), polyurethane (PU), glass (GS), and cotton cloth (CC)] by CD gas. The main aims of the study were to determine the sporicidal efficacy of CD gas and the effect of prehumidification before decontamination on sporicidal efficacy. Methods: Material coupons (1.2 cm diameter of SS, PS, and PU; 1.0 cm×1.0 cm for PVC, GS, and CC) were contaminated with 10 μl of Bacillus subtilis var. niger (ATCC 9372) spore suspension in mixed organic burden and then dried in a biosafety cabinet for 12 h. The spores were recovered by soaking the coupons in 5 ml of extraction liquid for 1 h and then vortexing the liquid for 1 min. Results: The log reductions in spore numbers on inoculated test materials exposed to CD gas [0.080% (volume ratio, v/v) for 3 h] were in the range of from 1.80 to 6.64. Statistically significant differences were found in decontamination efficacies on test material coupons of SS, PS, PU, and CC between with and without a 1-h prehumidification treatment. With the extraction method, there were no statistically significant differences in the recovery ratios between the porous and non-porous materials. Conclusions: The results reported from this study could provide information for developing decontamination technology based on CD gas for targeting surface microbial contamination. PMID:22467366

  10. Decontamination of Bacillus subtilis var.niger spores on selected surfaces by chlorine dioxide gas

    Institute of Scientific and Technical Information of China (English)

    Yan-ju LI; Neng ZHU; Hai-quan JIA; Jin-hui WU; Ying YI; Jian-cheng QI

    2012-01-01

    Objective:Chlorine dioxide (CD) gas has been used as a fumigant in the disinfection of biosafety laboratories.In this study,some experiments were conducted to assess the inactivation of spores inoculated on six materials [stainless steel (SS),painted steel (PS),polyvinyl chlorid (PVC),polyurethane (PU),glass (GS),and cotton cloth (CC)] by CD gas.The main aims of the study were to determine the sporicidal efficacy of CD gas and the effect of prehumidification before decontamination on sporicidal efficacy.Methods:Material coupons (1.2 cm diameter of SS,PS,and PU; 1.0 cm×1.0 cm for PVC,GS,and CC) were contaminated with 10 μl of Bacillus subtilis var.niger(ATCC 9372) spore suspension in mixed organic burden and then dried in a biosafety cabinet for 12 h.The spores were recovered by soaking the coupons in 5 ml of extraction liquid for 1 h and then vortexing the liquid for 1 min.Results:The log reductions in spore numbers on inoculated test materials exposed to CD gas [0.080% (volume ratio,v/v) for 3 h]were in the range of from 1.80 to 6.64.Statistically significant differences were found in decontamination efficacies on test material coupons of SS,PS,PU,and CC between with and without a 1-h prehumidification treatment.With the extraction method,there were no statistically significant differences in the recovery ratios between the porous and non-porous materials.Conclusions:The results reported from this study could provide information for developing decontamination technology based on CD gas for targeting surface microbial contamination.

  11. Adsorption of β-galactosidase of Alicyclobacillus acidocaldarius on wild type and mutants spores of Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Sirec Teja

    2012-08-01

    Full Text Available Abstract Background The Bacillus subtilis spore has long been used as a surface display system with potential applications in a variety of fields ranging from mucosal vaccine delivery, bioremediation and biocatalyst development. More recently, a non-recombinant approach of spore display has been proposed and heterologous proteins adsorbed on the spore surface. We used the well-characterized β-galactosidase from the thermoacidophilic bacterium Alicyclobacillus acidocaldarius as a model to study enzyme adsorption, to analyze whether and how spore-adsorption affects the properties of the enzyme and to improve the efficiency of the process. Results We report that purified β-galactosidase molecules were adsorbed to purified spores of a wild type strain of B. subtilis retaining ca. 50% of their enzymatic activity. Optimal pH and temperature of the enzyme were not altered by the presence of the spore, that protected the adsorbed β-galactosidase from exposure to acidic pH conditions. A collection of mutant strains of B. subtilis lacking a single or several spore coat proteins was compared to the isogenic parental strain for the adsorption efficiency. Mutants with an altered outermost spore layer (crust were able to adsorb 60-80% of the enzyme, while mutants with a severely altered or totally lacking outer coat adsorbed 100% of the β-galactosidase molecules present in the adsorption reaction. Conclusion Our results indicate that the spore surface structures, the crust and the outer coat layer, have an negative effect on the adhesion of the β-galactosidase. Electrostatic forces, previously suggested as main determinants of spore adsorption, do not seem to play an essential role in the spore-β-galactosidase interaction. The analysis of mutants with altered spore surface has shown that the process of spore adsorption can be improved and has suggested that such improvement has to be based on a better understanding of the spore surface structure

  12. Evaluating the transport of bacillus subtilis spores as a potential surrogate for Cryptosporidium parvum Oocysts

    Science.gov (United States)

    The USEPA has recommended the use of aerobic spores as an indicator for Cryptosporidium oocysts when determining groundwater under the direct influence of surface water. Surface properties, interaction energies, transport, retention, and release behavior of B. subtilis spores were measured over a r...

  13. Modelling the effect of sub(lethal) heat treatment of Bacillus subtilis spores on germination rate and outgrowth to exponentially growing vegetative cells

    NARCIS (Netherlands)

    Smelt, J.P.P.M.; Bos, A.P.; Kort, R.; Brul, S.

    2008-01-01

    Spores of Bacillus subtilis were subjected to relatively mild heat treatments in distilled water and properties of these spores were studied. These spores had lost all or part of their dipicolinic acid (DPA) depending on the severity of the heat treatment. Even after relatively mild heat treatments

  14. Response of Bacillus subtilis spores to dehydration and UV irradiation at extremely low temperatures.

    Science.gov (United States)

    Dose, K; Klein, A

    1996-02-01

    Spores of Bacillus subtilis have been exposed to the conditions of extreme dehydration (argon/silica gel; simulated space vacuum) for up to 12 weeks at 298 K and 80 K in the dark. The inactivation has been correlated with the production of DNA-double strand-breaks. The temperature-dependence of the rate constants for inactivation or production of DNA-double strand-breaks is surprisingly low. Controls kept in the frozen state at 250 K for the same period of time showed no sign of deterioration. In another series of experiments the spores have been UV irradiated (253.7 nm) at 298 K, 200 K and 80 K after exposure to dehydrating conditions for 3 days. Fluence-effect relationships for inactivation, production of DNA-double strand-breaks and DNA-protein cross-links are presented. The corresponding F37-values for inactivation and production of DNA lesions are significantly increased only at 80 K (factor of 4 to 5). The data indicate that the low temperatures that prevail in the outer parts of the Solar System or at the nightside of Mars or the Moon are not sufficiently low to crucially inhibit inactivation by dehydration. Our data place further constraints on the panspermia hypothesis.

  15. Novel Secretion Apparatus Maintains Spore Integrity and Developmental Gene Expression in Bacillus subtilis

    Science.gov (United States)

    Meisner, Jeffrey; Serrano, Monica; Henriques, Adriano O.; Moran, Charles P.; Rudner, David Z.

    2009-01-01

    Sporulation in Bacillus subtilis involves two cells that follow separate but coordinately regulated developmental programs. Late in sporulation, the developing spore (the forespore) resides within a mother cell. The regulation of the forespore transcription factor σG that acts at this stage has remained enigmatic. σG activity requires eight mother-cell proteins encoded in the spoIIIA operon and the forespore protein SpoIIQ. Several of the SpoIIIA proteins share similarity with components of specialized secretion systems. One of them resembles a secretion ATPase and we demonstrate that the ATPase motifs are required for σG activity. We further show that the SpoIIIA proteins and SpoIIQ reside in a multimeric complex that spans the two membranes surrounding the forespore. Finally, we have discovered that these proteins are all required to maintain forespore integrity. In their absence, the forespore develops large invaginations and collapses. Importantly, maintenance of forespore integrity does not require σG. These results support a model in which the SpoIIIA-SpoIIQ proteins form a novel secretion apparatus that allows the mother cell to nurture the forespore, thereby maintaining forespore physiology and σG activity during spore maturation. PMID:19609349

  16. Dna stability and survival of bacillus subtilis spores in extreme dryness

    Science.gov (United States)

    Dose, Klaus; Gill, Markus

    1995-06-01

    The inactivation of Bacillus subtilis spores during long-term exposure (up to several months) to extreme dryness (especially vacuum) is strain-dependent, through only to a small degree. During a first phase (lasting about four days) monolayers of spores lose about 20% of their viability, regardless of the strain studied. During this phase loss in viability can be equally attributed both to damages of hydrophobic structures (membranes and proteins) and DNA. During a second phase lasting for the remaining time of experimental observation (weeks, months and years) the loss in viability is slowed. A viability of 55% to 75% (depending on the strain) is attained after a total exposure of 36 days. The loss in viability during the second phase can be correlated with the occurrence of DNA double strand breaks. Also covalent DNA-protein cross-links are formed by vacuum exposure. If the protein moiety of these cross-links is degraded by proteinase K-treatment in vitro additional DNA double strand breaks result. The data are also discussed with respect to survival on Mars and in near Earth orbits.

  17. Investigation of UV-TiO2 photocatalysis and its mechanism in Bacillus subtilis spore inactivation.

    Science.gov (United States)

    Zhang, Yiqing; Zhou, Lingling; Zhang, Yongji

    2014-09-01

    The inactivation levels of Bacillus subtilis spores for various disinfection processes (ultraviolet (UV), TiO2 and UV-TiO2) were compared. The results showed that the inactivation effect of B. subtilis spores by UV treatment alone was far below that for bacteria without endospores. TiO2 alone in the dark, as a control experiment, exhibited almost no inactivation effect. Compared with UV treatment alone, the inactivation effect increased significantly with the addition of TiO2. Increases of the UV irradiance and TiO2 concentration both contributed to the increase of the inactivation effect. Lipid peroxidation was found to be the underlying mechanism of inactivation. Malondialdehyde (MDA), the degradation product of lipid peroxidation, was used as an index to determine the extent of the reaction. The MDA concentration surged surprisingly to 3.24nmol/mg dry cell with the combination disinfection for 600sec (0.10mW/cm(2) irradiance and 10mg/L TiO2). In contrast, for UV alone or TiO2 in the dark, the MDA concentration was 0.38 and 0.25nmol/mg dry cell, respectively, under the same conditions. This indicated that both UV and TiO2 were essential for lipid peroxidation. Changes in cell ultrastructure were observed by transmission electron microscopy. The cell membrane was heavily damaged and cellular contents were completely lysed with the UV-TiO2 process, suggesting that lipid peroxidation was the root of the enhancement in inactivation efficiency.

  18. Inactivation of Bacillus subtilis spores using various combinations of ultraviolet treatment with addition of hydrogen peroxide.

    Science.gov (United States)

    Zhang, Yiqing; Zhou, Lingling; Zhang, Yongji; Tan, Chaoqun

    2014-01-01

    This study aims at comparing the inactivation of Bacillus subtilis spores by various combinations of UV treatment and hydrogen peroxide (H2O2) addition. The combinations included sequential (UV-H2O2, H2O2-UV) and simultaneous (UV/H2O2) processes. Results showed that B. subtilis spores achieved a certain inactivation effect through UV treatment. However, hardly any inactivation effect by H2O2 alone was observed. H2O2 had a significant synergetic effect when combined with UV treatment, while high irradiance and H2O2 concentration both favored the reaction. When treated with 0.60 mm H2O2 and 113.0 μW/cm(2) UV irradiance for 6 min, the simultaneous UV/H2O2 treatment showed significantly improved disinfection effect (4.13 log) compared to that of UV-H2O2 (3.03 log) and H2O2-UV (2.88 log). The relationship between the inactivation effect and the exposure time followed a typical pseudo-first-order kinetics model. The pseudo-first-order rate constants were 0.478, 0.447 and 0.634 min(-1), for the UV-H2O2, H2O2-UV and UV/H2O2 processes, respectively, further confirming the optimal disinfection effect of the UV/H2O2 process. The disinfection could be ascribed to the OH radicals, as verified by the level of para-chlorobenzoic acid (pCBA).

  19. Atomic force microscopy imaging and single molecule recognition force spectroscopy of coat proteins on the surface of Bacillus subtilis spore.

    Science.gov (United States)

    Tang, Jilin; Krajcikova, Daniela; Zhu, Rong; Ebner, Andreas; Cutting, Simon; Gruber, Hermann J; Barak, Imrich; Hinterdorfer, Peter

    2007-01-01

    Coat assembly in Bacillus subtilis serves as a tractable model for the study of the self-assembly process of biological structures and has a significant potential for use in nano-biotechnological applications. In the present study, the morphology of B. subtilis spores was investigated by magnetically driven dynamic force microscopy (MAC mode atomic force microscopy) under physiological conditions. B. subtilis spores appeared as prolate structures, with a length of 0.6-3 microm and a width of about 0.5-2 microm. The spore surface was mainly covered with bump-like structures with diameters ranging from 8 to 70 nm. Besides topographical explorations, single molecule recognition force spectroscopy (SMRFS) was used to characterize the spore coat protein CotA. This protein was specifically recognized by a polyclonal antibody directed against CotA (anti-CotA), the antibody being covalently tethered to the AFM tip via a polyethylene glycol linker. The unbinding force between CotA and anti-CotA was determined as 55 +/- 2 pN. From the high-binding probability of more than 20% in force-distance cycles it is concluded that CotA locates in the outer surface of B. subtilis spores.

  20. The Synergistic Effect of High Pressure CO2 and Nisin on Inactivation of Bacillus subtilis Spores in Aqueous Solutions

    Science.gov (United States)

    Rao, Lei; Wang, Yongtao; Chen, Fang; Liao, Xiaojun

    2016-01-01

    The inactivation effects of high pressure CO2 + nisin (simultaneous treatment of HPCD and nisin, HPCD + nisin), HPCD→nisin (HPCD was followed by nisin), and nisin→HPCD (nisin was followed by HPCD) treatments on Bacillus subtilis spores in aqueous solutions were compared. The spores were treated by HPCD at 6.5 or 20 MPa, 84–86°C and 0–30 min, and the concentration of nisin was 0.02%. Treated spores were examined for the viability, the permeability of inner membrane (IM) using flow cytometry method and pyridine-2, 6-dicarboxylic acid (DPA) release, and structural damage by transmission electron microscopy. A synergistic effect of HPCD + nisin treatment on inactivation of the spores was found, and the inactivation efficiency of the spores was HPCD + nisin > HPCD→nisin or nisin→HPCD. Moreover, HPCD + nisin caused higher IM permeability and DPA release of the spores than HPCD. A possible action mode of nisin-enhanced inactivation of the spores was suggested as that HPCD firstly damaged the coat and cortex of spores, and nisin penetrated into and acted on the IM of spores, which increased the damage to the IM of spores, and resulted in higher inactivation of the spores. PMID:27708639

  1. Understanding of the importance of the spore coat structure and pigmentation in the Bacillus subtilis spore resistance to low-pressure plasma sterilization

    Science.gov (United States)

    Raguse, Marina; Fiebrandt, Marcel; Denis, Benjamin; Stapelmann, Katharina; Eichenberger, Patrick; Driks, Adam; Eaton, Peter; Awakowicz, Peter; Moeller, Ralf

    2016-07-01

    Low-pressure plasmas have been evaluated for their potential in biomedical and defense purposes. The sterilizing effect of plasma can be attributed to several active agents, including (V)UV radiation, charged particles, radical species, neutral and excited atoms and molecules, and the electric field. Spores of Bacillus subtilis were used as a bioindicator and a genetic model system to study the sporicidal effects of low-pressure plasma decontamination. Wild-type spores, spores lacking the major protective coat layers (inner, outer, and crust), pigmentation-deficient spores or spore impaired in encasement (a late step in coat assembly) were systematically tested for their resistance to low-pressure argon, hydrogen, and oxygen plasmas with and without admixtures. We demonstrate that low-pressure plasma discharges of argon and oxygen discharges cause significant physical damage to spore surface structures as visualized by atomic force microscopy. Spore resistance to low-pressure plasma was primarily dependent on the presence of the inner, and outer spore coat layers as well as spore encasement, with minor or less importance of the crust and spore pigmentation, whereas spore inactivation itself was strongly influenced by the gas composition and operational settings.

  2. Survival and Germinability of Bacillus subtilis Spores Exposed to Simulated Mars Solar Radiation: Implications for Life Detection and Planetary Protection

    Science.gov (United States)

    Tauscher, Courtney; Schuerger, Andrew C.; Nicholson, Wayne L.

    2006-08-01

    Bacterial spores have been considered as microbial life that could survive interplanetary transport by natural impact processes or human spaceflight activity. Deposition of terrestrial microbes or their biosignature molecules onto the surface of Mars could negatively impact life detection experiments and planetary protection measures. Simulated Mars solar radiation, particularly the ultraviolet component, has been shown to reduce spore viability, but its effect on spore germination and resulting production of biosignature molecules has not been explored. We examined the survival and germinability of Bacillus subtilis spores exposed to simulated martian conditions that include solar radiation. Spores of B. subtilis that contain luciferase resulting from expression of an sspB-luxAB gene fusion were deposited on aluminum coupons to simulate deposition on spacecraft surfaces and exposed to simulated Mars atmosphere and solar radiation. The equivalent of 42 min of simulated Mars solar radiation exposure reduced spore viability by nearly 3 logs, while germination-induced bioluminescence, a measure of germination metabolism, was reduced by less than 1 log. The data indicate that spores can retain the potential to initiate germination-associated metabolic processes and produce biological signature molecules after being rendered nonviable by exposure to Mars solar radiation.

  3. Bacillus subtilis spores expressing the VP28 antigen: a potential oral treatment to protect Litopenaeus vannamei against white spot syndrome.

    Science.gov (United States)

    Nguyen, Anh T V; Pham, Cuong K; Pham, Huong T T; Pham, Hang L; Nguyen, Anh H; Dang, Lua T; Huynh, Hong A; Cutting, Simon M; Phan, Tuan-Nghia

    2014-09-01

    The envelope protein VP28 of white spot syndrome virus (WSSV) is considered a candidate antigen for use in a potential vaccine to this important shrimp pathogen (the cause of white spot syndrome, WSS). Here, we used spores of Bacillus subtilis to display VP28 on the spore surface. Trials were conducted to evaluate their ability to protect shrimps against WSSV infection. The gene cotB-vp28 was integrated into the chromosome of the laboratory strain B. subtilis PY79, and expression of CotB-VP28 was detected by Western blotting and immunofluorescence. Expression of CotB-VP28 was equivalent to 1000 molecules per spore. PY79 and CotB-VP28 spores were mixed with pellets for feeding of whiteleg shrimps (Litopenaeus vannamei), followed by WSSV challenge. Superoxidase dismutase (SOD), phenoloxidase activities and mortality rates of the two shrimp groups were evaluated. Groups fed with PY79 and CotB-VP28 spores at day 7 had increased SOD activities of 29% and increased phenoloxidase activities of 15% and 33%, respectively, compared to those of the control group. Fourteen days postchallenge, 35% of vaccinated shrimps had died compared to 49% of those fed naked spores (PY79) and 66% untreated, unchallenged animals. These data suggest that spores expressing VP28 have potential as a prophylactic treatment of WSS.

  4. Carbon-13 (13C) labeling of Bacillus subtilis vegetative cells and spores: suitability for DNA stable isotope probing (DNA-SIP) of spores in soils.

    Science.gov (United States)

    Nicholson, Wayne L; Fedenko, Jeffrey; Schuerger, Andrew C

    2009-07-01

    To test the suitability of DNA stable isotope probing (DNA-SIP) for characterizing bacterial spore populations in soils, the properties of Bacillus subtilis cells and spores intensely labeled with [(13)C]glucose were characterized. Spore germination, vegetative growth rates, and sporulation efficiency were indistinguishable on glucose versus [(13)C]glucose, as were spore wet heat and UV resistance. Unlabeled and (13)C-labeled spores contained 1.0989 and 74.336 at.% (13)C, and exhibited wet densities of 1.356 and 1.365 g/ml, respectively. Chromosomal DNAs containing (12)C versus (13)C were readily separated by their different buoyant densities in cesium chloride/ethidium bromide gradients.

  5. SirA enforces diploidy by inhibiting the replication initiator DnaA during spore formation in Bacillus subtilis.

    Science.gov (United States)

    Wagner, Jennifer K; Marquis, Kathleen A; Rudner, David Z

    2009-09-01

    How cells maintain their ploidy is relevant to cellular development and disease. Here, we investigate the mechanism by which the bacterium Bacillus subtilis enforces diploidy as it differentiates into a dormant spore. We demonstrate that a sporulation-induced protein SirA (originally annotated YneE) blocks new rounds of replication by targeting the highly conserved replication initiation factor DnaA. We show that SirA interacts with DnaA and displaces it from the replication origin. As a result, expression of SirA during growth rapidly blocks replication and causes cell death in a DnaA-dependent manner. Finally, cells lacking SirA over-replicate during sporulation. These results support a model in which induction of SirA enforces diploidy by inhibiting replication initiation as B. subtilis cells develop into spores.

  6. Quantifying the effect of sorbic acid, heat and combination of both on germination and outgrowth of Bacillus subtilis spores at single cell resolution

    NARCIS (Netherlands)

    R. Pandey; G.H. Pieper; A. ter Beek; N.O.E. Vischer; J.P.P.M. Smelt; E.M.M. Manders; S. Brul

    2015-01-01

    Bacillus subtilis spores are a problem for the food industry as they are able to survive preservation processes. The spores often reside in food products, where their inherent protection against various stress treatments causes food spoilage. Sorbic acid is widely used as a weak acid preservative in

  7. Identification of Differentially Expressed Genes during Bacillus subtilis Spore Outgrowth in High-Salinity Environments Using RNA Sequencing

    Science.gov (United States)

    Nagler, Katja; Krawczyk, Antonina O.; De Jong, Anne; Madela, Kazimierz; Hoffmann, Tamara; Laue, Michael; Kuipers, Oscar P.; Bremer, Erhard; Moeller, Ralf

    2016-01-01

    In its natural habitat, the soil bacterium Bacillus subtilis often has to cope with fluctuating osmolality and nutrient availability. Upon nutrient depletion it can form dormant spores, which can revive to form vegetative cells when nutrients become available again. While the effects of salt stress on spore germination have been analyzed previously, detailed knowledge on the salt stress response during the subsequent outgrowth phase is lacking. In this study, we investigated the changes in gene expression during B. subtilis outgrowth in the presence of 1.2 M NaCl using RNA sequencing. In total, 402 different genes were upregulated and 632 genes were downregulated during 90 min of outgrowth in the presence of salt. The salt stress response of outgrowing spores largely resembled the osmospecific response of vegetative cells exposed to sustained high salinity and included strong upregulation of genes involved in osmoprotectant uptake and compatible solute synthesis. The σB-dependent general stress response typically triggered by salt shocks was not induced, whereas the σW regulon appears to play an important role for osmoadaptation of outgrowing spores. Furthermore, high salinity induced many changes in the membrane protein and transporter transcriptome. Overall, salt stress seemed to slow down the complex molecular reorganization processes (“ripening”) of outgrowing spores by exerting detrimental effects on vegetative functions such as amino acid metabolism. PMID:27766092

  8. Survivability of bare, individual Bacillus subtilis spores to high-velocity surface impact: Implications for microbial transfer through space

    Science.gov (United States)

    Barney, Brandon L.; Pratt, Sara N.; Austin, Daniel E.

    2016-06-01

    Laboratory experiments show that endospores of Bacillus subtilis survive impact against a solid surface at velocities as high as 299 ±28 m/s. During impact, spores experience and survive accelerations of at least 1010 m/s2. The spores were introduced into a vacuum chamber using an electrospray source and accelerated to a narrow velocity distribution by entrainment in a differentially pumped gas flow. Different velocity ranges were studied by modifying the gas flow parameters. The spores were electrically charged, allowing direct measurement of the velocity of each spore as it passed through an image charge detector prior to surface impact. Spores impacted a glass surface and were collected for subsequent analysis by culturing. Most spores survived impact at all measured velocities. These experiments differ fundamentally from other studies that show either shock or impact survivability of bacteria embedded within or on the surface of a projectile. Bacteria in the present experiments undergo a single interaction with a solid surface at the full impact velocity, in the absence of any other effects such as cushioning due to microbe agglomerations, deceleration due to air or vapor, or transfer of impact shock through solid or liquid media. During these full-velocity impact events, the spores experience extremely high decelerations. This study is the first reported instance of accelerations of this magnitude experienced during a bacteria impact event. These results are discussed in the context of potential transfer of viable microbes in space and other scenarios involving surface impacts at high velocities.

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

    NARCIS (Netherlands)

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

    2011-01-01

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

  10. Mucosal immunity induced by gliadin-presenting spores of Bacillus subtilis in HLA-DQ8-transgenic mice.

    Science.gov (United States)

    Bonavita, Roberta; Isticato, Rachele; Maurano, Francesco; Ricca, Ezio; Rossi, Mauro

    2015-06-01

    The induction of mucosal immunity requires efficient antigen delivery and adjuvant systems. Probiotic bacterial strains are considered to be very promising tools to address both of these needs. In particular, Bacillus subtilis spores are currently under investigation as a long-lived, protease-resistant adjuvant system for different antigens. Furthermore, a non-recombinant approach has been developed based on the stable adsorption of antigen on the spore surface. In the present study, we explored this strategy as a means of modulating the immune response to wheat gliadin, the triggering agent of celiac disease (CD), an enteropathy driven by inflammatory CD4(+) T cells. Gliadin adsorption was tested on untreated or autoclaved wild-type (wt) and mutant (cotH or cotE) spores. We found that gliadin was stably and maximally adsorbed by autoclaved wt spores. We then tested the immune properties of the spore-adsorbed gliadin in HLA-DQ8-transgenic mice, which express one of the two HLA heterodimers associated with CD. In vitro, spore-adsorbed gliadin was efficiently taken up by mouse dendritic cells (DCs). Interestingly, gliadin-pulsed DCs efficiently stimulated splenic CD4(+) T cells from mice immunised with spore-adsorbed gliadin. Nasal pre-dosing with spore-adsorbed gliadin failed to down-regulate the ongoing cellular response in gliadin-sensitised DQ8 mice. Notably, naïve mice inoculated intranasally with multiple doses of spore-adsorbed gliadin developed an intestinal antigen-specific CD4(+) T cell-mediated response. In conclusion, our data highlight the ability of spore-adsorbed gliadin to elicit a T-cell response in the gut that could be exploitable for developing immune strategies in CD.

  11. Gel-free proteomic identification of the Bacillus subtilis insoluble spore coat protein fraction

    NARCIS (Netherlands)

    W. Abhyankar; A. ter Beek; H. Dekker; R. Kort; S. Brul; C.G. de Koster

    2011-01-01

    Species from the genus Bacillus have the ability to form endospores, dormant cellular forms that are able to survive heat and acid preservation techniques commonly used in the food industry. Resistance characteristics of spores towards various environmental stresses are in part attributed to their c

  12. SirA enforces diploidy by inhibiting the replication initiator DnaA during spore formation in Bacillus subtilis

    OpenAIRE

    Jennifer K. Wagner; Marquis, Kathleen A.; Rudner, David Z.

    2009-01-01

    How cells maintain their ploidy is relevant to cellular development and disease. Here, we investigate the mechanism by which the bacterium Bacillus subtilis enforces diploidy as it differentiates into a dormant spore. We demonstrate that a sporulation-induced protein SirA (originally annotated YneE) blocks new rounds of replication by targeting the highly conserved replication initiation factor DnaA. We show that SirA interacts with DnaA and displaces it from the replication origin. As a resu...

  13. Bacillus subtilis spore survival and expression of germination-induced bioluminescence after prolonged incubation under simulated Mars atmospheric pressure and composition: implications for planetary protection and lithopanspermia

    Science.gov (United States)

    Nicholson, Wayne L.; Schuerger, Andrew C.

    2005-01-01

    Bacterial endospores in the genus Bacillus are considered good models for studying interplanetary transfer of microbes by natural or human processes. Although spore survival during transfer itself has been the subject of considerable study, the fate of spores in extraterrestrial environments has received less attention. In this report we subjected spores of a strain of Bacillus subtilis, containing luciferase resulting from expression of an sspB-luxAB gene fusion, to simulated martian atmospheric pressure (7-18 mbar) and composition (100% CO(2)) for up to 19 days in a Mars simulation chamber. We report here that survival was similar between spores exposed to Earth conditions and spores exposed up to 19 days to simulated martian conditions. However, germination-induced bioluminescence was lower in spores exposed to simulated martian atmosphere, which suggests sublethal impairment of some endogenous spore germination processes.

  14. A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores

    Science.gov (United States)

    Nerandzic, Michelle M.; Sankar C, Thriveen; Setlow, Peter; Donskey, Curtis J.

    2016-01-01

    Background. Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5–2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods. We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results. Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200–2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions. Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin. PMID:26885539

  15. A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores.

    Science.gov (United States)

    Nerandzic, Michelle M; Sankar C, Thriveen; Setlow, Peter; Donskey, Curtis J

    2016-01-01

    Background.  Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5-2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods.  We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results.  Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200-2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions.  Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin.

  16. Synergistic effect of sequential or combined use of ozone and UV radiation for the disinfection of Bacillus subtilis spores.

    Science.gov (United States)

    Jung, Yeon Jung; Oh, Byung Soo; Kang, Joon-Wun

    2008-03-01

    This study was performed to evaluate the inactivation efficiency or synergy of combined ozone and UV processes (combined ozone/UV process) or sequential processes (ozone-UV, UV-ozone) compared with individual unit processes and to investigate the specific roles of ozone, UV and the hydroxyl radical, which is formed as an intermediate in the combined ozone/UV process. The Bacillus subtilis spore, which has often been used as a surrogate microorganism for Cryptosporidium parvum oocysts, was used as a target microorganism. Compared to individual unit processes with ozone or UV, the inactivation of B. subtilis spores by the combined ozone/UV process was enhanced under identical conditions. To investigate the specific roles of ozone and UV in the combined ozone/UV process, sequential ozone-UV and UV-ozone processes were tested for degrees of inactivation. Additionally, the experiment was performed in the presence and absence of tert-butyl alcohol, which acted as a hydroxyl radical scavenger to assess the role of inactivation by the hydroxyl radical in the combined ozone/UV process. Among the five candidate processes, the greatest synergistic effect was observed in the combined ozone/UV process. From the comparison of five candidate processes, the hydroxyl radical and ozone were each determined to significantly enhance the overall inactivation efficiency in the combined ozone/UV process.

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

    Science.gov (United States)

    Brul, Stanley; van Beilen, Johan; Caspers, Martien; O'Brien, Andrea; de Koster, Chris; Oomes, Suus; Smelt, Jan; Kort, Remco; Ter Beek, Alex

    2011-04-01

    Bacterial spore formers are prime organisms of concern in the food industry. Spores from the genus Bacillus are extremely stress resistant, most notably exemplified by high thermotolerance. This sometimes allows surviving spores to germinate and grow out to vegetative cells causing food spoilage and possible intoxication. Similar issues though more pending toward spore toxigenicity are observed for the anaerobic Clostridia. The paper indicates the nature of stress resistance and highlights contemporary molecular approaches to analyze the mechanistic basis of it in Bacilli. A molecular comparison between a laboratory strain and a food borne isolate, very similar at the genomic level to the laboratory strain but generating extremely heat resistant spores, is discussed. The approaches cover genome-wide genotyping, proteomics and genome-wide expression analyses studies. The analyses aim at gathering sufficient molecular information to be able to put together an initial framework for dynamic modelling of spore germination and outgrowth behaviour. Such emerging models should be developed both at the population and at the single spore level. Tools and challenges in achieving the latter are succinctly discussed.

  18. The differential effects of heat-shocking on the viability of spores from Bacillus anthracis, Bacillus subtilis, and Clostridium sporogenes after treatment with peracetic acid- and glutaraldehyde-based disinfectants.

    Science.gov (United States)

    March, Jordon K; Pratt, Michael D; Lowe, Chinn-Woan; Cohen, Marissa N; Satterfield, Benjamin A; Schaalje, Bruce; O'Neill, Kim L; Robison, Richard A

    2015-10-01

    This study investigated (1) the susceptibility of Bacillus anthracis (Ames strain), Bacillus subtilis (ATCC 19659), and Clostridium sporogenes (ATCC 3584) spores to commercially available peracetic acid (PAA)- and glutaraldehyde (GA)-based disinfectants, (2) the effects that heat-shocking spores after treatment with these disinfectants has on spore recovery, and (3) the timing of heat-shocking after disinfectant treatment that promotes the optimal recovery of spores deposited on carriers. Suspension tests were used to obtain inactivation kinetics for the disinfectants against three spore types. The effects of heat-shocking spores after disinfectant treatment were also determined. Generalized linear mixed models were used to estimate 6-log reduction times for each spore type, disinfectant, and heat treatment combination. Reduction times were compared statistically using the delta method. Carrier tests were performed according to AOAC Official Method 966.04 and a modified version that employed immediate heat-shocking after disinfectant treatment. Carrier test results were analyzed using Fisher's exact test. PAA-based disinfectants had significantly shorter 6-log reduction times than the GA-based disinfectant. Heat-shocking B. anthracis spores after PAA treatment resulted in significantly shorter 6-log reduction times. Conversely, heat-shocking B. subtilis spores after PAA treatment resulted in significantly longer 6-log reduction times. Significant interactions were also observed between spore type, disinfectant, and heat treatment combinations. Immediately heat-shocking spore carriers after disinfectant treatment produced greater spore recovery. Sporicidal activities of disinfectants were not consistent across spore species. The effects of heat-shocking spores after disinfectant treatment were dependent on both disinfectant and spore species. Caution must be used when extrapolating sporicidal data of disinfectants from one spore species to another. Heat

  19. SporeWeb : an interactive journey through the complete sporulation cycle of Bacillus subtilis

    NARCIS (Netherlands)

    Eijlander, Robyn T.; Jong, Anne de; Krawczyk, Antonina O.; Holsappel, Siger; Kuipers, Oscar P.

    2014-01-01

    Bacterial spores are a continuous problem for both food-based and health-related industries. Decades of scientific research dedicated towards understanding molecular and gene regulatory aspects of sporulation, spore germination and spore properties have resulted in a wealth of data and information.

  20. Recombinant Bacillus subtilis spores expressing cholera toxin B subunit and Helicobacter pylori urease B confer protection against H. pylori in mice.

    Science.gov (United States)

    Zhou, Zhenwen; Dong, Hui; Huang, Yanmei; Yao, Shuwen; Liang, Bingshao; Xie, Yongqiang; Long, Yan; Mai, Jialiang; Gong, Sitang

    2017-01-01

    Helicobacter pylori infection is associated with chronic gastritis, peptic ulcers, gastric cancer and mucosa-associated lymphoid tissue lymphoma. The limitations of current therapies for H. pylori infection include poor compliance and antibiotic resistance. Therefore, an effective anti-H. pylori vaccine would be an alternative or complement to antibiotic treatment. Urease B (UreB) is considered an ideal vaccine antigen against H. pylori infection. In this study, cholera toxin B subunit (CTB), a mucosal adjuvant, was used to enhance the immunogenicity of a novel Bacillus subtilis spore vaccine expressing CTB-UreB, along with the B. subtilis spore coat protein CotC as a fusion protein. Oral administration of B. subtilis spores expressing CotC-UreB or CotC-CTB-UreB led to increased levels of UreB-specific IgG in serum and UreB-specific IgA in faeces, as well as elevated levels of IL-10 and IFN-γ in splenocytes. In addition, oral administration of CotC-UreB or CotC-CTB-UreB spores induced significant reductions (80.0 and 90.5 %, respectively) in gastric H. pylori bacterial load (1.11±0.36×105 and 0.53±0.21×105 c.f.u., respectively) compared to that of the CotC control group (5.56±1.64×105 c.f.u., P<0.01). Moreover, CotC-CTB-UreB spores were significantly more effective at reducing the bacterial load than CotC-UreB spores (P<0.05). These results indicate that CotC-CTB-UreB-expressing B. subtilis spores are a potential vaccine candidate for the control of H. pylori infection.

  1. Analysis of the Effects of a gerP Mutation on the Germination of Spores of Bacillus subtilis

    Science.gov (United States)

    2012-11-01

    defective in these spores. The original gerP mutation was in the gerPC gene, the third gene in the likely hexacistronic gerP operon ; a similar operon is...present in other Bacillales species. Mutations in individual genes of the B. subtilis gerP operon also reduce spore germination with nutrient germinants...but do not reduce spore viability, and deletion of the whole operon gives the same general phenotype as do mutations in individual gerP genes. The B

  2. 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

  3. 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...

  4. Protection of Penaeus monodon against white spot syndrome by continuous oral administration of a low concentration of Bacillus subtilis spores expressing the VP28 antigen.

    Science.gov (United States)

    Pham, K-C; Tran, H T T; Van Doan, C; Le, P H; Van Nguyen, A T; Nguyen, H A; Hong, H A; Cutting, S M; Phan, T-N

    2017-03-01

    In this study, Bacillus subtilis spores expressing a chimeric protein, CotB-VP28, were used as a probiotic vaccine to protect black tiger shrimps (Penaeus monodon) against white spot syndrome virus (WSSV) infection. Oral administration of pellets coated with CotB-VP28 spores (at ≥1 × 10(9 ) CFU per g pellet) to shrimps induced immune-relating phenoloxydase activity (PO) in shrimps after 14 days of feeding (prior challenge) and at day 3 post challenge (1·26 and 1·70 fold increase respectively). A 75% protection rate was obtained by continuous feeding of the spore-coated pellets at ≥1 × 10(9 ) CFU per g for 14 days prior to WSSV challenge and during all the postchallenge period. Even when the amount of CotB-VP28 spores in feed pellets was reduced down to ≥5 × 10(7)  CFU per g and ≥1 × 10(6)  CFU per g, relatively high protection rates of 70 and 67·5%, respectively, were still obtained. By contrast, feeding pellets without spores (untreated group) and with naked spores (PY79 group) at ≥1 × 10(9)  CFU per g could not protect shrimps against WSSV. These data suggest that supplementation of CotB-VP28 spores at low dose of ≥1 × 10(6)  CFU per g could be effective as a prophylactic treatment of WSS for black tiger shrimps.

  5. A microfluidic device for real-time monitoring of Bacillus subtilis bacterial spores during germination based on non-specific physicochemical interactions on the nanoscale level.

    Science.gov (United States)

    Zabrocka, L; Langer, K; Michalski, A; Kocik, J; Langer, J J

    2015-01-07

    A microfluidic device for studies on the germination of bacterial spores (e.g. Bacillus subtilis) based on non-specific interactions on the nanoscale is presented. A decrease in the population of spores during germination followed by the appearance of transition forms and an increase in the number of vegetative cells can be registered directly and simultaneously by using the microfluidic device, which is equipped with a conductive polymer layer (polyaniline) in the form of a nano-network. The lab-on-a-chip-type device, operating in a continuous flow regime, allows monitoring of germination of bacterial spores and analysis of the process in detail. The procedure is fast and accurate enough for quantitative real-time monitoring of the main steps of germination, including final transformation of the spores into vegetative cells. All of this is done without the use of biomarkers or any bio-specific materials, such as enzymes, antibodies and aptamers, and is simply based on an analysis of physicochemical interactions on the nanoscale level.

  6. 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 ...

  7. Spore UV and acceleration resistance of endolithic Bacillus pumilus and Bacillus subtilis isolates obtained from Sonoran desert basalt: implications for lithopanspermia.

    Science.gov (United States)

    Benardini, James N; Sawyer, John; Venkateswaran, Kasthuri; Nicholson, Wayne L

    2003-01-01

    Bacterial spores have been used as model systems for studying the theory of interplanetary transport of life by natural processes such as asteroidal or cometary impacts (i.e., lithopanspermia). Because current spallation theory predicts that near-surface rocks are ideal candidates for planetary ejection and surface basalts are widely distributed throughout the rocky planets, we isolated spore-forming bacteria from the interior of near-subsurface basalt rocks collected in the Sonoran desert near Tucson, Arizona. Spores were found to inhabit basalt at very low concentrations (basalt samples. Populations of purified spores prepared from the isolated strains were subjected to 254-nm UV and ballistics tests in order to assess their resistance to UV radiation and to extreme acceleration shock, two proposed lethal factors for spores during interplanetary transfer. Specific natural isolates of B. pumilus were found to be substantially more resistant to UV and extreme acceleration than were reference laboratory strains of B. subtilis, the benchmark organism, suggesting that spores of environmental B. pumilus isolates may be more likely to survive the rigors of interplanetary transfer.

  8. 枯草芽胞杆菌芽胞表面展示外源蛋白的研究进展%Research Progress on Bacillus subtilis Spore Display of Recombinant Proteins

    Institute of Scientific and Technical Information of China (English)

    余小霞; 田健; 伍宁丰

    2013-01-01

    Bacillus subtilis is Gram-positive bacteria with biological safy. It can form spores with strong stress resistance in poor nutrient environment. The spore of Bacillus subtilis consists of 3 parts including the core, cortex and spore coat protein. Recently, the Bacillus subtilis spore coat proteins, such as CotB, CotC, CotG, CotX and OxdD, have been successfully used as vectors to display the antigen proteins, enzymes or reporter protein on the spore surfaces. The recombinant proteins on Bacillus subtilis spores usually have many advantages, such as good stability, easy purification and safety. Therefore, they can be used in medicine, food and feed industry, and other fields. It has a great application prospect. This review introduced in detail the molecular characteristics of Bacillus subtilis spores, the construction process of the expressive system on spore surface and its application prospect. Thus, the paper provided a foundation for the basic and applied research about spore surface display system.%枯草芽胞杆菌是一种生物安全的革兰氏阳性细菌,在营养匮乏的环境下,可形成具有强抗逆性的芽胞。枯草芽胞杆菌的芽胞由核心、皮层、孢子外套蛋白三部分组成,目前已成功利用枯草芽胞杆菌芽胞外套蛋白CotB、CotC、CotG、CotX和OxdD为载体,将酶蛋白、抗原蛋白或荧光标记蛋白等展示于芽胞表面。芽胞表面展示的蛋白通常具有较好的稳定性、易于纯化和安全性好等优点,可应用于医药、食品及饲料工业等领域,具有较大的应用前景。详细介绍了枯草芽胞杆菌芽胞的分子特点及芽胞表面展示系统的构建过程及其应用前景,为芽胞表面展示载体的基础及应用研究奠定基础。

  9. Inactivation kinetics of Bacillus subtilis spores with ozone%臭氧灭活水中枯草芽孢杆菌的动力学

    Institute of Scientific and Technical Information of China (English)

    刘枫; 昌盛; 陈忠林

    2016-01-01

    以枯草芽孢杆菌(ATCC6633)的孢子作为难灭活微生物的代表,研究了消毒剂浓度和反应时间的乘积值(CT值)、pH值、温度对臭氧灭活水中芽孢效果的影响,并探讨了相关灭活反应的动力学特征.结果表明,臭氧灭活芽孢的过程可分为延滞期和灭活期,其灭活反应符合Chick-Watson延迟反应动力学模型.在半连续流反应模式下,当臭氧浓度在0.42-4.00 mg· L-1,反应时间0-20 min,pH值6-8,温度1-30℃范围内时,臭氧对芽孢的灭活效果与臭氧的CT值显著相关,与单独的臭氧浓度无关,CT值越高,所能达到的灭活率也越高.同时,温度对反应速率常数k影响较大,即随着温度的升高,灭活反应的延滞期CT1ag显著减小,反应速率常数k增大,臭氧对芽孢的灭活能力增强;而反应速率常数k在各pH值下基本不变,pH值对芽孢的灭活影响甚微.%In general,spores of Bacillus subtilis (ATCC6633) would be used as potential model for the resistant microorganisms.In this study,the inactivation kinetics of spores in drinking water by ozone was investigated,and the factors such as ozone such as the numerical value of the product of the concentration of ozone and the reaction time (CT) values,pH,and temperature which could influence the inactivation process were evaluated.Results showed that the inactivation process of spores with ozone was characterized by a lag phase followed by a logarithmic inactivation phase,and the delayed Chick-Watson model could well describe the inactivation process.In this study,the disinfection of Bacillus subtilis spores was performed in a semi-batch reactor under the conditions with the ozone concentration,reaction time,pH,and temperature ranged in 0.42-4.00 mg·L-1,0-20 min,6-8,and 1-30 ℃,respectively.It showed that the inactivation is independent of ozone dose and is obvious relative to the ozone CT values.A higher inactivation level of spores wou ld be achieved at higher CT values.In addition

  10. Survival of spores of the UV-resistant Bacillus subtilis strain MW01 after exposure to low-earth orbit and simulated martian conditions: data from the space experiment ADAPT on EXPOSE-E.

    Science.gov (United States)

    Wassmann, Marko; Moeller, Ralf; Rabbow, Elke; Panitz, Corinna; Horneck, Gerda; Reitz, Günther; Douki, Thierry; Cadet, Jean; Stan-Lotter, Helga; Cockell, Charles S; Rettberg, Petra

    2012-05-01

    In the space experiment "Molecular adaptation strategies of microorganisms to different space and planetary UV climate conditions" (ADAPT), bacterial endospores of the highly UV-resistant Bacillus subtilis strain MW01 were exposed to low-Earth orbit (LEO) and simulated martian surface conditions for 559 days on board the European Space Agency's exposure facility EXPOSE-E, mounted outside the International Space Station. The survival of B. subtilis MW01 spores from both assays (LEO and simulated martian conditions) was determined by a colony-formation assay after retrieval. It was clearly shown that solar extraterrestrial UV radiation (λ≥110 nm) as well as the martian UV spectrum (λ≥200 nm) was the most deleterious factor applied; in some samples only a few spore survivors were recovered from B. subtilis MW01 spores exposed in monolayers. However, if shielded from solar irradiation, about 8% of MW01 spores survived in LEO conditions, and 100% survived in simulated martian conditions, compared to the laboratory controls. The results demonstrate the effect of shielding against the high inactivation potential of extraterrestrial solar UV radiation, which limits the chances of survival of even the highly UV-resistant strain of B. subtilis MW01 in the harsh environments of outer space and the martian surface.

  11. Inactivation of vegetative cells, but not spores, of Bacillus anthracis, B. cereus, and B. subtilis on stainless steel surfaces coated with an antimicrobial silver- and zinc-containing zeolite formulation.

    Science.gov (United States)

    Galeano, Belinda; Korff, Emily; Nicholson, Wayne L

    2003-07-01

    Stainless steel surfaces coated with paints containing a silver- and zinc-containing zeolite (AgION antimicrobial) were assayed in comparison to uncoated stainless steel for antimicrobial activity against vegetative cells and spores of three Bacillus species, namely, B. anthracis Sterne, B. cereus T, and B. subtilis 168. Under the test conditions (25 degrees C and 80% relative humidity), the zeolite coating produced approximately 3 log(10) inactivation of vegetative cells within a 5- to 24-h period, but viability of spores of the three species was not significantly affected.

  12. Investigating synergism during sequential inactivation of MS-2 phage and Bacillus subtilis spores with UV/H2O2 followed by free chlorine.

    Science.gov (United States)

    Cho, Min; Gandhi, Varun; Hwang, Tae-Mun; Lee, Sangho; Kim, Jae-Hong

    2011-01-01

    A sequential application of UV as a primary disinfectant with and without H(2)O(2) addition followed by free chlorine as secondary, residual disinfectant was performed to evaluate the synergistic inactivation of selected indicator microorganisms, MS-2 bacteriophage and Bacillus subtilis spores. No synergism was observed when the UV irradiation treatment was followed by free chlorine, i.e., the overall level of inactivation was the same as the sum of the inactivation levels achieved by each disinfection step. With the addition of H(2)O(2) in the primary UV disinfection step, however, enhanced microbial inactivation was observed. The synergism was observed in two folds manners: (1) additional inactivation achieved by hydroxyl radicals generated from the photolysis of H(2)O(2) in the primary UV disinfection step, and (2) damage to microorganisms in the primary step which facilitated the subsequent chlorine inactivation. Addition of H(2)O(2) in the primary disinfection step was also found to be beneficial for the degradation of selected model organic pollutants including bisphenol-A (endocrine disruptor), geosmin (taste and odor causing compound) and 2,4-D (herbicide). The results suggest that the efficiency of UV/free chlorine sequential disinfection processes, which are widely employed in drinking water treatment, could be significantly enhanced by adding H(2)O(2) in the primary step and hence converting the UV process to an advanced oxidation process.

  13. Etude de la résistance à la chaleur des spores de Bacillus subtilis déshydratées

    OpenAIRE

    Hauck Tiburski, Julia

    2013-01-01

    In response to starvation, species from the genre Bacillus are able to form metabolicallydormant spores which are very resistant to multiple forms of stress. They are found in quitehigh concentrations in some dried foods which, upon rehydration, may lead to food deterioration or food-borne diseases. Moreover, their destruction is rather difficult and mostof the techniques commonly used to treat dry foods result in a very low spore inactivation.The aim of this work is to better understand the ...

  14. 傅里叶变换红外光谱对枯草芽孢杆菌的光学特性研究%Optical Properties Research of Bacillus Subtilis Spores by Fourier Transform Infrared Spectroscopy

    Institute of Scientific and Technical Information of China (English)

    冯明春; 徐亮; 高闽光; 焦洋; 魏秀丽; 金岭; 程巳阳; 李相贤; 冯书香

    2012-01-01

    使用傅里叶变换红外光谱(FTIR)技术,测量了两种不同浓度的枯草芽孢杆菌的红外透过率谱,根据朗伯-比尔定律计算出它们的质量消光截面,通过算出复折射率的虚部,再使用KK(Kramers-Kronig)关系,导出复折射率的实部,并对实验结果作了分析和讨论.通过研究枯草芽孢杆菌复折射率的测量和分析方法,对于进一步研究生物气溶胶的吸收和散射特性、拓宽生物气溶胶的测量和遥测技术方法,具有重要的意义.%The authors measured IR transmission spectra of two different concentrations of bacillus subtilis spores by using Fourier transform infrared spectroscopy (FTIR) technology. The mass extinction cross section k of bacillus subtilis spores was calculated according to Lambert-Beer law and the imaginary part n, of the complex refractive index was also calculated through k. The real part nr of the complex refractive index was derived from the KK (Kramers-Kronig) relationship and the experimental results were also analyzed and discussed with the study of measurement and analysis method of the complex refractive index on bacillus subtilis spores, it is of great significance to further research the absorption and scattering characteristics, and to broaden the measurement and remote sensing technology method of the biological aerosols.

  15. A method for the determination of bacterial spore DNA content based on isotopic labelling, spore germination and diphenylamine assay; ploidy of spores of several Bacillus species.

    Science.gov (United States)

    Hauser, P M; Karamata, D

    1992-01-01

    A reliable method for measuring the spore DNA content, based on radioactive DNA labelling, spore germination in absence of DNA replication and diphenylamine assay, was developed. The accuracy of the method, within 10-15%, is adequate for determining the number of chromosomes per spore, provided that the genome size is known. B subtilis spores were shown to be invariably monogenomic, while those of larger bacilli Bacillus megaterium, Bacillus cereus and Bacillus thuringiensis, often, if not invariably, contain two genomes. Attempts to modify the spore DNA content of B subtilis by altering the richness of the sporulation medium, the sporulation conditions (liquid or solid medium), or by mutation, were apparently unsuccessful. An increase of spore size with medium richness, not accompanied by an increase in DNA content, was observed. The implication of the apparently species-specific spore ploidy and the influence of the sporulation conditions on spore size and shape are discussed.

  16. 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.

  17. Hydrazine inactivates bacillus spores

    Science.gov (United States)

    Schubert, Wayne; Plett, G. A.; Yavrouian, A. H.; Barengoltz, J.

    2005-01-01

    Planetary Protection places requirements on the maximum number of viable bacterial spores that may be delivered by a spacecraft to another solar system body. Therefore, for such space missions, the spores that may be found in hydrazine are of concern. A proposed change in processing procedures that eliminated a 0.2 um filtration step propmpted this study to ensure microbial contamination issue existed, especially since no information was found in the literature to substantiate bacterial spore inactivation by hydrazine.

  18. 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.

  19. Monitoring Rates and Heterogeneity of High-Pressure Germination of Bacillus Spores by Phase-Contrast Microscopy of Individual Spores

    Science.gov (United States)

    2014-01-01

    SECURITY CLASSIFICATION OF: The germination of multiple individual Bacillus subtilis spores by a high pressure (HP) of 140-150 (unless noted...otherwise) megaPascals (MPa) that activates spore germinant receptors (GRs) was monitored by phase contrast microscopy in a diamond anvil cell. Major...conclusions were that: i) >95% of spores germinated in 40 min; ii) individual spore’s HP germination kinetics were very similar to those for nutrient

  20. 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.

  1. Research on Ozone Inactivation of Bacillus subtilis Spores%臭氧对枯草芽孢杆菌孢子的灭活研究

    Institute of Scientific and Technical Information of China (English)

    齐爱玲; 李继; 纪家林; 邹俐; 郭路路

    2011-01-01

    Cryptosporidium parvum oocysts bring challenges to drinking water safety due to their high resistance to the disinfectants, resulting in great potential harm to public health. This research rehtes to B. Subtilis spores(ATCC 6633)used as a surrogate microorganism to study the impact of turbidity and other water parameters as well as ozone concentration on the inactivation effect. The result showed that ozone inactivation of B. Subtilis spores was mainly associated with CT value rather than the concentration of ozone per se, and reduction of turbidity and organic matter in water increased inactivation efficiency. In addition, when water temperature became low it was necessary to increase CT value to ensure the disinfection.%隐孢子虫灭活困难,给饮用水安全带来了挑战.实验以枯草芽孢杆菌孢子(ATCC6633)作为隐孢子虫的指示菌,研究了臭氧浓度、浊度、有机物、温度等因素对臭氧灭活枯草芽孢杆菌孢子的影响.研究表明,臭氧对枯草芽孢杆菌孢子的灭活与CT值相关,臭氧浓度对消毒效果影响较小;降低浊度、有机物含量,能提高臭氧对枯草芽孢杆菌孢子的灭活效率;温度降低,为保证一定的消毒效率,所需的CT值越大.研究结果为水厂合理控制运行参数提供了借鉴.

  2. Effects of Mn2+ Levels on the Resistance Properties of Bacillus cereus Spores

    Science.gov (United States)

    2013-01-01

    In contrast, Bacillus subtilis spores with over a 200-fold range of protoplast Mn levels exhibited no significant differences in resistance to...Bacillus megaterium by wet heat. Lett. Appl . Microbiol. 50:507-514. Daly MJ (2012) Death by protein damage in irradiated cells. DNA Repair 11:12-21...levels on resistance of Bacillus megaterium spores to heat, radiation and hydrogen peroxide. J. Appl . Microbiol. 111:663-670. Ghosh S, Setlow P (2010

  3. Quantitative and Sensitive RNA Based Detection of Bacillus Spores

    Directory of Open Access Journals (Sweden)

    Ekaterina eOsmekhina

    2014-03-01

    Full Text Available The fast and reliable detection of bacterial spores is of great importance and still remains a challenge. Here we describe a direct RNA based diagnostic method for the specific detection of viable bacterial spores which does not depends on an enzymatic amplification step and therefore is directly appropriate for quantification. The procedure includes the following steps: (i heat activation of spores, (ii germination and enrichment cultivation, (iii cell lysis, and (iv analysis of 16S rRNA in crude cell lysates using a sandwich hybridization assay. The sensitivity of the method is dependent on the cultivation time and the detection limit; it is possible to detect 10 spores per ml when the RNA analysis is performed after 6 h of enrichment cultivation. At spore concentrations above 106 spores per ml the cultivation time can be shortened to 30 min. Total analysis times are in the range of 2 to 8 hours depending on the spore concentration in samples. The developed procedure is optimized at the example of Bacillus subtilis spores but should be applicable to other organisms. The new method can easily be modified for other target RNAs and is suitable for specific detection of spores from known groups of organisms.

  4. 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...

  5. Regulation of Growth of the Mother Cell and Chromosome Replication during Sporulation of Bacillus subtilis

    OpenAIRE

    Xenopoulos, Panagiotis; Piggot, Patrick J.

    2011-01-01

    During spore formation, Bacillus subtilis divides asymmetrically, resulting in two cells with different fates. Immediately after division, the transcription factor σF becomes active in the smaller prespore, followed by activation of σE in the larger mother cell. We recently showed that a delay in σE activation resulted in the novel phenotype of two spores (twins) forming within the same mother cell. Mother cells bearing twins are substantially longer than mother cells with single spores. Here...

  6. 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.

  7. Relevance of diffusion through bacterial spore coats/membranes and the associated concentration boundary layers in the initial lag phase of inactivation: a case study for Bacillus subtilis with ozone and monochloramine.

    Science.gov (United States)

    Fernando, W J N; Othman, R

    2006-02-01

    Disinfectants are generally used to inactivate microorganisms in solutions. The process of inactivation involves the disinfectant in the liquid diffusing towards the bacteria sites and thereafter reacting with bacteria at rates determined by the respective reaction rates. Such processes have demonstrated an initial lag phase followed by an active depletion phase of bacteria. This paper attempts to study the importance of the combined effects of diffusion of the disinfectant through the outer membrane of the bacteria and transport through the associated concentration boundary layers (CBLs) during the initial lag phase. Mathematical equations are developed correlating the initial concentration of the disinfectant with time required for reaching a critical concentration (C*) at the inner side of the membrane of the cell based on diffusion of disinfectant through the outer membranes of the bacteria and the formation of concentration boundary layers on both sides of the membranes. Experimental data of the lag phases of inactivation already available in the literature for inactivation of Bacillus subtilis spores with ozone and monochloramine are tested with the equations. The results seem to be in good agreement with the theoretical equations indicating the importance of diffusion process across the outer cell membranes and the resulting CBL's during the lag phase of disinfection.

  8. 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.

  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. 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.

  11. 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.

  12. Gas discharge plasmas are effective in inactivating Bacillus and Clostridium spores.

    Science.gov (United States)

    Tseng, Shawn; Abramzon, Nina; Jackson, James O; Lin, Wei-Jen

    2012-03-01

    Bacterial spores are the most resistant form of life and have been a major threat to public health and food safety. Nonthermal atmospheric gas discharge plasma is a novel sterilization method that leaves no chemical residue. In our study, a helium radio-frequency cold plasma jet was used to examine its sporicidal effect on selected strains of Bacillus and Clostridium. The species tested included Bacillus subtilis, Bacillus stearothermophilus, Clostridium sporogenes, Clostridium perfringens, Clostridium difficile, and Clostridium botulinum type A and type E. The plasmas were effective in inactivating selected Bacillus and Clostridia spores with D values (decimal reduction time) ranging from 2 to 8 min. Among all spores tested, C. botulinum type A and C. sporogenes were significantly more resistant to plasma inactivation than other species. Observations by phase contrast microscopy showed that B. subtilis spores were severely damaged by plasmas and the majority of the treated spores were unable to initiate the germination process. There was no detectable fragmentation of the DNA when the spores were treated for up to 20 min. The release of dipicolinic acid was observed almost immediately after the plasma treatment, indicating the spore envelope damage could occur quickly resulting in dipicolinic acid release and the reduction of spore resistance.

  13. 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.

  14. Complete Genome Sequence of Bacillus subtilis subsp. subtilis Strain 3NA

    OpenAIRE

    Reuß, Daniel R.; Schuldes, Jörg; Daniel, Rolf; Altenbuchner, Josef

    2015-01-01

    Bacillus subtilis 3NA reaches high cell densities during fed-batch fermentation and is an interesting target for further optimization as a production strain. Here, we announce the full genome of B. subtilis 3NA. The presence of specific Bacillus subtilis 168 and W23 genetic features suggests that 3NA is a hybrid of these strains.

  15. Effects of Bacillus subtilis KD1 on broiler intestinal flora.

    Science.gov (United States)

    Wu, B Q; Zhang, T; Guo, L Q; Lin, J F

    2011-11-01

    A novel Bacillus subtilis KD1 strain was isolated and identified from healthy broilers, and its phylogenetic classification was subsequently analyzed. To evaluate its probiotic availability, its growth characteristics and tolerance for the gut environment were evaluated in vitro. The results suggest that B. subtilis KD1 is superior in secreting neutral protease and is highly tolerant of gastric acid and bile salt. In the logarithmic growth phase, the neutral protease reached a maximum of 1,369.3 U/mL. When all live bacteria had become spores in the broth, B. subtilis KD1 was freeze dried and fed to broilers at 10(9), 5 × 10(9), and 10(10) bacilli/kg of feed. The animal trial results suggest that the addition of the new strain significantly improved intestinal flora by increasing lactobacilli and reducing Escherichia coli (P < 0.05) as compared with the control; hence, B. subtilis KD1 is a promising probiotic organism in broilers.

  16. Antimicrobial copper alloy surfaces are effective against vegetative but not sporulated cells of gram-positive Bacillus subtilis

    OpenAIRE

    San, Kaungmyat; Long, Janet; Michels, Corinne A.; Gadura, Nidhi

    2015-01-01

    This study explores the role of membrane phospholipid peroxidation in the copper alloy mediated contact killing of Bacillus subtilis, a spore-forming gram-positive bacterial species. We found that B. subtilis endospores exhibited significant resistance to copper alloy surface killing but vegetative cells were highly sensitive to copper surface exposure. Cell death and lipid peroxidation occurred in B. subtilis upon copper alloy surface exposure. In a sporulation-defective strain carrying a de...

  17. Inactivation and ultrastructure analysis of Bacillus spp. and Clostridium perfringens spores.

    Science.gov (United States)

    Brantner, Christine A; Hannah, Ryan M; Burans, James P; Pope, Robert K

    2014-02-01

    Bacterial endospores are resistant to many environmental factors from temperature extremes to ultraviolet irradiation and are generally more difficult to inactivate or kill than vegetative bacterial cells. It is often considered necessary to treat spores or samples containing spores with chemical fixative solutions for prolonged periods of time (e.g., 1-21 days) to achieve fixation/inactivation to enable electron microscopy (EM) examination outside of containment laboratories. Prolonged exposure to chemical fixatives, however, can alter the ultrastructure of spores for EM analyses. This study was undertaken to determine the minimum amount of time required to inactivate/sterilize and fix spore preparations from several bacterial species using a universal fixative solution for EM that maintains the ultrastructural integrity of the spores. We show that a solution of 4% paraformaldehyde with 1% glutaraldehyde inactivated spore preparations of Bacillus anthracis, Bacillus cereus, Bacillus megaterium, Bacillus thuringiensis, and Clostridium perfringens in 30 min, and Bacillus subtilis in 240 min. These results suggest that this fixative solution can be used to inactivate and fix spores from several major groups of bacterial spore formers after 240 min, enabling the fixed preparations to be removed from biocontainment and safely analyzed by EM outside of biocontainment.

  18. 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

  19. 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...

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. 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.

  5. Changes in Bacillus Spore Small Molecules, rRNA, Germination, and Outgrowth after Extended Sublethal Exposure to Various Temperatures: Evidence that Protein Synthesis Is Not Essential for Spore Germination.

    Science.gov (United States)

    Korza, George; Setlow, Barbara; Rao, Lei; Li, Qiao; Setlow, Peter

    2016-12-15

    rRNAs of dormant spores of Bacillus subtilis were >95% degraded during extended incubation at 50°C, as reported previously (E. Segev, Y. Smith, and S. Ben-Yehuda, Cell 148:139-114, 2012, doi:http://dx.doi.org/10.1016/j.cell.2011.11.059), and this was also true of spores of Bacillus megaterium Incubation of spores of these two species for ∼20 h at 75 to 80°C also resulted in the degradation of all or the great majority of the 23S and 16S rRNAs, although this rRNA degradation was slower than nonenzymatic hydrolysis of purified rRNAs at these temperatures. This rRNA degradation at high temperature generated almost exclusively oligonucleotides with minimal levels of mononucleotides. RNase Y, suggested to be involved in rRNA hydrolysis during B. subtilis spore incubation at 50°C, did not play a role in B. subtilis spore rRNA breakdown at 80°C. Twenty hours of incubation of Bacillus spores at 70°C also decreased the already minimal levels of ATP in dormant spores 10- to 30-fold, to ≤0.01% of the total free adenine nucleotide levels. Spores depleted of rRNA were viable and germinated relatively normally, often even faster than starting spores. Their return to vegetative growth was also similar to that of untreated spores for B. megaterium spores and slower for heat-treated B. subtilis spores; accumulation of rRNA took place only after completion of spore germination. These findings thus strongly suggest that protein synthesis is not essential for Bacillus spore germination.IMPORTANCE A recent report (L. Sinai, A. Rosenberg, Y. Smith, E. Segev, and S. Ben-Yehuda, Mol Cell 57:3486-3495, 2015, doi:http://dx.doi.org/10.1016/j.molcel.2014.12.019) suggested that protein synthesis is essential for early steps in the germination of dormant spores of Bacillus subtilis If true, this would be a paradigm shift in our understanding of spore germination. We now show that essentially all of the rRNA can be eliminated from spores of Bacillus megaterium or B. subtilis, and these

  6. Germination of Spores of Astrobiologically Relevant Bacillus Species in High-Salinity Environments

    Science.gov (United States)

    Nagler, Katja; Julius, Christina; Moeller, Ralf

    2016-07-01

    In times of increasing space exploration and search for extraterrestrial life, new questions and challenges for planetary protection, aiming to avoid forward contamination of different planets or moons with terrestrial life, are emerging. Spore-forming bacteria such as Bacillus species have a high contamination potential due to their spores' extreme resistance, enabling them to withstand space conditions. Spores require liquid water for their conversion into a growing cell (i.e., spore germination and subsequent growth). If present, water on extraterrestrial planets or moons is likely to be closely associated with salts (e.g., in salty oceans or brines), thus constituting high-salinity environments. Spores of Bacillus subtilis can germinate despite very high salt concentrations, although salt stress does exert negative effects on this process. In this study, germination and metabolic reactivation ("outgrowth") of spores of five astrobiologically relevant Bacillus species (B. megaterium, B. pumilus SAFR-032, B. nealsonii, B. mojavensis, and B. vallismortis) in high salinity (≤3.6 M NaCl) were investigated. Spores of different species exhibited different germination and outgrowth capabilities in high salinity, which strongly depended on germination conditions, especially the exact composition of the medium. In this context, a new "universal" germination trigger for Bacillus spores, named KAGE (KCl, L-alanine, D-glucose, ectoine), was identified, which will be very useful for future comparative germination and outgrowth studies on different Bacillus species. Overall, this study yielded interesting new insights on salt stress effects on spore germination and points out the difficulty of predicting the potential of spores to contaminate salty environments on extraterrestrial celestial bodies.

  7. 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.

  8. Ultraviolet irradiation of DNA complexed with. alpha. /. beta. -type small, acid-soluble proteins from spores of Bacillus or Clostridium species makes spore photoproduct but not thymine dimers

    Energy Technology Data Exchange (ETDEWEB)

    Nicholson, W.L.; Setlow, B.; Setlow, P. (Univ. of Connecticut Health Center, Farmington (United States))

    1991-10-01

    UV irradiation of complexes of DNA and an {alpha}/{beta}-type small, acid-soluble protein (SASP) from Bacillus subtilis spores gave decreasing amounts of pyrimidine dimers and increasing amounts of spore photoproduct as the SASP/DNA ratio was increased. The yields of pyrimidine dimers and spore photoproduct were < 0.2% and 8% of total thymine, respectively, when DNA saturated with SASP was irradiated at 254 nm with 30 kJ/m{sup 2}; in the absence of SASP the yields were reversed - 4.5% and 0.3%, respectively. Complexes of DNA with {alpha}/{beta}-type SASP from Bacillus cereus, Bacillus megaterium, or Clostridium bifermentans spores also gave spore photoproduct upon UV irradiation. However, incubation of these SASPs with DNA under conditions preventing complex formation or use of mutant SASPs that do not form complexes did not affect the photoproducts formed in vitro. These results suggest that the UV photochemistry of bacterial spore DNA in vivo is due to the binding of {alpha}/{beta}-type SASP, a binding that is known to cause a change in DNA conformation in vitro from the B form to the A form. The yields of spore photoproduct in vitro were significantly lower than in vivo, perhaps because of the presence of substances other than SASP in spores. It is suggested that as these factors diffuse out in the first minutes of spore germination, spore photoproduct yields become similar to those observed for irradiation of SASP/DNA complexes in vitro.

  9. Nanomechanical Characterization of Bacillus anthracis Spores by Atomic Force Microscopy

    OpenAIRE

    2016-01-01

    The study of structures and properties of bacterial spores is important to understanding spore formation and biological responses to environmental stresses. While significant progress has been made over the years in elucidating the multilayer architecture of spores, the mechanical properties of the spore interior are not known. Here, we present a thermal atomic force microscopy (AFM) study of the nanomechanical properties of internal structures of Bacillus anthracis spores. We developed a nan...

  10. The minimal Bacillus subtilis nonhomologous end joining repair machinery.

    Directory of Open Access Journals (Sweden)

    Miguel de Vega

    Full Text Available It is widely accepted that repair of double-strand breaks in bacteria that either sporulate or that undergo extended periods of stationary phase relies not only on homologous recombination but also on a minimal nonhomologous end joining (NHEJ system consisting of a dedicated multifunctional ATP-dependent DNA Ligase D (LigD and the DNA-end-binding protein Ku. Bacillus subtilis is one of the bacterial members with a NHEJ system that contributes to genome stability during the stationary phase and germination of spores, having been characterized exclusively in vivo. Here, the in vitro analysis of the functional properties of the purified B. subtilis LigD (BsuLigD and Ku (BsuKu proteins is presented. The results show that the essential biochemical signatures exhibited by BsuLigD agree with its proposed function in NHEJ: i inherent polymerization activity showing preferential insertion of NMPs, ii specific recognition of the phosphate group at the downstream 5' end, iii intrinsic ligase activity, iv ability to promote realignments of the template and primer strands during elongation of mispaired 3' ends, and v it is recruited to DNA by BsuKu that stimulates the inherent polymerization and ligase activities of the enzyme allowing it to deal with and to hold different and unstable DNA realignments.

  11. Bacillus subtilis FZB24 affects flower quantity and quality of saffron (Crocus sativus).

    Science.gov (United States)

    Sharaf-Eldin, Mahmoud; Elkholy, Shereen; Fernández, José-Antonio; Junge, Helmut; Cheetham, Ronald; Guardiola, José; Weathers, Pamela

    2008-08-01

    The effect of Bacillus subtilis FZB24 on saffron ( Crocus sativus L.) was studied using saffron corms from Spain and the powdered form of B. SUBTILIS FZB24(R). Corms were soaked in water or in B. subtilis FZB24 spore solution for 15 min before sowing. Some corms were further soil drenched with the spore solution 6, 10 or 14 weeks after sowing. Growth and saffron stigma chemical composition were measured. Compared to untreated controls, application of B. subtilis FZB24 significantly increased leaf length, flowers per corm, weight of the first flower stigma, total stigma biomass; microbe addition also significantly decreased the time required for corms to sprout and the number of shoot sprouts. Compared to the controls, picrocrocin, crocetin and safranal compounds were significantly increased when the plants were soil drenched with the spore solution 14 weeks after sowing; in contrast crocin was highest in untreated controls. Results of this study suggest that application of B. subtilis FZB24 may provide some benefit to saffron growers by speeding corm growth (earlier shoot emergence) and increasing stigma biomass yield by 12 %. While some treatment conditions also increased saffron chemical composition, these were generally not the same treatments that simultaneously improved growth yields and thus, more study is required.

  12. Inhibiting Inosine Hydrolase and Alanine Racemase to Enhance the Germination of Bacillus anthracis Sterne Spores: Potential Spore Decontamination Strategies

    Science.gov (United States)

    2015-06-19

    2015): << Inhibiting inosine hydrolase and alanine racemase to enhance the germination of Bacillus anthracis Sterne spores: potential spore...inosine hydrolase and alanine racemase to enhance the germination of Bacillus anthracis Sterne spores potential spore decontamination strategies 5a...EASIER, SAFER, and CHEAPER Inducing spore germination should make resulting bacteria much more susceptible to decontamination methods and will be

  13. 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.

  14. Isolation and Characterization of Phages Infecting Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Anna Krasowska

    2015-01-01

    Full Text Available Bacteriophages have been suggested as an alternative approach to reduce the amount of pathogens in various applications. Bacteriophages of various specificity and virulence were isolated as a means of controlling food-borne pathogens. We studied the interaction of bacteriophages with Bacillus species, which are very often persistent in industrial applications such as food production due to their antibiotic resistance and spore formation. A comparative study using electron microscopy, PFGE, and SDS-PAGE as well as determination of host range, pH and temperature resistance, adsorption rate, latent time, and phage burst size was performed on three phages of the Myoviridae family and one phage of the Siphoviridae family which infected Bacillus subtilis strains. The phages are morphologically different and characterized by icosahedral heads and contractile (SIOΦ, SUBω, and SPOσ phages or noncontractile (ARπ phage tails. The genomes of SIOΦ and SUBω are composed of 154 kb. The capsid of SIOΦ is composed of four proteins. Bacteriophages SPOσ and ARπ have genome sizes of 25 kbp and 40 kbp, respectively. Both phages as well as SUBω phage have 14 proteins in their capsids. Phages SIOΦ and SPOσ are resistant to high temperatures and to the acid (4.0 and alkaline (9.0 and 10.0 pH.

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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.

  20. 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

  1. 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-...

  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. 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...

  4. A novel antifungal protein of Bacillus subtilis B25.

    Science.gov (United States)

    Tan, Zhiqiong; Lin, Baoying; Zhang, Rongyi

    2013-01-01

    Bacillus subtilis B25 was isolated from banana rhizosphere soil. It has been confirmed for B25 to have stronger antagonism against Fusarium oxysporum f.sp.cubense, Additionally B25 has good inhibitory to plant pathogens, including Corynespora cassiicola, Alternaria solani, Botrytis cinerea and Colletotrichum gloeosporioides on potato dextrose agar (PDA) plates. The antagonistic substance can be extracted from cell-free culture broth supernatants by 70% (w/v) (NH4)2 SO4 saturation. Clear blank band was observed between the protein and a pathogen. The examination of antagonistic mechanism under light microscope showed that the antifungal protein of B25 appeared to inhibit pathogens by leading to mycelium and spores tumescence, distortion, abnormality. The isolation procedure comprised ion exchange chromatography on DEAE-Sephadex Fast Flow and gel filtration chromatography on SephadexG-100. The purified antifungal fraction showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The active fraction was identified by NanoLC-ESI-MS/MS The amino acid sequences of 17 peptides segments were obtained. The analysis of the protein suggested that it was a hypothetical protein (gi154685475), with a relative molecular mass of 38708.67 Da and isoelectric point (pI) of 5.63.

  5. Development of a heat-stable and orally delivered recombinant M2e-expressing B. subtilis spore-based influenza vaccine.

    Science.gov (United States)

    Zhao, Guangyu; Miao, Yu; Guo, Yan; Qiu, Hongjie; Sun, Shihui; Kou, Zhihua; Yu, Hong; Li, Junfeng; Chen, Yue; Jiang, Shibo; Du, Lanying; Zhou, Yusen

    2014-01-01

    Highly conserved ectodomain of influenza virus M2 protein (M2e) is an important target for the development of universal influenza vaccines. Today, the use of chemical or genetic fusion constructs have been undertaken to overcome the low immunogenicity of M2e in vaccine formulation. However, current M2e vaccines are neither orally delivered nor heat-stable. In this study, we evaluated the immune efficacy of an orally delivered recombinant M2e vaccine containing 3 molcules of M2e consensus sequence of influenza A viruses, termed RSM2e3. To accomplish this, CotB, a spore coat of Bacillus subtilis (B. subtilis), was used as a fusion partner, and heat-stable nonpathogenic B. subtilis spores were used as the carrier. Our results showed that CotB-M2e3 fusion had no effect on spore structure or function in the resultant recombinant RSM2e3 strain and that heterologous influenza virus M2e protein was successfully displayed on the surface of the recombinant RSM2e3 spore. Importantly, recombinant RSM2e3 spores elicited strong and long-term M2e-specific systemic and mucosal immune responses, completely protecting immunized mice from lethal challenge of A/PR/8/34(H1N1) influenza virus. Taken together, our study forms a solid basis for the development of a novel orally delivered and heat-stable influenza vaccine based on B. subtilis spore surface display.

  6. 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...

  7. Comparison of hand hygiene procedures for removing Bacillus cereus spores.

    Science.gov (United States)

    Sasahara, Teppei; Hayashi, Shunji; Hosoda, Kouichi; Morisawa, Yuji; Hirai, Yoshikazu

    2014-01-01

    Bacillus cereus is a spore-forming bacterium. B. cereus occasionally causes nosocomial infections, in which hand contamination with the spores plays an important role. Therefore, hand hygiene is the most important practice for controlling nosocomial B. cereus infections. This study aimed to determine the appropriate hand hygiene procedure for removing B. cereus spores. Thirty volunteers' hands were experimentally contaminated with B. cereus spores, after which they performed 6 different hand hygiene procedures. We compared the efficacy of the procedures in removing the spores from hands. The alcohol-based hand-rubbing procedures scarcely removed them. The soap washing procedures reduced the number of spores by more than 2 log10. Extending the washing time increased the spore-removing efficacy of the washing procedures. There was no significant difference in efficacy between the use of plain soap and antiseptic soap. Handwashing with soap is appropriate for removing B. cereus spores from hands. Alcohol-based hand-rubbing is not effective.

  8. Electron Beam Irradiation Dose Dependently Damages the Bacillus Spore Coat and Spore Membrane

    Directory of Open Access Journals (Sweden)

    S. E. Fiester

    2012-01-01

    Full Text Available Effective control of spore-forming bacilli begs suitable physical or chemical methods. While many spore inactivation techniques have been proven effective, electron beam (EB irradiation has been frequently chosen to eradicate Bacillus spores. Despite its widespread use, there are limited data evaluating the effects of EB irradiation on Bacillus spores. To study this, B. atrophaeus spores were purified, suspended in sterile, distilled water, and irradiated with EB (up to 20 kGy. Irradiated spores were found (1 to contain structural damage as observed by electron microscopy, (2 to have spilled cytoplasmic contents as measured by spectroscopy, (3 to have reduced membrane integrity as determined by fluorescence cytometry, and (4 to have fragmented genomic DNA as measured by gel electrophoresis, all in a dose-dependent manner. Additionally, cytometry data reveal decreased spore size, increased surface alterations, and increased uptake of propidium iodide, with increasing EB dose, suggesting spore coat alterations with membrane damage, prior to loss of spore viability. The present study suggests that EB irradiation of spores in water results in substantial structural damage of the spore coat and inner membrane, and that, along with DNA fragmentation, results in dose-dependent spore inactivation.

  9. Protection against UV disinfection of E. coli bacteria and B. subtilis spores ingested by C. elegans nematodes.

    Science.gov (United States)

    Bichai, Françoise; Barbeau, Benoit; Payment, Pierre

    2009-08-01

    Nematodes, which occur abundantly in granular media filters of drinking water treatment plants and in distribution systems, can ingest and transport pathogenic bacteria and provide them protection against chemical disinfectants. However, protection against UV disinfection had not been investigated to date. In this study, Caenorhabditis elegans nematodes (wild-type strain N2) were allowed to feed on Escherichia coli OP50 and Bacillus subtilis spores before being exposed to 5 and 40 mJ/cm(2) UV fluences, using a collimated beam apparatus (LP, 254 nm). Sonication (15 W, 60s) was used to extract bacteria from nematode guts following UV exposure in order to assess the amount of ingested bacteria that resisted the UV treatment using a standard culture method. Bacteria located inside the gut of C. elegans were shown to benefit from a significant protection against UV. Approximately 15% of the applied UV fluence of 40 mJ/cm(2) (as typically used in WTP) was found to reach the bacteria located inside nematode guts based on the inactivation of recovered bacteria (2.7 log reduction of E. coli bacteria and 0.7 log reduction of B. subtilis spores at 40 mJ/cm(2)). To our knowledge, this study is the first demonstration of the protection effect of bacterial internalization by higher organisms against UV treatment, using the specific case of E. coli and B. subtilis spores ingested by C. elegans.

  10. 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.

  11. Scientific Opinion on the safety and efficacy of Bacillus subtilis PB6 (Bacillus subtilis as a feed additive for turkeys for fattening and turkeys reared for breeding

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2013-04-01

    Full Text Available Bacillus subtilis PB6 is the trade name for a feed additive based on viable spores of a strain of Bacillus subtilis. This species is considered by EFSA to be suitable for the qualified presumption of safety approach to establishing safety for the target species, consumers and the environment. This approach requires the identity of the active agent to be established and the absence of toxigenic potential and resistance to antibiotics of human or veterinary clinical significance to be demonstrated. EFSA considered these issues and reported the results in a previous opinion on the use of the product in chickens for fattening. The applicant is now requesting the authorisation of the additive in diets for turkeys for fattening and turkeys reared for breeding at a dose of 1 × 108 CFU/kg complete feedingstuffs. In the course of the former assessment, safety for users was also examined. In the view of the FEEDAP Panel, the use with these additional avian species will not introduce hazards not already considered. Therefore, in the current assessment, the FEEDAP Panel has considered only the efficacy data for turkeys for fattening and turkeys reared for breeding. Based on results of three trials carried out in turkeys for fattening, the Panel concluded that B. subtilis PB6 has the potential to improve the zootechnical performance parameters at the dose of 1 × 108 CFU/kg feed. This conclusion can be extended to turkeys reared for breeding.

  12. 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.

  13. 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...

  14. 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...

  15. Evaluation of in situ valine production by Bacillus subtilis in young pigs

    DEFF Research Database (Denmark)

    Nørgaard, Jan Værum; Canibe, Nuria; Assadi Soumeh, Elham;

    2016-01-01

    Mutants of Bacillus subtilis can be developed to overproduce Val in vitro. It was hypothesized that addition of Bacillus subtilis mutants to pig diets can be a strategy to supply the animal with Val. The objective was to investigate the effect of Bacillus subtilis mutants on growth performance an...

  16. Safety assessment of Bacillus subtilis CU1 for use as a probiotic in humans.

    Science.gov (United States)

    Lefevre, Marie; Racedo, Silvia M; Denayrolles, Muriel; Ripert, Gabrielle; Desfougères, Thomas; Lobach, Alexandra R; Simon, Ryan; Pélerin, Fanny; Jüsten, Peter; Urdaci, Maria C

    2017-02-01

    Bacillus subtilis CU1 is a recently described probiotic strain with beneficial effects on immune health in elderly subjects. The following work describes a series of studies supporting the safety of the strain for use as an ingredient in food and supplement preparations. Using a combination of 16S rDNA and gyrB nucleotide analyses, the species was identified as a member of the Bacillus subtilis complex (B. subtilis subsp. spizizenii). Further characterization of the organism at the strain level was achieved using random amplified polymorphic DNA polymerase chain reaction (RAPD PCR) and pulsed field gel electrophoresis (PFGE) analyses. B. subtilis CU1 did not demonstrate antibiotic resistance greater than existing regulatory cutoffs against clinically important antibiotics, did not induce hemolysis or produce surfactant factors, and was absent of toxigenic activity in vitro. Use of B. subtilis CU1 as a probiotic has recently been evaluated in a 16-week randomized, double-blind, placebo-controlled, parallel-arm study, in which 2 × 10(9) spores per day of B. subtilis CU1 were administered for a total 40 days to healthy elderly subjects (4 consumption periods of 10 days separated by 18-day washouts). This work describes safety related endpoints not previously reported. B. subtilis CU1 was safe and well-tolerated in the clinical subjects without undesirable physiological effects on markers of liver and kidney function, complete blood counts, hemodynamic parameters, and vital signs.

  17. Direct investigation of viscosity of an atypical inner membrane of Bacillus spores: a molecular rotor/FLIM study.

    Science.gov (United States)

    Loison, Pauline; Hosny, Neveen A; Gervais, Patrick; Champion, Dominique; Kuimova, Marina K; Perrier-Cornet, Jean-Marie

    2013-11-01

    We utilize the fluorescent molecular rotor Bodipy-C12 to investigate the viscoelastic properties of hydrophobic layers of bacterial spores Bacillus subtilis. The molecular rotor shows a marked increase in fluorescence lifetime, from 0.3 to 4ns, upon viscosity increase from 1 to 1500cP and can be incorporated into the hydrophobic layers within the spores from dormant state through to germination. We use fluorescence lifetime imaging microscopy to visualize the viscosity inside different compartments of the bacterial spore in order to investigate the inner membrane and relate its compaction to the extreme resistance observed during exposure of spores to toxic chemicals. We demonstrate that the bacterial spores possess an inner membrane that is characterized by a very high viscosity, exceeding 1000cP, where the lipid bilayer is likely in a gel state. We also show that this membrane evolves during germination to reach a viscosity value close to that of a vegetative cell membrane, ca. 600cP. The present study demonstrates quantitative imaging of the microscopic viscosity in hydrophobic layers of bacterial spores Bacillus subtilis and shows the potential for further investigation of spore membranes under environmental stress.

  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. Transformation of Bacillus Subtilis with cloned thymidylate synthetases

    Energy Technology Data Exchange (ETDEWEB)

    Rubin, Edward M.

    1980-01-01

    Bacillus subtilis carries two genes, thyA and thyB, each encoding different protein products, with thymidylate synthetase (TSase) activity. Either of these genes alone is sufficient for thymidine independence in B. subtilis. In addition there exist two B. subtilis temperate bacteriophages which upon infection of thymine requiring auxotrophs results in conversion of the organism to thymine independence. Chimeric plasmids selected for Thy/sup +/ transforming activity in E. coli were constructed and then used as a source of defined highly enriched DNA with which to transform competent B. subtilis. These plasmids were studied for their: (1) abiility to transform B. subtilis to thymine independence; (2) site of integration within the B. subtilis chromosome upon transformation; (3) phenotype of Thy/sup +/ plasmid generated transformants; and (4) nucleotide sequence homology among the cloned DNA fragments conferring thymine independence. Plasmids containing the two bacteriophage thy genes displayed the phenotype associated with thyA, whereas the plasmids containing the cloned B. subtilis chromosomal genes displayed the phenotype associated with thyB. Utilizing similar technology, the ability of an entirely foreign hybred bacterial plasmiid to transform B. subtilis was examined. In this case the gene from E. coli encoding thymidylate synthetase was cloned in the plasmid pBR322. The resulting chimeric plasmid was effective in transforming both E. coli and B. subtilis to thymine prototrophy. Uncloned linear E. coli chromosomal DNA was unable to transform thymine requiring strains of B. subtilis to thymine independence. Although the Thy/sup +/ transformants of E. coli contained plasmid DNA, the Thy/sup +/ transformants derived from the transformation of B. subtilis did not contain detectable extrachromosomal DNA. Instead the DNA from the chimeric plasmid was integrated into the chromosome of B. subtilis. (ERB)

  20. Bacillus atrophaeus Outer Spore Coat Assembly and Ultrastructure

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Leighton, T J; Wheeler, K E; Pitesky, M E; Malkin, A J

    2005-11-21

    Our previous atomic force microscopy (AFM) studies successfully visualized native Bacillus atrophaeus spore coat ultrastructure and surface morphology. We have shown that the outer spore coat surface is formed by a crystalline array of {approx}11 nm thick rodlets, having a periodicity of {approx}8 nm. We present here further AFM ultrastructural investigations of air-dried and fully hydrated spore surface architecture. In the rodlet layer, planar and point defects, as well as domain boundaries, similar to those described for inorganic and macromolecular crystals, were identified. For several Bacillus species, rodlet structure assembly and architectural variation appear to be a consequence of species-specific nucleation and crystallization mechanisms that regulate the formation of the outer spore coat. We propose a unifying mechanism for nucleation and self-assembly of this crystalline layer on the outer spore coat surface.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. 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...

  7. 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.

  8. 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

  9. 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

  10. 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

  11. [Asymmetric biosynthesis of d-pseudoephedrine by recombinant Bacillus subtilis].

    Science.gov (United States)

    Peng, Yanhong; Zhang, Liang; Ding, Zhongyang; Wang, Zhengxiang; Shi, Guiyang

    2011-07-01

    In order to successfully express the carbonyl reductase gene mldh in Bacillus subtilis and complete coenzyme regeneration by B. subtilis glucose dehydrogenase, the promoter PrpsD and the terminator TrpsD from B. subtilis rpsD gene were used as the expression cassette to be a recombinant plasmid pHY300plk-PrpsD-TrpsD. After that, the carbonyl reductase gene mldh was inserted into the previous plasmid and a plasmid pHY300plk-PrpsD-mldh-TrpsD was achieved, followed by transformed into B. subtilis Wb600 to obtain a recombinant B. subtilis Wb600 (pHY300plk-PrpsD-mldh-TrpsD). Subsequently, the results for whole-cell biotransformation from recombinant B. subtilis showed that it could be used to catalyze MAK (1-phenyl- 1-keto-2-methylaminopropane) to d-pseudoephedrine in the presence of glucose. The yield of d-pseudoephedrine could be up to 97.5 mg/L and the conversion rate of MAK was 24.1%. This study indicates the possibility of biotransformation production of d-pseudoephedrine from recombinant B. subtilis.

  12. Detection of Bacillus spores within 15 minutes by surface-enhanced Raman spectroscopy

    Science.gov (United States)

    Shende, Chetan; Inscore, Frank; Huang, Hermes; Farquharson, Stuart; Sengupta, Atanu

    2012-06-01

    Since the distribution of Bacillus anthracis causing spores through the US Postal System, there has been a persistent fear that biological warfare agents (BWAs) will be used by terrorists against our military abroad and our civilians at home. Despite the substantial effort to develop BWA analyzers, they remain either too slow, produce high falsealarm rates, lack sensitivity, or cannot be fielded. Consequently there remains a need for a portable analyzer that can overcome these limitations as expressed at the 2011 Biological Weapons Convention. To meet this need we have been developing a sample system that selectively binds BWAs and produce surface-enhanced Raman (SER) spectra using portable Raman spectrometers. Here we describe the use of a short peptide ligand functionalized on silver nanoparticles to selectively capture Bacillus cereus spores (a surrogate of B. anthracis) and their subsequent detection by SER spectroscopy. This technique was used to specifically detect B. cereus spores over closely related species like B. subtilis belonging to the same genus within 15 minutes. Sensitivity of the method was demonstrated by detecting 104 B. cereus spores/mL of water. The technology, once developed should prove invaluable for rapid monitoring of BWAs, which will immensely help first responders and emergency personnel in implementing appropriate counter measures.

  13. Reconstruction of the Regulatory Network for Bacillus subtilis and Reconciliation with Gene Expression Data

    Science.gov (United States)

    Faria, José P.; Overbeek, Ross; Taylor, Ronald C.; Conrad, Neal; Vonstein, Veronika; Goelzer, Anne; Fromion, Vincent; Rocha, Miguel; Rocha, Isabel; Henry, Christopher S.

    2016-01-01

    We introduce a manually constructed and curated regulatory network model that describes the current state of knowledge of transcriptional regulation of Bacillus subtilis. The model corresponds to an updated and enlarged version of the regulatory model of central metabolism originally proposed in 2008. We extended the original network to the whole genome by integration of information from DBTBS, a compendium of regulatory data that includes promoters, transcription factors (TFs), binding sites, motifs, and regulated operons. Additionally, we consolidated our network with all the information on regulation included in the SporeWeb and Subtiwiki community-curated resources on B. subtilis. Finally, we reconciled our network with data from RegPrecise, which recently released their own less comprehensive reconstruction of the regulatory network for B. subtilis. Our model describes 275 regulators and their target genes, representing 30 different mechanisms of regulation such as TFs, RNA switches, Riboswitches, and small regulatory RNAs. Overall, regulatory information is included in the model for ∼2500 of the ∼4200 genes in B. subtilis 168. In an effort to further expand our knowledge of B. subtilis regulation, we reconciled our model with expression data. For this process, we reconstructed the Atomic Regulons (ARs) for B. subtilis, which are the sets of genes that share the same “ON” and “OFF” gene expression profiles across multiple samples of experimental data. We show how ARs for B. subtilis are able to capture many sets of genes corresponding to regulated operons in our manually curated network. Additionally, we demonstrate how ARs can be used to help expand or validate the knowledge of the regulatory networks by looking at highly correlated genes in the ARs for which regulatory information is lacking. During this process, we were also able to infer novel stimuli for hypothetical genes by exploring the genome expression metadata relating to experimental

  14. Comprehensive Assignment of Mass Spectral Signatures from Individual Bacillus atrophaeus Spores in Matrix-Free Bioaerosol Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, A; Pitesky, M; Steele, P; Tobias, H; Fergenson, D P; Horn, J; Russell, S C; Czerwieniec, G; Lebrilla, C; Gard, E E; Frank, M

    2004-10-22

    We have conducted studies to fully characterize the mass spectral signature of individual Bacillus atrophaeus, previously known as Bacillus subtilis var niger or Bacillus globigii, spores obtained in matrix-free bioaerosol mass spectrometry (BAMS). Mass spectra of spores grown in unlabeled, {sup 13}C-labeled and {sup 15}N-labeled growth media are used to determine the number of carbon and nitrogen atoms associated with each mass peak. To determine the parent ion structure associated with fragment ions present in the spore spectra, the mass-to-charge (m/z) fragmentation pattern of several chemical standards was obtained. Our results agree with prior assignments of dipicolinic acid, amino acids and calcium complex ions made in the spore mass spectra. Identity of several previously unidentified mass peaks, key to recognition of Bacillus spore by matrix-free BAMS, is revealed. Specifically, a set of fragment peaks in the negative polarity is shown to be consistent with the fragmentation pattern of purine nucleobase containing compounds. The identity of m/z=+74, a marker peak that helps discriminate Bacillus atrophaeus from Bacillus thuringiensis spores grown in rich medium, is surprisingly a non-description, viz. [N{sub 1}C{sub 4}H{sub 12}]{sup +}. A probable precursor molecule for the [N{sub 1}C{sub 4}H{sub 12}]{sup +} ion observed in spore spectra is trimethyl glycine ({sup +}N(CH{sub 3}){sub 3}CH{sub 2}COOH) that produces a m/z=74 peak in presence of dipicolinic acid.

  15. Presence survival spores of Bacillus thuringiensis varieties in grain warehouse

    Directory of Open Access Journals (Sweden)

    Sánchez-Yáñez Juan Manuel

    2016-08-01

    Full Text Available Genus Bacillus thuringiensis (Bt synthesized spores and crystals toxic to pest-insects in agriculture. Bt is comospolitan then possible to isolate some subspecies or varieties from warehouse. The aims of study were: i to isolate Bt varieties from grain at werehouse ii to evaluate Bt toxicity on Spodoptera frugiperda and Shit-ophilus zeamaisese iii to analyze Bt spores persistence in Zea mays grains at werehouse compared to same Bt on grains exposed to sun radiation. Results showed that at werehouse were recovered more than one variety of Bt spores. According to each isolate Bt1 o Bt2 were toxic to S. frugiperda or S. zeamaisese. One those Bt belong to var morrisoni. At werehouse these spores on Z. mays grains surviving more time, while the same spores exposed to boicide sun radiation they died.

  16. Decontamination Options for Drinking Water Contaminated with Bacillus anthracis Spores

    Energy Technology Data Exchange (ETDEWEB)

    Raber, E; Burklund, A

    2010-02-16

    Five parameters were evaluated with surrogates of Bacillus anthracis spores to determine effective decontamination options for use in a contaminated drinking water supply. The parameters were: (1) type of Bacillus spore surrogate (B. thuringiensis or B. atrophaeus); (2) spore concentration in suspension (10{sup 2} to 10{sup 6} spores/ml); (3) chemical characteristics of decontaminant [sodium dicholor-s-triazinetrione dihydrate (Dichlor), hydrogen peroxide, potassium peroxymonosulfate (Oxone), sodium hypochlorite, and VirkonS{reg_sign}]; (4) decontaminant concentration (0.01% to 5%); and (5) decontaminant exposure time (10 min to 24 hr). Results from 162 suspension tests with appropriate controls are reported. Hydrogen peroxide at a concentration of 5%, and Dichlor and sodium hypochlorite at a concentration of 2%, were effective at spore inactivation regardless of spore type tested, spore exposure time, or spore concentration evaluated. This is the first reported study of Dichlor as an effective decontaminant for B. anthracis spore surrogates. Dichlor's desirable characteristics of high oxidation potential, high level of free chlorine, and more neutral pH than that of other oxidizers evaluated appear to make it an excellent alternative. All three oxidizers were effective against B. atrophaeus spores in meeting EPA's biocide standard of greater than a 6 log kill after a 10-minute exposure time and at lower concentrations than typically reported for biocide use. Solutions of 5% VirkonS{reg_sign} and Oxone were less effective decontaminants than other options evaluated in this study and did not meet the EPA's efficacy standard for biocides. Differences in methods and procedures reported by other investigators make quantitative comparisons among studies difficult.

  17. Diverse supramolecular structures formed by self-assembling proteins of the B acillus subtilis spore coat

    OpenAIRE

    2015-01-01

    Summary Bacterial spores (endospores), such as those of the pathogens C lostridium difficile and B acillus anthracis, are uniquely stable cell forms, highly resistant to harsh environmental insults. B acillus subtilis is the best studied spore‐former and we have used it to address the question of how the spore coat is assembled from multiple components to form a robust, protective superstructure. B . subtilis coat proteins (CotY, CotE, CotV and CotW) expressed in E scherichia coli can arrange...

  18. Weak organic acid stress in Bacillus subtilis

    NARCIS (Netherlands)

    ter Beek, A.S.

    2009-01-01

    Weak organic acids are commonly used food preservatives that protect food products from bacterial contamination. A variety of spore-forming bacterial species pose a serious problem to the food industry by causing extensive food spoilage or even food poisoning. Understanding the mechanisms of bacteri

  19. 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.

  20. 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...

  1. Complete Genome Sequence of Bacillus subtilis Strain 29R7-12, a Piezophilic Bacterium Isolated from Coal-Bearing Sediment 2.4 Kilometers below the Seafloor

    Science.gov (United States)

    Wei, Yuli; Cao, Junwei; Kato, Chiaki; Cui, Weicheng

    2017-01-01

    ABSTRACT Here, we report the genome sequence of Bacillus subtilis strain 29R7-12, a piezophilic bacterium isolated from coal-bearing sediment down to ~2.4 km below the ocean floor in the northwestern Pacific. The strain is a Gram-positive spore-forming bacterium, closely related to Bacillus subtilis within the phylum Firmicutes. This is the first complete genome sequence of a Bacillus subtilis strain from the deep biosphere. The genome sequence will provide a valuable resource for comparative studies of microorganisms from the surface and subsurface environments. PMID:28232436

  2. Detection of Bacillus anthracis Spores Using Peptide Functionalized SERS-Active Substrates

    Directory of Open Access Journals (Sweden)

    Atanu Sengupta

    2012-01-01

    Full Text Available The need for portable technologies that can rapidly identify biological warfare agents (BWAs in the field remains an international priority as expressed at the 2011 Biological Weapons Convention. In recent years, the ability of surface-enhanced Raman spectroscopy (SERS to rapidly detect various BWAs at very low concentrations has been demonstrated. However, in the specific case of Bacillus anthracis, differentiation at the species level is required since other bacilli are common in the environment, representing potential false-positive responses. To overcome this limitation, we describe the use of a peptide attached to the SERS-active metal that selectively binds Bacillus anthracis-Sterne as the target analyte. Using this approach, 109  B. anthracis-Sterne spores/mL produced an intense dipicolinic acid spectrum upon the addition of acetic acid, while the same concentration and treatment of B. cereus and B. subtilis did not.

  3. 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.

  4. 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.''

  5. Bacillus subtilis Hfq: A role in chemotaxis and motility

    Indian Academy of Sciences (India)

    CHANDRAKANT B JAGTAP; PRADEEP KUMAR; K KRISHNAMURTHY RAO

    2016-09-01

    Hfq is a global post-transcriptional regulator that modulates the translation and stability of target mRNAs and therebyregulates pleiotropic functions, such as growth, stress, virulence and motility, in many Gram-negative bacteria.However, comparatively little is known about the regulation and function(s) of Hfq in Gram-positive bacteria.Recently, in Bacillus subtilis, a role for Hfq in stationary phase survival has been suggested, although the possibilityof Hfq having an additional role(s) cannot be ruled out. In this study we show that an ortholog of Hfq in B. subtilis isregulated by the stress sigma factor, σB, in addition to the stationary phase sigma factor, σH. We further demonstratethat Hfq positively regulates the expression of flagellum and chemotaxis genes (fla/che) that control chemotaxis andmotility, thus assigning a new function for Hfq in B. subtilis.

  6. Effect of Bacillus subtilis microecological probiotics on livestock breeding

    Directory of Open Access Journals (Sweden)

    Xiaohui ZHOU

    2016-10-01

    Full Text Available As a kind of green and healthy microecologics, Bacillus subtilis could balance the intestinal flora, promote the nutrient absorption and enhance immunity. Microecologics is one of the ideal antibiotics alternative, which are effective in preventing and treating animal disease and promoting the growth and development of the animal. Because of its advantages, such as no toxin side effect and no residual or drug-resistant, microecologics has been used in livestock breeding widely. Here, we concluded the characteristics and mechanism of Bacillus subtilis,elaborated application of microecologics on livestock breeding, discussed its problems and suggested its solved methods. In the end, the future of microecologics was expected in order to provide a reference for subsequent livestock breeding.

  7. Biodegradation of furfural by Bacillus subtilis strain DS3.

    Science.gov (United States)

    Zheng, Dan; Bao, Jianguo; Lu, Jueming; Lv, Quanxi

    2015-07-01

    An aerobic bacterial strain DS3, capable of growing on furfural as sole carbon source, was isolated from actived sludge of wastewater treatment plant in a diosgenin factory after enrichment. Based on morphological physiological tests as well as 16SrDNA sequence and Biolog analyses it was identified as Bacillus subtilis. The study revealed that strain DS3 utilized furfural, as analyzed by high-performance liquid chromatography (HPLC). Under following conditions: pH 8.0, temperature 35 degrees C, 150 rpm and 10% inoculum, strain DS3 showed 31.2% furfural degradation. Furthermore, DS3 strain was found to tolerate furfural concentration as high as 6000 mg(-1). The ability of Bacillus subtilis strain DS3 to degrade furfural has been demonstrated for the first time in the present study.

  8. Bacillus subtilis chromosome organization oscillates between two distinct patterns

    OpenAIRE

    Wang, Xindan; Montero Llopis, Paula; Rudner, David Z.

    2014-01-01

    In bacteria, faithful and efficient DNA segregation is intimately linked to the spatial organization of the chromosome. Two distinct organization patterns have been described for bacterial chromosomes (ori-ter and left-ori-right) that appear to arise from distinct segregation mechanisms. Here, we show that the Bacillus subtilis chromosome oscillates between them during a replication–segregation cycle. Our data further suggest that the highly conserved condensin complex and the parABS partitio...

  9. Genome engineering using a synthetic gene circuit in Bacillus subtilis.

    Science.gov (United States)

    Jeong, Da-Eun; Park, Seung-Hwan; Pan, Jae-Gu; Kim, Eui-Joong; Choi, Soo-Keun

    2015-03-31

    Genome engineering without leaving foreign DNA behind requires an efficient counter-selectable marker system. Here, we developed a genome engineering method in Bacillus subtilis using a synthetic gene circuit as a counter-selectable marker system. The system contained two repressible promoters (B. subtilis xylA (Pxyl) and spac (Pspac)) and two repressor genes (lacI and xylR). Pxyl-lacI was integrated into the B. subtilis genome with a target gene containing a desired mutation. The xylR and Pspac-chloramphenicol resistant genes (cat) were located on a helper plasmid. In the presence of xylose, repression of XylR by xylose induced LacI expression, the LacIs repressed the Pspac promoter and the cells become chloramphenicol sensitive. Thus, to survive in the presence of chloramphenicol, the cell must delete Pxyl-lacI by recombination between the wild-type and mutated target genes. The recombination leads to mutation of the target gene. The remaining helper plasmid was removed easily under the chloramphenicol absent condition. In this study, we showed base insertion, deletion and point mutation of the B. subtilis genome without leaving any foreign DNA behind. Additionally, we successfully deleted a 2-kb gene (amyE) and a 38-kb operon (ppsABCDE). This method will be useful to construct designer Bacillus strains for various industrial applications.

  10. Role of YpeB in Cortex Hydrolysis during Germination of Bacillus anthracis Spores

    OpenAIRE

    2014-01-01

    The infectious agent of the disease anthrax is the spore of Bacillus anthracis. Bacterial spores are extremely resistant to environmental stresses, which greatly hinders spore decontamination efforts. The spore cortex, a thick layer of modified peptidoglycan, contributes to spore dormancy and resistance by maintaining the low water content of the spore core. The cortex is degraded by germination-specific lytic enzymes (GSLEs) during spore germination, rendering the cells vulnerable to common ...

  11. Metabolic engineering of Bacillus subtilis for terpenoid production.

    Science.gov (United States)

    Guan, Zheng; Xue, Dan; Abdallah, Ingy I; Dijkshoorn, Linda; Setroikromo, Rita; Lv, Guiyuan; Quax, Wim J

    2015-11-01

    Terpenoids are the largest group of small-molecule natural products, with more than 60,000 compounds made from isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP). As the most diverse group of small-molecule natural products, terpenoids play an important role in the pharmaceutical, food, and cosmetic industries. For decades, Escherichia coli (E. coli) and Saccharomyces cerevisiae (S. cerevisiae) were extensively studied to biosynthesize terpenoids, because they are both fully amenable to genetic modifications and have vast molecular resources. On the other hand, our literature survey (20 years) revealed that terpenoids are naturally more widespread in Bacillales. In the mid-1990s, an inherent methylerythritol phosphate (MEP) pathway was discovered in Bacillus subtilis (B. subtilis). Since B. subtilis is a generally recognized as safe (GRAS) organism and has long been used for the industrial production of proteins, attempts to biosynthesize terpenoids in this bacterium have aroused much interest in the scientific community. This review discusses metabolic engineering of B. subtilis for terpenoid production, and encountered challenges will be discussed. We will summarize some major advances and outline future directions for exploiting the potential of B. subtilis as a desired "cell factory" to produce terpenoids.

  12. Anaerobic growth of a "strict aerobe" (Bacillus subtilis).

    Science.gov (United States)

    Nakano, M M; Zuber, P

    1998-01-01

    There was a long-held belief that the gram-positive soil bacterium Bacillus subtilis is a strict aerobe. But recent studies have shown that B. subtilis will grow anaerobically, either by using nitrate or nitrite as a terminal electron acceptor, or by fermentation. How B. subtilis alters its metabolic activity according to the availability of oxygen and alternative electron acceptors is but one focus of study. A two-component signal transduction system composed of a sensor kinase, ResE, and a response regulator, ResD, occupies an early stage in the regulatory pathway governing anaerobic respiration. One of the essential roles of ResD and ResE in anaerobic gene regulation is induction of fnr transcription upon oxygen limitation. FNR is a transcriptional activator for anaerobically induced genes, including those for respiratory nitrate reductase, narGHJI.B. subtilis has two distinct nitrate reductases, one for the assimilation of nitrate nitrogen and the other for nitrate respiration. In contrast, one nitrite reductase functions both in nitrite nitrogen assimilation and nitrite respiration. Unlike many anaerobes, which use pyruvate formate lyase, B. subtilis can carry out fermentation in the absence of external electron acceptors wherein pyruvate dehydrogenase is utilized to metabolize pyruvate.

  13. Production of Bioactive Compounds by Bacillus subtilis against Sclerotium rolfsii

    Directory of Open Access Journals (Sweden)

    Nalisha, I.

    2006-01-01

    Full Text Available This study aims to investigate the characteristic of bioactive compound produced by Bacillus subtilis against Sclerotium rolfsii and the influence of additive supplements on the antagonistic activity of B. subtilis. The fact that B. subtilis produced an antifungal substance which has inhibitory effect on wide range of fungi, including S. rolfsii, is well known. To learn the effect of pH, temperature and light condition on the production of antifungal compound, B. subtilis was inoculated in Potato Dextrose Broth at various initial pH, temperatures and light conditions, respectively. This antagonist was found to produce antifungal compound that stable at 80C with 58.3 % inhibition on S. rolfsii. The activity was constant within a wide range of pH (3–11. However, treatment with pH11 lead to higher antifungal activity (31.57 % inhibition and it was also found to produce substance that can endure dark condition (46.24 % inhibition with fungicidal effect on S. rolfsii. A series of experiments also been carried out to enhance the antifungal production by supplementing different carbon source preparation into bacterial liquid culture. B. subtilis were grown in minimal medium containing 1 % of oil palm root, Ganoderma lucidum or chitin, respectively prior to bioassay. Crude culture from oil palm root supplemented culture shown significantly reduction in S. rolfsii growth compared to other carbon source crude culture or the antagonism alone, suggesting that this approach may provide improved biocontrol efficiency.

  14. Tracking the Elusive Function of Bacillus subtilis Hfq.

    Science.gov (United States)

    Rochat, Tatiana; Delumeau, Olivier; Figueroa-Bossi, Nara; Noirot, Philippe; Bossi, Lionello; Dervyn, Etienne; Bouloc, Philippe

    2015-01-01

    RNA-binding protein Hfq is a key component of the adaptive responses of many proteobacterial species including Escherichia coli, Salmonella enterica and Vibrio cholera. In these organisms, the importance of Hfq largely stems from its participation to regulatory mechanisms involving small non-coding RNAs. In contrast, the function of Hfq in Gram-positive bacteria has remained elusive and somewhat controversial. In the present study, we have further addressed this point by comparing growth phenotypes and transcription profiles between wild-type and an hfq deletion mutant of the model Gram-positive bacterium, Bacillus subtilis. The absence of Hfq had no significant consequences on growth rates under nearly two thousand metabolic conditions and chemical treatments. The only phenotypic difference was a survival defect of B. subtilis hfq mutant in rich medium in stationary phase. Transcriptomic analysis correlated this phenotype with a change in the levels of nearly one hundred transcripts. Albeit a significant fraction of these RNAs (36%) encoded sporulation-related functions, analyses in a strain unable to sporulate ruled out sporulation per se as the basis of the hfq mutant's stationary phase fitness defect. When expressed in Salmonella, B. subtilis hfq complemented the sharp loss of viability of a degP hfq double mutant, attenuating the chronic σE-activated phenotype of this strain. However, B. subtilis hfq did not complement other regulatory deficiencies resulting from loss of Hfq-dependent small RNA activity in Salmonella indicating a limited functional overlap between Salmonella and B. subtilis Hfqs. Overall, this study confirmed that, despite structural similarities with other Hfq proteins, B. subtilis Hfq does not play a central role in post-transcriptional regulation but might have a more specialized function connected with stationary phase physiology. This would account for the high degree of conservation of Hfq proteins in all 17 B. subtilis strains whose

  15. 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...

  16. Multigeneration Cross-Contamination of Mail with Bacillus anthracis Spores.

    Directory of Open Access Journals (Sweden)

    Jason Edmonds

    Full Text Available The release of biological agents, including those which could be used in biowarfare or bioterrorism in large urban areas, has been a concern for governments for nearly three decades. Previous incidents from Sverdlosk and the postal anthrax attack of 2001 have raised questions on the mechanism of spread of Bacillus anthracis spores as an aerosol or contaminant. Prior studies have demonstrated that Bacillus atrophaeus is easily transferred through simulated mail handing, but no reports have demonstrated this ability with Bacillus anthracis spores, which have morphological differences that may affect adhesion properties between spore and formite. In this study, equipment developed to simulate interactions across three generations of envelopes subjected to tumbling and mixing was used to evaluate the potential for cross-contamination of B. anthracis spores in simulated mail handling. In these experiments, we found that the potential for cross-contamination through letter tumbling from one generation to the next varied between generations while the presence of a fluidizer had no statistical impact on the transfer of material. Likewise, the presence or absence of a fluidizer had no statistically significant impact on cross-contamination levels or reaerosolization from letter opening.

  17. Multigeneration Cross-Contamination of Mail with Bacillus anthracis Spores

    Science.gov (United States)

    Edmonds, Jason; Lindquist, H. D. Alan; Sabol, Jonathan; Martinez, Kenneth; Shadomy, Sean; Cymet, Tyler; Emanuel, Peter

    2016-01-01

    The release of biological agents, including those which could be used in biowarfare or bioterrorism in large urban areas, has been a concern for governments for nearly three decades. Previous incidents from Sverdlosk and the postal anthrax attack of 2001 have raised questions on the mechanism of spread of Bacillus anthracis spores as an aerosol or contaminant. Prior studies have demonstrated that Bacillus atrophaeus is easily transferred through simulated mail handing, but no reports have demonstrated this ability with Bacillus anthracis spores, which have morphological differences that may affect adhesion properties between spore and formite. In this study, equipment developed to simulate interactions across three generations of envelopes subjected to tumbling and mixing was used to evaluate the potential for cross-contamination of B. anthracis spores in simulated mail handling. In these experiments, we found that the potential for cross-contamination through letter tumbling from one generation to the next varied between generations while the presence of a fluidizer had no statistical impact on the transfer of material. Likewise, the presence or absence of a fluidizer had no statistically significant impact on cross-contamination levels or reaerosolization from letter opening. PMID:27123934

  18. Homolactic fermentation from glucose and cellobiose using Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Martinez Alfredo

    2009-04-01

    Full Text Available Abstract Backgroung Biodegradable plastics can be made from polylactate, which is a polymer made from lactic acid. This compound can be produced from renewable resources as substrates using microorganisms. Bacillus subtilis is a Gram-positive bacterium recognized as a GRAS microorganism (generally regarded as safe by the FDA. B. subtilis produces and secretes different kind of enzymes, such as proteases, cellulases, xylanases and amylases to utilize carbon sources more complex than the monosaccharides present in the environment. Thus, B. subtilis could be potentially used to hydrolyze carbohydrate polymers contained in lignocellulosic biomass to produce chemical commodities. Enzymatic hydrolysis of the cellulosic fraction of agroindustrial wastes produces cellobiose and a lower amount of glucose. Under aerobic conditions, B. subtilis grows using cellobiose as substrate. Results In this study, we proved that under non-aerated conditions, B. subtilis ferments cellobiose to produce L-lactate with 82% of the theoretical yield, and with a specific rate of L-lactate production similar to that one obtained fermenting glucose. Under fermentative conditions in a complex media supplemented with glucose, B. subtilis produces L-lactate and a low amount of 2,3-butanediol. To increase the L-lactate production of this organism, we generated the B subtilis CH1 alsS- strain that lacks the ability to synthesize 2,3-butanediol. Inactivation of this pathway, that competed for pyruvate availability, let a 15% increase in L-lactate yield from glucose compared with the parental strain. CH1 alsS- fermented 5 and 10% of glucose to completion in mineral medium supplemented with yeast extract in four and nine days, respectively. CH1 alsS- produced 105 g/L of L-lactate in this last medium supplemented with 10% of glucose. The L-lactate yield was up to 95% using mineral media, and the optical purity of L-lactate was of 99.5% since B. subtilis has only one gene (lctE that

  19. Studies on the bacterial spore coat 6 effects of alkali extraction on the spore of Bacillus thiaminolyticus.

    Science.gov (United States)

    Minami, J; Ichikawa, T; Kondo, M

    1977-01-01

    Thin sections of the spore of Bacillus thiaminolyticus Matsukawa and Misawa show a characteristic surface structure with five ridges, and a series of three district layers. The outer layer of the spore coat was peeled off by SDS sonic treatment, and than the middle layer was solubilized by alkali extraction of the SDS sonic-treated spore. The spores subjected to these treatments were still refractile, heat resistant, and contained dipicolinic acid, but lost their resistance to mechanical shock.

  20. Control of Initiation of DNA Replication in Bacillus subtilis and Escherichia coli.

    Science.gov (United States)

    Jameson, Katie H; Wilkinson, Anthony J

    2017-01-10

    Initiation of DNA Replication is tightly regulated in all cells since imbalances in chromosomal copy number are deleterious and often lethal. In bacteria such as Bacillus subtilis and Escherichia coli, at the point of cytokinesis, there must be two complete copies of the chromosome to partition into the daughter cells following division at mid-cell during vegetative growth. Under conditions of rapid growth, when the time taken to replicate the chromosome exceeds the doubling time of the cells, there will be multiple initiations per cell cycle and daughter cells will inherit chromosomes that are already undergoing replication. In contrast, cells entering the sporulation pathway in B. subtilis can do so only during a short interval in the cell cycle when there are two, and only two, chromosomes per cell, one destined for the spore and one for the mother cell. Here, we briefly describe the overall process of DNA replication in bacteria before reviewing initiation of DNA replication in detail. The review covers DnaA-directed assembly of the replisome at oriC and the multitude of mechanisms of regulation of initiation, with a focus on the similarities and differences between E. coli and B. subtilis.

  1. Control of Initiation of DNA Replication in Bacillus subtilis and Escherichia coli

    Directory of Open Access Journals (Sweden)

    Katie H. Jameson

    2017-01-01

    Full Text Available Initiation of DNA Replication is tightly regulated in all cells since imbalances in chromosomal copy number are deleterious and often lethal. In bacteria such as Bacillus subtilis and Escherichia coli, at the point of cytokinesis, there must be two complete copies of the chromosome to partition into the daughter cells following division at mid-cell during vegetative growth. Under conditions of rapid growth, when the time taken to replicate the chromosome exceeds the doubling time of the cells, there will be multiple initiations per cell cycle and daughter cells will inherit chromosomes that are already undergoing replication. In contrast, cells entering the sporulation pathway in B. subtilis can do so only during a short interval in the cell cycle when there are two, and only two, chromosomes per cell, one destined for the spore and one for the mother cell. Here, we briefly describe the overall process of DNA replication in bacteria before reviewing initiation of DNA replication in detail. The review covers DnaA-directed assembly of the replisome at oriC and the multitude of mechanisms of regulation of initiation, with a focus on the similarities and differences between E. coli and B. subtilis.

  2. MstX and a putative potassium channel facilitate biofilm formation in Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Matthew E Lundberg

    Full Text Available Biofilms constitute the predominant form of microbial life and a potent reservoir for innate antibiotic resistance in systemic infections. In the spore-forming bacterium Bacillus subtilis, the transition from a planktonic to sessile state is mediated by mutually exclusive regulatory pathways controlling the expression of genes required for flagellum or biofilm formation. Here, we identify mstX and yugO as novel regulators of biofilm formation in B. subtilis. We show that expression of mstX and the downstream putative K+ efflux channel, yugO, is necessary for biofilm development in B. subtilis, and that overexpression of mstX induces biofilm assembly. Transcription of the mstX-yugO operon is under the negative regulation of SinR, a transcription factor that governs the switch between planktonic and sessile states. Furthermore, mstX regulates the activity of Spo0A through a positive autoregulatory loop involving KinC, a histidine kinase that is activated by potassium leakage. The addition of potassium abrogated mstX-mediated biofilm formation. Our findings expand the role of Spo0A and potassium homeostasis in the regulation of bacterial development.

  3. 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.

  4. High-Resolution Spore Coat Architecture and Assembly of Bacillus Spores

    Energy Technology Data Exchange (ETDEWEB)

    Malkin, A J; Elhadj, S; Plomp, M

    2011-03-14

    Elucidating the molecular architecture of bacterial and cellular surfaces and its structural dynamics is essential to understanding mechanisms of pathogenesis, immune response, physicochemical interactions, environmental resistance, and provide the means for identifying spore formulation and processing attributes. I will discuss the application of in vitro atomic force microscopy (AFM) for studies of high-resolution coat architecture and assembly of several Bacillus spore species. We have demonstrated that bacterial spore coat structures are phylogenetically and growth medium determined. We have proposed that strikingly different species-dependent coat structures of bacterial spore species are a consequence of sporulation media-dependent nucleation and crystallization mechanisms that regulate the assembly of the outer spore coat. Spore coat layers were found to exhibit screw dislocations and two-dimensional nuclei typically observed on inorganic and macromolecular crystals. This presents the first case of non-mineral crystal growth patterns being revealed for a biological organism, which provides an unexpected example of nature exploiting fundamental materials science mechanisms for the morphogenetic control of biological ultrastructures. We have discovered and validated, distinctive formulation-specific high-resolution structural spore coat and dimensional signatures of B. anthracis spores (Sterne strain) grown in different formulation condition. We further demonstrated that measurement of the dimensional characteristics of B. anthracis spores provides formulation classification and sample matching with high sensitivity and specificity. I will present data on the development of an AFM-based immunolabeling technique for the proteomic mapping of macromolecular structures on the B. anthracis surfaces. These studies demonstrate that AFM can probe microbial surface architecture, environmental dynamics and the life cycle of bacterial and cellular systems at near

  5. Enhanced secretion of natto phytase by Bacillus subtilis.

    Science.gov (United States)

    Tsuji, Shogo; Tanaka, Kosei; Takenaka, Shinji; Yoshida, Ken-ichi

    2015-01-01

    Phytases comprise a group of phosphatases that trim inorganic phosphates from phytic acid (IP6). In this study, we aimed to achieve the efficient secretion of phytase by Bacillus subtilis. B. subtilis laboratory standard strain 168 and its derivatives exhibit no phytase activity, whereas a natto starter secretes phytase actively. The natto phytase gene was cloned into strain RIK1285, a protease-defective derivative of 168, to construct a random library of its N-terminal fusions with 173 different signal peptides (SPs) identified in the 168 genome. The library was screened to assess the efficiency of phytase secretion based on clear zones around colonies on plates, which appeared when IP6 was hydrolyzed. The pbp SP enhanced the secretion of the natto phytase most efficiently, i.e. twice that of the original SP. Thus, the secreted natto phytase was purified and found to remove up to 3 phosphates from IP6.

  6. An improved protocol for harvesting Bacillus subtilis colony biofilms.

    Science.gov (United States)

    Fuchs, Felix Matthias; Driks, Adam; Setlow, Peter; Moeller, Ralf

    2017-03-01

    Bacterial biofilms cause severe problems in medicine and industry due to the high resistance to disinfectants and environmental stress of organisms within biofilms. Addressing challenges caused by biofilms requires full understanding of the underlying mechanisms for bacterial resistance and survival in biofilms. However, such work is hampered by a relative lack of systems for biofilm cultivation that are practical and reproducible. To address this problem, we developed a readily applicable method to culture Bacillus subtilis biofilms on a membrane filter. The method results in biofilms with highly reproducible characteristics, and which can be readily analyzed by a variety of methods with little further manipulation. This biofilm preparation method simplifies routine generation of B. subtilis biofilms for molecular and cellular analysis, and could be applicable to other microbial systems.

  7. Effect of the electroimmobilization process on Bacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Mogilevich, N.F.; Garbara, S.V.

    1980-11-01

    The culture of Bacillus subtilis 21 was subjected to the action of nonuniform electric field, and the effect of the latter on the bacterial survival and biochemical activity was studied. The action of the field on the cells was shown to depend on the material of a load on which the culture was immobilized. The studied properties of Bac. subtilis 21 did not change when the culture was immobilized on cellulose fiber. About 50 to 60% of the cells died on silica gel under the action of the field; the respiration activity and the rate of hexamethylene diamine destruction did not change. Almost all of the bacterial cells lost their viability upon electroimmobilization on ion-exchange resins. The destructive properties of the culture retained by the field exceed the activity of the control variants.

  8. 14C Analysis of Protein Extracts from Bacillus Spores

    Science.gov (United States)

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

    2014-01-01

    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 F14C (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 F14C values of biological materials and has been used to date the synthesis of biomaterials over the bomb pulse era (1955 to present). The F14C 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 F14C 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 14C 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 14C bomb-pulse dating. Since media is contemporary, 14C bomb-pulse dating of isolated soluble proteins can be used to distinguish between historical archives of bioagents and those produced from recent media. PMID:24814329

  9. Biocontrol Activity of Bacillus subtilis Isolated from Agaricus bisporus Mushroom Compost Against Pathogenic Fungi.

    Science.gov (United States)

    Liu, Can; Sheng, Jiping; Chen, Lin; Zheng, Yanyan; Lee, David Yue Wei; Yang, Yang; Xu, Mingshuang; Shen, Lin

    2015-07-08

    Bacillus subtilis strain B154, isolated from Agaricus bisporus mushroom compost infected by red bread mold, exhibited antagonistic activities against Neurospora sitophila. Antifungal activity against phytopathogenic fungi was also observed. The maximum antifungal activity was reached during the stationary phase. This antifungal activity was stable over a wide pH and temperature range and was not affected by proteases. Assay of antifungal activity in vitro indicated that a purified antifungal substance could strongly inhibit mycelia growth and spore germination of N. sitophila. In addition, treatment with strain B154 in A. bisporus mushroom compost infected with N. sitophila significantly increased the yield of bisporus mushrooms. Ultraviolet scan spectroscopy, tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis, matrix-associated laser desorption ionization time-of-flight mass spectrometry, and electrospray ionization tandem mass spectrometry analyses revealed a molecular weight consistent with 1498.7633 Da. The antifungal compound might belong to a new type of lipopeptide fengycin.

  10. Spore Cortex Hydrolysis Precedes Dipicolinic Acid Release during Clostridium difficile Spore Germination

    OpenAIRE

    2015-01-01

    Bacterial spore germination is a process whereby a dormant spore returns to active, vegetative growth, and this process has largely been studied in the model organism Bacillus subtilis. In B. subtilis, the initiation of germinant receptor-mediated spore germination is divided into two genetically separable stages. Stage I is characterized by the release of dipicolinic acid (DPA) from the spore core. Stage II is characterized by cortex degradation, and stage II is activated by the DPA released...

  11. [Expression of N domain of chromogranin A in Bacillus subtilis and its antifungal activity].

    Science.gov (United States)

    Li, Rui-Fang; Lou, Jin-Xian; Zhang, Tian-Yuan

    2004-03-01

    Chromogranin A (CGA) is a soluble protein existed in most secreted cells and neurons. It was recently found that the bovine CGA N terminal region has vasoinhibitory, antibacterial and antifungal activities. Since the need for effective antifungal agents increases in parallel with the expanding number of immunocompromised patients at risk for fungal infections, it becomes imperative to find antifungal compounds with low toxicity toward mammalian cells. To study the antifungal activity of CGA N terminal region, the DNA fragment encoding for the N terminal 1-76 amino acid sequence (CGA1-76) of human CGA was amplified by PCR technique. After DNA sequence analysis, the amplified DNA fragment was cloned into the Bacillus subtilis inducible and expression vector pSBPTQ constructed in this study and the resultant plasmid pSVTQ was then transformed into triple-protease deficient Bacillus subtilis strain DB403 competent cells. The transformants was screened on LB plates containing 10 microg/mL kanamycin. The positive transformant DB403 (pSVTQ) was grown on kanmycin containing 2 x MSR medium and sucrose was added to 2% final concentration for induction after 2h cultivation. The culture supernatant was used to run SDS-PAGE. The result of SDS-PAGE showed that the CGA1-76 was expressed by sucrose induction and the expressed product secreted into the medium with a yield of 5 mg/L. The expressed product reacts specifically with mouse anti CGA47-68 monoclonal antibody. The antifungal activity of the expressed product was examined by adding the culture supernatant to the fungal spore or Candida albican suspensions at appropriate proportion and found that the recombinant human CGA1-76 produced in Bacillus subtilis inhibits the growth of Fusarium sp. Alternaria sp. and Candida albican at the concerntration of 4 micromol/L. These results demonstrate that human CGA1-76 has expressed in Bacillus subtilis and the expressed product is immunogenic and has the antifungal activity.

  12. 40 CFR 180.1209 - Bacillus subtilis strain QST 713; 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 strain QST 713... RESIDUES IN FOOD Exemptions From Tolerances § 180.1209 Bacillus subtilis strain QST 713; exemption from the... the microbial pesticide Bacillus subtilis strain QST 713 when used in or on all food commodities....

  13. 40 CFR 180.1243 - Bacillus subtilis var. amyloliquefaciens strain FZB24; exemption from the requirement of a...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus subtilis var... EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1243 Bacillus subtilis... the requirement of a tolerance for residues of the Bacillus subtilis var. amyloliquefaciens...

  14. Molecular cloning with bifunctional plasmid vectors in Bacillus subtilis: isolation of a spontaneous mutant of Bacillus subtilis with enhanced transformability for Escherichia coli-propagated chimeric plasmid DNA.

    OpenAIRE

    Ostroff, G. R.; Pène, J. J.

    1983-01-01

    Hybrid plasmid DNA cloned in Escherichia coli undergoes deletions when returned to competent Bacillus subtilis, even in defined restriction and modification mutants of strain 168. We have isolated a mutant of B. subtilis MI112 which is stably transformed at high frequency by chimeric plasmid DNA propagated in E. coli.

  15. Achieving Consistent Multiple Daily Low-Dose Bacillus anthracis Spore Inhalation Exposures in the Rabbit Model

    Science.gov (United States)

    2012-06-13

    daily low-dose Bacillus anthracis spore inhalation exposures in the rabbit model Roy E. Barnewall 1, Jason E. Comer 1, Brian D. Miller 1, BradfordW...multiple exposure days. Keywords: Bacillus anthracis , inhalation exposures, low-dose, subchronic exposures, spores, anthrax, aerosol system INTRODUCTION... Bacillus Anthracis Spore Inhalation Exposures In The Rabbit Model 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) 5d

  16. Unlocking the Sporicidal Potential of Ethanol: Induced Sporicidal Activity of Ethanol against Clostridium difficile and Bacillus Spores under Altered Physical and Chemical Conditions

    Science.gov (United States)

    Nerandzic, Michelle M.; Sunkesula, Venkata C. K.; C., Thriveen Sankar; Setlow, Peter; Donskey, Curtis J.

    2015-01-01

    Background Due to their efficacy and convenience, alcohol-based hand sanitizers have been widely adopted as the primary method of hand hygiene in healthcare settings. However, alcohols lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We hypothesized that sporicidal activity could be induced in alcohols through alteration of physical or chemical conditions that have been shown to degrade or allow penetration of spore coats. Principal Findings Acidification, alkalinization, and heating of ethanol induced rapid sporicidal activity against C. difficile, and to a lesser extent Bacillus thuringiensis and Bacillus subtilis. The sporicidal activity of acidified ethanol was enhanced by increasing ionic strength and mild elevations in temperature. On skin, sporicidal ethanol formulations were as effective as soap and water hand washing in reducing levels of C. difficile spores. Conclusions These findings demonstrate that novel ethanol-based sporicidal hand hygiene formulations can be developed through alteration of physical and chemical conditions. PMID:26177038

  17. 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.

  18. Inside the Meteorite — Bacterial Spore Survival After Exposure to Galactic Cosmic Radiation

    Science.gov (United States)

    Moeller, R.; Berger, T.; Matthiä, D.; Okayasu, R.; Kato, T.; Kitamura, H.; Reitz, G.

    2010-04-01

    Based on their unique resistance to various space parameters, bacterial spores are one of the model systems used for astrobiological studies. In our research, we studied the response of Bacillus subtilis spores to the exposure of galactic cosmic radiation.

  19. Inhibition of Bacillus cereus spore outgrowth and multiplication by chitosan.

    Science.gov (United States)

    Mellegård, Hilde; From, Cecilie; Christensen, Bjørn E; Granum, Per E

    2011-10-03

    Bacillus cereus is an endospore-forming bacterium able to cause food-associated illness. Different treatment processes are used in the food industry to reduce the number of spores and thereby the potential of foodborne disease. Chitosan is a polysaccharide with well-documented antibacterial activity towards vegetative cells. The activity against bacterial spores, spore germination and subsequent outgrowth and growth (the latter two events hereafter denoted (out)growth), however, is poorly documented. By using six different chitosans with defined macromolecular properties, we evaluated the effect of chitosan on Bacillus cereus spore germination and (out)growth using optical density assays and a dipicolinic acid release assay. (Out)growth was inhibited by chitosan, but germination was not. The action of chitosan was found to be concentration-dependent and also closely related to weight average molecular weight (M(w)) and fraction of acetylation (F(A)) of the biopolymer. Chitosans of low acetylation (F(A)=0.01 or 0.16) inhibited (out)growth more effectively than higher acetylated chitosans (F(A)=0.48). For the F(A)=0.16 chitosans with medium (56.8kDa) and higher M(w) (98.3kDa), a better (out)growth inhibition was observed compared to low M(w) (10.6kDa) chitosan. The same trend was not evident with chitosans of 0.48 acetylation, where the difference in activity between the low (19.6kDa) and high M(w) (163.0kDa) chitosans was only minor. In a spore test concentration corresponding to 10(2)-10(3)CFU/ml (spore numbers relevant to food), less chitosan was needed to suppress (out)growth compared to higher spore numbers (equivalent to 10(8)CFU/ml), as expected. No major differences in chitosan susceptibility between three different strains of B. cereus were detected. Our results contribute to a better understanding of chitosan activity towards bacterial spore germination and (out)growth.

  20. PRODUKSI ANTIBIOTIKA OLEH Bacillus subtilis M10 DALAM MEDIA UREA-SORBITOL

    Directory of Open Access Journals (Sweden)

    Supartono Supartono

    2012-04-01

    Full Text Available PRODUCTION OF ANTIBIOTICS BY Bacillus subtilis M10 IN UREA-SORBITOL MEDIUM. Infection diseases still become the main health problems that suffered by people in Indonesia. Besides, there were many pathogen bacteria found to be resistant to the some antibiotics. Therefore, the efforts to get a new antibiotic require to be done continuously. A new local strain of Bacillus subtilis BAC4 has been known producing an antibiotic that inhibit Serratia marcescens ATCC 27117 growth. To make efficient the local strain, mutation on Bacillus subtilis BAC4 was done by using acridine orange and a mutant cell of Bacillus subtilis M10 that overproduction for producing antibiotic was obtained. Nevertheless, the production kinetics of antibiotic by this mutant has not been reported. The objective of this research was to study the production kinetics of antibiotic by Bacillus subtilis M10 mutant. The production of antibiotic was conducted using batch fermentation and antibiotic assay was performed with agar absorption method using Serratia marcescens ATCC 27117 as bacteria assay. Research result provided that Bacillus subtilis M10 mutant with overproduction of antibiotic produced an antibiotic since 8th hour’s fermentation and optimum of it production was at 14th hours after inoculation.  Penyakit infeksi masih menjadi masalah yang utama diderita oleh masyarakat Indonesia. Di samping itu, banyak bakteri patogen yang ditemukan resisten terhadap beberapa antibiotika. Oleh karena itu, upaya-upaya untuk mendapatkan antibiotika baru perlu dilakukan secara terus-menerus. Suatu galur lokal baru Bacillus subtilis BAC4 teridentifikasi memproduksi senyawa antibiotika yang menghambat pertumbuhan Serratia marcescens ATCC27117. Untuk memberdayakan galur tersebut, terhadap Bacillus subtilis BAC4 dilakukan mutasi dengan larutan akridin oranye dan diperoleh mutan Bacillus subtilis M10 yang memproduksi antibiotika berlebihan. Namun, kinetika produksi antibiotika oleh Bacillus

  1. 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.

  2. Adhesion of Spores of Bacillus thuringiensis on a Planar Surface

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Eunhyea [Georgia Institute of Technology; Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Lee, Ida [University of Tennessee, Knoxville (UTK); Joy, David Charles [ORNL; Palumbo, Anthony Vito [ORNL; Tsouris, Costas [ORNL

    2010-01-01

    Adhesion of spores of Bacillus thuringiensis (Bt) and spherical silica particles on surfaces was experimentally and theoretically investigated in this study. Topography analysis via atomic force microscopy (AFM) and electron microscopy indicates that Bt spores are rod shaped, {approx}1.3 {mu}m in length and {approx}0.8 {mu}m in diameter. The adhesion force of Bt spores and silica particles on gold-coated glass was measured at various relative humidity (RH) levels by AFM. It was expected that the adhesion force would vary with RH because the individual force components contributing to the adhesion force depend on RH. The adhesion force between a particle and a planar surface in atmospheric environments was modeled as the contribution of three major force components: capillary, van der Waals, and electrostatic interaction forces. Adhesion force measurements for Bt spore (silica particle) and the gold surface system were comparable with calculations. Modeling results show that there is a critical RH value, which depends on the hydrophobicity of the materials involved, below which the water meniscus does not form and the contribution of the capillary force is zero. As RH increases, the van der Waals force decreases while the capillary force increases to a maximum value.

  3. Isolation and Identification of the Antimicrobial Substance Produced by Bacillus subtilis fmbR%Bacillus subtilis fmbR抗菌物质的分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    别小妹; 陆兆新; 吕凤霞; 赵海珍; 杨胜远; 孙力军

    2006-01-01

    [目的]对Bacillus subtilis fmbR产生的抗菌物质进行分离和鉴定研究,以确定抗菌物质的组成和结构.[方法]采用HPLC和TLC层析对Bacillus subtilis fmbR抗菌物质进行分离纯化,通过ESI-MS和MALDI-MS分析对抗菌物质的组成和结构进行初步鉴定.[结果]HPLC层析表明了Bacillus subtilis fmbR抗菌物质含有保留时间与surfactin相似的成分.TLC层析和原位酸解证明了Bacillus subtilis fmbR抗菌物质含有闭合肽键类的物质,其中之一为相对迁移率Rf与标样surfactin相近的组分.采用ESI-MS分析检测到Bacillus subtilis fmbR抗菌物质含有分子量与surfactinA相同的m/z1009.1、m/z1023.2 和m/z1037.0等3种同系物;通过MALDI-MS分析获得[M+H]+为m/z 3403.95抗菌物质,该物质分子量与Bacillus subtilis 168产生的细菌素subtilosin的m/z3403.3 相同.[结论]Bacillus subtilis fmbR抗菌物质由C13~C15的3种surfactinA同系物和一种羊毛硫抗生素subtilosin组成.

  4. Bacillus subtilis homologs of MviN (MurJ), the putative Escherichia coli lipid II flippase, are not essential for growth.

    Science.gov (United States)

    Fay, Allison; Dworkin, Jonathan

    2009-10-01

    Although peptidoglycan synthesis is one of the best-studied metabolic pathways in bacteria, the mechanism underlying the membrane translocation of lipid II, the undecaprenyl-disaccharide pentapeptide peptidoglycan precursor, remains mysterious. Recently, it was proposed that the essential Escherichia coli mviN gene encodes the lipid II flippase. Bacillus subtilis contains four proteins that are putatively homologous to MviN, including SpoVB, previously reported to be necessary for spore cortex peptidoglycan synthesis during sporulation. MviN complemented the sporulation defect of a DeltaspoVB mutation, and SpoVB and another of the B. subtilis homologs, YtgP, complemented the growth defect of an E. coli strain depleted for MviN. Thus, these B. subtilis proteins are likely to be MviN homologs. However, B. subtilis strains lacking these four proteins have no defects in growth, indicating that they likely do not serve as lipid II flippases in this organism.

  5. [Features of Bacillus subtilis IMB B-7023 and its streptomycin-resistant strain].

    Science.gov (United States)

    Roĭ, A A; Iatsenko, I P; Gordienko, A S; Kurdish, I K

    2011-01-01

    Features of phosphate-mobilizing bacteria Bacillus subtilis IMB B-7023 and its streptomycin-resistant strain were investigated. While cultivated in medium with glucose and glycerophosphate, the growth rate of the antibiotic-marked strain was approximately similar to this parameter for Bacillus subtilis IMB B-7023 but cell sizes were 1.3-fold less. Both strains significantly stimulated the germinating of plant seeds, attached to their roots, and insignificantly differed in antagonistic activity toward phytopathogens and quantitative content of cell fatty acids and phosphatase activity. Streptomycin-resistant strain may be used for monitoring of Bacillus subtilis introduced to agroecosystem.

  6. Isolation and characterization of radioresistant mutants in Bacillus subtilis and Bacillus thuringiensis

    Energy Technology Data Exchange (ETDEWEB)

    Kalinin, V.L.; Petrov, V.N.; Petrova, T.M. (AN SSSR, Leningrad. Inst. Yadernoj Fiziki)

    Vegetative cells of Bac. thuringiensis var. galleriae (the wild-type strain 351) are much more sensitive to lethal effects of UV light and /sup 60/Co-..gamma..-rays than those of Bac. subtilis (the wild-type strain 168). This difference is less pronounced for spores of these strains. By means of repeated ..gamma..-irradiation-regrowth cycles radioresistant mutants of Bac. thuringiensis Gamsup(r) 14 and Bac. subtilis Gamsup(r) 9 were selected. The vegetative cells of these mutants are correspondingly 19 times and 3.9 times more resistant to lethal effects of ..gamma..-radiation than the cells of the parental strains. The resistance of the Gamsup(r) mutant cells to lethal effects of UV light and H/sub 2/O/sub 2/ is also increased. The spores of the Gamsup(r) 14 mutant are 1.5-1.7 times more resistant to ..gamma..-radiation and UV light than the wild-type spores. The radioresistant mutants and the parental strains do not vary in their capacity for host-cell reactivation of UV- or ..gamma..-irradiated phages Tg13 and 105.

  7. Influence of nano-MgO particle size on bactericidal action against Bacillus subtilis var. niger

    Institute of Scientific and Technical Information of China (English)

    HUANG Lei; LI Dianqing; LIN Yanjun; David G.Evans; DUAN Xue

    2005-01-01

    Nano-MgO with various particle sizes, synthesized by different methods using Mg(NO3)2·6H2O, Na2CO3, Na2SO4, urea and ammonia solution as reactants, was used to carry out bactericidal experiments on Bacillus subtilis var. niger. The results were compared with the effect of TiO2, a common kind of photocatalytic material. The materials were characterized by X-ray Diffraction (XRD), Transmission Electron Microscopy (TEM), low temperature N2 adsorption-desorption measurements and FT-IR, and the results showed that the bactericidal ability of MgO increases with decreasing particle size. Nano-MgO and an interior wall-paint containing the material have better bactericidal effects than nono-TiO2 in both presence and absence of light. The bactericidal mechanism is discussed. The surface of MgO can generate high concentrations of which is highly active and can react with the peptide linkages in the coating walls of the spores. The spores are destroyed by the resulting damage to their structure.

  8. Purification and characterization of a novel antifungal protein from Bacillus subtilis strain B29"

    Institute of Scientific and Technical Information of China (English)

    Jing LI; Qian YANG; Li-hua ZHAO; Shu-mei ZHANG; Yu-xia WANG; Xiao-yu ZHAO

    2009-01-01

    An antifungal protein was isolated from a culture of Bacillus subtilis strain B29. The isolation procedure comprised ion exchange chromatography on diethylaminoethyl (DEAE)-52 cellulose and gel filtration chromatography on Bio-Gel P-100.The protein was absorbed on DEAE-cellulose and Bio-Gel P-100. The purified antifungal fraction was designated as B29I, with a molecular mass of 42.3 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), pl value 5.69 by isoelectric focusing (IEF)-PAGE, and 97.81% purity by high performance liquid chromatography (HPLC). B29I exhibited in-hibitory activity on mycelial growth in Fusarium oxysporum, Rhizoctonia solani, Fusarium moniliforme, and Sclerotinia scle-rotiorum. The 50% inhibitory concentrations (IC50) of its antifungal activity toward Fusarium oxysporum and Rhizoctonia solani were 45 and 112 μmol/L, respectively. B291 also demonstrated an inhibitory effect on conidial spore germination of Fusarium oxysporum and suppression of germ-tube elongation, and induced distortion, tumescence, and rupture of a portion of the germi-nated spores.

  9. Bacillus tequilensis sp. nov., isolated from a 2000-year-old Mexican shaft-tomb, is closely related to Bacillus subtilis.

    Science.gov (United States)

    Gatson, Joshua W; Benz, Bruce F; Chandrasekaran, Chitra; Satomi, Masataka; Venkateswaran, Kasthuri; Hart, Mark E

    2006-07-01

    A Gram-positive, spore-forming bacillus was isolated from a sample taken from an approximately 2000-year-old shaft-tomb located in the Mexican state of Jalisco, near the city of Tequila. Tentative identification using conventional biochemical analysis consistently identified the isolate as Bacillus subtilis. DNA isolated from the tomb isolate, strain 10b(T), and closely related species was used to amplify a Bacillus-specific portion of the highly conserved 16S rRNA gene and an internal region of the superoxide dismutase gene (sodA(int)). Trees derived from maximum-likelihood methods applied to the sodA(int) sequences yielded non-zero branch lengths between strain 10b(T) and its closest relative, whereas a comparison of a Bacillus-specific 546 bp amplicon of the 16S rRNA gene demonstrated 99 % similarity with B. subtilis. Although the 16S rRNA gene sequences of strain 10b(T) and B. subtilis were 99 % similar, PFGE of NotI-digested DNA of strain 10b(T) revealed a restriction profile that was considerably different from those of B. subtilis and other closely related species. Whereas qualitative differences in whole-cell fatty acids were not observed, significant quantitative differences were found to exist between strain 10b(T) and each of the other closely related Bacillus species examined. In addition, DNA-DNA hybridization studies demonstrated that strain 10b(T) had a relatedness value of less than 70 % with B. subtilis and other closely related species. Evidence from the sodA(int) sequences, whole-cell fatty acid profiles and PFGE analysis, together with results from DNA-DNA hybridization studies, justify the classification of strain 10b(T) as representing a distinct species, for which the name Bacillus tequilensis sp. nov. is proposed. The type strain is 10b(T) (=ATCC BAA-819(T)=NCTC 13306(T)).

  10. 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

  11. Safety evaluation of a xylanase expressed in Bacillus subtilis.

    Science.gov (United States)

    Harbak, L; Thygesen, H V

    2002-01-01

    A programme of studies was conducted to establish the safety of a xylanase expressed in a self-cloned strain of Bacillus subtilis to be used as a processing aid in the baking industry. To assess acute and subchronic oral toxicity, rat feeding studies were conducted. In addition, the potential of the enzyme to cause mutagenicity and chromosomal aberrations was assessed in microbial and tissue culture in vitro studies. Acute and subchronic oral toxicity was not detected at the highest dose recommended by OECD guidelines. There was no evidence of mutagenic potential or chromosomal aberrations. Furthermore, the organism used for production of the xylanase is already accepted as safe by several major national regulatory agencies.

  12. The ESX system in Bacillus subtilis mediates protein secretion.

    Directory of Open Access Journals (Sweden)

    Laura A Huppert

    Full Text Available Esat-6 protein secretion systems (ESX or Ess are required for the virulence of several human pathogens, most notably Mycobacterium tuberculosis and Staphylococcus aureus. These secretion systems are defined by a conserved FtsK/SpoIIIE family ATPase and one or more WXG100 family secreted substrates. Gene clusters coding for ESX systems have been identified amongst many organisms including the highly tractable model system, Bacillus subtilis. In this study, we demonstrate that the B. subtilis yuk/yue locus codes for a nonessential ESX secretion system. We develop a functional secretion assay to demonstrate that each of the locus gene products is specifically required for secretion of the WXG100 virulence factor homolog, YukE. We then employ an unbiased approach to search for additional secreted substrates. By quantitative profiling of culture supernatants, we find that YukE may be the sole substrate that depends on the FtsK/SpoIIIE family ATPase for secretion. We discuss potential functional implications for secretion of a unique substrate.

  13. Inactivation of Bacillus Subtilis by Atomic Oxygen Radical Anion

    Institute of Scientific and Technical Information of China (English)

    LI Longchun; WANG Lian; YU Zhou; LV Xuanzhong; LI Quanxin

    2007-01-01

    UAtomic oxygen radical anion (O- ) is one of the most active oxygen species, and has extremely high oxidation ability toward small-molecules of hydrocarbons. However, to our knowledge, little is known about the effects of O- on cells of micro-organisms. This work showed that O- could quickly react with the Bacillus subtilis cells and seriously damage the cell walls a s well as their other contents, leading to a fast and irreversible inactivation. SEM micrographs revealed that the cell structures were dramatically destroyed by their exposure to O-. The inactivation efficiencies of B. subtilis depend on the O-- intensity, the initial population of cells and the treatment temperature, but not on the pH in the range of our investigation. For a cell concentration of 106 cfu/ml, the number of survived cells dropped from 106 cfu/ml to 103 cfu/ml after about five-minute irradiation by an O- flux in an intensity of 233 nA/cm2 under a dry argon environment (30 ℃, 1 atm, exposed size: 1.8 cm2). The inactivation mechanism of micro-organisms induced by O- is also discussed.

  14. Inhibition of Bacillus subtilis growth and sporulation by threonine.

    Science.gov (United States)

    Lamb, D H; Bott, K F

    1979-01-01

    A 1-mg/ml amount of threonine (8.4 mM) inhibited growth and sporulation of Bacillus subtilis 168. Inhibition of sporulation was efficiently reversed by valine and less efficiently by pyruvate, arginine, glutamine, and isoleucine. Inhibition of vegetative growth was reversed by asparate and glutamate as well as by valine, arginine, or glutamine. Cells in minimal growth medium were inhibited only transiently by very high concentrations of threonine, whereas inhibition of sporulation was permanent. Addition of threonine prevented the normal increase in alkaline phosphatase and reduced the production of extracellular protease by about 50%, suggesting that threonine blocked the sporulation process relatively early. 2-Ketobutyrate was able to mimic the effect of threonine on sporulation. Sporulation in a strain selected for resistance to azaleucine was partially resistant. Seventy-five percent of the mutants selected for the ability to grow vegetatively in the presence of high threonine concentrations were found to be simultaneously isoleucine auxotrophs. In at least one of these mutants, the threonine resistance phenotpye could not be dissociated from the isoleucine requirement by transformation. This mutation was closely linked to a known ilvA mutation (recombination index, 0.16). This strain also had reduced intracellular threonine deaminase activity. These results suggest that threonine inhibits B. subtilis by causing valine starvation.

  15. Comparative proteome analysis of two antagonist Bacillus subtilis strains.

    Science.gov (United States)

    Zhang, C X; Zhao, X; Han, F; Yang, M F; Chen, H; Chida, T; Shen, S H

    2009-04-01

    Natural wild-type strains of Bacillus subtilis are extensively used in agriculture as biocontrol agents for plants. This study examined two antagonist B. subtilis strains, KB-1111 and KB-1122, and the results illustrated that KB-1122 was a more potent inhibitor of the indicator pathogen than KB- 1111. Thus, to investigate the intrinsic differences between the two antagonist strains under normal culture conditions, samples of KB-1111 and KB-1122 were analyzed using MALDI-TOF-MS. The main differences were related to 20 abundant intracellular and 17 extracellular proteins. When searching the NCBI database, a number of the differentially expressed proteins were identified, including 11 cellular proteins and 10 secretory proteins. Among these proteins, class III stress-response-related ATPase, aconitate hydratase, alpha-amylase precursor, and a secretory protein, endo-1, 4-beta-glucanase, were differentially expressed by the two strains. These results are useful to comprehend the intrinsic differences between the antagonism of KB-1111 and KB-1122.

  16. The Exosporium Layer of Bacterial Spores: a Connection to the Environment and the Infected Host

    OpenAIRE

    2015-01-01

    Much of what we know regarding bacterial spore structure and function has been learned from studies of the genetically well-characterized bacterium Bacillus subtilis. Molecular aspects of spore structure, assembly, and function are well defined. However, certain bacteria produce spores with an outer spore layer, the exosporium, which is not present on B. subtilis spores. Our understanding of the composition and biological functions of the exosporium layer is much more limited than that of oth...

  17. Setting risk-informed environmental standards for Bacillus anthracis spores.

    Science.gov (United States)

    Hong, Tao; Gurian, Patrick L; Ward, Nicholas F Dudley

    2010-10-01

    In many cases, human health risk from biological agents is associated with aerosol exposures. Because air concentrations decline rapidly after a release, it may be necessary to use concentrations found in other environmental media to infer future or past aerosol exposures. This article presents an approach for linking environmental concentrations of Bacillus. anthracis (B. anthracis) spores on walls, floors, ventilation system filters, and in human nasal passages with human health risk from exposure to B. anthracis spores. This approach is then used to calculate example values of risk-informed concentration standards for both retrospective risk mitigation (e.g., prophylactic antibiotics) and prospective risk mitigation (e.g., environmental clean up and reoccupancy). A large number of assumptions are required to calculate these values, and the resulting values have large uncertainties associated with them. The values calculated here suggest that documenting compliance with risks in the range of 10(-4) to 10(-6) would be challenging for small diameter (respirable) spore particles. For less stringent risk targets and for releases of larger diameter particles (which are less respirable and hence less hazardous), environmental sampling would be more promising.

  18. Regiospecific Addition of Uracil to Acrylates Catalyzed by Alkaline Protease from Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    Ying CAI; Jian Yi WU; Na WANG; Xiao Feng SUN; Xian Fu LIN

    2004-01-01

    Michael addition reactions of uracil to acrylates were catalyzed by an alkaline protease from Bacillus subtilis in dimethyl sulfoxide at 55 ℃ for 72 h. The adducts were determined by TLC, IR and 1H NMR.

  19. Bacillus subtilis HJ18-4 from traditional fermented soybean food inhibits Bacillus cereus growth and toxin-related genes.

    Science.gov (United States)

    Eom, Jeong Seon; Lee, Sun Young; Choi, Hye Sun

    2014-11-01

    Bacillus subtilis HJ18-4 isolated from buckwheat sokseongjang, a traditional Korean fermented soybean food, exhibits broad-spectrum antimicrobial activity against foodborne pathogens, including Bacillus cereus. In this study, we investigated the antibacterial efficacy and regulation of toxin gene expression in B. cereus by B. subtilis HJ18-4. Expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM) was downregulated by B. subtilis HJ18-4, which also exhibited strong antibacterial activity against B. cereus. We also found that water extracts of soy product fermented with B. subtilis HJ18-4 significantly inhibited the growth of B. cereus and toxin expression. These results indicate that B. subtilis HJ18-4 could be used as an antimicrobial agent to control B. cereus in the fermented soybean food industry. Our findings also provide an opportunity to develop an efficient biological control agent against B. cereus.

  20. Biosynthesis of Active Bacillus subtilis Urease in the Absence of Known Urease Accessory Proteins

    OpenAIRE

    Kim, Jong Kyong; Mulrooney, Scott B.; Hausinger, Robert P.

    2005-01-01

    Bacillus subtilis contains urease structural genes but lacks the accessory genes typically required for GTP-dependent incorporation of nickel. Nevertheless, B. subtilis was shown to possess a functional urease, and the recombinant enzyme conferred low levels of nickel-dependent activity to Escherichia coli. Additional investigations of the system lead to the suggestion that B. subtilis may use unidentified accessory proteins for in vivo urease activation.

  1. NanoSIMS analysis of Bacillus spores for forensics

    Energy Technology Data Exchange (ETDEWEB)

    Weber, P K; Davisson, M L; Velsko, S P

    2010-02-23

    The threat associated with the potential use of radiological, nuclear, chemical and biological materials in terrorist acts has resulted in new fields of forensic science requiring the application of state-of-the-science analytical techniques. Since the anthrax letter attacks in the United States in the fall of 2001, there has been increased interest in physical and chemical characterization of bacterial spores. While molecular methods are powerful tools for identifying genetic differences, other methods may be able to differentiate genetically identical samples based on physical and chemical properties, as well as provide complimentary information, such as methods of production and approximate date of production. Microanalysis has the potential to contribute significantly to microbial forensics. Bacillus spores are highly structured, consisting of a core, cortex, coat, and in some species, an exosporium. This structure provides a template for constraining elemental abundance differences at the nanometer scale. The primary controls on the distribution of major elements in spores are likely structural and physiological. For example, P and Ca are known to be abundant in the spore core because that is where P-rich nucleic acids and Cadipicolinic acid are located, respectively. Trace elements are known to bind to the spore coat but the controls on these elements are less well understood. Elemental distributions and abundances may be directly related to spore production, purification and stabilization methodologies, which are of particular interest for forensic investigation. To this end, we are developing a high-resolution secondary ion mass spectrometry method using a Cameca NanoSIMS 50 to study the distribution and abundance of trace elements in bacterial spores. In this presentation we will review and compare methods for preparing and analyzing samples, as well as review results on the distribution and abundance of elements in bacterial spores. We use NanoSIMS to

  2. [Joint cultivation of Bacillus subtilis and Escherichia coli strains promising for obtaining complex probiotic].

    Science.gov (United States)

    Tsaruk'ianova, I G; Osadchaia, A I

    2007-01-01

    The ability of joint cultivation of Bacillus subtilis UCM B-5007 and Escherichia coli M-17 in subsurface conditions has been studied. These strains are available for creation of a new complex probiotic. Symbiotic relationships between these microorganisms were proved. Bacillus subtilis and Escherichia coli strains use different growth "strategy". The most optimum ratio of cultures (1:1) for growth, biomass accumulation, and for antagonism to test-cultures has been chosen.

  3. Tryptophan provision by dietary supplementation of a Bacillus subtilis mutant strain in piglets

    DEFF Research Database (Denmark)

    Torres-Pitarch, A; Nielsen, B.; Canibe, Nuria

    2015-01-01

    Supplementing Bacillus (B.) subtilis mutants selected to overproduce a specific amino acid (AA) may be an alternative method to provide essential AA in pig diets. Two experiments on a B. subtilis strain selected to overproduce Trp were conducted using 8-kg pigs fed Trp-deficient diets for 20 d. B...

  4. Biochemical properties and three-dimensional structures of two extracellular lipolytic enzymes from Bacillus subtilis

    NARCIS (Netherlands)

    Eggert, Thorsten; Pouderoyen, Gertie van; Pencreac’h, Gaëlle; Douchet, Isabelle; Verger, Robert; Dijkstra, Bauke W.; Jaeger, Karl-Erich

    2002-01-01

    This article reviews our present knowledge on the extracellular lipolytic enzymes LipA and LipB from Bacillus subtilis. Growth of B. subtilis to the late logarithmic growth phase results in a total lipolytic activity of 12–18 units per liter of culture supernatant. Immunodetection with LipA- and Lip

  5. Genetic or chemical protease inhibition causes significant changes in the Bacillus subtilis exoproteome

    NARCIS (Netherlands)

    Westers, Lidia; Westers, Helga; Zanen, Geeske; Antelmann, Haike; Hecker, Michael; Noone, David; Devine, Kevin M.; van Dijl, Jan Maarten; Quax, Wim J.

    2008-01-01

    Bacillus subtilis is a prolific producer of enzymes and biopharmaceuticals. However, the susceptibility of heterologous proteins to degradation by (extracellular) proteases is a major limitation for use of B. subtilis as a protein cell factory. An increase in protein production levels has previously

  6. Non-functional expression of Escherichia coli signal peptidase I in Bacillus subtilis

    NARCIS (Netherlands)

    van Dijl, J M; de Jong, A; Smith, H; Bron, S; Venema, G

    1991-01-01

    The Escherichia coli lep gene, encoding signal peptidase I (SPase I) was provided with Bacillus subtilis transcription/translation signals and expressed in this organism. When present on a low-copy-number plasmid, the amount of E. coli SPase I produced (per mg cell protein) in B. subtilis was half t

  7. Complete genome sequence of Bacillus subtilis SG6 antagonistic against Fusarium graminearum.

    Science.gov (United States)

    Zhao, Yueju; Sangare, Lancine; Wang, Yao; Folly, Yawa Minnie Elodie; Selvaraj, Jonathan Nimal; Xing, Fuguo; Zhou, Lu; Wang, Yan; Liu, Yang

    2015-01-20

    Bacillus subtilis SG6 exhibited a high antifungal effect on the mycelium growth, sporulation and DON production of F. graminearum and significantly reduced disease incidence, Fusarium head blight (FHB) index and DON in the field. Here, we present the complete genome sequence of B. subtilis SG6, providing insights into the genomic basis of its effects and facilitating its application in FHB control.

  8. Metabolic protein interactions in Bacillus subtilis studied at the single cell level

    NARCIS (Netherlands)

    Detert Oude Weme, Ruud Gerardus Johannes

    2015-01-01

    We have investigated protein-protein interactions in live Bacillus subtilis cells (a bacterium). B. subtilis’ natural habitat is the soil and the roots of plants, but also the human microbiota. B. subtilis is used worldwide as a model organism. Unlike eukaryotic cells, bacteria do not have organelle

  9. The Rok protein of Bacillus subtilis represses genes for cell surface and extracellular functions

    NARCIS (Netherlands)

    Albano, M; Smits, WK; Ho, LTY; Kraigher, B; Mandic-Mulec, [No Value; Kuipers, OP; Dubnau, D; Smits, Wiep Klaas; Ho, Linh T.Y.; Mandic-Mulec, Ines

    2005-01-01

    Rok is a repressor of the transcriptional activator ComK and is therefore an important regulator of competence in Bacillus subtilis (T. T. Hoa, P. Tortosa, M. Albano, and D. Dubnau, Mol. Microbiol. 43:15-26, 2002). To address the wider role of Rok in the physiology of B. subtilis, we have used a com

  10. NONFUNCTIONAL EXPRESSION OF ESCHERICHIA-COLI SIGNAL PEPTIDASE-I IN BACILLUS-SUBTILIS

    NARCIS (Netherlands)

    VANDIJL, JM; DEJONG, A; SMITH, H; BRON, S; VENEMA, G; van Dijl, Jan Maarten

    1991-01-01

    The Escherichia coli lep gene, encoding signal peptidase I (SPase I) was provided with Bacillus subtilis transcription/translation signals and expressed in this organism. When present on a low-copy-number plasmid, the amount of E. coli SPase I produced (per mg cell protein) in B. subtilis was half t

  11. Observations on the migration of bacillus spores outside a contaminated facility during a decontamination efficacy study

    Science.gov (United States)

    Silvestri, Erin E.; Perkins, Sarah; Lordo, Robert; Kovacik, William; Nichols, Tonya L.; Bowling, Charlena Yoder; Griffin, Dale W.; Schaefer, Frank W.

    2015-01-01

    The potential for an intentional wide-area or indoor release of Bacillus anthracis spores remains a concern, but the fate and transport of B. anthracis spores in indoor and outdoor environments are not well understood. Some studies have examined the possibility of spore transport within ventilation systems and in buildings and transport into a building following an outdoor release. Little research exists regarding the potential for spores to migrate to the outside of a building following an indoor release.

  12. Impact of Spore Biology on the Rate of Kill and Suppression of Resistance in Bacillus anthracis▿

    OpenAIRE

    Drusano, G L; Okusanya, O. O.; Okusanya, A. O.; van Scoy, B.; Brown, D L; Fregeau, C.; Kulawy, R.; Kinzig, M; Sörgel, F; Heine, H. S.; Louie, A

    2009-01-01

    Bacillus anthracis is complex because of its spore form. The spore is invulnerable to antibiotic action. It also has an impact on the emergence of resistance. We employed the hollow-fiber infection model to study the impacts of different doses and schedules of moxifloxacin on the total-organism population, the spore population, and the subpopulations of vegetative- and spore-phase organisms that were resistant to moxifloxacin. We then generated a mathematical model of the impact of moxifloxac...

  13. Isolation of a new Mexican strain of Bacillus subtilis with antifungal and antibacterial activities.

    Science.gov (United States)

    Basurto-Cadena, M G L; Vázquez-Arista, M; García-Jiménez, J; Salcedo-Hernández, R; Bideshi, D K; Barboza-Corona, J E

    2012-01-01

    Although several strains of B. subtilis with antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a native B. subtilis strain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis 21) demonstrated in vitro antagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected with Rhizoctonia solani and Fusarium verticillioides and treated with B. subtilis 21 produced augment in the number of leaves per plant and an increment in the length of healthy leaves in comparison with untreated plants. In addition, B. subtilis 21 showed activity against pathogenic bacteria. Secreted proteins by B. subtilis 21 were studied, detecting the presence of proteases and bacteriocin-like inhibitor substances that could be implicated in its antagonistic activity. Chitinases and zwittermicin production could not be detected. Then, B. subtilis 21 could potentially be used to control phytopathogenic fungi that infect strawberry plants.

  14. Isolation of a New Mexican Strain of Bacillus subtilis with Antifungal and Antibacterial Activities

    Directory of Open Access Journals (Sweden)

    M. G. L. Basurto-Cadena

    2012-01-01

    Full Text Available Although several strains of B. subtilis with antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a native B. subtilis strain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis 21 demonstrated in vitro antagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected with Rhizoctonia solani and Fusarium verticillioides and treated with B. subtilis 21 produced augment in the number of leaves per plant and an increment in the length of healthy leaves in comparison with untreated plants. In addition, B. subtilis 21 showed activity against pathogenic bacteria. Secreted proteins by B. subtilis 21 were studied, detecting the presence of proteases and bacteriocin-like inhibitor substances that could be implicated in its antagonistic activity. Chitinases and zwittermicin production could not be detected. Then, B. subtilis 21 could potentially be used to control phytopathogenic fungi that infect strawberry plants.

  15. Genomics, evolution, and crystal structure of a new family of bacterial spore kinases

    OpenAIRE

    2009-01-01

    Bacterial spore formation is a complex process of fundamental relevance to biology and human disease. The spore coat structure is complex and poorly understood, and the roles of many of the protein components remain unclear. We describe a new family of spore coat proteins, the bacterial spore kinases (BSKs), and the first crystal structure of a BSK, YtaA (CotI) from Bacillus subtilis. BSKs are widely distributed in spore-forming Bacillus and Clostridium species, and have a dynamic evolutionar...

  16. Acid and base stress and transcriptomic responses in Bacillus subtilis.

    Science.gov (United States)

    Wilks, Jessica C; Kitko, Ryan D; Cleeton, Sarah H; Lee, Grace E; Ugwu, Chinagozi S; Jones, Brian D; BonDurant, Sandra S; Slonczewski, Joan L

    2009-02-01

    Acid and base environmental stress responses were investigated in Bacillus subtilis. B. subtilis AG174 cultures in buffered potassium-modified Luria broth were switched from pH 8.5 to pH 6.0 and recovered growth rapidly, whereas cultures switched from pH 6.0 to pH 8.5 showed a long lag time. Log-phase cultures at pH 6.0 survived 60 to 100% at pH 4.5, whereas cells grown at pH 7.0 survived acid or base induced adaptation to a more extreme acid or base, respectively. Expression indices from Affymetrix chip hybridization were obtained for 4,095 protein-encoding open reading frames of B. subtilis grown at external pH 6, pH 7, and pH 9. Growth at pH 6 upregulated acetoin production (alsDS), dehydrogenases (adhA, ald, fdhD, and gabD), and decarboxylases (psd and speA). Acid upregulated malate metabolism (maeN), metal export (czcDO and cadA), oxidative stress (catalase katA; OYE family namA), and the SigX extracytoplasmic stress regulon. Growth at pH 9 upregulated arginine catabolism (roc), which generates organic acids, glutamate synthase (gltAB), polyamine acetylation and transport (blt), the K(+)/H(+) antiporter (yhaTU), and cytochrome oxidoreductases (cyd, ctaACE, and qcrC). The SigH, SigL, and SigW regulons were upregulated at high pH. Overall, greater genetic adaptation was seen at pH 9 than at pH 6, which may explain the lag time required for growth shift to high pH. Low external pH favored dehydrogenases and decarboxylases that may consume acids and generate basic amines, whereas high external pH favored catabolism-generating acids.

  17. Evaluating Composite Sampling Methods of Bacillus spores at Low Concentrations

    Energy Technology Data Exchange (ETDEWEB)

    Hess, Becky M.; Amidan, Brett G.; Anderson, Kevin K.; Hutchison, Janine R.

    2016-10-13

    Restoring facility operations after the 2001 Amerithrax attacks took over three months to complete, highlighting the need to reduce remediation time. The most time intensive tasks were environmental sampling and sample analyses. Composite sampling allows disparate samples to be combined, with only a single analysis needed, making it a promising method to reduce response times. We developed a statistical experimental design to test three different composite sampling methods: 1) single medium single pass composite: a single cellulose sponge samples multiple coupons; 2) single medium multi-pass composite: a single cellulose sponge is used to sample multiple coupons; and 3) multi-medium post-sample composite: a single cellulose sponge samples a single surface, and then multiple sponges are combined during sample extraction. Five spore concentrations of Bacillus atrophaeus Nakamura spores were tested; concentrations ranged from 5 to 100 CFU/coupon (0.00775 to 0.155CFU/cm2, respectively). Study variables included four clean surface materials (stainless steel, vinyl tile, ceramic tile, and painted wallboard) and three grime coated/dirty materials (stainless steel, vinyl tile, and ceramic tile). Analysis of variance for the clean study showed two significant factors: composite method (p-value < 0.0001) and coupon material (p-value = 0.0008). Recovery efficiency (RE) was higher overall using the post-sample composite (PSC) method compared to single medium composite from both clean and grime coated materials. RE with the PSC method for concentrations tested (10 to 100 CFU/coupon) was similar for ceramic tile, painted wall board, and stainless steel for clean materials. RE was lowest for vinyl tile with both composite methods. Statistical tests for the dirty study showed RE was significantly higher for vinyl and stainless steel materials, but significantly lower for ceramic tile. These results suggest post-sample compositing can be used to reduce sample analysis time when

  18. 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 ℃ .

  19. 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℃.

  20. Diverse supramolecular structures formed by self‐assembling proteins of the B acillus subtilis spore coat

    Science.gov (United States)

    Jiang, Shuo; Wan, Qiang; Krajcikova, Daniela; Tang, Jilin; Tzokov, Svetomir B.; Barak, Imrich

    2015-01-01

    Summary Bacterial spores (endospores), such as those of the pathogens C lostridium difficile and B acillus anthracis, are uniquely stable cell forms, highly resistant to harsh environmental insults. B acillus subtilis is the best studied spore‐former and we have used it to address the question of how the spore coat is assembled from multiple components to form a robust, protective superstructure. B . subtilis coat proteins (CotY, CotE, CotV and CotW) expressed in E scherichia coli can arrange intracellularly into highly stable macro‐structures through processes of self‐assembly. Using electron microscopy, we demonstrate the capacity of these proteins to generate ordered one‐dimensional fibres, two‐dimensional sheets and three‐dimensional stacks. In one case (CotY), the high degree of order favours strong, cooperative intracellular disulfide cross‐linking. Assemblies of this kind could form exquisitely adapted building blocks for higher‐order assembly across all spore‐formers. These physically robust arrayed units could also have novel applications in nano‐biotechnology processes. PMID:25872412

  1. Evaluation of bottlenecks in the late stages of protein secretion in Bacillus subtilis

    NARCIS (Netherlands)

    Bolhuis, A; Tjalsma, H; Smith, H.E; Meima, R.; Venema, G; Bron, S; van Dijl, J.M

    1999-01-01

    Despite a high capacity for secretion of homologous proteins, the secretion of heterologous proteins by Bacillus subtilis is frequently inefficient. In the present studies, we have investigated and compared bottlenecks in the secretion of four heterologous proteins: Bacillus lichenifomis alpha-amyla

  2. Subunit II of Bacillus subtilis cytochrome c oxidase is a lipoprotein

    NARCIS (Netherlands)

    Bengtsson, J; Tjalsma, H; Rivolta, C; Hederstedt, L

    1999-01-01

    The sequence of the N-terminal end of the deduced ctaC gene product of Bacillus species has the features of a bacterial lipoprotein. CtaC is the subunit II of cytochrome caa(3), which is a cytochrome c oxidase. Using Bacillus subtilis mutants blocked in lipoprotein synthesis, we show that CtaC is a

  3. The structure-function relationship of the lipases from Pseudomonas aeruginosa and Bacillus subtilis

    NARCIS (Netherlands)

    Misset, Onno; Gerritse, Gijs; Jaeger, Karl-Erich; Winkler, Ulrich; Colson, Charles; Schanck, Karin; Lesuisse, Emmanuel; Dartois, Véronique; Blaauw, Mieke; Ransac, Stéphane; Dijkstra, Bauke W.

    1994-01-01

    Within the BRIDGE T-project on lipases we investigate the structure-function relationships of the lipases from Bacillus subtilis and Pseudomonas aeruginosa. Construction of an overproducing Bacillus strain allowed the purification of > 100 mg lipase from 30 I culture supernatant. After testing a lar

  4. 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

  5. Weak solutions for a bioconvection model related to Bacillus subtilis

    CERN Document Server

    Vorotnikov, Dmitry

    2012-01-01

    We consider the initial-boundary value problem for the coupled Navier-Stokes-Keller-Segel-Fisher-Kolmogorov-Petrovskii-Piskunov system in two- and three-dimensional domains. The problem describes oxytaxis and growth of Bacillus subtilis in moving water. We prove existence of global weak solutions to the problem. We distinguish between two cases determined by the cell diffusion term and the space dimension, which are referred as the supercritical and subcritical ones. At the first case, the choice of the growth function enjoys wide range of possibilities: in particular, it can be zero. Our results are new even at the absence of the growth term. At the second case, the restrictions on the growth function are less relaxed: for instance, it cannot be zero but can be Fisher-like. In the case of linear cell diffusion, the solution is regular and unique provided the domain is the whole plane. In addition, we study the long-time behaviour of the problem, find dissipative estimates, and construct attractors.

  6. Biodegradation of pendimethalin by Bacillus subtilis Y3.

    Science.gov (United States)

    Ni, Haiyan; Yao, Li; Li, Na; Cao, Qin; Dai, Chen; Zhang, Jun; He, Qin; He, Jian

    2016-03-01

    A bacterium strain Y3, capable of efficiently degrading pendimethalin, was isolated from activated sludge and identified as Bacillus subtilis according to its phenotypic features and 16S rRNA phylogenetic analysis. This strain could grow on pendimethalin as a sole carbon source and degrade 99.5% of 100mg/L pendimethalin within 2.5days in batch liquid culture, demonstrating a greater efficiency than any other reported strains. Three metabolic products, 6-aminopendimethalin, 5-amino-2-methyl-3-nitroso-4-(pentan-3-ylamino) benzoic acid, and 8-amino-2-ethyl-5-(hydroxymethyl)-1,2-dihydroquinoxaline-6-carboxylic acid, were identified by HPLC-MS/MS, and a new microbial degradation pathway was proposed. A nitroreductase catalyzing nitroreduction of pendimethalin to 6-aminopendimethalin was detected in the cell lysate of strain Y3. The cofactor was nicotinamide adenine dinucleotide phosphate (NADPH) or more preferably nicotinamide adenine dinucleotide (NADH). The optimal temperature and pH for the nitroreductase were 30°C and 7.5, respectively. Hg(2+), Ni(2+), Pb(2+), Co(2+), Mn(2+) Cu(2+), Ag(+), and EDTA severely inhibited the nitroreductase activity, whereas Fe(2+), Mg(2+), and Ca(2+) enhanced it. This study provides an efficient pendimethalin-degrading microorganism and broadens the knowledge of the microbial degradation pathway of pendimethalin.

  7. Bacillus subtilis chromosome organization oscillates between two distinct patterns.

    Science.gov (United States)

    Wang, Xindan; Montero Llopis, Paula; Rudner, David Z

    2014-09-02

    Bacterial chromosomes have been found to possess one of two distinct patterns of spatial organization. In the first, called "ori-ter" and exemplified by Caulobacter crescentus, the chromosome arms lie side-by-side, with the replication origin and terminus at opposite cell poles. In the second, observed in slow-growing Escherichia coli ("left-ori-right"), the two chromosome arms reside in separate cell halves, on either side of a centrally located origin. These two patterns, rotated 90° relative to each other, appear to result from different segregation mechanisms. Here, we show that the Bacillus subtilis chromosome alternates between them. For most of the cell cycle, newly replicated origins are maintained at opposite poles with chromosome arms adjacent to each other, in an ori-ter configuration. Shortly after replication initiation, the duplicated origins move as a unit to midcell and the two unreplicated arms resolve into opposite cell halves, generating a left-ori-right pattern. The origins are then actively segregated toward opposite poles, resetting the cycle. Our data suggest that the condensin complex and the parABS partitioning system are the principal driving forces underlying this oscillatory cycle. We propose that the distinct organization patterns observed for bacterial chromosomes reflect a common organization-segregation mechanism, and that simple modifications to it underlie the unique patterns observed in different species.

  8. The sodium effect of Bacillus subtilis growth on aspartate.

    Science.gov (United States)

    Whiteman, P; Marks, C; Freese, E

    1980-08-01

    aspH mutants of Bacillus subtilis have a constitutive aspartase activity and grow well on aspartate as sole carbon source. aspH aspT mutants, which are deficient in high affinity aspartate transport as a result of the aspT mutation, grow as well as aspH mutants in medium containing high concentrations of aspartate and Na+. This Na+ effect is not due to an enhancement of aspartate transport but is the result of increased cellular metabolism. The ability to grow rapidly in sodium aspartate is induced by prior growth in the presence of Na+. In potassium aspartate, the addition of arginine, citrulline, ornithine, delta 1-pyrroline-5-carboxylase or proline instead of Na+ also allows rapid growth; but in a mutant deficient in ornithine--oxo-acid aminotransferase, only pyrroline-carboxylate or proline can replace Na+. The amino acid pool of cells growing slowly in potassium aspartate contains proline at a low concentration which increases upon addition of proline (but not Na+) to the medium. Thus, Na+ addition does not increase the synthesis of proline, but proline or pyrroline-carboxylate acts similarly to Na+ either in preventing some inhibitory effect (by aspartate or the accumulating NH4+) or in overcoming some deficiency (e.g. in further proline metabolism.

  9. Two purine nucleoside phosphorylases in Bacillus subtilis. Purification and some properties of the adenosine-specific phosphorylase

    DEFF Research Database (Denmark)

    Jensen, Kaj Frank

    1978-01-01

    Two purine nucleoside phosphorylases (purine-nucleoside:orthophosphate ribosyltransferase, EC 2.4.2.1) were purified from vegetative Bacillus subtilis cells. One enzyme, inosine-guanosine phosphorylase, showed great similarity to the homologous enzyme of Bacillus cereus. It appeared...

  10. The Adsorption Properties of Bacillus atrophaeus Spore on Functionalized Carbon Nanotubes

    Directory of Open Access Journals (Sweden)

    P. Cortes

    2010-01-01

    Full Text Available An equilibrium study of Bacillus atrophaeus (B.a spores on functionalized Single-Wall Carbon Nanotubes (SWCNTs has been performed in order to characterize the adsorption properties of the spores/nanotubes complex. The carbon nanotubes here investigated were subjected to a two-step purification and functionalization treatment in order to introduce chemical groups on their basal planes. The inclusion of carboxyl functional groups on the nanotubes was corroborated by Raman and infrared spectroscopy. These carboxyl groups appear to enhance the nanotube-B.a. interaction by reacting with the proteinaceous pili appendages present on the spore surface. The adsorption data demonstrate that bacillus spores diffuse faster on functionalized carbon nanotubes than on as-received and purified nanomaterials. Transmission Electron Microscopy also shows that the chemically treated nanotubes resulted in a swollen nano-network which seems to further enhance the bacillus adsorption due to a more extensive spore-nanotube contact area.

  11. Inactivation of E. coli, B. subtilis spores, and MS2, T4, and T7 phage using UV/H2O2 advanced oxidation.

    Science.gov (United States)

    Mamane, Hadas; Shemer, Hilla; Linden, Karl G

    2007-07-31

    The goal of this study was to evaluate the potential of an advanced oxidation process (AOP) for microbiocidal and virucidal inactivation. The viruses chosen for this study were bacteriophage MS2, T4, and T7. In addition, Bacillus subtilis spores and Escherichia coli were studied. By using H(2)O(2) in the presence of filtered ultraviolet (UV) irradiation (UV/H(2)O(2)) to generate wavelengths above 295nm, the direct UV photolysis disinfection mechanism was minimized, while disinfection by H(2)O(2) was also negligible. Virus T4 and E. coli in phosphate buffered saline (PBS) were sensitive to >295nm filtered UV irradiation (without H(2)O(2)), while MS2 was very resistant. Addition of H(2)O(2) at 25mg/l in the presence of filtered UV irradiation over a 15min reaction time did not result in any additional disinfection of virus T4, while an additional one log inactivation for T7 and 2.5 logs for MS2 were obtained. With E. coli, only a slight additional effect was observed when H(2)O(2) was added. B. subtilis spores did not show any inactivation at any of the conditions used in this study. The OH radical exposure (CT value) was calculated to present the relationship between the hydroxyl radical dose and microbial inactivation.

  12. Inactivation of E. coli, B. subtilis spores, and MS2, T4, and T7 phage using UV/H{sub 2}O{sub 2} advanced oxidation

    Energy Technology Data Exchange (ETDEWEB)

    Mamane, Hadas [School of Mechanical Engineering and Porter School of Environmental Studies, Tel-Aviv University, Tel-Aviv 69978 (Israel)]. E-mail: hadasmg@post.tau.ac.il; Shemer, Hilla [Department of Civil and Environmental Engineering, Duke University, Durham, NC 27708-0287 (United States); Linden, Karl G. [Department of Civil and Environmental Engineering, Duke University, Durham, NC 27708-0287 (United States)

    2007-07-31

    The goal of this study was to evaluate the potential of an advanced oxidation process (AOP) for microbiocidal and virucidal inactivation. The viruses chosen for this study were bacteriophage MS2, T4, and T7. In addition, Bacillus subtilis spores and Escherichia coli were studied. By using H{sub 2}O{sub 2} in the presence of filtered ultraviolet (UV) irradiation (UV/H{sub 2}O{sub 2}) to generate wavelengths above 295 nm, the direct UV photolysis disinfection mechanism was minimized, while disinfection by H{sub 2}O{sub 2} was also negligible. Virus T4 and E. coli in phosphate buffered saline (PBS) were sensitive to >295 nm filtered UV irradiation (without H{sub 2}O{sub 2}), while MS2 was very resistant. Addition of H{sub 2}O{sub 2} at 25 mg/l in the presence of filtered UV irradiation over a 15 min reaction time did not result in any additional disinfection of virus T4, while an additional one log inactivation for T7 and 2.5 logs for MS2 were obtained. With E. coli, only a slight additional effect was observed when H{sub 2}O{sub 2} was added. B. subtilis spores did not show any inactivation at any of the conditions used in this study. The OH radical exposure (CT value) was calculated to present the relationship between the hydroxyl radical dose and microbial inactivation.

  13. An exogenous surfactant-producing Bacillus subtilis facilitates indigenous microbial enhanced oil recovery

    Directory of Open Access Journals (Sweden)

    Peike eGao

    2016-02-01

    Full Text Available This study used an exogenous lipopeptide-producing Bacillus subtilis to strengthen the indigenous microbial enhanced oil recovery (IMEOR process in a water-flooded reservoir in the laboratory. The microbial processes and driving mechanisms were investigated in terms of the changes in oil properties and the interplay between the exogenous Bacillus subtilis and indigenous microbial populations. The exogenous Bacillus subtilis is a lipopeptide producer, with a short growth cycle and no oil-degrading ability. The Bacillus subtilis facilitates the IMEOR process through improving oil emulsification and accelerating microbial growth with oil as the carbon source. Microbial community studies using quantitative PCR and high-throughput sequencing revealed that the exogenous Bacillus subtilis could live together with reservoir microbial populations, and did not exert an observable inhibitory effect on the indigenous microbial populations during nutrient stimulation. Core-flooding tests showed that the combined exogenous and indigenous microbial flooding increased oil displacement efficiency by 16.71%, compared with 7.59% in the control where only nutrients were added, demonstrating the application potential in enhanced oil recovery in water-flooded reservoirs, in particular, for reservoirs where IMEOR treatment cannot effectively improve oil recovery.

  14. Removal of Bacillus anthracis sterne spore from commercial unpasteurized liquid egg white using crossflow microfiltration

    Science.gov (United States)

    Current pasteurization technology used by the egg industry is ineffective for destruction of spores such as those of Bacillus anthracis (BA). The validity of a cross-flow microfiltration (MF) process for separation of the attenuated strain of BA (Sterne) spores from commercial unpasteurized liquid ...

  15. Regulation of the Spore Cortex Lytic Enzyme SleB in Bacillus anthracis

    OpenAIRE

    2014-01-01

    Bacillus anthracis is the causative agent of the disease anthrax and poses a threat due to its potential to be used as a biological weapon. The spore form of this bacterium is an extremely resistant structure, making spore decontamination exceptionally challenging. During spore germination, nutrient germinants interact with Ger receptors, triggering a cascade of events. A crucial event in this process is degradation of the cortex peptidoglycan by germination-specific lytic enzymes (GSLEs),...

  16. GLYCOGEN IN BACILLUS-SUBTILIS - MOLECULAR CHARACTERIZATION OF AN OPERON ENCODING ENZYMES INVOLVED IN GLYCOGEN BIOSYNTHESIS AND DEGRADATION

    NARCIS (Netherlands)

    KIEL, JAKW; BOELS, JM; BELDMAN, G; VENEMA, G

    1994-01-01

    Although it has never been reported that Bacillus subtilis is capable of accumulating glycogen, we have isolated a region from the chromosome of B. subtilis containing a glycogen operon. The operon is located directly downstream from trnB, which maps at 275 degrees on the B. subtilis chromosome. It

  17. Effect of Riboflavin Operon Dosage on Riboflavin Productivity in Bacillus Subtilis

    Institute of Scientific and Technical Information of China (English)

    CHEN Tao; CHEN Xun; WANG Jingyu; ZHAO Xueming

    2005-01-01

    After deregulating the purine and riboflavin synthesis in the Gram-positive bacterium Bacillus subtilis,it is critical to amplify riboflavin operon with appropriate dosage in the host strain for remarkable increase of riboflavin production.Bacillus subtilis RH13, a riboflavin-producing strain, was selected as host strain in the construction of engineering strains by protoplast fusion. The integrative plasmid pRB63 and autonomous plasmid pRB49, pRB62 containing riboflavin operon of B.subtilis 24 were constructed and transformed into the host strain respectively. Increasing one operon copy in B.subtilis RH13 results in about 0.4 g/L improvement in riboflavin yield and the appropriate number of operon copies was about 7-8. Amplifying more riboflavin operons is of no use for further improvement of yield of riboflavin. Furthermore, excessive operon dosage results in metabolic unbalance and is fatal to the host cells producing riboflavin.

  18. Germinant-enhanced decontamination of Bacillus spores adhered to iron and cement-mortar drinking water infrastructures.

    Science.gov (United States)

    Szabo, Jeffrey G; Muhammad, Nur; Heckman, Lee; Rice, Eugene W; Hall, John

    2012-04-01

    Germination was evaluated as an enhancement to decontamination methods for removing Bacillus spores from drinking water infrastructure. Germinating spores before chlorinating cement mortar or flushing corroded iron was more effective than chlorinating or flushing alone.

  19. Integrative bacterial artificial chromosomes for DNA integration into the Bacillus subtilis chromosome.

    Science.gov (United States)

    Juhas, Mario; Ajioka, James W

    2016-06-01

    Bacillus subtilis is a well-characterized model bacterium frequently used for a number of biotechnology and synthetic biology applications. Novel strategies combining the advantages of B. subtilis with the DNA assembly and editing tools of Escherichia coli are crucial for B. subtilis engineering efforts. We combined Gibson Assembly and λ red recombineering in E. coli with RecA-mediated homologous recombination in B. subtilis for bacterial artificial chromosome-mediated DNA integration into the well-characterized amyE target locus of the B. subtilis chromosome. The engineered integrative bacterial artificial chromosome iBAC(cav) can accept any DNA fragment for integration into B. subtilis chromosome and allows rapid selection of transformants by B. subtilis-specific antibiotic resistance and the yellow fluorescent protein (mVenus) expression. We used the developed iBAC(cav)-mediated system to integrate 10kb DNA fragment from E. coli K12 MG1655 into B. subtilis chromosome. iBAC(cav)-mediated chromosomal integration approach will facilitate rational design of synthetic biology applications in B. subtilis.

  20. Impact of spore biology on the rate of kill and suppression of resistance in Bacillus anthracis.

    Science.gov (United States)

    Drusano, G L; Okusanya, O O; Okusanya, A O; van Scoy, B; Brown, D L; Fregeau, C; Kulawy, R; Kinzig, M; Sörgel, F; Heine, H S; Louie, A

    2009-11-01

    Bacillus anthracis is complex because of its spore form. The spore is invulnerable to antibiotic action. It also has an impact on the emergence of resistance. We employed the hollow-fiber infection model to study the impacts of different doses and schedules of moxifloxacin on the total-organism population, the spore population, and the subpopulations of vegetative- and spore-phase organisms that were resistant to moxifloxacin. We then generated a mathematical model of the impact of moxifloxacin, administered by continuous infusion or once daily, on vegetative- and spore-phase organisms. The ratio of the rate constant for vegetative-phase cells going to spore phase (K(vs)) to the rate constant for spore-phase cells going to vegetative phase (K(sv)) determines the rate of organism clearance. The continuous-infusion drug profile is more easily sensed as a threat; the K(vs)/K(sv) ratio increases at lower drug exposures (possibly related to quorum sensing). This movement to spore phase protects the organism but makes the emergence of resistance less likely. Suppression of resistance requires a higher level of drug exposure with once-daily administration than with a continuous infusion, a difference that is related to vegetative-to-spore (and back) transitioning. Spore biology has a major impact on drug therapy and resistance suppression. These findings explain why all drugs of different classes have approximately the same rate of organism clearance for Bacillus anthracis.

  1. Menaquinone and iron are essential for complex colony development in Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Gidi Pelchovich

    Full Text Available Cells of undomesticated species of Bacillus subtilis frequently form complex colonies during spreading on agar surfaces. Given that menaquinone is involved in another form of coordinated behavior, namely, sporulation, we looked for a possible role for menaquinone in complex colony development (CCD in the B. subtilis strain NCIB 3610. Here we show that inhibition of menaquinone biosynthesis in B. subtilis indeed abolished its ability to develop complex colonies. Additionally some mutations of B. subtilis which confer defective CCD could be suppressed by menaquinone derivatives. Several such mutants mapped to the dhb operon encoding the genes responsible for the biosynthesis of the iron siderophore, bacillibactin. Our results demonstrate that both menaquinone and iron are essential for CCD in B. subtilis.

  2. Bacillus subtilis biofilm extends Caenorhabditis elegans longevity through downregulation of the insulin-like signalling pathway

    Science.gov (United States)

    Donato, Verónica; Ayala, Facundo Rodríguez; Cogliati, Sebastián; Bauman, Carlos; Costa, Juan Gabriel; Leñini, Cecilia; Grau, Roberto

    2017-01-01

    Beneficial bacteria have been shown to affect host longevity, but the molecular mechanisms mediating such effects remain largely unclear. Here we show that formation of Bacillus subtilis biofilms increases Caenorhabditis elegans lifespan. Biofilm-proficient B. subtilis colonizes the C. elegans gut and extends worm lifespan more than biofilm-deficient isogenic strains. Two molecules produced by B. subtilis — the quorum-sensing pentapeptide CSF and nitric oxide (NO) — are sufficient to extend C. elegans longevity. When B. subtilis is cultured under biofilm-supporting conditions, the synthesis of NO and CSF is increased in comparison with their production under planktonic growth conditions. We further show that the prolongevity effect of B. subtilis biofilms depends on the DAF-2/DAF-16/HSF-1 signalling axis and the downregulation of the insulin-like signalling (ILS) pathway. PMID:28134244

  3. The Bacillus subtilis Acyl Lipid Desaturase Is a Δ5 Desaturase

    Science.gov (United States)

    Altabe, Silvia G.; Aguilar, Pablo; Caballero, Gerardo M.; de Mendoza, Diego

    2003-01-01

    Bacillus subtilis was recently reported to synthesize unsaturated fatty acids (UFAs) with a double bond at positions Δ5, Δ7, and Δ9 (M. H. Weber, W. Klein, L. Muller, U. M. Niess, and M. A. Marahiel, Mol. Microbiol. 39:1321-1329, 2001). Since this finding would have considerable importance in the double-bond positional specificity displayed by the B. subtilis acyl lipid desaturase, we have attempted to confirm this observation. We report that the double bond of UFAs synthesized by B. subtilis is located exclusively at the Δ5 position, regardless of the growth temperature and the length chain of the fatty acids. PMID:12730185

  4. Expressão de uricase de Bacillus subtilis em Escherichia coli

    OpenAIRE

    Pfrimer, Pollyanna

    2007-01-01

    A uricase, uma enzima que converte ácido úrico em alantoína, é comumente usada em kits comerciais de dosagem de ácido úrico. Com a finalidade de produzir esta enzima em grande escala por técnicas de Engenharia Genética, o gene da uricase de Bacillus subtilis subtilis foi clonado e expresso em Escherichia coli. Para tanto, foi feita uma PCR utilizando-se como template o DNA cromossomal de B. subtilis e primers específicos para amplificação do gene pucLM. O amplicon foi sub-clonado seguindo-se ...

  5. Nanoscale structural and mechanical analysis of Bacillus anthracis spores inactivated with rapid dry heating.

    Science.gov (United States)

    Xing, Yun; Li, Alex; Felker, Daniel L; Burggraf, Larry W

    2014-03-01

    Effective killing of Bacillus anthracis spores is of paramount importance to antibioterrorism, food safety, environmental protection, and the medical device industry. Thus, a deeper understanding of the mechanisms of spore resistance and inactivation is highly desired for developing new strategies or improving the known methods for spore destruction. Previous studies have shown that spore inactivation mechanisms differ considerably depending upon the killing agents, such as heat (wet heat, dry heat), UV, ionizing radiation, and chemicals. It is believed that wet heat kills spores by inactivating critical enzymes, while dry heat kills spores by damaging their DNA. Many studies have focused on the biochemical aspects of spore inactivation by dry heat; few have investigated structural damages and changes in spore mechanical properties. In this study, we have inactivated Bacillus anthracis spores with rapid dry heating and performed nanoscale topographical and mechanical analysis of inactivated spores using atomic force microscopy (AFM). Our results revealed significant changes in spore morphology and nanomechanical properties after heat inactivation. In addition, we also found that these changes were different under different heating conditions that produced similar inactivation probabilities (high temperature for short exposure time versus low temperature for long exposure time). We attributed the differences to the differential thermal and mechanical stresses in the spore. The buildup of internal thermal and mechanical stresses may become prominent only in ultrafast, high-temperature heat inactivation when the experimental timescale is too short for heat-generated vapor to efficiently escape from the spore. Our results thus provide direct, visual evidences of the importance of thermal stresses and heat and mass transfer to spore inactivation by very rapid dry heating.

  6. 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.

  7. Modeling Radiation Effectiveness for Inactivation of Bacillus Spores

    Science.gov (United States)

    2015-09-17

    will undergo germination which is the first step in the process by which bacteria transforms from a dormant spore into a vegetative cell [28]. The...order to survive a dose of radiation, a spore must repair its damaged DNA during germination . The DNA repair process is dependent on reactions catalyzed...in the next section. 2.2 Life Cycle of a Bacterial Spore A dormant spore is formed via a multi- step process called sporulation (refer to Figure 2.5

  8. Current Physical and SDS Extraction Methods Do Not Efficiently Remove Exosporium Proteins from Bacillus anthracis spores

    Science.gov (United States)

    Thompson, Brian M.; Binkley, Jana M.; Stewart, George C.

    2011-01-01

    Biochemical studies of the outermost spore layers of the Bacillus cereus family are hindered by difficulties in efficient dispersal of the external spore layers and difficulties in dissociating protein complexes that comprise the exosporium layer. Detergent and physical methods have been utilized to disrupt the exosporium layer. Herein we compare commonly used SDS extraction buffers used to extract spore proteins and demonstrate the incomplete extractability of the exosporium layer by these methods. Sonication and bead beating methods for exosporium layer removal were also examined. A combination of genetic and physical methods is the most effective for isolating proteins found in the spore exosporium. PMID:21338631

  9. Use of bacillus subtilis strains to inhibit postharvest pathogenic fungi; Attivita` antagonista di alcuni ceppi di bacillus subtilis nei confronti di funghi patogeni

    Energy Technology Data Exchange (ETDEWEB)

    Arras, G.; Gambella, F.; Demontis, S.; Petretto, A.

    1995-09-01

    An isolate (87) of the bacillus subtilis strains isolated from cold stored citrus fruit 13 proved to inhibit the growth in vitro of the penicillium italicum used in the experiment (from 50.6% to 92.2%) and to inhibit botrytis cinerea (from 65.3% to 95.9%). A further test, superimposing on plates containing PDA strains Nos. 13, 173, and 160, totally inhibited the fungi. Tested in vivo on artificially bruised oranges, they significantly inhibited two fungi.

  10. A Safety and Environmental Assessment of the Biological Simulants Bacillus subtilis and Newcastle Disease Virus. Volume 1: Discussion

    Science.gov (United States)

    1993-01-01

    Vander Snoeck, P., Daneau, R.D., and Meunier, F. "Nosocomial Bacteremia Caused by Bacillus species", Clin. Micro bioi. Infect. Dis., 7, pp. 783...between B. cereus and B. subtilis existed in diagnostic laboratories before that time (Gordon 1973). B. subtilis, as well as other Bacillus species...or other interventions, which may have introduced the organism to sensitive tissue. Richard et al. (1988) described 11 cases of Bacillus bacteremias

  11. 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

  12. WprA基因在Bacillus subtilis WB800中的克隆表达%Clonging and Expression of a WprA gene in Bacillus subtilis WB800

    Institute of Scientific and Technical Information of China (English)

    柴海云; 崔堂兵

    2012-01-01

    A fibrinolytic enzyme gene (WprA) was cloned from Bacillus subtilis 168. To efficiently express WprA in Bacillus subtilis WB800, WprA gene was inserted into pBE3 to yield a nove vector pBE-WprA. Then the vector pBE-WprA was transformed and expressed in Bacillus subtilis WB800. Results showed WprA gene was efficiently expressed during the exponential and stationary growth stages, and WprA was secreted into the medium.%对源自Bacillus subtilis 168的具有纤溶活性的基因序列(WprA)进行克隆,然后将WprA基因克隆到大肠杆菌-枯草杆菌穿梭载体pBE3中,构建表达载体pBE-WprA,将重组载体转化到Bacillus subtilis WB800中,实现了WprA基因在Bacillus subtilis WB800中的表达.结果表明,WprA基因在Bacillus subtilis WB800中的对数生长期和平衡期均获得了表达,且产物被分泌到胞外.

  13. Anthrax Toxins in Context of Bacillus anthracis Spores and Spore Germination.

    Science.gov (United States)

    Cote, Christopher K; Welkos, Susan L

    2015-08-17

    The interaction of anthrax toxin or toxin components with B. anthracis spores has been demonstrated. Germinating spores can produce significant amounts of toxin components very soon after the initiation of germination. In this review, we will summarize the work performed that has led to our understanding of toxin and spore interactions and discuss the complexities associated with these interactions.

  14. In vitro and in vivo analyses of the Bacillus anthracis spore cortex lytic protein SleL

    OpenAIRE

    2012-01-01

    The bacterial endospore is the most resilient biological structure known. Multiple protective integument layers shield the spore core and promote spore dehydration and dormancy. Dormancy is broken when a spore germinates and becomes a metabolically active vegetative cell. Germination requires the breakdown of a modified layer of peptidoglycan (PG) known as the spore cortex. This study reports in vitro and in vivo analyses of the Bacillus anthracis SleL protein. SleL is a spore cortex lytic en...

  15. Nonribosomal peptide synthase gene clusters for lipopeptide biosynthesis in Bacillus subtilis 916 and their phenotypic functions.

    Science.gov (United States)

    Luo, Chuping; Liu, Xuehui; Zhou, Huafei; Wang, Xiaoyu; Chen, Zhiyi

    2015-01-01

    Bacillus cyclic lipopeptides (LPs) have been well studied for their phytopathogen-antagonistic activities. Recently, research has shown that these LPs also contribute to the phenotypic features of Bacillus strains, such as hemolytic activity, swarming motility, biofilm formation, and colony morphology. Bacillus subtilis 916 not only coproduces the three families of well-known LPs, i.e., surfactins, bacillomycin Ls (iturin family), and fengycins, but also produces a new family of LP called locillomycins. The genome of B. subtilis 916 contains four nonribosomal peptide synthase (NRPS) gene clusters, srf, bmy, fen, and loc, which are responsible for the biosynthesis of surfactins, bacillomycin Ls, fengycins, and locillomycins, respectively. By studying B. subtilis 916 mutants lacking production of one, two, or three LPs, we attempted to unveil the connections between LPs and phenotypic features. We demonstrated that bacillomycin Ls and fengycins contribute mainly to antifungal activity. Although surfactins have weak antifungal activity in vitro, the strain mutated in srfAA had significantly decreased antifungal activity. This may be due to the impaired productions of fengycins and bacillomycin Ls. We also found that the disruption of any LP gene cluster other than fen resulted in a change in colony morphology. While surfactins and bacillomycin Ls play very important roles in hemolytic activity, swarming motility, and biofilm formation, the fengycins and locillomycins had little influence on these phenotypic features. In conclusion, B. subtilis 916 coproduces four families of LPs which contribute to the phenotypic features of B. subtilis 916 in an intricate way.

  16. Natural products from Bacillus subtilis with antimicrobial properties☆

    Institute of Scientific and Technical Information of China (English)

    Tao Wang; Yafei Liang; Mianbin Wu; Zhengjie Chen; Jianping Lin; Lirong Yang

    2015-01-01

    Bacil us subtilis produces many chemical y-diverse secondary metabolites of interest to chemists and biologists. Based on this, this review gives a detalled overview of the natural components produced by B. subtilis including cyclic lipopeptides, polypeptides, proteins (enzymes), and non-peptide products. Their structures, bioactive ac-tivities and the relevant variants as novel lead structures for drug discovery are also described. The challenging effects of fermentation metabolites, isolation and purification, as wel as the overproduction of bioactive com-pounds from B. subtilis by metabolic engineering, were also highlighted. Systematical y exploring biosynthetic routes and the functions of secondary metabolites from B. subtilis may not only be beneficial in improving yields of the products, but also in helping them to be used in food industry and public medical service on a large-scale.

  17. Evaluation of surface sampling method performance for Bacillus Spores on clean and dirty outdoor surfaces.

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, Mollye C.; Einfeld, Wayne; Boucher, Raymond M.; Brown, Gary Stephen; Tezak, Matthew Stephen

    2011-06-01

    Recovery of Bacillus atrophaeous spores from grime-treated and clean surfaces was measured in a controlled chamber study to assess sampling method performance. Outdoor surfaces investigated by wipe and vacuum sampling methods included stainless steel, glass, marble and concrete. Bacillus atrophaeous spores were used as a surrogate for Bacillus anthracis spores in this study designed to assess whether grime-coated surfaces significantly affected surface sampling method performance when compared to clean surfaces. A series of chamber tests were carried out in which known amounts of spores were allowed to gravitationally settle onto both clean and dirty surfaces. Reference coupons were co-located with test coupons in all chamber experiments to provide a quantitative measure of initial surface concentrations of spores on all surfaces, thereby allowing sampling recovery calculations. Results from these tests, carried out under both low and high humidity conditions, show that spore recovery from grime-coated surfaces is the same as or better than spore recovery from clean surfaces. Statistically significant differences between method performance for grime-coated and clean surfaces were observed in only about half of the chamber tests conducted.

  18. A microbial transformation using Bacillus subtilis B7-S to produce natural vanillin from ferulic acid.

    Science.gov (United States)

    Chen, Peng; Yan, Lei; Wu, Zhengrong; Li, Suyue; Bai, Zhongtian; Yan, Xiaojuan; Wang, Ningbo; Liang, Ning; Li, Hongyu

    2016-02-04

    Bacillus subtilis strain B7-S screened from18 strains is an aerobic, endospore-forming, model organism of Gram-positive bacteria which is capable to form vanillin during ferulic acid bioconversion. The bioconversion of ferulic acid to vanillin by Bacillus subtilis B7-S (B. subtilis B7-S) was investigated. Based on our results, the optimum bioconversion conditions for the production of vanillin by B. subtilis B7-S can be summarized as follows: temperature 35 °C; initial pH 9.0; inoculum volume 5%; ferulic acid concentration 0.6 g/L; volume of culture medium 20%; and shaking speed 200 r/min. Under these conditions, several repeated small-scale batch experiments showed that the maximum conversion efficiency was 63.30% after 3 h of bioconversion. The vanillin products were confirmed by spectral data achieved from UV-vis, inductively coupled plasma atomic emission spectroscope (ICP-AES) and Fourier transform infrared spectrometer (FT-IR) spectra. Scanning electron microscopy (SEM) and transmission electron spectroscopy (TEM) results confirmed that the cell surface of B. subtilis plays a role in the induction of ferulic acid tolerance. These results demonstrate that B. subtilis B7-S has the potential for use in vanillin production through bioconversion of ferulic acid.

  19. The LuxS based quorum sensing governs lactose induced biofilm formation by Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Danielle eDuanis-Assaf

    2016-01-01

    Full Text Available Bacillus species present a major concern in the dairy industry as they can form biofilms in pipelines and on surfaces of equipment and machinery used in the entire line of production. These biofilms represent a continuous hygienic problem and can lead to serious economic losses due to food spoilage and equipment impairment. Biofilm formation by Bacillus subtilis is apparently dependent on LuxS quorum sensing (QS by Autoinducer-2 (AI-2. However, the link between sensing environmental cues and AI-2 induced biofilm formation remains largely unknown. The aim of this study is to investigate the role of lactose, the primary sugar in milk, on biofilm formation by B. subtilis and its possible link to QS processes. Our phenotypic analysis shows that lactose induces formation of biofilm bundles as well as formation of colony type biofilms. Furthermore, using reporter strain assays, we observed an increase in AI-2 production by B. subtilis in response to lactose in a dose dependent manner. Moreover, we found that expression of eps and tapA operons, responsible for extracellular matrix synthesis in B. subtilis, were notably up-regulated in response to lactose. Importantly, we also observed that LuxS is essential for B. subtilis biofilm formation in the presence of lactose. Overall, our results suggest that lactose may induce biofilm formation by B. subtilis through the LuxS pathway.

  20. Regulation and expression of the metal citrate transporter CitM of Bacillus subtilis

    NARCIS (Netherlands)

    de Jonge - Warner, Jessica

    2002-01-01

    The main topic of this thesis is the regulation of transcription of the citM gene of Bacillus subtilis, encoding the major metal citrate transporter. CitM belongs to a small family of secondary transport proteins, the MeCit family, that is comprised of 12 members. CitM mediates the transport of meta

  1. Condition-Dependent Transcriptome Reveals High-Level Regulatory Architecture in Bacillus subtilis

    NARCIS (Netherlands)

    Nicolas, Pierre; Maeder, Ulrike; Dervyn, Etienne; Rochat, Tatiana; Leduc, Aurelie; Pigeonneau, Nathalie; Bidnenko, Elena; Marchadier, Elodie; Hoebeke, Mark; Aymerich, Stephane; Becher, Doerte; Bisicchia, Paola; Botella, Eric; Delumeau, Olivier; Doherty, Geoff; Denham, Emma L.; Fogg, Mark J.; Fromion, Vincent; Goelzer, Anne; Hansen, Annette; Haertig, Elisabeth; Harwood, Colin R.; Homuth, Georg; Jarmer, Hanne; Jules, Matthieu; Klipp, Edda; Le Chat, Ludovic; Lecointe, Francois; Lewis, Peter; Liebermeister, Wolfram; March, Anika; Mars, Ruben A. T.; Nannapaneni, Priyanka; Noone, David; Pohl, Susanne; Rinn, Bernd; Ruegheimer, Frank; Sappa, Praveen K.; Samson, Franck; Schaffer, Marc; Schwikowski, Benno; Steil, Leif; Stuelke, Joerg; Wiegert, Thomas; Devine, Kevin M.; Wilkinson, Anthony J.; van Dijl, Jan Maarten; Hecker, Michael; Voelker, Uwe; Bessieres, Philippe; Noirot, Philippe

    2012-01-01

    Bacteria adapt to environmental stimuli by adjusting their transcriptomes in a complex manner, the full potential of which has yet to be established for any individual bacterial species. Here, we report the transcriptomes of Bacillus subtilis exposed to a wide range of environmental and nutritional

  2. Identification of a Bacillus subtilis secretion mutant using a beta-galactosidase screening procedure

    DEFF Research Database (Denmark)

    Jacobs, M F; Borchert, T V; Kontinen, V P

    1995-01-01

    High-level synthesis of exportable beta-galactosidase (LacZ) fusion proteins in Bacillus subtilis results in a lethal phenotype, and has been suggested as a tool for the selection of secretion mutants. We tested a plasmid-based, inducible lacZ fusion gene system for this purpose, but frequent...

  3. Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins

    NARCIS (Netherlands)

    Marciniak, Bogumila C.; Trip, Hein; van-der Veek, Patricia J.; Kuipers, Oscar P.; Marciniak, Bogumiła C.

    2012-01-01

    Background: Bacillus subtilis is a favorable host for the production of industrially relevant proteins because of its capacity of secreting proteins into the medium to high levels, its GRAS (Generally Recognized As Safe) status, its genetic accessibility and its capacity to grow in large fermentatio

  4. Regioselective Synthesis of Polymerizable Vinyl Guaifenesin Esters Catalyzed by an Alkaline Protease of Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    Na WANG; Qi WU; Jian Ming XU; Xiu Ming JIANG; Xian Fu LIN

    2004-01-01

    Three polymerizable vinyl guaifenesin esters with different acyl donor carbon chain lengths (C4,C6,C10) were regioselectivly synthesized by an alkaline protease from Bacillus subtilis in pyridine at 50°C for 1, 3, 5 days respectively.

  5. Thermal Regulation of Membrane Lipid Fluidity by a Two-Component System in "Bacillus Subtilis"

    Science.gov (United States)

    Bredeston, L. M.; Marciano, D.; Albanesi, D.; De Mendoza, D.; Delfino, J. M.

    2011-01-01

    This article describes a simple and robust laboratory exercise on the regulation of membrane unsaturated fatty acid composition in bacteria by a decrease in growth temperature. We take advantage of the well characterized Des pathway of "Bacillus subtilis", composed of a [delta]5-desaturase (encoded by the "des" gene) and the canonical…

  6. Condition-dependent transcriptome reveals high-level regulatory architecture in Bacillus subtilis

    DEFF Research Database (Denmark)

    Nicolas, Pierre; Mäder, Ulrike; Dervyn, Etienne

    2012-01-01

    Bacteria adapt to environmental stimuli by adjusting their transcriptomes in a complex manner, the full potential of which has yet to be established for any individual bacterial species. Here, we report the transcriptomes of Bacillus subtilis exposed to a wide range of environmental and nutritional...

  7. CLONING, SEQUENCING, AND EXPRESSION OF BACILLUS-SUBTILIS GENES INVOLVED IN ATP-DEPENDENT NUCLEASE SYNTHESIS

    NARCIS (Netherlands)

    KOOISTRA, J; VENEMA, G

    1991-01-01

    The genes encoding the subunits of the Bacillus subtilis ATP-dependent nuclease (add genes) have been cloned. The genes were located on an 8.8-kb SalI-SmaI chromosomal DNA fragment. Transformants of a recBCD deletion mutant of Escherichia coli with plasmid pGV1 carrying this DNA fragment showed ATP-

  8. THE BACILLUS-SUBTILIS ADDAB GENES ARE FULLY FUNCTIONAL IN ESCHERICHIA-COLI

    NARCIS (Netherlands)

    KOOISTRA, J; HAIJEMA, BJ; VENEMA, G

    1993-01-01

    An Escherichia coli recBCD deletion mutant was transformed with plasmids containing the Bacillus subtilis add genes. The transformants had relatively high ATP-dependent exonuclease- and ATP-dependent helicase activities, and their viability, the ability to repair u.v.-damaged DNA and the recombinati

  9. Biochemical Characterization of the C-4-Dicarboxylate Transporter DctA from Bacillus subtilis

    NARCIS (Netherlands)

    Groeneveld, Maarten; Detert Oude Weme, Ruud; Duurkens, Ria H.; Slotboom, Dirk Jan

    2010-01-01

    Bacterial secondary transporters of the DctA family mediate ion-coupled uptake of C-4-dicarboxylates. Here, we have expressed the DctA homologue from Bacillus subtilis in the Gram-positive bacterium Lactococcus lactis. Transport of dicarboxylates in vitro in isolated membrane vesicles was assayed. W

  10. Clp-dependent proteolysis down-regulates central metabolic pathways in glucose-starved Bacillus subtilis

    NARCIS (Netherlands)

    Gerth, Ulf; Kock, Holger; Kusters, Ilja; Michalik, Stephan; Switzer, Robert L.; Hecker, Michael

    2008-01-01

    Entry into stationary phase in Bacillus subtilis is linked not only to a redirection of the gene expression program but also to posttranslational events such as protein degradation. Using S-35-labeled methionine pulse-chase labeling and two-dimensional polyacrylamide gel electrophoresis we monitored

  11. Expression of Bacillus subtilis levanase gene in Lactobacillus plantarum and Lactobacillus casei

    NARCIS (Netherlands)

    Wanker, E.; Leer, R.J.; Pouwels, P.H.; Schwab, H.

    1995-01-01

    Two Lactobacillus-Escherichia coli shuttle vectors, harbouring the levanase gene from Bacillus subtilis under the control of its own promoter (pLPEW1) or behind the E. coli tac promoter (pE-SIEW2), were constructed. Lactobacillus plantarum showed the same growth characteristics on selective plates a

  12. Influence of physical, chemical and inducer treatments on menaquinone-7 biosynthesis by Bacillus subtilis MTCC 2756

    Directory of Open Access Journals (Sweden)

    Alka Puri

    2015-06-01

    Full Text Available Effects of physical and chemical treatment on nutrient mobility, their utilization for menaquinone-7 (MK-7 biosynthesis; growth of microbial cells has been investigated in the present research. Bacillus subtilis MTCC 2756 fermented medium was supplied with 1-naphthol and hypoxanthine resulted in a significant increase in MK-7 production. Ultrasonication, electric shock, heat shock, and tween 80 were used for inducer uptake by Bacillus subtilis and menaquinone-7 production. Induction of Bacillus subtilis (at 16 hours of fermentation using 1-naphthol (2 mg/ml, along with tween 80 (0.1% was found to increase the MK-7 production by 3 fold i.e. 14.4 µg/ml as compared to the untreated fermentation medium. The ultrasonicated (ultrasonic power 33 W, treatment time 4 min and frequency 36 KHz microbial cells yielded higher biomass and 2.5 fold increase in the MK-7 production i.e.10.3 µg/ml than control. 1-naphthol along with physical or chemical treatment is required for maximum MK-7 production by Bacillus subtilis.

  13. Primary structure of the tms and prs genes of Bacillus subtilis

    DEFF Research Database (Denmark)

    Nilsson, Dan; Hove-Jensen, Bjarne; Arnvig, Kirsten

    1989-01-01

    The nucleotide sequence was determined of a 3211 nucleotide pair EcoRI-PvuII DNA fragment containing the tms and prs genes as well as a part of the ctc gene of Bacillus subtilis. The prs gene encodes phosphoribosylpyrophosphate (PRPP) synthetase, whereas the functioning of the tms and ctc gene pr...

  14. The complete genome sequence of the Gram-positive bacterium Bacillus subtilis

    NARCIS (Netherlands)

    Kunst, F; Ogasawara, N; Moszer, [No Value; Albertini, AM; Alloni, G; Azevedo, [No Value; Bertero, MG; Bessieres, P; Bolotin, A; Borchert, S; Borriss, R; Boursier, L; Brans, A; Brignell, SC; Bron, S; Brouillet, S; Bruschi, CV; Caldwell, B; Capuano, [No Value; Carter, NM; Choi, SK; Codani, JJ; Connerton, IF; Cummings, NJ; Daniel, RA; Denizot, F; Devine, KM; Dusterhoft, A; Ehrlich, SD; Emmerson, PT; Entian, KD; Errington, J; Fabret, C; Ferrari, E; Foulger, D; Fujita, M; Fujita, Y; Fuma, S; Galizzi, A; Galleron, N; Ghim, SY; Glaser, P; Goffeau, A; Golightly, EJ; Grandi, G; Guiseppi, G; Guy, BJ; Haga, K; Haiech, J; Harwood, CR; Henaut, A; Hilbert, H; Holsappel, S; Hosono, S; Hullo, MF; Itaya, M; Jones, L; Joris, B; Karamata, D; Kasahara, Y; KlaerrBlanchard, M; Klein, C; Kobayashi, Y; Koetter, P; Koningstein, G; Krogh, S; Kumano, M; Kurita, K; Lapidus, A; Lardinois, S; Lauber, J; Lazarevic, [No Value; Lee, SM; Levine, A; Liu, H; Masuda, S; Mauel, C; Medigue, C; Medina, N; Mellado, RP; Mizuno, M; Moestl, D; Nakai, S; Noback, M; Noone, D; OReilly, M; Ogawa, K; Ogiwara, A; Oudega, B; Park, SH; Parro, [No Value; Pohl, TM; Portetelle, D; Porwollik, S; Prescott, AM; Presecan, E; Pujic, P; Purnelle, B; Rapoport, G; Rey, M; Reynolds, S; Rieger, M; Rivolta, C; Rocha, E; Roche, B; Rose, M; Sadaie, Y; Sato, T; Scanlan, E; Schleich, S; Schroeter, R; Scoffone, F; Sekiguchi, J; Sekowska, A; Seror, SJ; Serror, P; Shin, BS; Soldo, B; Sorokin, A; Tacconi, E; Takagi, T; Takahashi, H; Takemaru, K; Takeuchi, M; Tamakoshi, A; Tanaka, T; Terpstra, P; Tognoni, A; Tosato, [No Value; Uchiyama, S; Vandenbol, M; Vannier, F; Vassarotti, A; Viari, A; Wambutt, R; Wedler, E; Wedler, H; Weitzenegger, T; Winters, P; Wipat, A; Yamamoto, H; Yamane, K; Yasumoto, K; Yata, K; Yoshida, K; Yoshikawa, HF; Zumstein, E; Yoshikawa, H; Danchin, A

    1997-01-01

    Bacillus subtilis is the best-characterized member of the Gram-positive bacteria. Its genome of 4,214,810 base pairs comprises 4,100 protein-coding genes. Of these protein-coding genes, 53% are represented once, while a quarter of the genome corresponds to several gene families that have been greatl

  15. Cellular lysis in Bacillus subtilis; the affect of multiple extracellular protease deficiencies

    NARCIS (Netherlands)

    Stephenson, K; Bron, S; Harwood, CR

    1999-01-01

    Cellular lysis properties of strains of Bacillus subtilis deficient in the synthesis of extracellular proteases was investigated. In all cases, extracellular protease deficiency was found to increase the extent of cellular lysis of batch cultured strains following the transition to stationary phase,

  16. In vitro characterization of the Bacillus subtilis protein tyrosine phosphatase YwqE

    DEFF Research Database (Denmark)

    Mijakovic, Ivan; Musumeci, Lucia; Tautz, Lutz;

    2005-01-01

    Both gram-negative and gram-positive bacteria possess protein tyrosine phosphatases (PTPs) with a catalytic Cys residue. In addition, many gram-positive bacteria have acquired a new family of PTPs, whose first characterized member was CpsB from Streptococcus pneumoniae. Bacillus subtilis contains...

  17. Bacillus subtilis SpoIIIJ and YqjG function in membrane protein biogenesis.

    NARCIS (Netherlands)

    Saller, Manfred J.; Fusetti, Fabrizia; Driessen, Arnold J. M.

    2009-01-01

    In all domains of life Oxa1p-like proteins are involved in membrane protein biogenesis. Bacillus subtilis, a model organism for gram-positive bacteria, contains two Oxa1p homologs: SpoIIIJ and YqjG. These molecules appear to be mutually exchangeable, although SpoIIIJ is specifically required for spo

  18. Engineering of quorum-sensing systems for improved production of alkaline protease by Bacillus subtilis

    NARCIS (Netherlands)

    Tjalsma, H; Koetje, EJ; Kiewiet, R; Kuipers, OP; Kolkman, M; van der Laan, J; Daskin, R; Ferrari, E; Bron, S

    2004-01-01

    Aim: Engineering of Rap-Phr quorum-sensing systems of Bacillus subtilis and subsequent evaluation of the transcription of the aprE gene, encoding a major extracellular alkaline protease. Methods and Results: Addition of synthetic Phr pentapeptides to the growth medium, or overproduction of pre-Phr p

  19. Transcriptome analysis documents induced competence of Bacillus subtilis during nitrogen limiting conditions

    DEFF Research Database (Denmark)

    Jarmer, Hanne Østergaard; Berka, R.; Knudsen, Steen;

    2002-01-01

    DNA microarrays were used to analyze the changes in gene expression in Bacillus subtilis strain 168 when nitrogen limiting (glutamate) and nitrogen excess (ammonium plus glutamate) growth conditions were compared. Among more than 100 genes that were significantly induced during nitrogen starvation...

  20. Influence of high-pressure-low-temperature treatment on the inactivation of Bacillus subtilis cells.

    NARCIS (Netherlands)

    T. Shen; G. Urrutia Benet; S. Brul; D. Knorr

    2005-01-01

    High pressure inactivation processes, especially at subzero temperatures, were performed on Bacillus subtilis vegetative cells at various pressure, temperature and time combinations. Whilst atmospheric pressure, lowering the temperature for various periods to as low as 45 -C was found to have minor

  1. A novel screening system for secretion of heterologous proteins in Bacillus subtilis

    NARCIS (Netherlands)

    Trip, Hein; van der Veek, Patricia J.; Renniers, Ton C.; Meima, Rob; Sagt, Cees M.; Mohrmann, Lisette; Kuipers, Oscar P.

    2011-01-01

    High-level production of secretory proteins in Bacillus subtilis leads to a stress response involving the two-component system CssRS and its target genes htrA and htrB. Here, we used this sensing system in a reporter strain in which gfp is under control of P(htrA), the secretion stress responsive pr

  2. Homogeneity and heterogeneity in amylase production by Bacillus subtilis under different growth conditions

    NARCIS (Netherlands)

    Ploss, Tina N.; Reilman, Ewoud; Monteferrante, Carmine G.; Denham, Emma L.; Piersma, Sjouke; Lingner, Anja; Vehmaanpera, Jari; Lorenz, Patrick; van Dijl, Jan Maarten

    2016-01-01

    Background: Bacillus subtilis is an important cell factory for the biotechnological industry due to its ability to secrete commercially relevant proteins in large amounts directly into the growth medium. However, hyper-secretion of proteins, such as alpha-amylases, leads to induction of the secretio

  3. Probing the enantioselectivity of Bacillus subtilis esterase BS2 for tert. alcohols

    NARCIS (Netherlands)

    Wiggers, Michiel; Holt, Jarle; Kourist, Robert; Bartsch, Sebastian; Arends, Isabel W. C. E.; Minnaard, Adriaan J.; Bornscheuer, Uwe T.; Hanefeld, Ulf

    2009-01-01

    The activity and enantioselectivity of several mutants of the esterase BS2 from Bacillus subtilis have been investigated. In the enzymatic hydrolysis of alpha,alpha-disubstituted cyanohydrin acetates, a class of tert. alcohol esters, they were active but not selective. In contrast to this result sim

  4. Bacillus subtilis as a tool for screening soil metagenomic libraries for antimicrobial activities.

    Science.gov (United States)

    Biver, Sophie; Steels, Sébastien; Portetelle, Daniel; Vandenbol, Micheline

    2013-06-28

    Finding new antimicrobial activities by functional metagenomics has been shown to depend on the heterologous host used to express the foreign DNA. Therefore, efforts are devoted to developing new tools for constructing metagenomic libraries in shuttle vectors replicatable in phylogenetically distinct hosts. Here we evaluated the use of the Escherichia coli-Bacillus subtilis shuttle vector pHT01 to construct a forest-soil metagenomic library. This library was screened in both hosts for antimicrobial activities against four opportunistic bacteria: Proteus vulgaris, Bacillus cereus, Staphylococcus epidermidis, and Micrococcus luteus. A new antibacterial activity against B. cereus was found upon screening in B. subtilis. The new antimicrobial agent, sensitive to proteinase K, was not active when the corresponding DNA fragment was expressed in E. coli. Our results validate the use of pHT01 as a shuttle vector and B. subtilis as a host to isolate new activities by functional metagenomics.

  5. The role of a purine-specific nucleoside hydrolase in spore germination of Bacillus thuringiensis.

    Science.gov (United States)

    Liang, Liang; He, Xihong; Liu, Gang; Tan, Huarong

    2008-05-01

    A homologous gene (iunH) of a putative nucleoside hydrolase (NH), which had been identified from the exosporia of Bacillus cereus and Bacillus anthracis spores, was cloned from Bacillus thuringiensis subsp. kurstaki. Disruption of iunH did not affect the vegetative growth and sporulation of Bacillus thuringiensis, but promoted both inosine- and adenosine-induced spore germination. The inosine- or adenosine-induced germination rate decreased when the wild-type iunH gene was overexpressed in Bacillus thuringiensis. The iunH gene product was characterized as a purine-specific NH. The kinetic parameters of IunH with inosine as substrate were K(m)=399+/-115 microM, k(cat)=48.9+/-8.5 s(-1) and k(cat)/K(m)=1.23 x 10(5) M(-1) s(-1). The optimal pH and temperature for IunH were found to be pH 6 and 80 degrees C. Meanwhile, the specific activity of inosine hydrolase in intact spores of the wild-type strain with inosine as substrate was 2.89+/-0.23x10(-2) micromol min(-1) (mg dry wt)(-1). These results indicate that IunH is important in moderating inosine- or adenosine-induced germination of Bacillus thuringiensis spores.

  6. Fluctuations in spo0A transcription control rare developmental transitions in Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Nicolas Mirouze

    2011-04-01

    Full Text Available Phosphorylated Spo0A is a master regulator of stationary phase development in the model bacterium Bacillus subtilis, controlling the formation of spores, biofilms, and cells competent for transformation. We have monitored the rate of transcription of the spo0A gene during growth in sporulation medium using promoter fusions to firefly luciferase. This rate increases sharply during transient diauxie-like pauses in growth rate and then declines as growth resumes. In contrast, the rate of transcription of an rRNA gene decreases and increases in parallel with the growth rate, as expected for stable RNA synthesis. The growth pause-dependent bursts of spo0A transcription, which reflect the activity of the spo0A vegetative promoter, are largely independent of all known regulators of spo0A transcription. Evidence is offered in support of a "passive regulation" model in which RNA polymerase stops transcribing rRNA genes during growth pauses, thus becoming available for the transcription of spo0A. We show that the bursts are followed by the production of phosphorylated Spo0A, and we propose that they represent initial responses to stress that bring the average cell closer to the thresholds for transition to bimodally expressed developmental responses. Measurement of the numbers of cells expressing a competence marker before and after the bursts supports this hypothesis. In the absence of ppGpp, the increase in spo0A transcription that accompanies the entrance to stationary phase is delayed and sporulation is markedly diminished. In spite of this, our data contradicts the hypothesis that sporulation is initiated when a ppGpp-induced depression of the GTP pool relieves repression by CodY. We suggest that, while the programmed induction of sporulation that occurs in stationary phase is apparently provoked by increased flux through the phosphorelay, bet-hedging stochastic transitions to at least competence are induced by bursts in transcription.

  7. Whole genome assembly of a natto production strain Bacillus subtilis natto from very short read data

    Directory of Open Access Journals (Sweden)

    Fujiyama Asao

    2010-04-01

    Full Text Available Abstract Background Bacillus subtilis natto is closely related to the laboratory standard strain B. subtilis Marburg 168, and functions as a starter for the production of the traditional Japanese food "natto" made from soybeans. Although re-sequencing whole genomes of several laboratory domesticated B. subtilis 168 derivatives has already been attempted using short read sequencing data, the assembly of the whole genome sequence of a closely related strain, B. subtilis natto, from very short read data is more challenging, particularly with our aim to assemble one fully connected scaffold from short reads around 35 bp in length. Results We applied a comparative genome assembly method, which combines de novo assembly and reference guided assembly, to one of the B. subtilis natto strains. We successfully assembled 28 scaffolds and managed to avoid substantial fragmentation. Completion of the assembly through long PCR experiments resulted in one connected scaffold for B. subtilis natto. Based on the assembled genome sequence, our orthologous gene analysis between natto BEST195 and Marburg 168 revealed that 82.4% of 4375 predicted genes in BEST195 are one-to-one orthologous to genes in 168, with two genes in-paralog, 3.2% are deleted in 168, 14.3% are inserted in BEST195, and 5.9% of genes present in 168 are deleted in BEST195. The natto genome contains the same alleles in the promoter region of degQ and the coding region of swrAA as the wild strain, RO-FF-1. These are specific for γ-PGA production ability, which is related to natto production. Further, the B. subtilis natto strain completely lacked a polyketide synthesis operon, disrupted the plipastatin production operon, and possesses previously unidentified transposases. Conclusions The determination of the whole genome sequence of Bacillus subtilis natto provided detailed analyses of a set of genes related to natto production, demonstrating the number and locations of insertion sequences that B

  8. Association and decontamination of Bacillus spores in a simulated drinking water system.

    Science.gov (United States)

    Morrow, J B; Almeida, J L; Fitzgerald, L A; Cole, K D

    2008-12-01

    The objective of this work was to elucidate the disinfectant susceptibility of Bacillus anthracis Sterne (BA) and a commercial preparation of Bacillus thuringiensis (BT) spores associated with a simulated drinking water system. Biofilms composed of indigenous water system bacteria were accumulated on copper and polyvinyl chloride (PVC) pipe material surfaces in a low-flow pipe loop and uniformly mixed tank reactor (CDC biofilm reactor). Application of a distributed shear during spore contact resulted in approximately a 1.0 and 1.6 log10 increase in the number of spores associated with copper and PVC surfaces, respectively. Decontamination of spores in both free suspension and after association with biofilm-conditioned pipe materials was attempted using free chlorine and monochloramine. Associated spores required 5- to 10-fold higher disinfectant concentrations to observe the same reduction of viable spores as in suspension. High disinfectant concentrations (103 mg/L free chlorine and 49 mg/L monochloramine) yielded less than a 2-log10 reduction in viable associated spores after 60 min. Spores associated with biofilms on copper surfaces consistently yielded higher Ct values than PVC.

  9. The RecA-Dependent SOS Response Is Active and Required for Processing of DNA Damage during Bacillus subtilis Sporulation.

    Science.gov (United States)

    Ramírez-Guadiana, Fernando H; Barajas-Ornelas, Rocío Del Carmen; Corona-Bautista, Saúl U; Setlow, Peter; Pedraza-Reyes, Mario

    2016-01-01

    The expression of and role played by RecA in protecting sporulating cells of Bacillus subtilis from DNA damage has been determined. Results showed that the DNA-alkylating agent Mitomycin-C (M-C) activated expression of a PrecA-gfpmut3a fusion in both sporulating cells' mother cell and forespore compartments. The expression levels of a recA-lacZ fusion were significantly lower in sporulating than in growing cells. However, M-C induced levels of ß-galactosidase from a recA-lacZ fusion ~6- and 3-fold in the mother cell and forespore compartments of B. subtilis sporangia, respectively. Disruption of recA slowed sporulation and sensitized sporulating cells to M-C and UV-C radiation, and the M-C and UV-C sensitivity of sporangia lacking the transcriptional repair-coupling factor Mfd was significantly increased by loss of RecA. We postulate that when DNA damage is encountered during sporulation, RecA activates the SOS response thus providing sporangia with the repair machinery to process DNA lesions that may compromise the spatio-temporal expression of genes that are essential for efficient spore formation.

  10. The RecA-Dependent SOS Response Is Active and Required for Processing of DNA Damage during Bacillus subtilis Sporulation.

    Directory of Open Access Journals (Sweden)

    Fernando H Ramírez-Guadiana

    Full Text Available The expression of and role played by RecA in protecting sporulating cells of Bacillus subtilis from DNA damage has been determined. Results showed that the DNA-alkylating agent Mitomycin-C (M-C activated expression of a PrecA-gfpmut3a fusion in both sporulating cells' mother cell and forespore compartments. The expression levels of a recA-lacZ fusion were significantly lower in sporulating than in growing cells. However, M-C induced levels of ß-galactosidase from a recA-lacZ fusion ~6- and 3-fold in the mother cell and forespore compartments of B. subtilis sporangia, respectively. Disruption of recA slowed sporulation and sensitized sporulating cells to M-C and UV-C radiation, and the M-C and UV-C sensitivity of sporangia lacking the transcriptional repair-coupling factor Mfd was significantly increased by loss of RecA. We postulate that when DNA damage is encountered during sporulation, RecA activates the SOS response thus providing sporangia with the repair machinery to process DNA lesions that may compromise the spatio-temporal expression of genes that are essential for efficient spore formation.

  11. Bacillus spore classification via surface-enhanced Raman spectroscopy and principal component analysis.

    Science.gov (United States)

    Guicheteau, J; Argue, L; Emge, D; Hyre, A; Jacobson, M; Christesen, S

    2008-03-01

    Surface-enhanced Raman spectroscopy (SERS) can provide rapid fingerprinting of biomaterial in a nondestructive manner. The adsorption of colloidal silver to biological material suppresses native biofluorescence while providing electromagnetic surface enhancement of the normal Raman signal. This work validates the applicability of qualitative SER spectroscopy for analysis of bacterial species by utilizing principal component analysis (PCA) to show discrimination of biological threat simulants, based upon multivariate statistical confidence limits bounding known data clusters. Gram-positive Bacillus spores (Bacillus atrophaeus, Bacillus anthracis, and Bacillus thuringiensis) are investigated along with the Gram-negative bacterium Pantoea agglomerans.

  12. Effects of salinomycin and Bacillus subtilis on growth performance and immune responses in broiler chickens.

    Science.gov (United States)

    Lee, Kyung-Woo; Lillehoj, Hyun S; Jang, Seung I; Lee, Sung-Hyen

    2014-10-01

    The present study was undertaken to compare the effect of salinomycin and Bacillus subtilis on growth performance, serum antibody levels against Clostridium spp. and Eimeria spp., and cytokine mRNA expression levels in broiler chickens raised in the used litter. Broiler chickens fed a diet containing salinomycin showed lower (P salinomycin-fed or control diet-fed chickens. None of the dietary treatments affected (P > 0.05) serum antibody levels against Clostridium perfringens toxins. Both salinomycin and B.subtilis significantly lowered (P Salinomycin, but not B. subtilis, significantly modulated (P salinomycin and B. subtilis affected serum anticoccidial antibody and intestinal cytokine expression, but failed to improve growth performance in broiler chickens. Further study is warranted to investigate the mode of action of salinomycin on host immune response and growth performance in broiler chickens.

  13. Fed-Batch Biomolecule Production by Bacillus subtilis: A State of the Art Review.

    Science.gov (United States)

    Öztürk, Sibel; Çalık, Pınar; Özdamar, Tunçer H

    2016-04-01

    Bacillus subtilis is a highly promising production system for various biomolecules. This review begins with the algorithm of fed-batch operations (FBOs) and then illustrates the approaches to design the initial production medium and/or feed stream. Additionally, the feeding strategies developed with or without feedback control for fed-batch B. subtilis fermentations were compiled with a special emphasis on recombinant protein (r-protein) production. For biomolecule production by wild-type B. subtilis, due to the different intracellular production patterns, no consensus exists on the FBO strategy that gives the maximum productivity, whereas for r-protein production appropriate feeding strategies vary depending on the promoter used. Thus, we conclude that the B. subtilis community is still seeking an approved strong promoter and generalized FBO strategies.

  14. Growth of and valine production by a Bacillus subtilis mutant in the small intestine of pigs

    DEFF Research Database (Denmark)

    Canibe, Nuria; Poulsen, Henrik Vestergaard; Nørgaard, Jan Værum;

    2016-01-01

    :Lys of 0.63:1 (Neg), 2) the Neg diet with added Bacillus subtilis-valine (1.28 × 108 cfu/g feed) (+Bac), and 3) the Neg diet with added L-Val to a Val:Lys of 0.69:1 (+Val). Eighteen gilts (6 on each treatment) with initial weights of ∼15 kg were fed the diets for 23 d before the animals were euthanized...... and samples from the small intestine were obtained. The number of B. subtilis cfu in digesta was higher in the +Bac group than in the Neg group (P ... concentrations were measured in the +Bac group. Short-term in vitro incubations of digesta showed a decrease (P ≤ 0.03) in the number of B. subtilis cfu over time for the +Bac group and no difference in the rate of Val production compared to that in the Neg group. In conclusion, more B. subtilis cfu were present...

  15. Investigating the efficacy of Bacillus subtilis SM21 on controlling Rhizopus rot in peach fruit.

    Science.gov (United States)

    Wang, Xiaoli; Wang, Jing; Jin, Peng; Zheng, Yonghua

    2013-06-17

    The efficacy of Bacillus subtilis SM21 on controlling Rhizopus rot caused by Rhizopus stolonifer in postharvest peach fruit and the possible mechanisms were investigated. The results indicated B. subtilis SM21 treatment reduced lesion diameter and disease incidence by 37.2% and 26.7% on the 2nd day of inoculation compared with the control. The in vitro test showed significant inhibitory effect of B. subtilis SM21 on mycelial growth of R. stolonifer with an inhibition rate of 48.9%. B. subtilis SM21 treatment significantly enhanced activities of chitinase and β-1,3-glucanase, and promoted accumulation of H2O2. Total phenolic content and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity were also increased by this treatment. Transcription of seven defense related genes was much stronger in fruit treated with B. subtilis SM21 or those both treated with B. subtilis SM21 and inoculated with R. stolonifer compared with fruit inoculated with R. stolonifer alone. These results suggest that B. subtilis SM21 can effectively inhibit Rhizopus rot caused by R. stolonifer in postharvest peach fruit, possibly by directly inhibiting growth of the pathogen, and indirectly inducing disease resistance in the fruit.

  16. Production of Enzymes from Agroindustrial Wastes by Biosurfactant-Producing Strains of Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Francisco Fábio Cavalcante Barros

    2013-01-01

    Full Text Available Bacteria in the genus Bacillus are the source of several enzymes of current industrial interest. Hydrolases, such as amylases, proteases, and lipases, are the main enzymes consumed worldwide and have applications in a wide range of products and industrial processes. Fermentation processes by Bacillus subtilis using cassava wastewater as a substrate are reported in the technical literature; however, the same combination of microorganisms and this culture medium is limited or nonexistent. In this paper, the amylase, protease, and lipase production of ten Bacillus subtilis strains previously identified as biosurfactant producers in cassava wastewater was evaluated. The LB1a and LB5a strains were selected for analysis using a synthetic medium and cassava wastewater and were identified as good enzyme producers, especially of amylases and proteases. In addition, the enzymatic activity results indicate that cassava wastewater was better than the synthetic medium for the induction of these enzymes.

  17. Decontamination options for Bacillus anthracis-contaminated drinking water determined from spore surrogate studies.

    Science.gov (United States)

    Raber, Ellen; Burklund, Alison

    2010-10-01

    Five parameters were evaluated with surrogates of Bacillus anthracis spores to determine effective decontamination alternatives for use in a contaminated drinking water supply. The parameters were as follows: (i) type of Bacillus spore surrogate (B. thuringiensis or B. atrophaeus), (ii) spore concentration in suspension (10(2) and 10(6) spores/ml), (iii) chemical characteristics of the decontaminant (sodium dichloro-S-triazinetrione dihydrate [Dichlor], hydrogen peroxide, potassium peroxymonosulfate [Oxone], sodium hypochlorite, and VirkonS), (iv) decontaminant concentration (0.01% to 5%), and (v) exposure time to decontaminant (10 min to 1 h). Results from 138 suspension tests with appropriate controls are reported. Hydrogen peroxide at a concentration of 5% and Dichlor or sodium hypochlorite at a concentration of 2% were highly effective at spore inactivation regardless of spore type tested, spore exposure time, or spore concentration evaluated. This is the first reported study of Dichlor as an effective decontaminant for B. anthracis spore surrogates. Dichlor's desirable characteristics of high oxidation potential, high level of free chlorine, and a more neutral pH than that of other oxidizers evaluated appear to make it an excellent alternative. All three oxidizers were effective against B. atrophaeus spores in meeting the EPA biocide standard of greater than a 6-log kill after a 10-min exposure time and at lower concentrations than typically reported for biocide use. Solutions of 5% VirkonS and Oxone were less effective as decontaminants than other options evaluated in this study and did not meet the EPA's efficacy standard for a biocide, although they were found to be as effective for concentrations of 10(2) spores/ml. Differences in methods and procedures reported by other investigators make quantitative comparisons among studies difficult.

  18. The characterisation of Bacillus spores occurring in the manufacturing of (low acid) canned products.

    Science.gov (United States)

    Oomes, S J C M; van Zuijlen, A C M; Hehenkamp, J O; Witsenboer, H; van der Vossen, J M B M; Brul, S

    2007-11-30

    Spore-forming bacteria can be a problem in the food industry, especially in the canning industry. Spores present in ingredients or present in the processing environment severely challenge the preservation process since their thermal resistance may be very high. We therefore asked the question which bacterial spore formers are found in a typical soup manufacturing plant, where they originate from and what the thermal resistance of their spores is. To answer these questions molecular techniques for bacterial species and strain identification were used as well as a protocol for the assessment of spore heat stress resistance based on the Kooiman method. The data indicate the existence and physiological cause of the high thermal resistance of spores of many of the occurring species. In particular it shows that ingredients used in soup manufacturing are a rich source of high thermal resistant spores and that sporulation in the presence of ingredients rich in divalent metal ions exerts a strong influence on spore heat resistance. It was also indicated that Bacillus spores may well be able to germinate and resporulate during manufacturing i.e. through growth and sporulation in line. Both these spores and those originating from the ingredients were able to survive certain thermal processing settings. Species identity was confirmed using fatty acid analysis, 16SrRNA gene sequencing and DNA-DNA hybridisation. Finally, molecular typing experiments using Ribotyping and AFLP analysis show that strains within the various Bacillus species can be clustered according to the thermal resistance properties of their spores. AFLP performed slightly better than Ribotyping. The data proofed to be useful for the generation of strain specific probes. Protocols to validate these probes in routine identification and innovation aimed at tailor made heat processing in soup manufacturing have been formulated.

  19. Mutagenesis and ultraviolet inactivation of transforming DNA of ``Haemophilus influenzae`` complexed with a ``Bacillus subtilis`` protein that alter DNA conformation

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, Jane K. [Brookhaven National Lab., Upton, NY (United States); Setlow, Barbara C.; Setlow, Peter [Connecticut Univ., Farmington, CT (United States)

    1996-12-31

    The wild-type ``Bacillus subtilis`` spore protein, SspC{sup wt}, binds to DNA ``in vitro`` and ``in vivo`` and changes the conformation of DNA from B to A. Synthesis of the cloned SspC{sup wt} gene in ``Escherichia coli`` also causes large increases in mutation frequency. Binding of SspC{sup wt} to transforming DNA from ``Haemophilus influenzae`` made the DNA resistant to ultraviolet (UV) radiation. The mutant protein, SspC{sup ala}, which does not bind DNA, did not change the UV resistance. The UV sensitivity of the DNA/SspC{sup wt} complex was not increased when the recipients of the DNA were defective in excision of pyrimidine dimers. These data indicate that the ``H. influenzae`` excision mechanism does not operate on the spore photoproduct formed by UV irradiation of the complex. Selection for the streptomycin- or erythromycin-resistance markers on the transforming DNA evidenced significant mutations at loci closely linked to these, but not at other loci. SspC{sup wt} apparently entered the cell attached to the transforming DNA, and caused mutations in adjacent loci. The amount of such mutations decreased when the transforming DNA was UV irradiated, because UV unlinks linked markers. (author). 22 refs, 4 figs, 4 tabs.

  20. The Adsorption Properties of Bacillus atrophaeus Spores on Single-Wall Carbon Nanotubes

    Directory of Open Access Journals (Sweden)

    P. Cortes

    2009-01-01

    Full Text Available An adsorption equilibrium and a kinetic study of Bacillus atrophaeus on Single-Wall Carbon Nanotubes (SWCNTs were here performed to provide the basis for developing biosensor devices for detecting threatening micro-organisms in water supply systems. B. atrophaeus spores and carbon nanotubes were subjected to a batch adsorption process to document their equilibria and kinetics. Here, commercial nanotubes were either studied as received or were acid-purified before adsorption experiments. The Bacillus spores appear to show higher affinity towards the purified nanotubes than to the as-received nanomaterial. The effective diffusivity of the spores onto the purified nanotubes was found to be approximately 30 percent higher than onto the as-received nanotubes. It seems that the removal of amorphous carbon from the as-received nanotubes through a purification process yielded an intimate nantoubes-spore interaction as revealed by transmission electron microscopy. Freundlich model successfully correlated the adsorption equilibrium data for the nanotubes-spore interaction. Transmission electron micrographs showed extensive contact between the Bacillus and the purified nanotubes, but the association appeared less intimate between the spores and the as-received nanotubes.

  1. Role of YpeB in cortex hydrolysis during germination of Bacillus anthracis spores.

    Science.gov (United States)

    Bernhards, Casey B; Popham, David L

    2014-10-01

    The infectious agent of the disease anthrax is the spore of Bacillus anthracis. Bacterial spores are extremely resistant to environmental stresses, which greatly hinders spore decontamination efforts. The spore cortex, a thick layer of modified peptidoglycan, contributes to spore dormancy and resistance by maintaining the low water content of the spore core. The cortex is degraded by germination-specific lytic enzymes (GSLEs) during spore germination, rendering the cells vulnerable to common disinfection techniques. This study investigates the relationship between SleB, a GSLE in B. anthracis, and YpeB, a protein necessary for SleB stability and function. The results indicate that ΔsleB and ΔypeB spores exhibit similar germination phenotypes and that the two proteins have a strict codependency for their incorporation into the dormant spore. In the absence of its partner protein, SleB or YpeB is proteolytically degraded soon after expression during sporulation, rather than escaping the developing spore. The three PepSY domains of YpeB were examined for their roles in the interaction with SleB. YpeB truncation mutants illustrate the necessity of a region beyond the first PepSY domain for SleB stability. Furthermore, site-directed mutagenesis of highly conserved residues within the PepSY domains resulted in germination defects corresponding to reduced levels of both SleB and YpeB in the mutant spores. These results identify residues involved in the stability of both proteins and reiterate their codependent relationship. It is hoped that the study of GSLEs and interacting proteins will lead to the use of GSLEs as targets for efficient activation of spore germination and facilitation of spore cleanup.

  2. Role of YpeB in Cortex Hydrolysis during Germination of Bacillus anthracis Spores

    Science.gov (United States)

    Bernhards, Casey B.

    2014-01-01

    The infectious agent of the disease anthrax is the spore of Bacillus anthracis. Bacterial spores are extremely resistant to environmental stresses, which greatly hinders spore decontamination efforts. The spore cortex, a thick layer of modified peptidoglycan, contributes to spore dormancy and resistance by maintaining the low water content of the spore core. The cortex is degraded by germination-specific lytic enzymes (GSLEs) during spore germination, rendering the cells vulnerable to common disinfection techniques. This study investigates the relationship between SleB, a GSLE in B. anthracis, and YpeB, a protein necessary for SleB stability and function. The results indicate that ΔsleB and ΔypeB spores exhibit similar germination phenotypes and that the two proteins have a strict codependency for their incorporation into the dormant spore. In the absence of its partner protein, SleB or YpeB is proteolytically degraded soon after expression during sporulation, rather than escaping the developing spore. The three PepSY domains of YpeB were examined for their roles in the interaction with SleB. YpeB truncation mutants illustrate the necessity of a region beyond the first PepSY domain for SleB stability. Furthermore, site-directed mutagenesis of highly conserved residues within the PepSY domains resulted in germination defects corresponding to reduced levels of both SleB and YpeB in the mutant spores. These results identify residues involved in the stability of both proteins and reiterate their codependent relationship. It is hoped that the study of GSLEs and interacting proteins will lead to the use of GSLEs as targets for efficient activation of spore germination and facilitation of spore cleanup. PMID:25022853

  3. Quantification of the impact of single and multiple mild stresses on outgrowth heterogeneity of Bacillus cereus spores

    NARCIS (Netherlands)

    Melis, van C.C.J.; Besten, den H.M.W.; Nierop Groot, M.N.; Abee, T.

    2014-01-01

    Outgrowth heterogeneity of bacterial spore populations complicates both prediction and efficient control of spore outgrowth. In this study, the impact of mild preservation stresses on outgrowth of Bacillus cereus ATCC 14579 spores was quantified during the first stages of outgrowth. Heterogeneity in

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2011-01-01

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

  5. Aktifitas Antimikroba Ekstrak Angsana (Pterocarpus indicus terhadap Bacillus subtilis dan Klebsiella pneumoniae

    Directory of Open Access Journals (Sweden)

    TULUS JUNANTO

    2008-11-01

    Full Text Available Indonesia has much kind of plants, which have medicinal properties and used to cure various diseases. Angsana (Pterocarpus indicus is one of tree plant that has many used, one of them as city ornamental tree. The aim of the research was to know the antimicrobial effect off crude extract angsana against Bacillus subtilis and Klebsiella pneumoniae. Crude extract angsana is made in maceration with methanol, chloroform, and hexane. The part of angsana is leaf, stem bark and root. Diffusion method is used to test antimicrobial activity. Effect of antimicrobial is shown by halo zone. The minimum inhibitory concentrations (MMICs of methanol crude extract of leaf is 250 µg//µl, methanol crude extract of stem bark and root are 100 µg/µl and 100 µg/µll for K. pneumoniae. MICs of methanol crude extract of stem bark and root are 100 µg/µµl and 1000 µµg/µl for Bacillus subtilis. MICs of chloroform crude extract off stem bark and root are 1000 µg/µl and 500 µg/µl for KK. pneumoniae. MICs of chloroform cru de extract of stem bark and root are 550 µg/µl and 550 µg/µl for B. subtilis. MICs of hexane crude extract of stem bark is 500 µg/µl and 1000 µg/µl for K. pneumoniae and B. subtilis, respectively. Crude extract of leaf, stem bark and root of angsana could inhibit growth of B. subtilis and K. pneumoniae bacteria.

  6. Self-cloning significantly enhances the production of catalase in Bacillus subtilis WSHDZ-01.

    Science.gov (United States)

    Xu, Sha; Guo, Yaqiong; Du, Guocheng; Zhou, Jingwen; Chen, Jian

    2014-08-01

    The katA gene that encodes catalase (CAT) in Bacillus subtilis WSHDZ-01 was overexpressed in B. subtilis WB600 and B. subtilis WSHDZ-01. The CAT yield in both transformed strains was significantly improved compared to that in the wild-type WSHDZ-01 in shake flask culture. When cultured in a 3-L stirred tank reactor (STR), the recombinant CAT activity in B. subtilis WSHDZ-01 could be improved by 419 %, reaching up to 39,117 U/mL and was 8,149.4 U/mg dry cell weight, which is the highest activity reported in Bacillus sp. However, the recombinant CAT in B. subtilis WB600 cultured in a 3-L STR was not significantly improved by any of the common means for process optimization, and the highest CAT activity was 3,673.5 U/mg dry cell weight. The results suggest that self-cloning of the complete expression cassette in the original strain is a reasonable strategy to improve the yield of wild-type enzymes.

  7. 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.

  8. Rapid Detection of Bacillus anthracis Spores Using Immunomagnetic Separation and Amperometry

    Science.gov (United States)

    Waller, David F.; Hew, Brian E.; Holdaway, Charlie; Jen, Michael; Peckham, Gabriel D.

    2016-01-01

    Portable detection and quantitation methods for Bacillus anthracis (anthrax) spores in pure culture or in environmental samples are lacking. Here, an amperometric immunoassay has been developed utilizing immunomagnetic separation to capture the spores and remove potential interferents from test samples followed by amperometric measurement on a field-portable instrument. Antibody-conjugated magnetic beads and antibody-conjugated glucose oxidase were used in a sandwich format for the capture and detection of target spores. Glucose oxidase activity of spore pellets was measured indirectly via amperometry by applying a bias voltage after incubation with glucose, horseradish peroxidase, and the electron mediator 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid). Target capture was mediated by polyclonal antisera, whereas monoclonal antibodies were used for signal generation. This strategy maximized sensitivity (500 target spores, 5000 cfu/mL), while also providing a good specificity for Bacillus anthracis spores. Minimal signal deviation occurs in the presence of environmental interferents including soil and modified pH conditions, demonstrating the strengths of immunomagnetic separation. The simultaneous incubation of capture and detection antibodies and rapid substrate development (5 min) result in short sample-to-signal times (less than an hour). With attributes comparable or exceeding that of ELISA and LFDs, amperometry is a low-cost, low-weight, and practical method for detecting anthrax spores in the field. PMID:27999382

  9. The structure of the transposable genetic element ISBsu2 from the cryptic plasmid p1516 of a soil Bacillus subtilis strain and the presence of homologues of this element in the chromosomes of various Bacillus subtilis strains

    NARCIS (Netherlands)

    Holsappel, S; Gagarina, EY; Poluektova, EU; Nezametdinova, VZ; Gel'fand, MS; Prozorov, AA; Bron, S

    2003-01-01

    A cryptic plasmid from a soil strain of Bacillus subtilis was found to contain a sequence having features of an IS element. Homologous sequences were also found in the chromosome of this strain and in the chromosomes of some other B. subtilis strains.

  10. From Gene Regulation to Gene Function: Regulatory Networks in Bacillus Subtilis

    Directory of Open Access Journals (Sweden)

    Ivan Moszer

    2006-04-01

    Full Text Available Bacillus subtilis is a sporulating Gram-positive bacterium that lives primarily in the soil and associated water sources. The publication of the B. subtilis genome sequence and subsequent systematic functional analysis and gene regulation programmes, together with an extensive understanding of its biochemistry and physiology, makes this micro-organism a prime candidate in which to model regulatory networks in silico. In this paper we discuss combined molecular biological and bioinformatical approaches that are being developed to model this organism’s responses to changes in its environment.

  11. Identification of a Bacillus subtilis secretion mutant using a ß-galactosidase screening procedure

    DEFF Research Database (Denmark)

    Jacobs, Myra F.; Andersen, Jens Bo; Borchert, Torben V.;

    1995-01-01

    High-level synthesis of exportable beta-galactosidase (LacZ) fusion proteins in Bacillus subtilis results in a lethal phenotype, and has been suggested as a tool for the selection of secretion mutants. We tested a plasmid-based, inducible lacZ fusion gene system for this purpose, but frequent...... mutations in cis, which reduced expression of the fusion gene, forced abandonment of the induction-selection strategy. Instead, after modification of the indicator plasmid, a screening procedure for increased basal LacZ activity levels was adopted. This led to the identification of a conditional B. subtilis...

  12. Sampling and quantitative analysis of clean B. subtilis spores at sub-monolayer coverage by reflectance fourier transform infrared microscopy using gold-coated filter substrates.

    Science.gov (United States)

    Brooke, Heather; Perkins, David L; Setlow, Barbara; Setlow, Peter; Bronk, Burt V; Myrick, Michael L

    2008-08-01

    A study was conducted to determine the concentration dependency of the mid-infrared (MIR) absorbance of bacterial spores. A range of concentrations of Bacillus subtilis endospores filtered across gold-coated filter membranes were analyzed by Fourier transform infrared (FT-IR) reflectance microscopy. Calibration curves were derived from the peak absorbances associated with Amide A, Amide I, and Amide II vibrational frequencies by automatic baseline fitting to remove most of the scattering contribution. Linear relationships (R2 >or= 0.99) were observed between the concentrations of spores and the baseline-corrected peak absorbance for each frequency studied. Detection limits for our sampled area of 100 x100 microm2 were determined to be 79, 39, and 184 spores (or 7.92 x 10(5), 3.92 x 10(5), and 1.84 x 10(6) spores/cm2) for the Amide A, Amide I, and Amide II peaks, respectively. Absorbance increased linearly above the scattering baseline with particle surface concentration up to 0.9 monolayer (ML) coverage, with the monolayer density calculated to be approximately 1.17 x 10(8) spores/cm2. Scattering as a function of surface concentration, as estimated from extinction values at wavelengths exhibiting low absorbance, becomes nonlinear at a much lower surface concentration. The apparent scattering cross-section per spore decreased monotonically as concentrations increased toward 1.2 ML, while the absolute scattering decreased between 0.9 ML and 1.2 ML coverage. Calculations suggest that transverse spatial coherence effects are the origin of this nonlinearity, while the onset of nonlinearity in the baseline-corrected absorption is probably due to multiple scattering effects, which appear at a high surface concentration. Absorption cross-sections at peaks of the three bands were measured to be (2.15 +/- 0.05) x 10(-9), (1.48 +/- 0.03) x 10(-9), and (0.805 +/- 0.023) x 10(-9) cm2, respectively. These values are smaller by a factor of 2-4 than expected from the literature

  13. Novel strategies for enhanced removal of persistent Bacillus anthracis surrogates and Clostridium difficile spores from skin.

    Directory of Open Access Journals (Sweden)

    Michelle M Nerandzic

    Full Text Available BACKGROUND: Removing spores of Clostridium difficile and Bacillus anthracis from skin is challenging because they are resistant to commonly used antimicrobials and soap and water washing provides only modest efficacy. We hypothesized that hygiene interventions incorporating a sporicidal electrochemically generated hypochlorous acid solution (Vashe(® would reduce the burden of spores on skin. METHODS: Hands of volunteers were inoculated with non-toxigenic C. difficile spores or B. anthracis spore surrogates to assess the effectiveness of Vashe solution for reducing spores on skin. Reduction in spores was compared for Vashe hygiene interventions versus soap and water (control. To determine the effectiveness of Vashe solution for removal of C. difficile spores from the skin of patients with C. difficile infection (CDI, reductions in levels of spores on skin were compared for soap and water versus Vashe bed baths. RESULTS: Spore removal from hands was enhanced with Vashe soak (>2.5 log10 reduction versus soap and water wash or soak (~2.0 log10 reduction; P3.5 log10 spores from hands (P<0.01 compared to washing or soaking alone. Bed baths using soap and water (N =26 patients did not reduce the percentage of positive skin cultures for CDI patients (64% before versus 57% after bathing; P =0.5, whereas bathing with Vashe solution (N =21 patients significantly reduced skin contamination (54% before versus 8% after bathing; P =0.0001. Vashe was well-tolerated with no evidence of adverse effects on skin. CONCLUSIONS: Vashe was safe and effective for reducing the burden of B. anthracis surrogates and C. difficile spores on hands. Bed baths with Vashe were effective for reducing C. difficile on skin. These findings suggest a novel strategy to reduce the burden of spores on skin.

  14. Adaptation of Bacillus subtilis carbon core metabolism to simultaneous nutrient limitation and osmotic challenge : a multi-omics perspective

    NARCIS (Netherlands)

    Kohlstedt, Michael; Sappa, Praveen K; Meyer, Hanna; Maaß, Sandra; Zaprasis, Adrienne; Hoffmann, Tamara; Becker, Judith; Steil, Leif; Hecker, Michael; van Dijl, Jan Maarten; Lalk, Michael; Mäder, Ulrike; Stülke, Jörg; Bremer, Erhard; Völker, Uwe; Wittmann, Christoph

    2014-01-01

    The Gram-positive bacterium Bacillus subtilis encounters nutrient limitations and osmotic stress in its natural soil ecosystem. To ensure survival and sustain growth, highly integrated adaptive responses are required. Here, we investigated the system-wide response of B.subtilis to different, simulta

  15. Isolation and characterization of Bacillus subtilis genes involved in siderophore biosynthesis: relationship between B. subtilis sfpo and Escherichia coli entD genes.

    OpenAIRE

    Grossman, T H; Tuckman, M; Ellestad, S; Osburne, M S

    1993-01-01

    In response to iron deprivation, Bacillus subtilis secretes a catecholic siderophore, 2,3-dihydroxybenzoyl glycine, which is similar to the precursor of the Escherichia coli siderophore enterobactin. We isolated two sets of B. subtilis DNA sequences that complemented the mutations of several E. coli siderophore-deficient (ent) mutants with defective enterobactin biosynthesis enzymes. One set contained DNA sequences that complemented only an entD mutation. The second set contained DNA sequence...

  16. Analysis of a Novel Spore Antigen in Bacillus anthracis That Contributes to Spore Opsonization

    Science.gov (United States)

    2008-01-01

    identity with homologues in B. cereus and Bacillus thuringiensis (99 and 94 %, respectively). In addition, a small ORF (BA5270) was located immediately...N. R. (1962). Field evaluation of a human anthrax vaccine. Am J Public Health 52, 632–645. Brossier, F. & Mock, M. (2001). Toxins of Bacillus ...authors (2007). The complete genome sequence of Bacillus thuringiensis Al Hakam. J Bacteriol 189, 3680–3681. Clements, M. O. & Moir, A. (1998). Role of

  17. Comparative analysis of Bacillus weihenstephanensis KBAB4 spores obtained at different temperatures

    NARCIS (Netherlands)

    Garcia, D.; Voort, van der M.; Abee, T.

    2010-01-01

    The impact of Bacillus weihenstephanensis KBAB4 sporulation temperature history was assessed on spore heat resistance, germination and outgrowth capacity at a temperature range from 7 to 30 °C. Sporulation rate and efficiency decreased at low temperature, as cells sporulated at 12, 20 and 30 °C with

  18. Soya bean tempe extracts show antibacterial activity against Bacillus cereus cells and spores

    NARCIS (Netherlands)

    Roubos-van den Hil, P.J.; Dalmas, E.; Nout, M.J.R.; Abee, T.

    2010-01-01

    Aims: Tempe, a Rhizopus ssp.-fermented soya bean food product, was investigated for bacteriostatic and/or bactericidal effects against cells and spores of the food-borne pathogen Bacillus cereus. Methods and results: Tempe extract showed a high antibacterial activity against B. cereus ATCC 14579 bas

  19. Characterization of a spore-specific protein of the Bacillus cereus group

    NARCIS (Netherlands)

    From, C.; Voort, van der M.; Abee, T.; Granum, P.E.

    2012-01-01

    Bc1245 is a monocistronic chromosomal gene of Bacillus cereus ATCC 14579 encoding a putative protein of 143 amino acids identified in this study to have a spore-related function in B. cereus. Bc1245 is highly conserved in the genome of members of the B. cereus group, indicating an important function

  20. Germination and outgrowth of spores of Bacillus cereus group members: diversity and role of germinant receptors.

    Science.gov (United States)

    Abee, Tjakko; Groot, Masja Nierop; Tempelaars, Marcel; Zwietering, Marcel; Moezelaar, Roy; van der Voort, Menno

    2011-04-01

    Bacillus cereus is a gram-positive, facultative anaerobic, endospore-forming toxicogenic human pathogen. Endospores are highly specialized, metabolically dormant cell types that are resistant to extreme environmental conditions, including heat, dehydration and other physical stresses. B. cereus can enter a range of environments, and can in its spore form, survive harsh conditions. If these conditions become favorable, spores can germinate and grow out and reach considerable numbers in a range of environments including processed foods. Certainly the last decade, when consumer preferences have shifted to mildly processed food, new opportunities arose for spore-forming spoilage and pathogenic organisms. Only rigorous methods have been shown to be capable of destroying all spores present in food, thus a shift toward e.g., milder heat preservation strategies, may result in low but significant amounts of viable spores in food products. Hence, the need for a mild spore destruction strategy is eminent including control of spore outgrowth. Consequently, there is a large interest in triggering spore germination in foodstuffs, since germinated spores have lost the extreme resistance of dormant spores and are relatively easy to kill. Another option could be to prevent germination so that no dangerous levels can be reached. This contribution will focus on germination and outgrowth characteristics of B. cereus and other members of the B. cereus group, providing an overview of the niches these spore-formers can occupy, the signals that trigger germination, and how B. cereus copes with these wake-up calls in different environments including foods, during food processing and upon interaction with the human host.

  1. Morphological and mechanical imaging of Bacillus cereus spore formation at the nanoscale.

    Science.gov (United States)

    Wang, Congzhou; Stanciu, Cristina; Ehrhardt, Christopher J; Yadavalli, Vamsi K

    2015-04-01

    Bacteria from the genus Bacillus are able to transform into metabolically dormant states called (endo) spores in response to nutrient deprivation and other harsh conditions. These morphologically distinct spores are fascinating constructs, amongst the most durable cells in nature, and have attracted attention owing to their relevance in food-related illnesses and bioterrorism. Observing the course of bacterial spore formation (sporulation) spatially, temporally and mechanically, from the vegetative cell to a mature spore, is critical for a better understanding of this process. Here, we present a fast and versatile strategy for monitoring both the morphological and mechanical changes of Bacillus cereus bacteria at the nanoscale using atomic force microscopy. Through a strategy of imaging and nanomechanical mapping, we show the morphogenesis of the endospore and released mature endospore. Finally, we investigate individual spores to characterize their surface mechanically. The progression in elasticity coupled with a similarity of characteristic distributions between the incipient endospores and the formed spores show these distinct stages. Taken together, our data demonstrates the power of atomic force microscopy applied in microbiology for probing this important biological process at the single cell scale.

  2. Thermal inactivation of Bacillus anthracis surrogate spores in a bench-scale enclosed landfill gas flare.

    Science.gov (United States)

    Tufts, Jenia A McBrian; Rosati, Jacky A

    2012-02-01

    A bench-scale landfill flare system was designed and built to test the potential for landfilled biological spores that migrate from the waste into the landfill gas to pass through the flare and exit into the environment as viable. The residence times and temperatures of the flare were characterized and compared to full-scale systems. Geobacillus stearothermophilus and Bacillus atrophaeus, nonpathogenic spores that may serve as surrogates for Bacillus anthracis, the causative agent for anthrax, were investigated to determine whether these organisms would be inactivated or remain viable after passing through a simulated landfill flare. High concentration spore solutions were aerosolized, dried, and sent through a bench-scale system to simulate the fate of biological weapon (BW)-grade spores in a landfill gas flare. Sampling was conducted downstream of the flare using a bioaerosol collection device containing sterile white mineral oil. The samples were cultured, incubated for seven days, and assessed for viability. Results showed that the bench-scale system exhibited good similarity to the real-world conditions of an enclosed standard combustor flare stack with a single orifice, forced-draft diffusion burner. All spores of G. stearothermophilus and B. atrophaeus were inactivated in the flare, indicating that spores that become re-entrained in landfill gas may not escape the landfill as viable, apparently becoming completely inactivated as they exit through a landfill flare.

  3. Bacillus globigii bugbeads: a model simulant of a bacterial spore.

    Science.gov (United States)

    Farrell, Svetlana; Halsall, H Brian; Heineman, William R

    2005-01-15

    Nonpathogenic microorganisms are often used as simulants of biological pathogens during the initial phase of detection method development. While these simulants approximate the size, shape, and cellular organization of the microorganism of interest, they do not resemble its surface protein content, a factor particularly important in methods based on immunorecognition. Here, we develop and detect an artificial bacterial spore--B. globigii (BG) Bugbead-a particle mimicking the antigenic surface of BG spores. Two methods of spore protein extraction were compared both quantitatively (by protein concentration assay) and qualitatively (by SDS-PAGE and Western blot): extraction by mechanical disruption and extraction by chemical decoating. The former method was more efficient in producing more protein and a greater number of antigens. BG Bugbeads were made by conjugating the extracted proteins to 0.8-microm carboxyl-coated polystyrene particles via carbodiimide coupling. BG Bugbeads were successfully detected by a bead-based enzyme-labeled immunoassay with fluorescence detection with a detection limit of 6.9 x 10(3) particles/mL. Formation of the Bugbead-capture bead complex was confirmed by ESEM. The concept of a harmless artificial spore can be applied to developing improved simulants for pathogenic spore-forming microorganisms such as B. anthracis, C. botulinum, and B. cereus, which can to be used for method validation, instrument calibration, and troubleshooting.

  4. Adsorption of Cu2+, Zn2+ and Cd2+ on Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A process of biosorption of Cu2+, Zn2+ and Cd2+ on Bacillus subtilis was investigated.The experiments show that the process of biosorption is quite fast. The maximum adsorption was reached after 5 min and hardly changed with time. The experimental data was analyzed using four sorption kinetic models: the pseudo-first-order, the Ritchie second-order, the modified second-order and the Elovich equations, which helped to determine the best-fit equation for the sorption of metal ions onto biomass. The results show that both the Ritchie second-order and modified secondorder equations can fit the experimental data. The Langmuir model is able to accurately describe adsorption of Cu2+ and Zn2+ on B. subtilis. The experimental data points of adsorption Cd2+ and Zn2+ on B. subtilis are described by Freundlich isotherms model.

  5. Association of Eu(III) and Cm(III) with Bacillus subtilis and Halobacterium salinarum

    Energy Technology Data Exchange (ETDEWEB)

    Ozaki, Takuo; Kimura, Takaumi; Ohnuki, Toshihiko; Yoshida, Zenko [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment; Gillow, Jeffrey B.; Francis, Arokiasamy J. [Brookhaven National Laboratory, Upton, NY (United States)

    2002-11-01

    Adsorption behavior of Eu(III) and Cm(III) by Bacillus subtilis and Halobacterium salinarum was investigated. Both microorganisms showed almost identical pH dependence on the distribution ratio (K{sub d}) of the metals examined, i.e., K{sub d} of Eu(III) and Cm(III) increased with an increase of pH. The coordination state of Eu(III) adsorbed on the microorganisms was studied by time-resolved laser-induced fluorescence spectroscopy (TRLFS). The coordination states of Eu(III) adsorbed on the B. subtilis and H. salinarum was of different characteristics. H. salinarum exhibited more outer-spherical interaction with Eu(III) than B. subtilis. (author)

  6. Reconstruction of the Regulatory Network for Bacillus subtilis and Reconciliation with Gene Expression Data

    Energy Technology Data Exchange (ETDEWEB)

    Faria, José P.; Overbeek, Ross; Taylor, Ronald C.; Conrad, Neal; Vonstein, Veronika; Goelzer, Anne; Fromion, Vincent; Rocha, Miguel; Rocha, Isabel; Henry, Christopher S.

    2016-03-18

    We introduce a manually constructed and curated regulatory network model that describes the current state of knowledge of transcriptional regulation of B. subtilis. The model corresponds to an updated and enlarged version of the regulatory model of central metabolism originally proposed in 2008. We extended the original network to the whole genome by integration of information from DBTBS, a compendium of regulatory data that includes promoters, transcription factors (TFs), binding sites, motifs and regulated operons. Additionally, we consolidated our network with all the information on regulation included in the SporeWeb and Subtiwiki community-curated resources on B. subtilis. Finally, we reconciled our network with data from RegPrecise, which recently released their own less comprehensive reconstruction of the regulatory network for B. subtilis. Our model describes 275 regulators and their target genes, representing 30 different mechanisms of regulation such as TFs, RNA switches, Riboswitches and small regulatory RNAs. Overall, regulatory information is included in the model for approximately 2500 of the ~4200 genes in B. subtilis 168. In an effort to further expand our knowledge of B. subtilis regulation, we reconciled our model with expression data. For this process, we reconstructed the Atomic Regulons (ARs) for B. subtilis, which are the sets of genes that share the same “ON” and “OFF” gene expression profiles across multiple samples of experimental data. We show how atomic regulons for B. subtilis are able to capture many sets of genes corresponding to regulated operons in our manually curated network. Additionally, we demonstrate how atomic regulons can be used to help expand or validate the knowledge of the regulatory networks by looking at highly correlated genes in the ARs for which regulatory information is lacking. During this process, we were also able to infer novel stimuli for hypothetical genes by exploring the genome expression metadata

  7. Expression, purification and preliminary crystallographic characterization of FlhF from Bacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Bange, Gert; Petzold, Georg; Wild, Klemens; Sinning, Irmgard, E-mail: irmi.sinning@bzh.uni-heidelberg.de [Heidelberg University Biochemistry Centre (BZH), INF 328, 69120 Heidelberg (Germany)

    2007-05-01

    Preliminary crystallographic data are reported for the third SRP GTPase FlhF from Bacillus subtilis. The Gram-positive bacterium Bacillus subtilis contains three proteins belonging to the signal recognition particle (SRP) type GTPase family. The well characterized signal sequence-binding protein SRP54 and the SRP receptor protein FtsY are universally conserved components of the SRP system of protein transport. The third member, FlhF, has been implicated in the placement and assembly of polar flagella. This article describes the overexpression and preliminary X-ray crystallographic analysis of an FlhF fragment that corresponds to the well characterized GTPase domains in SRP54 and FtsY. Three crystal forms are reported with either GDP or GMPPNP and diffract to a resolution of about 3 Å.

  8. Improved production, characterization and flocculation properties of poly (-glutamic acid produced from Bacillus Subtilis

    Directory of Open Access Journals (Sweden)

    Bhunia B

    2012-04-01

    Full Text Available Bacillus subtilis 2063 produced extracellular biopolymer whichshowed excellent flocculation activity. The biopolymer wasconfirmed as poly (γ-glutamic acid (PGA by using productcharacterization. HPLC profile showed that molecular weight ofPGA was found to be 5.8×106 Da. Improved production,Characterization and flocculation properties of PGA produced byBacillus species were studied. PGA produced by B. subtilis wasdevoid of any polysaccharides. The flocculating activity wasmarkedly stimulated by the addition of cations. The pH of reaction mixture also influenced the flocculating activity. Glycerol and ammonium chloride were found to be most useful carbon and nitrogen sources. An overall 4.24-fold increase in protease production was achieved in the design medium composed with Glycerol and ammonium chloride as a carbon and nitrogen sources as compared with basal media. PGA production increased significantly with optimized medium (21.42 gl-1 when compared with basal medium (5.06 gl-1.

  9. Five new amicoumacins isolated from a marine-derived Bacterium bacillus subtilis

    KAUST Repository

    Li, Yongxin

    2012-02-03

    Four novel amicoumacins, namely lipoamicoumacins A-D (1-4), and one new bacilosarcin analog (5) were isolated from culture broth of a marine-derived bacterium Bacillus subtilis, together with six known amicoumacins. Their structures were elucidated on the basis of extensive spectroscopic (2D NNR, IR, CD and MS) analysis and in comparison with data in literature. 2012 by the authors; licensee MDPI.

  10. REMOVAL OF PHOSPHATE FROM RHIZOSPHERE SOIL USING Bacillus subtilis AND Enterobacter aerogenes

    Directory of Open Access Journals (Sweden)

    Andrew J.

    2014-03-01

    Full Text Available The addition of phosphorus is one of the major environmental problems because of its leading contribution to the increased eutrophication process of lakes and other natural waters. The eutrophication is the process where excessive nutrients in a lake or other body of water usually caused by runoff of nutrients (animal waste, fertilizers, and sewage from the land which causes a dense growth of plant life, the decomposition of the plants depletes the supply of oxygen which leads to the death of animal life. Microbial process is widely used for the removal of phosphorus from soil and wastewater to avoid eutrophication. The most efficient phosphate reducers chosen were namely Bacillus subtilis and Enterobacter aerogenes. The Mineral Salt Medium and the carbon sources (glucose, sucrose, lactose and starch at 0.5% and 0.7% were prepared. On the removal of phosphate by Bacillus subtilis and Enterobacter aerogenes it was found that the Bacillus subtilis was giving the maximum bacterial growth and was observed to be in lactose 0.107 OD at 0.7% concentration for 72th hour. In the case of Enterobacter aerogenes the maximum bacterial growth was found to be in sucrose 0.133 OD at 0.7% concentration at 72 hr. The pH change in the medium was found to be in both the isolates with different carbon sources but in overall the constant pH was at 7. Among the two organisms, Bacillus subtilis showed the maximum removal of phosphate 83% as starch as carbon source at 0.5% concentration whereas Enterobacter aerogenes showed 77.4% of phosphate removal at 0.5% concentration as glucose as carbon source. Therefore, these bacterial isolates can be used in the remediation of phosphate contaminated environments.

  11. Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis

    OpenAIRE

    Márcia Aiko Shirakawa; Maria Alba Cincotto; Daniel Atencio; Gaylarde,Christine C.; John,Vanderley M.

    2011-01-01

    The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS), as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in...

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

    OpenAIRE

    2014-01-01

    Author Summary DNA replication must be coordinated with cellular physiology to ensure proper genome inheritance. Model bacteria such as the soil-dwelling Bacillus subtilis can achieve a wide range of growth rates in response to nutritional and chemical signals. In order to match the rate of DNA synthesis to the rate of nutrient-mediated cell growth, bacteria regulate the initiation frequency of DNA replication. This control of bacterial DNA replication initiation was first observed over forty...

  13. The Crystal Structure of Bacillus subtilis Lipase : A Minimal α/β Hydrolase Fold Enzyme

    NARCIS (Netherlands)

    Pouderoyen, Gertie van; Eggert, Thorsten; Jaeger, Karl-Erich; Dijkstra, Bauke W.

    2001-01-01

    The X-ray structure of the lipase LipA from Bacillus subtilis has been determined at 1.5 Å resolution. It is the first structure of a member of homology family I.4 of bacterial lipases. The lipase shows a compact minimal α/β hydrolase fold with a six-stranded parallel β-sheet flanked by five α-helic

  14. Bacillus cereus spores and cereulide in food-borne illness

    OpenAIRE

    Shaheen, Ranad

    2009-01-01

    B. cereus is a gram-positive bacterium that possesses two different forms of life:the large, rod-shaped cells (ca. 0.002 mm by 0.004 mm) that are able to propagate and the small (0.001 mm), oval shaped spores. The spores can survive in almost any environment for up to centuries without nourishment or water. They are insensitive towards most agents that normally kill bacteria: heating up to several hours at 90 ºC, radiation, disinfectants and extreme alkaline (≥ pH 13) and acid (≀ pH 1) e...

  15. Graphical procedure for comparing thermal death of Bacillus stearothermophilus spores in saturated and superheated steam.

    Science.gov (United States)

    SHULL, J J; ERNST, R R

    1962-09-01

    The thermal death curve of dried spores of Bacillus stearothermophilus in saturated steam was characterized by three phases: (i) a sharp initial rise in viable count; (ii) a low rate of death which gradually increased; and (iii) logarithmic death at maximal rate. The first phase was a reflection of inadequate heat activation of the spore population. The second and third phases represented the characteristic thermal death curve of the spores in saturated steam. A jacketed steam sterilizer, equipped with a system for initial evacuation of the chamber, was examined for superheat during normal operation. Measurements of spore inactivation and temperature revealed superheat in surface layers of fabrics being processed in steam at 121 C. The high temperature of the fabric surfaces was attributed to absorption of excess heat energy from superheated steam. The superheated steam was produced at the beginning of the normal sterilizing cycle by transfer of heat from the steam-heated jacket to saturated steam entering the vessel.

  16. Isolation, purification and characterization of Bacillus subtilis Phytase from Holiwood Gresik

    Directory of Open Access Journals (Sweden)

    Leny Yuanita

    2012-01-01

    Full Text Available The aim of the research were isolation, purification and characterization of Bacillus subtilis phytase from Holiwood Gresik. The research was done in two stages; the first include enzyme isolation, precipitation with amonium sulphate, dialysis, gel filtration chromatography, SDS-PAGE analysis, while second determining optimum pH, optimum temperature, the effect of pH and temperature to enzim stability, the values of KM and Vmax Bacillus subtilis phytase from Holiwood Gresik. The first stage research design were One Shot Case Study and Post Test Only Control Group Design, while the second stage were Post Test Only Control Group Design and Factorial Design. The data being analyzed by one-way and two-way Anova. The results of research showed that Bacillus subtilis phytase has the molecular mass of 36.5 kDa, optimum pH at 6.5–7.0, optimum temperature at 41°C and it was found to be stable for 30 minute incubation at pH 7or 30° C with 2% or 3% lost of its activity respectively. KM value was 0.62 mM and VMax 0.393 mmol/ml/minute.

  17. Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis

    Directory of Open Access Journals (Sweden)

    Márcia Aiko Shirakawa

    2011-06-01

    Full Text Available The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS, as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in the two media after incubation with and without the bacterial inoculum was determined by atomic absorption analysis. Modified B4 gave the best results and in this medium Pseudomonas putida INQCS 113 produced the highest calcium carbonate precipitation, followed by Lysinibacillus sphaericus INQCS 414; the lowest precipitation was produced by Bacillus subtilis INQCS 328. In this culture medium XRD analysis showed that Pseudomonas putida and Bacillus subtilis precipitated calcite and vaterite polymorphs while Lysinibacillus sphaericus produced only vaterite. The shape and size of the crystals were affected by culture medium, bacterial strain and culture conditions, static or shaken. In conclusion, of the three strains Pseudomonas putida INQCS 113 in modified B4 medium gave the best results precipitating 96% of the calcium, this strain thus has good potential for use on building materials.

  18. Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis.

    Science.gov (United States)

    Shirakawa, Márcia Aiko; Cincotto, Maria Alba; Atencio, Daniel; Gaylarde, Christine C; John, Vanderley M

    2011-04-01

    The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS), as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in the two media after incubation with and without the bacterial inoculum was determined by atomic absorption analysis. Modified B4 gave the best results and in this medium Pseudomonas putida INQCS 113 produced the highest calcium carbonate precipitation, followed by Lysinibacillus sphaericus INQCS 414; the lowest precipitation was produced by Bacillus subtilis INQCS 328. In this culture medium XRD analysis showed that Pseudomonas putida and Bacillus subtilis precipitated calcite and vaterite polymorphs while Lysinibacillus sphaericus produced only vaterite. The shape and size of the crystals were affected by culture medium, bacterial strain and culture conditions, static or shaken. In conclusion, of the three strains Pseudomonas putida INQCS 113 in modified B4 medium gave the best results precipitating 96% of the calcium, this strain thus has good potential for use on building materials.

  19. Directed natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis

    KAUST Repository

    Li, Yongxin

    2015-03-24

    Bacilli are ubiquitous low G+C environmental Gram-positive bacteria that produce a wide assortment of specialized small molecules. Although their natural product biosynthetic potential is high, robust molecular tools to support the heterologous expression of large biosynthetic gene clusters in Bacillus hosts are rare. Herein we adapt transformation-associated recombination (TAR) in yeast to design a single genomic capture and expression vector for antibiotic production in Bacillus subtilis. After validating this direct cloning plug-and-playa approach with surfactin, we genetically interrogated amicoumacin biosynthetic gene cluster from the marine isolate Bacillus subtilis 1779. Its heterologous expression allowed us to explore an unusual maturation process involving the N-acyl-asparagine pro-drug intermediates preamicoumacins, which are hydrolyzed by the asparagine-specific peptidase into the active component amicoumacin A. This work represents the first direct cloning based heterologous expression of natural products in the model organism B. subtilis and paves the way to the development of future genome mining efforts in this genus.

  20. Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis

    Science.gov (United States)

    Shirakawa, Márcia Aiko; Cincotto, Maria Alba; Atencio, Daniel; Gaylarde, Christine C.; John, Vanderley M.

    2011-01-01

    The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS), as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in the two media after incubation with and without the bacterial inoculum was determined by atomic absorption analysis. Modified B4 gave the best results and in this medium Pseudomonas putida INQCS 113 produced the highest calcium carbonate precipitation, followed by Lysinibacillus sphaericus INQCS 414; the lowest precipitation was produced by Bacillus subtilis INQCS 328. In this culture medium XRD analysis showed that Pseudomonas putida and Bacillus subtilis precipitated calcite and vaterite polymorphs while Lysinibacillus sphaericus produced only vaterite. The shape and size of the crystals were affected by culture medium, bacterial strain and culture conditions, static or shaken. In conclusion, of the three strains Pseudomonas putida INQCS 113 in modified B4 medium gave the best results precipitating 96% of the calcium, this strain thus has good potential for use on building materials. PMID:24031661

  1. Study of the catalytic properties of bacillus subtilis proteases Estudio de las propiedades catalíticas de las proteasas bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Salcedo L.

    1998-06-01

    Full Text Available The catalytic properties of proteases isolated from the filtrate of submerged fermentation of Bacillus subtilis were investigated. Proteases present in the filtrate were determined to be of the serine protease type based on the use of specific protease inhibitors; ethylenediamintetraacetic acid (EDTA was used as a metalloprotease inhibitor, and phenylmethylsulfonylfluoride (PMSF was used as a serine protease inhibitor. Protease activity was highly stable in alkaline solutions and at high temperatures as well as in the presence of detergents. We propose that this protease preparation be used as biocomponent in detergent production.Se investigaron las propiedades catalíticas de las proteasas obtenidas del filtrado de cultivo de la bacteria Bacillus subtilis. Utilizando inhibidores específicos de proteasas se determinó que las proteasas presentes en el filtrado pertenecían al grupo de las serina proteasas. Se utilizó ácido etilendiaminatetraacético (EDTA como inhibidor de metaloproteasas, y fenilmetilsulfonil fluoruro (FMSF como inhibidor de serina proteasas. La actividad proteolítica fue altamente estable en soluciones alcalinas y a altas temperaturas, además tolero la presencia de detergentes. Se propone que estas proteasas sean utilizadas en calidad de biocomponente para la producción de detergentes.

  2. 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

  3. A Study on Effect of different culture media on amylase enzyme production by a native strain of Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    ziba Akbari

    2015-12-01

    Full Text Available Introduction: Amylases are among the most important enzymes and have great significance in present-day biotechnology. Amylase with commercial applications is mainly derived from the genus Bacillus. The main purpose of this study is identification and isolatation amylase enzyme producer Bacillus, determining the amylase enzyme activity and affecting a number of culture medium on amylase enzyme production. Materials and methods: Soil, water and wastewater samples were collected from agricultural area, choghakhor lake in chahar mahal e bakhtiari province and from food factory in Esfahan. Bacillus isolates were screened for amylolytic properties by starch hydrolysis test on starch agar plate. Amylase producing Bacillus were identified biochemical tests and molecular experiments. Amylase enzyme activity of isolates was measured using di-nitro salicylic acid (DNS method. Enzyme production was studied in variose medium culture TSB, NB, Yeast extract, molases and milk medium. Results: The enzyme amylase-producing strains, one sample showed was the highest amylase activity. The Bacillus has been detected as a member of Bacillus subtilis according to Bergey's Manual of Systematic Bacteriology and molecular recognition. The enzyme activity of Bacillus subtilis was measured 7/21 (U/ml in production media. Trough medium culture maximum amylase production for Bacillus subtilis was achieved in molases medium. Discussion and conclusion: In this study, Bacillus subtilis strains isolated from wastewater of a significant amount of enzyme producing 7/21 (U/ml as indicated. Among the medium-amylase from Bacillus subtilis highest enzyme activity was observed in beet molasses. According to this study, the use of Bacillus strains is an efficient way to achieve the amylase enzyme.

  4. Differential actions of chlorhexidine on the cell wall of Bacillus subtilis and Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Hon-Yeung Cheung

    Full Text Available Chlorhexidine is a chlorinated phenolic disinfectant used commonly in mouthwash for its action against bacteria. However, a comparative study of the action of chlorhexidine on the cell morphology of gram-positive and gram-negative bacteria is lacking. In this study, the actions of chlorhexidine on the cell morphology were identified with the aids of electron microscopy. After exposure to chlorhexidine, numerous spots of indentation on the cell wall were found in both Bacillus subtilis and Escherichia coli. The number of indentation spots increased with time of incubation and increasing chlorhexidine concentration. Interestingly, the dented spots found in B. subtilis appeared mainly at the hemispherical caps of the cells, while in E. coli the dented spots were found all over the cells. After being exposed to chlorhexidine for a prolonged period, leakage of cellular contents and subsequent ghost cells were observed, especially from B subtilis. By using 2-D gel/MS-MS analysis, five proteins related to purine nucleoside interconversion and metabolism were preferentially induced in the cell wall of E. coli, while three proteins related to stress response and four others in amino acid biosynthesis were up-regulated in the cell wall materials of B. subtilis. The localized morphological damages together with the biochemical and protein analysis of the chlorhexidine-treated cells suggest that chlorhexidine may act on the differentially distributed lipids in the cell membranes/wall of B. subtilis and E. coli.

  5. Antagonistic action of Bacillus subtilis strain SG6 on Fusarium graminearum.

    Science.gov (United States)

    Zhao, Yueju; Selvaraj, Jonathan Nimal; Xing, Fuguo; Zhou, Lu; Wang, Yan; Song, Huimin; Tan, Xinxin; Sun, Lichao; Sangare, Lancine; Folly, Yawa Minnie Elodie; Liu, Yang

    2014-01-01

    Fusarium graminearum causes Fusarium head blight (FHB), a devastating disease that leads to extensive yield and quality loss of wheat and barley. Bacteria isolated from wheat kernels and plant anthers were screened for antagonistic activity against F. graminearum. Based on its in vitro effectiveness, strain SG6 was selected for characterization and identified as Bacillus subtilis. B. subtilis SG6 exhibited a high antifungal effect on the mycelium growth, sporulation and DON production of F. graminearum with the inhibition rate of 87.9%, 95.6% and 100%, respectively. In order to gain insight into biological control effect in situ, we applied B. subtilis SG6 at anthesis through the soft dough stage of kernel development in field test. It was revealed that B. subtilis SG6 significantly reduced disease incidence (DI), FHB index and DON (P ≤ 0.05). Further, ultrastructural examination shows that B. subtilis SG6 strain induced stripping of F. graminearum hyphal surface by destroying the cellular structure. When hypha cell wall was damaged, the organelles and cytoplasm inside cell would exude, leading to cell death. The antifungal activity of SG6 could be associated with the coproduction of chitinase, fengycins and surfactins.

  6. Antagonistic action of Bacillus subtilis strain SG6 on Fusarium graminearum.

    Directory of Open Access Journals (Sweden)

    Yueju Zhao

    Full Text Available Fusarium graminearum causes Fusarium head blight (FHB, a devastating disease that leads to extensive yield and quality loss of wheat and barley. Bacteria isolated from wheat kernels and plant anthers were screened for antagonistic activity against F. graminearum. Based on its in vitro effectiveness, strain SG6 was selected for characterization and identified as Bacillus subtilis. B. subtilis SG6 exhibited a high antifungal effect on the mycelium growth, sporulation and DON production of F. graminearum with the inhibition rate of 87.9%, 95.6% and 100%, respectively. In order to gain insight into biological control effect in situ, we applied B. subtilis SG6 at anthesis through the soft dough stage of kernel development in field test. It was revealed that B. subtilis SG6 significantly reduced disease incidence (DI, FHB index and DON (P ≤ 0.05. Further, ultrastructural examination shows that B. subtilis SG6 strain induced stripping of F. graminearum hyphal surface by destroying the cellular structure. When hypha cell wall was damaged, the organelles and cytoplasm inside cell would exude, leading to cell death. The antifungal activity of SG6 could be associated with the coproduction of chitinase, fengycins and surfactins.

  7. Improved proteomic analysis following trichloroacetic acid extraction of Bacillus anthracis spore proteins.

    Science.gov (United States)

    Deatherage Kaiser, Brooke L; Wunschel, David S; Sydor, Michael A; Warner, Marvin G; Wahl, Karen L; Hutchison, Janine R

    2015-11-01

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Analysis of cellular proteins is dependent upon efficient extraction from bacterial samples, which can be challenging with increasing complexity and refractory characteristics. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrichment for certain classes of proteins. The method presented here is technically simple, does not require specialized equipment such as a mechanical disrupter, and is effective for protein extraction of the particularly challenging sample type of Bacillus anthracis Sterne spores. The ability of Trichloroacetic acid (TCA) extraction to isolate proteins from spores and enrich for spore-specific proteins was compared to the traditional mechanical disruption method of bead beating. TCA extraction improved the total average number of proteins identified within a sample as compared to bead beating (547 vs 495, respectively). Further, TCA extraction enriched for 270 spore proteins, including those typically identified by first isolating the spore coat and exosporium layers. Bead beating enriched for 156 spore proteins more typically identified from whole spore proteome analyses. The total average number of proteins identified was equal using TCA or bead beating for easily lysed samples, such as B. anthracis vegetative cells. As with all assays, supplemental methods such as implementation of an alternative preparation method may simplify sample preparation and provide additional insight to the protein biology of the organism being studied.

  8. Maximum shields: the assembly and function of the bacterial spore coat.

    Science.gov (United States)

    Driks, Adam

    2002-06-01

    Spores produced by bacilli and clostridia are surrounded by a multilayered protein shell called the coat. As the armor-like appearance of the coat suggests, this structure, along with others within the spore, confers the remarkable resistance properties that make Bacillus anthracis spores such potent biological weapons. Here, I review recent studies of coat assembly in the model organism Bacillus subtilis, and explore the implications of these findings for coat assembly in B. anthracis and for defense against biological weapons.

  9. Storage Effects on Sample Integrity of Environmental Surface Sampling Specimens with Bacillus anthracis Spores.

    Science.gov (United States)

    Perry, K Allison; O'Connell, Heather A; Rose, Laura J; Noble-Wang, Judith A; Arduino, Matthew J

    The effect of packaging, shipping temperatures and storage times on recovery of Bacillus anthracis. Sterne spores from swabs was investigated. Macrofoam swabs were pre-moistened, inoculated with Bacillus anthracis spores, and packaged in primary containment or secondary containment before storage at -15°C, 5°C, 21°C, or 35°C for 0-7 days. Swabs were processed according to validated Centers for Disease Control/Laboratory Response Network culture protocols, and the percent recovery relative to a reference sample (T0) was determined for each variable. No differences were observed in recovery between swabs held at -15° and 5°C, (p ≥ 0.23). These two temperatures provided significantly better recovery than swabs held at 21°C or 35°C (all 7 days pooled, p ≤ 0.04). The percent recovery at 5°C was not significantly different if processed on days 1, 2 or 4, but was significantly lower on day 7 (day 2 vs. 7, 5°C, 10(2), p=0.03). Secondary containment provided significantly better percent recovery than primary containment, regardless of storage time (5°C data, p ≤ 0.008). The integrity of environmental swab samples containing Bacillus anthracis spores shipped in secondary containment was maintained when stored at -15°C or 5°C and processed within 4 days to yield the optimum percent recovery of spores.

  10. The high-resolution architecture and structural dynamics of Bacillus spores

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Leighton, T J; Wheeler, K E; Malkin, A J

    2004-05-06

    The capability to image single microbial cell surfaces at nanometer scale under native conditions would profoundly impact mechanistic and structural studies of pathogenesis, immunobiology, environmental resistance and biotransformation. We report here that advances in atomic force microscopy (AFM) have allowed us to directly visualize high-resolution native structures of bacterial endospores, including the exosporium and spore coats of four Bacillus species in air and water environments. The dimensions of individual Bacillus atrophaeus spores were found to decrease reversibly by 12% in response to a change in the environment from aqueous to aerial phase. Intraspecies spore size distribution analyses revealed that spore length could vary by a factor of 2 while the absolute deviation is 7 - 13% in length and 4 - 6 % in width. AFM analysis also demonstrated that the mechanisms of spore coat self-assembly are similar to those described for inorganic and macromolecular crystallization. These results establish AFM as a powerful new tool for the analysis of molecular architecture and variability as a function of spatial, temporal and developmental organizational scales.

  11. Identification and validation of specific markers of Bacillus anthracis spores by proteomics and genomics approaches.

    Science.gov (United States)

    Chenau, Jérôme; Fenaille, François; Caro, Valérie; Haustant, Michel; Diancourt, Laure; Klee, Silke R; Junot, Christophe; Ezan, Eric; Goossens, Pierre L; Becher, François

    2014-03-01

    Bacillus anthracis is the causative bacteria of anthrax, an acute and often fatal disease in humans. The infectious agent, the spore, represents a real bioterrorism threat and its specific identification is crucial. However, because of the high genomic relatedness within the Bacillus cereus group, it is still a real challenge to identify B. anthracis spores confidently. Mass spectrometry-based tools represent a powerful approach to the efficient discovery and identification of such protein markers. Here we undertook comparative proteomics analyses of Bacillus anthracis, cereus and thuringiensis spores to identify proteoforms unique to B. anthracis. The marker discovery pipeline developed combined peptide- and protein-centric approaches using liquid chromatography coupled to tandem mass spectrometry experiments using a high resolution/high mass accuracy LTQ-Orbitrap instrument. By combining these data with those from complementary bioinformatics approaches, we were able to highlight a dozen novel proteins consistently observed across all the investigated B. anthracis spores while being absent in B. cereus/thuringiensis spores. To further demonstrate the relevance of these markers and their strict specificity to B. anthracis, the number of strains studied was extended to 55, by including closely related strains such as B. thuringiensis 9727, and above all the B. cereus biovar anthracis CI, CA strains that possess pXO1- and pXO2-like plasmids. Under these conditions, the combination of proteomics and genomics approaches confirms the pertinence of 11 markers. Genes encoding these 11 markers are located on the chromosome, which provides additional targets complementary to the commonly used plasmid-encoded markers. Last but not least, we also report the development of a targeted liquid chromatography coupled to tandem mass spectrometry method involving the selection reaction monitoring mode for the monitoring of the 4 most suitable protein markers. Within a proof

  12. Processing, Assembly and Localization of a Bacillus anthracis Spore Protein

    Science.gov (United States)

    2010-01-01

    10.1099/ mic .0.033407-0 174 033407 Printed in Great Britain Approved for public release. Distribution is unlimited Report Documentation Page Form...on LB agar plates to assay viable cells. In vivo challenges. Female Hartley guinea pigs (350–400 g) were obtained from Charles River Laboratories...Guinea pigs were challenged intramuscularly (Fellows et al., 2001) by injection of 200 ml of heat-activated spores suspended in water. The animals were

  13. Characteristics and antimicrobial activity of Bacillus subtilis strains isolated from soil.

    Science.gov (United States)

    Todorova, Sevdalina; Kozhuharova, Lubka

    2010-07-01

    Antagonistic Bacillus strains were isolated from soil and analyzed for the purpose of determining whether they could be used as natural biological agents. Primary in vitro screening for antagonism of the isolates was performed against five phytopathogenic mould fungi. Strains TS 01 and ZR 02 exhibited the most pronounced inhibitory effects. They were identified as Bacillus subtilis on the basis of their morphological, cultural and physiology-biochemical properties as well as their hierarchical cluster analysis conducted by means of computer program SPSS. The antimicrobial activity of the strains from cultural medium and sterile filtrate were determined in vitro against a great number of predominantly phytopathogenic fungi and bacteria. TS 01 and ZR 02 strains exhibited very broad and at the same time degree varying antibiotic spectra of activities against both Gram-positive and Gram-negative microorganisms. Many of them were tested against sensitivity to the antimicrobial action of B. subtilis for the very first time. B. subtilis TS 01 and ZR 02 showed highest antifungal activity (sterile zone in diameter over 37 mm) against Alternaria solani, Botrytis cinerea, Monilia linhartiana 869, Phytophthora cryptogea 759/1 and Rhizoctonia sp. The most sensitive bacterial species were found to be Pseudomonas syringae pv. tomato Ro and Xanthomonas campestris with sterile zones 48.0 and 50.0 mm in diameter, respectively. The latter draws a conclusion that the isolated and identified Bacillus subtilis strains are promising natural biocontrol agents and should be further studied and tested for control of numerous plant diseases.

  14. The Exosporium Layer of Bacterial Spores: a Connection to the Environment and the Infected Host.

    Science.gov (United States)

    Stewart, George C

    2015-12-01

    Much of what we know regarding bacterial spore structure and function has been learned from studies of the genetically well-characterized bacterium Bacillus subtilis. Molecular aspects of spore structure, assembly, and function are well defined. However, certain bacteria produce spores with an outer spore layer, the exosporium, which is not present on B. subtilis spores. Our understanding of the composition and biological functions of the exosporium layer is much more limited than that of other aspects of the spore. Because the bacterial spore surface is important for the spore's interactions with the environment, as well as being the site of interaction of the spore with the host's innate immune system in the case of spore-forming bacterial pathogens, the exosporium is worthy of continued investigation. Recent exosporium studies have focused largely on members of the Bacillus cereus family, principally Bacillus anthracis and Bacillus cereus. Our understanding of the composition of the exosporium, the pathway of its assembly, and its role in spore biology is now coming into sharper focus. This review expands on a 2007 review of spore surface layers which provided an excellent conceptual framework of exosporium structure and function (A. O. Henriques and C. P. Moran, Jr., Annu Rev Microbiol 61:555-588, 2007, http://dx.doi.org/10.1146/annurev.micro.61.080706.093224). That review began a process of considering outer spore layers as an integrated, multilayered structure rather than simply regarding the outer spore components as independent parts.

  15. The use of germinants to potentiate the sensitivity of Bacillus anthracis spores to peracetic acid

    Directory of Open Access Journals (Sweden)

    Ozgur eCelebi

    2016-01-01

    Full Text Available Elimination of Bacillus anthracis spores from the environment is a difficult and costly process due in part to the toxicity of current sporicidal agents. For this reason we investigated the ability of the spore germinants L-alanine (100 mM and inosine (5 mM to reduce the concentration of peracetic acid (PAA required to inactivate B.anthracis spores. While L-alanine significantly enhanced (p=0.0085 the bactericidal activity of 500 ppm PAA the same was not true for inosine suggesting some form of negative interaction. In contrast the germinant combination proved most effective at 100 ppm PAA (p=0.0009. To determine if we could achieve similar results in soil we treated soil collected from the burial site of an anthrax infected animal which had been supplemented with spores of the Sterne strain of B.anthracis to increase the level of contamination to 104 spores/g. Treatment with germinants followed one hour later by 5000 ppm PAA eliminated all of the spores. In contrast direct treatment of the animal burial site using this approach delivered using a back pack sprayer had no detectable effect on the level of B.anthracis contamination or on total culturable bacterial numbers over the course of the experiment. It did trigger a significant, but temporary, reduction (p<0.0001 in the total spore count suggesting that germination had been triggered under real world conditions. In conclusion, we have shown that the application of germinants increase the sensitivity of bacterial spores to PAA. While the results of the single field trial were inconclusive, the study highlighted the potential of this approach and the challenges faced when attempting to perform real world studies on B.anthracis spores contaminated sites.

  16. The Use of Germinants to Potentiate the Sensitivity of Bacillus anthracis Spores to Peracetic Acid.

    Science.gov (United States)

    Celebi, Ozgur; Buyuk, Fatih; Pottage, Tom; Crook, Ant; Hawkey, Suzanna; Cooper, Callum; Bennett, Allan; Sahin, Mitat; Baillie, Leslie

    2016-01-01

    Elimination of Bacillus anthracis spores from the environment is a difficult and costly process due in part to the toxicity of current sporicidal agents. For this reason we investigated the ability of the spore germinants L-alanine (100 mM) and inosine (5 mM) to reduce the concentration of peracetic acid (PAA) required to inactivate B. anthracis spores. While L-alanine significantly enhanced (p = 0.0085) the bactericidal activity of 500 ppm PAA the same was not true for inosine suggesting some form of negative interaction. In contrast the germinant combination proved most effective at 100 ppm PAA (p = 0.0009). To determine if we could achieve similar results in soil we treated soil collected from the burial site of an anthrax infected animal which had been supplemented with spores of the Sterne strain of B. anthracis to increase the level of contamination to 10(4) spores/g. Treatment with germinants followed 1 h later by 5000 ppm PAA eliminated all of the spores. In contrast direct treatment of the animal burial site using this approach delivered using a back pack sprayer had no detectable effect on the level of B. anthracis contamination or on total culturable bacterial numbers over the course of the experiment. It did trigger a significant, but temporary, reduction (p < 0.0001) in the total spore count suggesting that germination had been triggered under real world conditions. In conclusion, we have shown that the application of germinants increase the sensitivity of bacterial spores to PAA. While the results of the single field trial were inconclusive, the study highlighted the potential of this approach and the challenges faced when attempting to perform real world studies on B. anthracis spores contaminated sites.

  17. SKPDT is a signaling peptide that stimulates sporulation and cry1Aa expression in Bacillus thuringiensis but not in Bacillus subtilis.

    Science.gov (United States)

    Aceves-Diez, Angel E; Robles-Burgueño, Refugio; de la Torre, Mayra

    2007-08-01

    We have identified and characterized in the supernatant of the transition phase of Bacillus thuringiensis var. kurstaki the peptide SKPDT. This peptide was previously identified by in silico analysis by Pottathil and Lazazzera (Front Biosci 8:32-45 2003) as a putative signaling peptide (NprRB) of the Phr family in B. thuringiensis. The chemically synthesized NprRB did not affect the growth kinetics of B. thuringiensis var. kurstaki but stimulated the sporulation, spore release, and transcription of cry1Aa when added to cultures during the transition phase. In fact, when the peptide (100 nM) was added to a culture in transition phase, the transcription of cry1Aa was stimulated almost threefold, mainly from the late promoter BtII, which requires the late-stage sporulation-specific transcription factor sigma (K). On the other hand, NprRB did not have any effect on B. subtilis. Thus, SKPDT seems to be a signaling peptide specific for B. thuringiensis.

  18. Endospore production allows using spray-drying as a possible formulation system of the biocontrol agent Bacillus subtilis CPA-8.

    Science.gov (United States)

    Yánez-Mendizabal, V; Viñas, I; Usall, J; Cañamás, T; Teixidó, N

    2012-04-01

    The role of endospore production by Bacillus subtilis CPA-8 on survival during spray-drying was investigated by comparison with a non-spore-forming biocontrol agent Pantoea agglomerans CPA-2. Endospore formation promoted heat resistance in CPA-8 depending on growth time (72 h cultures were more resistant than 24 h ones). The survival of CPA-8 and CPA-2 after spray-drying was determined after being grown in optimised media for 24 and 72 h. Spray-dried 72 h CPA-8 had the best survival (32%), while CPA-2 viability was less than 2%. CPA-8 survival directly related with its ability to produce endospores. Spray-dried CPA-8 reduced Monilinia fructicola conidia germination similarly to fresh cells, demonstrating that spray-drying did not adversely affect biocontrol efficacy. Endospore production thus improves CPA-8 resistance to spray-drying. These results can provide a reliable basis for optimising of the spray-drying formulation process for CPA-8 and other microorganisms.

  19. Single mutations introduced in the essential ribosomal proteins L3 and S10 cause a sporulation defect in Bacillus subtilis.

    Science.gov (United States)

    Akanuma, Genki; Suzuki, Shota; Yano, Koichi; Nanamiya, Hideaki; Natori, Yousuke; Namba, Eri; Watanabe, Kazuya; Tagami, Kazumi; Takeda, Takuya; Iizuka, Yuka; Kobayashi, Ako; Ishizuka, Morio; Yoshikawa, Hirofumi; Kawamura, Fujio

    2013-01-01

    We introduced single mutations into the rplC and rpsJ genes, which encode the essential ribosomal proteins L3 (RplC) and S10 (RpsJ), respectively, and are located in the S10 gene cluster of the gram-positive, endospore-forming bacterium Bacillus subtilis, and examined whether these mutations affected their growth rate, sporulation, competence development and 70S ribosome formation. Mutant cells harboring the G52D mutation in the L3 ribosomal protein, which is located at the peptidyl transferase center of 50S, accumulated 30S subunit at 45°C, probably due to a defect in 50S formation, and exhibited a reduction in the sporulation frequency at high temperature. On the other hand, mutant cells harboring the H56R mutation in the S10 protein, which is located near the aminoacyl-tRNA site of 30S, showed severe growth defect and deficiency in spore formation, and also exhibited significant delay in competence development.

  20. The Molecular Timeline of a Reviving Bacterial Spore

    OpenAIRE

    2015-01-01

    Summary The bacterial spore can rapidly convert from a dormant to a fully active cell. Here we study this remarkable cellular transition in Bacillus subtilis and reveal the identity of the newly synthesized proteins throughout spore revival. Our analysis uncovers a highly ordered developmental program that correlates with the spore morphological changes and reveals the spatial and temporal molecular events fundamental to reconstruct a cell. As opposed to current knowledge, we found that trans...

  1. Persistence strategies of Bacillus cereus spores isolated from dairy silo tanks.

    Science.gov (United States)

    Shaheen, Ranad; Svensson, Birgitta; Andersson, Maria A; Christiansson, Anders; Salkinoja-Salonen, Mirja

    2010-05-01

    Survival of Bacillus cereus spores of dairy silo tank origin was investigated under conditions simulating those in operational dairy silos. Twenty-three strains were selected to represent all B. cereus isolates (n = 457) with genotypes (RAPD-PCR) that frequently colonised the silo tanks of at least two of the sampled eight dairies. The spores were studied for survival when immersed in liquids used for cleaning-in-place (1.0% sodium hydroxide at pH 13.1, 75 degrees C; 0.9% nitric acid at pH 0.8, 65 degrees C), for adhesion onto nonliving surfaces at 4 degrees C and for germination and biofilm formation in milk. Four groups with different strategies for survival were identified. First, high survival (log 15 min kill steel from cold water. Third, a cereulide producing group with spores characterised by slow germination in rich medium and well preserved viability when exposed to heating at 90 degrees C. Fourth, spores capable of germinating at 8 degrees C and possessing the cspA gene. There were indications that spores highly resistant to hot 1% sodium hydroxide may be effectively inactivated by hot 0.9% nitric acid. Eight out of the 14 dairy silo tank isolates possessing hot-alkali resistant spores were capable of germinating and forming biofilm in whole milk, not previously reported for B. cereus.

  2. Spatially resolved characterization of water and ion incorporation in Bacillus spores.

    Science.gov (United States)

    Ghosal, Sutapa; Leighton, Terrance J; Wheeler, Katherine E; Hutcheon, Ian D; Weber, Peter K

    2010-05-01

    We present the first direct visualization and quantification of water and ion uptake into the core of individual dormant Bacillus thuringiensis subsp. israelensis (B. thuringiensis subsp. israelensis) endospores. Isotopic and elemental gradients in the B. thuringiensis subsp. israelensis spores show the permeation and incorporation of deuterium in deuterated water (D(2)O) and solvated ions throughout individual spores, including the spore core. Under hydrated conditions, incorporation into a spore occurs on a time scale of minutes, with subsequent uptake of the permeating species continuing over a period of days. The distribution of available adsorption sites is shown to vary with the permeating species. Adsorption sites for Li(+), Cs(+), and Cl(-) are more abundant within the spore outer structures (exosporium, coat, and cortex) relative to the core, while F(-) adsorption sites are more abundant in the core. The results presented here demonstrate that elemental abundance and distribution in dormant spores are influenced by the ambient environment. As such, this study highlights the importance of understanding how microbial elemental and isotopic signatures can be altered postproduction, including during sample preparation for analysis, and therefore, this study is immediately relevant to the use of elemental and isotopic markers in environmental microbiology and microbial forensics.

  3. Evaluation of a Stochastic Inactivation Model for Heat-Activated Spores of Bacillus spp. ▿

    Science.gov (United States)

    Corradini, Maria G.; Normand, Mark D.; Eisenberg, Murray; Peleg, Micha

    2010-01-01

    Heat activates the dormant spores of certain Bacillus spp., which is reflected in the “activation shoulder” in their survival curves. At the same time, heat also inactivates the already active and just activated spores, as well as those still dormant. A stochastic model based on progressively changing probabilities of activation and inactivation can describe this phenomenon. The model is presented in a fully probabilistic discrete form for individual and small groups of spores and as a semicontinuous deterministic model for large spore populations. The same underlying algorithm applies to both isothermal and dynamic heat treatments. Its construction does not require the assumption of the activation and inactivation kinetics or knowledge of their biophysical and biochemical mechanisms. A simplified version of the semicontinuous model was used to simulate survival curves with the activation shoulder that are reminiscent of experimental curves reported in the literature. The model is not intended to replace current models to predict dynamic inactivation but only to offer a conceptual alternative to their interpretation. Nevertheless, by linking the survival curve's shape to probabilities of events at the individual spore level, the model explains, and can be used to simulate, the irregular activation and survival patterns of individual and small groups of spores, which might be involved in food poisoning and spoilage. PMID:20453137

  4. Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins

    Directory of Open Access Journals (Sweden)

    Marciniak Bogumiła C

    2012-05-01

    Full Text Available Abstract Background Bacillus subtilis is a favorable host for the production of industrially relevant proteins because of its capacity of secreting proteins into the medium to high levels, its GRAS (Generally Recognized As Safe status, its genetic accessibility and its capacity to grow in large fermentations. However, production of heterologous proteins still faces limitations. Results This study aimed at the identification of bottlenecks in secretory protein production by analyzing the response of B. subtilis at the transcriptome level to overproduction of eight secretory proteins of endogenous and heterologous origin and with different subcellular or extracellular destination: secreted proteins (NprE and XynA of B. subtilis, Usp45 of Lactococcus lactis, TEM-1 β-lactamase of Escherichia coli, membrane proteins (LmrA of L. lactis and XylP of Lactobacillus pentosus and lipoproteins (MntA and YcdH of B. subtilis. Responses specific for proteins with a common localization as well as more general stress responses were observed. The latter include upregulation of genes encoding intracellular stress proteins (groES/EL, CtsR regulated genes. Specific responses include upregulation of the liaIHGFSR operon under Usp45 and TEM-1 β-lactamase overproduction; cssRS, htrA and htrB under all secreted proteins overproduction; sigW and SigW-regulated genes mainly under membrane proteins overproduction; and ykrL (encoding an HtpX homologue specifically under membrane proteins overproduction. Conclusions The results give better insights into B. subtilis responses to protein overproduction stress and provide potential targets for genetic engineering in order to further improve B. subtilis as a protein production host.

  5. The two authentic methionine aminopeptidase genes are differentially expressed in Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Wang YiPing

    2005-10-01

    Full Text Available Abstract Background Two putative methionine aminopeptidase genes, map (essential and yflG (non-essential, were identified in the genome sequence of Bacillus subtilis. We investigated whether they can function as methionine aminopeptidases and further explored possible reasons for their essentiality or dispensability in B. subtilis. Results In silico analysis of MAP evolution uncovered a coordinated pattern of MAP and deformylase that did not correlate with the pattern of 16S RNA evolution. Biochemical assays showed that both MAP (MAP_Bs and YflG (YflG_Bs from B. subtilis overproduced in Escherichia coli and obtained as pure proteins exhibited a methionine aminopeptidase activity in vitro. Compared with MAP_Bs, YflG_Bs was approximately two orders of magnitude more efficient when assayed on synthetic peptide substrates. Both map and yflG genes expressed in multi-copy plasmids could complement the function of a defective map gene in the chromosomes of both E. coli and B. subtilis. In contrast, lacZ gene transcriptional fusions showed that the promoter activity of map was 50 to 100-fold higher than that of yflG. Primer extension analysis detected the transcription start site of the yflG promoter. Further work identified that YvoA acted as a possible weak repressor of yflG expression in B. subtilis in vivo. Conclusion Both MAP_Bs and YflG_Bs are functional methionine aminopeptidases in vitro and in vivo. The high expression level of map and low expression level of yflG may account for their essentiality and dispensality in B. subtilis, respectively, when cells are grown under laboratory conditions. Their difference in activity on synthetic substrates suggests that they have different protein targets in vivo.

  6. Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide

    Directory of Open Access Journals (Sweden)

    Mehrdad Moosazadeh Moghaddam

    2013-01-01

    Full Text Available Introduction: Some of bacterial species are able to uptake DNA molecule from environment, the yield of this process depends on some conditions such as plasmid size and host type. In the case of Bacillus subtilis, DNA uptake has low efficacy. Using Spizizen minimal medium is common method in plasmid transformation into B. subtilis, but rate of this process is not suitable and noteworthy. The aim of this study was investigation of novel method for improvement of DNA transformation into B. subtilis based on CM11 cationic peptide as a membrane permeable agent.Materials and methods: In this study, for optimization of pWB980 plasmid transformation into B. subtilis, the CM11 cationic peptide was used. For this purpose, B. subtilis competent cell preparation in the present of different concentration of peptide was implemented by two methods. In the first method, after treatment of bacteria with different amount of peptide for 14h, plasmid was added. In the second method, several concentration of peptide with plasmid was exposed to bacteria simultaneously. Bacteria that uptake DNA were screened on LB agar medium containing kanamycin. The total transformed bacteria per microgram of DNA was calculated and compared with the control.Results: Plasmid transformation in best conditions was 6.5 folds higher than the control. This result was statistically significant (P value <0.001.Discussion and conclusion: This study showed that CM11 cationic peptide as a membrane permeable agent was able to increase plasmid transformation rate into B. subtilis. This property was useful for resolution of low transformation efficacy.

  7. Dry heat exposures of surface exposed and embedded Bacillus spores

    Science.gov (United States)

    Schubert, Wayne

    Dry heat microbial reduction (DHMR) is the primary technique used to reduce the microbial load of spacecraft and component parts. Often, manufacturing procedures require heating flight hardware to high temperatures for purposes other than planetary protection DHMR. The existing specifications, however, do not allow for additional planetary protection bioburden reduction credit if the hardware is exposed without controlled relative humidity. The intent of this study was to provide adequate data on the DHMR technique to support modification of four aspects of current requirements; expansion of acceptable time and temperature combinations used for spacecraft dry heat microbial reduction processes above 125° C, determining the effect that humidity has on spore lethality as a function of temperature, understanding the lethality for spores with exceptionally high thermal resistance and to investigate the extended exposure requirement for materials that might contain embedded microorganisms. Spores from two bacterial species were tested, B. atrophaeus ATCC 9372 and B. sp. ATCC 29669, under three conditions encompassing 5 temperature points. Embedded experiments utilized a silicone rubber polymer that is commonly used on robotic spacecraft, and surface exposed experiments were performed under both ambient and vacuum-controlled humidity conditions. The results obtained support the use of DHMR protocols that extend the maximum temperature range from 125° C to 170° C, with either controlled or ambient humidity. If implemented, this will give projects bioburden reduction credit for shorter treatments at extended temperatures, and allow spacecraft to be processed in more readily available and less expensive facilities that do not have humidity control, with significant cost and schedule benefits. The study also demonstrated that the required heating time for materials presumed to have embedded bioburden is conservative.

  8. Development of a Rapid and Sensitive Immunoassay for Detection and Subsequent Recovery of Bacillus anthracis Spores in Environmental Samples

    Science.gov (United States)

    Bacillus anthracis is considered a major threat as an agent of bioterrorism. B. anthracis spores are readily dispersed as aerosols, are very persistent, and are resistant to normal disinfection treatments. Immunoassays have been developed to rapidly detect B. anthracis spores at high concentration...

  9. Sporulation environment of emetic toxin-producing Bacillus cereus strains determines spore size, heat resistance and germination capacity

    NARCIS (Netherlands)

    Voort, van der M.; Abee, T.

    2013-01-01

    Aim Heat resistance, germination and outgrowth capacity of Bacillus cereus spores in processed foods are major factors in causing the emetic type of gastrointestinal disease. In this study, we aim to identify the impact of different sporulation conditions on spore properties of emetic toxin-producin

  10. Partial biochemical characterization of crude extract extracellular chitinase enzyme from Bacillus subtilis B 298

    Science.gov (United States)

    Lestari, P.; Prihatiningsih, N.; Djatmiko, H. A.

    2017-02-01

    Extraction and characterization of extracellular chitinase from Bacillus subtilis B 298 have been done. Growth curve determination of B. subtilis B 298, production curve determination of crude extract chitinase from B. subtilis B 298, and partial biochemical characterization of crude extract chitinase have been achieved in this study. Optimum growth of B. subtilis B 298 was achieved at logarithmic phase within 9 hours incubation time, so it was used as inoculum for enzyme production. According to production curve of the enzyme, it was known that incubation time which gave the highest chitinase activity of 15 hours with activity of 6.937 U/mL respectively. Effect of various temperatures on chitinase activity showed that optimum activity was achieved at 40°C with an activity of 5.764 U/mL respectively. Meanwhile, the optimum pH for chitinase activity was achieved at pH of 5.0 with an activity of 6.813 U/mL respectively. This enzyme was then classified as metalloenzyme due to the decline of the activity by EDTA addition. All divalent cations tested acted as inhibitors.

  11. Identification and characterization of the vanillin dehydrogenase YfmT in Bacillus subtilis 3NA.

    Science.gov (United States)

    Graf, Nadja; Wenzel, Marian; Altenbuchner, Josef

    2016-04-01

    With vanillin as one of the most important flavoring agents, many efforts have been made to optimize its biotechnological production from natural abundant substrates. However, its toxicity against the hosts results in rather low yields and product concentrations. Bacillus subtilis as a soil-dwelling bacterium is a possible lignin-derived compound-degrading microorganism. Therefore, its vanillin and ferulic acid metabolism was investigated. With a rather high tolerance for vanillin up to 20 mM, it is a promising candidate to produce natural vanillin. In this study, the well-studied phenolic acid decarboxylases PadC and BsdBCD could be ascribed to function as the only enzymes in B. subtilis 3NA converting ferulic acid to 4-vinylguaiacol and vanillic acid to guaiacol, respectively. As vanillin also becomes converted to guaiacol, a previous conversion to vanillic acid was assumed. Usage of bioinformatic tools revealed YfmT, which could be shown to function as the only vanillin dehydrogenase in B. subtilis 3NA. Thus, YfmT was further characterized regarding its temperature and pH optima as well as its substrate range. Vanillin and ferulic acid metabolic routes in the tested B. subtilis strain were revealed, a direct conversion of ferulic acid to vanillin, however, could not be found.

  12. Bacteriocin from Bacillus subtilis as a novel drug against diabetic foot ulcer bacterial pathogens

    Institute of Scientific and Technical Information of China (English)

    Baby Joseph; Berlina Dhas; Vimalin Hena; Justin Raj

    2013-01-01

    Objective:To isolate and identify Bacillus subtilis (B. subtilis) from soil and to characterize and partially purify the bacteriocin. To evaluate the antimicrobial activity against four diabetic foot ulcer bacterial pathogens. Methods:Genotypic identification was done based on Bergey’s manual of systemic bacteriology. Antimicrobial susceptibility test was done by Kirby-Bauer disc diffusion method. Colonies were identified by colony morphology and biochemical characterization and also compared with MTCC 121 strain. Further identification was done by 16S rRNA sequencing. Inhibitory activities of partially purified bacteriocin on all the DFU isolates were done by agar well diffusion method. The strain was identified to produce bacteriocin by stab overlay assay. Bacteriocin was extracted by organic solvent extraction using chloroform, further purified by HPLC and physical, and chemical characterization was performed. Results: The four isolates showed high level of resistance to amoxyclav and sensitivity to ciprofloxacin. HPLC purification revealed that the extracts are bacteriocin. The phylogenetic tree analysis results showed that the isolate was 99%related to B. subtilis BSF01. The results reveled activity to all the four isolates and high level of activity was seen in case of Klebsiella sp. Conclusions:Partially purified bacteriocin was found to have antimicrobial activity against the four diabetic foot ulcer bacterial pathogens, which can thus be applied as a better drug molecule on further studies. The strain B. subtilis are found to be safe for use and these antimicrobial peptides can be used as an antimicrobial in humans to treat DFU bacterial pathogens.

  13. Soluble Expression of (+)-γ-Lactamase in Bacillus subtilis for the Enantioselective Preparation of Abacavir Precursor.

    Science.gov (United States)

    Xue, Tian-Yun; Xu, Guo-Chao; Han, Rui-Zhi; Ni, Ye

    2015-07-01

    Chiral Vince lactam (γ-lactam) is an important precursor of many carbocyclic nucleoside analogues and pharmaceuticals. Here, a (+)-γ-lactamase encoding gene delm from Delftia sp. CGMCC 5755 was identified through genome hunting. To achieve its soluble and functional expression, Escherichia coli and Bacillus subtilis expression systems were introduced. Compared with E. coli system, recombinant (+)-γ-lactamase showed improved protein solubility and catalytic activity in B. subtilis 168. Reaction conditions for enantioselective resolution of γ-lactam were optimized to be at 30 °C, pH 9.0, and 300 rpm when employing the recombinant B. subtilis 168/pMA5-delm whole cells. Kinetic analysis showed that the apparent V max and K m were 0.595 mmol/(min · gDCW) and 378 mmol/L, respectively. No obvious substrate inhibition was observed. In a 500-mL reaction system, enantioselective resolution of 100 g/L γ-lactam was achieved with 10 g/L dry cells, resulting in 55.2 % conversion and 99 % ee of (-)-γ-lactam. All above suggested that recombinant B. subtilis 168/pMA5-delm could potentially be applied in the preparation of optically pure (-)-γ-lactam.

  14. Growth of and valine production by a Bacillus subtilis mutant in the small intestine of pigs

    DEFF Research Database (Denmark)

    Canibe, Nuria; Poulsen, Henrik Vestergaard; Nørgaard, Jan Værum;

    2016-01-01

    :Lys of 0.63:1 (Neg), 2) the Neg diet with added Bacillus subtilis-valine (1.28 × 108 cfu/g feed) (+Bac), and 3) the Neg diet with added L-Val to a Val:Lys of 0.69:1 (+Val). Eighteen gilts (6 on each treatment) with initial weights of ∼15 kg were fed the diets for 23 d before the animals were euthanized...... and samples from the small intestine were obtained. The number of B. subtilis cfu in digesta was higher in the +Bac group than in the Neg group (P cfu were detected in the Neg group, whereas numbers between 3.4 and 4.4 log cfu/g and numerically higher Val and Lys...... concentrations were measured in the +Bac group. Short-term in vitro incubations of digesta showed a decrease (P ≤ 0.03) in the number of B. subtilis cfu over time for the +Bac group and no difference in the rate of Val production compared to that in the Neg group. In conclusion, more B. subtilis cfu were present...

  15. The Adsorption Properties of Bacillus atrophaeus Spores on Single-Wall Carbon Nanotubes

    OpenAIRE

    Cortes, P; S. Deng; Smith, G. B.

    2009-01-01

    An adsorption equilibrium and a kinetic study of Bacillus atrophaeus on Single-Wall Carbon Nanotubes (SWCNTs) were here performed to provide the basis for developing biosensor devices for detecting threatening micro-organisms in water supply systems. B. atrophaeus spores and carbon nanotubes were subjected to a batch adsorption process to document their equilibria and kinetics. Here, commercial nanotubes were either studied as received or were acid-purified before adsorption experiments. The ...

  16. Spore and crystal formation in Bacillus thuringiensis var thuringiensis during growth in cystine and cysteine.

    OpenAIRE

    Rajalakshmi, S.; Shethna, YI

    1980-01-01

    The effect of the addition of different concentratons of cystine and cysteine on sporulation and parasporal crystal formation in Bacillus thuringiensis var. thuringiensis was studied. The effect was well pronounced when the systine/cysteine additions were made after the stationary phase. Heat stable spores and crystals were formed when the culture was provided with a low concentration of cystine/cysteine (0.05 per cent w/v). At a moderate concentration of cystine or cysteine (0.15%), only ...

  17. Computational fluid dynamics modeling of Bacillus anthracis spore deposition in rabbit and human respiratory airways

    Energy Technology Data Exchange (ETDEWEB)

    Kabilan, S.; Suffield, S. R.; Recknagle, K. P.; Jacob, R. E.; Einstein, D. R.; Kuprat, A. P.; Carson, J. P.; Colby, S. M.; Saunders, J. H.; Hines, S. A.; Teeguarden, J. G.; Straub, T. M.; Moe, M.; Taft, S. C.; Corley, R. A.

    2016-09-01

    Three-dimensional computational fluid dynamics and Lagrangian particle deposition models were developed to compare the deposition of aerosolized Bacillus anthracis spores in the respiratory airways of a human with that of the rabbit, a species commonly used in the study of anthrax disease. The respiratory airway geometries for each species were derived respectively from computed tomography (CT) and µCT images. Both models encompassed airways that extended from the external nose to the lung with a total of 272 outlets in the human model and 2878 outlets in the rabbit model. All simulations of spore deposition were conducted under transient, inhalation–exhalation breathing conditions using average species-specific minute volumes. Two different exposure scenarios were modeled in the rabbit based upon experimental inhalation studies. For comparison, human simulations were conducted at the highest exposure concentration used during the rabbit experimental exposures. Results demonstrated that regional spore deposition patterns were sensitive to airway geometry and ventilation profiles. Due to the complex airway geometries in the rabbit nose, higher spore deposition efficiency was predicted in the nasal sinus compared to the human at the same air concentration of anthrax spores. In contrast, higher spore deposition was predicted in the lower conducting airways of the human compared to the rabbit lung due to differences in airway branching pattern. This information can be used to refine published and ongoing biokinetic models of inhalation anthrax spore exposures, which currently estimate deposited spore concentrations based solely upon exposure concentrations and inhaled doses that do not factor in species-specific anatomy and physiology for deposition.

  18. Degradation of polyester polyurethane by a newly isolated soil bacterium, Bacillus subtilis strain MZA-75.

    Science.gov (United States)

    Shah, Ziaullah; Krumholz, Lee; Aktas, Deniz Fulya; Hasan, Fariha; Khattak, Mutiullah; Shah, Aamer Ali

    2013-11-01

    A polyurethane (PU) degrading bacterial strain MZA-75 was isolated from soil through enrichment technique. The bacterium was identified through 16S rRNA gene sequencing, the phylogenetic analysis indicated the strain MZA-75 belonged to genus Bacillus having maximum similarity with Bacillus subtilis strain JBE0016. The degradation of PU films by strain MZA-75 in mineral salt medium (MSM) was analyzed by scanning electron microscopy (SEM), fourier transform infra-red spectroscopy (FT-IR) and gel permeation chromatography (GPC). SEM revealed the appearance of widespread cracks on the surface. FTIR spectrum showed decrease in ester functional group. Increase in polydispersity index was observed in GPC, which indicates chain scission as a result of microbial treatment. CO2 evolution and cell growth increased when PU was used as carbon source in MSM in Sturm test. Increase in both cell associated and extracellular esterases was observed in the presence of PU indicated by p-Nitrophenyl acetate (pNPA) hydrolysis assay. Analysis of cell free supernatant by gas chromatography-mass spectrometry (GC-MS) revealed that 1,4-butanediol and adipic acid monomers were produced. Bacillus subtilis strain MZA-75 can degrade the soft segment of polyester polyurethane, unfortunately no information about the fate of hard segment could be obtained. Growth of strain MZA-75 in the presence of these metabolites indicated mineralization of ester hydrolysis products into CO2 and H2O.

  19. Duodenal histology and carcass quality of feedlot cattle supplemented with calcium butyrate and Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Thiago Simas de Oliveira Moreira

    2016-01-01

    Full Text Available The experiment was carried out at the Comigo Technology Center, in Rio Verde, State of Goiás, Brazil, with the objective of evaluating the effects of supplementation with calcium butyrate, as a growth promoting agent for the duodenal mucosa and Bacillus subtilis as a probiotic performance enhancer in feedlot cattle. Calcium butyrate (5 and 10 g per animal per day and Bacillus (10 g per animal per day were added to a basal diet. There were used 85 Nelore bulls, with average weight of 315 ± 7 kg. The experiment lasted 118 days, including the adaptation period, until slaughter at 30 months of age. Diets were distributed in a completely randomized design with four treatments, where: T1 = control (basal diet; T2 = basal diet + 5 g calcium butyrate; T3 = basal diet + 10 g calcium butyrate and T4 = basal diet + 10 g calcium butyrate + 10 g probiotic with four replications and five to six animals per replication. It was used a forage: concentrate ratio of 30:70, the roughage used was the corn silage. Height and width measurements of intestinal villi were taken, and carcass and meat quality were evaluated. The supplementation of calcium butyrate and Bacillus subtilis positively influenced (p < 0.05 the carcass marbling level and calcium butyrate increased the villus height in the small intestine.

  20. Bacillus spores as building blocks for stimuli-responsive materials and nanogenerators

    Science.gov (United States)

    Sahin, Ozgur; Chen, Xi

    2014-03-01

    Materials that mechanically respond to external chemical stimuli have applications in a wide range of fields. Inspired by biological systems, stimuli-responsive materials that can oscillate, transport fluid, mimic homeostasis, and undergo complex changes in shape have been previously demonstrated. However, the effectiveness of synthetic stimuli-responsive materials in generating work is limited when compared to mechanical actuators. During studies of bacterial sporulation, we have found that the mechanical response of Bacillus spores to water gradients exhibits an energy density of more than 10 MJ/m3, which is two orders of magnitude higher than synthetic water-responsive materials. We also identified mutations that can approximately double the energy density of the spores, and found that spores can self-assemble into dense, submicron-thick monolayers on substrates such as silicon microcantilevers and elastomer sheets, creating self-assembled actuators that can remotely generate electrical power from an evaporating body of water. The energy conversion mechanism of Bacillus spores may facilitate synthetic stimuli-responsive materials with significantly higher energy densities. We acknowledge support from the U.S. Dept. of Energy Early Career Research Program, the Wyss Institute for Biologically Inspired Engineering, and the Rowland Institute at Harvard.

  1. Disinfection methods for spores of Bacillus atrophaeus, B. anthracis, Clostridium tetani, C. botulinum and C. difficile.

    Science.gov (United States)

    Oie, Shigeharu; Obayashi, Akiko; Yamasaki, Hirofumi; Furukawa, Hiroyuki; Kenri, Tsuyoshi; Takahashi, Motohide; Kawamoto, Keiko; Makino, Sou-ichi

    2011-01-01

    To evaluate disinfection methods for environments contaminated with bioterrorism-associated microorganism (Bacillus anthracis), we performed the following experiments. First, the sporicidal effects of sodium hypochlorite on spores of five bacterial species were evaluated. Bacillus atrophaeus was the most resistant to hypochlorite, followed in order by B. anthracis, Clostridium botulinum and Clostridium tetani, and Clostridium difficile. Subsequently, using B. atrophaeus spores that were the most resistant to hypochlorite, the sporicidal effects of hypochlorite at lower pH by adding vinegar were evaluated. Hypochlorite containing vinegar had far more marked sporicidal effects than hypochlorite alone. Cleaning with 0.5% (5000 ppm) hypochlorite containing vinegar inactivated B. atrophaeus spores attached to vinyl chloride and plywood plates within 15 s, while that not containing vinegar did not inactivate spores attached to cement or plywood plates even after 1 h. Therefore, the surfaces of cement or plywood plates were covered with gauze soaked in 0.5% hypochlorite containing vinegar, and the sporicidal effects were evaluated. B. atrophaeus spores attached to plywood plates were not inactivated even after 6 h, but those attached to cement plates were inactivated within 5 min. On the other hand, covering the surfaces of plywood plates with gauze soaked in 0.3% peracetic acid and gauze soaked in 2% glutaral inactivated B. atrophaeus spores within 5 min and 6 h, respectively. These results suggest that hypochlorite containing vinegar is effective for disinfecting vinyl chloride, tile, and cement plates contaminated with B. anthracis, and peracetic acid is effective for disinfecting plywood plates contaminated with such microorganism.

  2. Kinetics of p-aminoazobenzene degradation by Bacillus subtilis under denitrifying conditions

    Energy Technology Data Exchange (ETDEWEB)

    Zissi, U.S.; Kornaros, M.E.; Lyberatos, G.C.

    1999-05-01

    Bacillus subtilis is an organism capable of degrading an azo dye, such as p-aminoazobenzene (pAAB), under both aerobic and anoxic conditions. In both cases, pAAB is co-metabolized with a main carbon source and under anoxic conditions denitrification is observed. Kinetic experiments were carried out with a pure culture of B. subtilis and a mathematical model that accurately describes both biodegradation of pAAB under anoxic conditions and the denitrification process under both carbon- and nitrate- or nitrite-limited conditions is developed. Presence of pAAB in culture medium causes an inhibition of bacterial growth and of nitrite accumulation. Bacterial growth and pAAB degradation rates are found to be slower under anoxic conditions compared to the corresponding rates under aerobic conditions.

  3. Engineering of Bacillus subtilis for the Production of 2,3-Butanediol from Sugarcane Molasses.

    Science.gov (United States)

    Deshmukh, Apoorva Nandkumar; Nipanikar-Gokhale, Padmaja; Jain, Rishi

    2016-05-01

    2,3-butanediol is known to be a platform chemical with several potential industrial applications. Sustainable industrial scale production can be attained by using a sugarcane molasses based fermentation process using Bacillus subtilis. However, the accumulation of acetoin needs to be reduced to improve process efficiency. In this work, B. subtilis was genetically modified in order to increase the yield of 2,3-butanediol. Metabolic engineering strategies such as cofactor engineering and overexpression of the key enzyme butanediol dehydrogenase were attempted. Both the strategies individually led to a statistically significant increase in the 2,3-butanediol yields for sugarcane molasses based fermentation. Cofactor engineering led to a 26 % increase in 2,3-butanediol yield and overexpression of bdhA led to a 11 % increase. However, the combination of the two strategies did not lead to a synergistic increase in 2,3-butanediol yield.

  4. Rice Seed Priming with Picomolar Rutin Enhances Rhizospheric Bacillus subtilis CIM Colonization and Plant Growth.

    Directory of Open Access Journals (Sweden)

    Akanksha Singh

    Full Text Available The effect of rutin, a bioflavonoid on the growth and biofilm formation of Bacillus subtilis strain CIM was investigated. In addition to swimming, swarming, and twitching potentials of B. subtilis CIM (BS, one picomolar (1 pM of rutin was also observed to boost the biofilm forming ability of the bacterium. Bio-priming of rice seeds with BS and rutin not only augmented root and shoot lengths but also the photosynthetic pigments like chlorophyll and carotenoid. Similarly, high accumulation of phenolic and flavonoid contents was observed in the leaves. Fluorescent microscopic images revealed that BS plus rutin enhanced callose deposition in the leaves. It was also established that the least formation of reactive oxygen species in BS plus rutin treated rice plants was due to higher free radicals scavenging activity and total antioxidant potential. The results highlight chemo attractant nature of BS towards rutin, which by enhancing biofilm formation and root colonization indirectly strengthened the plants' defensive state.

  5. Rice Seed Priming with Picomolar Rutin Enhances Rhizospheric Bacillus subtilis CIM Colonization and Plant Growth.

    Science.gov (United States)

    Singh, Akanksha; Gupta, Rupali; Pandey, Rakesh

    2016-01-01

    The effect of rutin, a bioflavonoid on the growth and biofilm formation of Bacillus subtilis strain CIM was investigated. In addition to swimming, swarming, and twitching potentials of B. subtilis CIM (BS), one picomolar (1 pM) of rutin was also observed to boost the biofilm forming ability of the bacterium. Bio-priming of rice seeds with BS and rutin not only augmented root and shoot lengths but also the photosynthetic pigments like chlorophyll and carotenoid. Similarly, high accumulation of phenolic and flavonoid contents was observed in the leaves. Fluorescent microscopic images revealed that BS plus rutin enhanced callose deposition in the leaves. It was also established that the least formation of reactive oxygen species in BS plus rutin treated rice plants was due to higher free radicals scavenging activity and total antioxidant potential. The results highlight chemo attractant nature of BS towards rutin, which by enhancing biofilm formation and root colonization indirectly strengthened the plants' defensive state.

  6. Efficacy of Bacillus subtilis V26 as a biological control agent against Rhizoctonia solani on potato.

    Science.gov (United States)

    Ben Khedher, Saoussen; Kilani-Feki, Olfa; Dammak, Mouna; Jabnoun-Khiareddine, Hayfa; Daami-Remadi, Mejda; Tounsi, Slim

    2015-12-01

    The aim of this study is to evaluate the efficacy of the strain Bacillus subtilis V26, a local isolate from the Tunisian soil, to control potato black scurf caused by Rhizoctonia solani. The in vitro antifungal activity of V26 significantly inhibited R. solani growth compared to the untreated control. Microscopic observations revealed that V26 caused considerable morphological deformations of the fungal hyphae such as vacuolation, protoplast leakage and mycelia crack. The most effective control was achieved when strain V26 was applied 24h prior to inoculation (protective activity) in potato slices. The antagonistic bacterium V26 induced significant suppression of root canker and black scurf tuber colonization compared to untreated controls with a decrease in incidence disease of 63% and 81%, respectively, and promoted plant growth under greenhouse conditions on potato plants. Therefore, B. subtilis V26 has a great potential to be commercialized as a biocontrol agent against R. solani on potato crops.

  7. Influence of Silica Nanoparticles on Antioxidant Potential of Bacillus subtilis IMV B-7023

    Science.gov (United States)

    Skorochod, Iryna O.; Roy, Alla O.; Kurdish, Ivan K.

    2016-03-01

    It was found that if introduced into a nutrient medium of 0.05-1 g/L nano-SiO2, the oxidant activity (OA) of the culture medium (CM) of bacilli increased by 43.2-60.1 % and the antioxidant activity (AA) decreased by 4.5-11.8 %. SiO2 nanoparticles had different effects on antiradical activity (ARA) of the CM of Bacillus subtilis IMV B-7023. In particular, nano-SiO2 had no significant effect on the ability of the CM of bacilli to inactivate the 2.2-diphenyl-1-picrylhydrazyl (DPPH·) free radical. However, for the content of the nanomaterial of 0.01-1 g/L decreased hydroxyl radical scavenging in the CM of B. subtilis IMV B-7023 on 7.2-17.6 % compared with a control. Low doses of silica nanoparticles stimulated the reducing power of the CM of bacteria and then highly suppressed it.

  8. Antagonismo de Trichoderma SPP. E Bacillus subtilis (UFV3918 a Fusarium sambucinum em Pinus elliottii engelm

    Directory of Open Access Journals (Sweden)

    Caciara Gonzatto Maciel

    2014-06-01

    Full Text Available Pinus elliottii é uma espécie de importância no setor florestal e apresenta vulnerabilidade na qualidade sanitária de suas sementes, especialmente pela associação de Fusarium spp., responsável por perdas de plântulas no viveiro. Este trabalho teve como objetivo avaliar a ação antagonista in vitro e in vivo dos agentes Trichoderma spp. e Bacillus subtilis (UFV3918 no controle de Fusarium sambucinum, responsável por danos em plântulas de Pinus elliottii. O controle in vitro foi avaliado através da inibição do crescimento micelial (confronto pareado de culturas, após a incubação a 25±2 ºC e fotoperíodo de 12 h. Para os testes in vivo (desenvolvidos em condições de viveiro, as sementes inicialmente foram inoculadas com o patógeno e, na sequência, microbiolizadas com os agentes antagônicos, para posterior semeadura. Utilizaram-se as técnicas de contato com o biocontrolador em meio BDA por 48 h e peliculização, como formas de microbiolização. Tanto Trichoderma spp. quanto Bacillus subtilis (UFV3918 foram eficientes no controle in vitro de F. sambucinum, e no teste de biocontrole in vivo o produto Bacillus subtilis (UFV3918 destacou-se, reduzindo as perdas de plântulas causadas pelo patógeno, assim como potencializando as variáveis de comprimento de plântula, massa verde e massa seca.

  9. Ability of Bacillus subtilis protoplasts to repair irradiated bacteriophage deoxyribonucleic acid via acquired and natural enzymatic systems.

    OpenAIRE

    Yasbin, R E; Andersen, B J; Sutherland, B M

    1981-01-01

    A novel form of "enzyme therapy" was achieved by utilizing protoplasts of Bacillus subtilis. Photoreactivating enzyme of Escherichia coli was successfully inserted into the protoplasts of B. subtilis treated with polyethylene glycol. This enzyme was used to photoreactivate ultraviolet-damaged bacteriophage deoxyribonucleic acid (DNA). Furthermore, in polyethylene glycol-treated protoplasts, ultraviolet-irradiated transfecting bacteriophage DNA was shown to be a functional substrate for the ho...

  10. Enhancement of Bacillus subtilis Lipopeptide Biosurfactants Production through Optimization of Medium Composition and Adequate Control of Aeration

    OpenAIRE

    Dhouha Ghribi; Semia Ellouze-Chaabouni

    2011-01-01

    Interest in biosurfactants has increased considerably in recent years, as they are potentially used in many commercial applications in petroleum, pharmaceuticals, biomedical, and food processing industries. Since improvement of their production was of great importance to reduce the final coast, cultural conditions were analyzed to optimize biosurfactants production from Bacillus subtilis SPB1 strain. A high yield of biosurfactants was obtained from a culture of B. subtilis using carbohydrate ...

  11. Biodegradation of crude oil by a defined co-culture of indigenous bacterial consortium and exogenous Bacillus subtilis.

    Science.gov (United States)

    Tao, Kaiyun; Liu, Xiaoyan; Chen, Xueping; Hu, Xiaoxin; Cao, Liya; Yuan, Xiaoyu

    2017-01-01

    The aim of this work was to study biodegradation of crude oil by defined co-cultures of indigenous bacterial consortium and exogenous Bacillus subtilis. Through residual oil analysis, it is apparent that the defined co-culture displayed a degradation ratio (85.01%) superior to indigenous bacterial consortium (71.32%) after 7days of incubation when ratio of inoculation size of indigenous bacterial consortium and Bacillus subtilis was 2:1. Long-chain n-alkanes could be degraded markedly by Bacillus subtilis. Result analysis of the bacterial community showed that a decrease in bacterial diversity in the defined co-culture and the enrichment of Burkholderiales order (98.1%) degrading hydrocarbons. The research results revealed that the promising potential of the defined co-culture for application to degradation of crude oil.

  12. Production and purification of a maltose-producing amylase from Bacillus subtilis IMD 198

    Energy Technology Data Exchange (ETDEWEB)

    Fogarty, W.M.; Bourke, E.J.

    1983-09-01

    A strain of Bacillus subtilis (IMD 198) isolated from peat degraded starch to maltose with little production of glucose and other products. Highest levels of enzyme were achieved in a salts medium containing soya bean meal and starch. The enzyme was purified by precipitation with isopropanol, adsorption on calcium phosphate gel and fractionation on DEAE- and CM-cellulose ion-exchange resins. The latter chromatographic procedure removed a contaminating activity that produced dextrins as end-products from starch or amylose. The action pattern of the purified, major enzyme activity indicates that it may be beta-amylase. 52 references.

  13. PRODUCTION OPTIMIZATION OF EXTRACELLULAR L-ASPARAGINASE THROUGH SOLID- STATE FERMENTATION BY ISOLATED BACILLUS SUBTILIS.

    Directory of Open Access Journals (Sweden)

    Susmita Shukla

    2013-02-01

    Full Text Available L-asparaginase has been used as anti-tumor agent for the treatment of acute lymphoblastic leukemia and food processing aid to reduce the formation of cancer causing acrylamide. Extracellular Lasparaginase production was optimized through solid state fermentation using ground nut cake by isolated Bacillus subtilis. which was not reported in literature.Optimum production of L-asparaginase enzyme (18.4U/ml was obtained after 48h of incubation at 370C moisture content of 70% and at pH 7.

  14. Protease obtention using Bacillus subtilis 3411 and amaranth seed meal medium at different aeration rates

    Directory of Open Access Journals (Sweden)

    Pastor Maria Delia

    2001-01-01

    Full Text Available The influence of the addition of Amaranthus cruenthus seed meal to the medium, as nutrient and growth factor, on protease production by Bacillus subtilis 3411 was studied. Tests were carried out in a rotary shaker and in mechanically stirred fermenters. The influence of aeration was also evaluated. The addition of amaranth in a concentration of 20 g/L resulted in 400% increase in protease production. Aeration up to 750 r.p.m. and 1 L/L.min had a favorable effect.

  15. Enhancing Production of Alkaline Polygalacturonate Lyase from Bacillus subtilis by Fed-Batch Fermentation

    OpenAIRE

    Mouyong Zou; Fenfen Guo; Xuezhi Li; Jian Zhao; Yinbo Qu

    2014-01-01

    Alkaline polygalacturonate lyase (PGL, EC 4.2.2.2) is an enzyme used in many industries. We developed a fed-batch fermentation process that combines the enzymatic pretreatment of the carbon source with controlling the pH of the fermentative broth to enhance the PGL production from Bacillus subtilis 7-3-3 to decrease the production cost. Maintaining the fermentation broth at pH 6.5 prior to feeding with ammonia and at pH 6.0 after feeding significantly improved PGL activity (743.5 U mL-1) comp...

  16. [Anaerobic solid-phase fermentation of plant substrates by Bacillus subtilis].

    Science.gov (United States)

    Ushakova, N A; Brodskiĭ, E S; Kozlova, A A; Nifatov, A V

    2009-01-01

    Solid-phase growth of Bacillus subtilis 8130 on cellulose-rich plant substrates (presscakes or pulp) under hypoxic conditions was accompanied by cellulose depolymerization, protein hydrolysis, and degradation of other plant components, including some processes of mixed-type carbohydrate fermentation. The bacterial fermentation yielded propionic, butyric, and hexanoic acids and butyric acid derivatives. The bacterial metabolism and fermentation degree can be characterized by the proportions of fatty acids in the reaction mixture. The product of sea buckthorn cake fermentation has a good sorption quality.

  17. Pathway engineering of Bacillus subtilis for microbial production of N-acetylglucosamine.

    Science.gov (United States)

    Liu, Yanfeng; Liu, Long; Shin, Hyun-dong; Chen, Rachel R; Li, Jianghua; Du, Guocheng; Chen, Jian

    2013-09-01

    Glucosamine (GlcN) and its acetylated derivative, N-acetylglucosamine (GlcNAc), are widely used in nutraceutical and pharmaceutical industries. Currently, GlcN and GlcNAc are mainly produced by hydrolysis from crab and shrimp shells, which can cause severe environmental pollution and carries the potential risk of allergic reactions. In this study, we attempted to achieve microbial production of GlcNAc by pathway engineering of Bacillus subtilis 168. Specifically, glmS (encoding GlcN-6-phosphate synthase) from B. subtilis 168 and GNA1 (encoding GlcNAc-6-phosphate N-acetyltransferase) from Saccharomyces cerevisiae S288C were firstly co-overexpressed in B. subtilis; the level of GlcNAc reached 240mg/L in shake flask culture. Next, nagP, encoding the GlcNAc-specific enzyme of phosphotransferase system, was deleted to block the importation of extracellular GlcNAC, thus improving GlcNAc production to 615mg/L in shake flask culture. Then, nagA (encoding GlcNAc-6-phosphate deacetylase), gamA (encoding GlcN-6-phosphate deaminase), and nagB (encoding GlcN-6-phosphate deaminase) were deleted to block the catabolism of intracellular GlcNAc, thereby further increasing the GlcNAc titer to 1.85g/L in shake flask culture. Finally, microbial production of GlcNAc by the engineered B. subtilis 168 was conducted in a 3-L fed-batch bioreactor, and the GlcNAc titer reached 5.19g/L, which was 2.8-fold of that in shake flask culture. This is the first report regarding the pathway engineering of B. subtilis for microbial production of GlcNAc, and provides a good starting point for further metabolic engineering to achieve the industrial production of GlcNAc by a generally regarded as safe strain.

  18. Localization of the Bacillus subtilis murB gene within the dcw cluster is important for growth and sporulation.

    Science.gov (United States)

    Real, Gonçalo; Henriques, Adriano O

    2006-03-01

    The Bacillus subtilis murB gene, encoding UDP-N-acetylenolpyruvoylglucosamine reductase, a key enzyme in the peptidoglycan (PG) biosynthetic pathway, is embedded in the dcw (for "division and cell wall") cluster immediately upstream of divIB. Previous attempts to inactivate murB were unsuccessful, suggesting its essentiality. Here we show that the cell morphology, growth rate, and resistance to cell wall-active antibiotics of murB conditional mutants is a function of the expression level of murB. In one mutant, in which murB was insertionally inactivated in a merodiploid bearing a second xylose-inducible PxylA-murB allele, DivIB levels were reduced and a normal growth rate was achieved only if MurB levels were threefold that of the wild-type strain. However, expression of an extra copy of divIB restored normal growth at wild-type levels of MurB. In contrast, DivIB levels were normal in a second mutant containing an in-frame deletion of murB (DeltamurB) in the presence of the PxylA-murB gene. Furthermore, this strain grew normally with wild-type levels of MurB. During sporulation, the levels of MurB were highest at the time of synthesis of the spore cortex PG. Interestingly, the DeltamurB PxylA-murB mutant did not sporulate efficiently even at high concentrations of inducer. Since high levels of inducer did not interfere with sporulation of a murB(+)PxylA-murB strain, it appears that ectopic expression of murB fails to support efficient sporulation. These data suggest that coordinate expression of divIB and murB is important for growth and sporulation. The genetic context of the murB gene within the dcw cluster is unique to the Bacillus group and, taken together with our data, suggests that in these species it contributes to the optimal expression of cell division and PG biosynthetic functions during both vegetative growth and spore development.

  19. Mapping of Proteomic Composition on the Surfaces of Bacillus spores by Atomic Force Microscopy-based Immunolabeling

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Malkin, A J

    2008-06-02

    Atomic force microscopy provides a unique capability to image high-resolution architecture and structural dynamics of pathogens (e.g. viruses, bacteria and bacterial spores) at near molecular resolution in native conditions. Further development of atomic force microscopy in order to enable the correlation of pathogen protein surface structures with specific gene products is essential to understand the mechanisms of the pathogen life cycle. We have applied an AFM-based immunolabeling technique for the proteomic mapping of macromolecular structures through the visualization of the binding of antibodies, conjugated with nanogold particles, to specific epitopes on Bacillus spore surfaces. This information is generated while simultaneously acquiring the surface morphology of the pathogen. The immunospecificity of this labeling method was established through the utilization of specific polyclonal and monoclonal antibodies that target spore coat and exosporium epitopes of Bacillus atrophaeus and Bacillus anthracis spores.

  20. Computational Fluid Dynamics Modeling of Bacillus anthracis Spore Deposition in Rabbit and Human Respiratory Airways

    Energy Technology Data Exchange (ETDEWEB)

    Kabilan, Senthil; Suffield, Sarah R.; Recknagle, Kurtis P.; Jacob, Rick E.; Einstein, Daniel R.; Kuprat, Andrew P.; Carson, James P.; Colby, Sean M.; Saunders, James H.; Hines, Stephanie; Teeguarden, Justin G.; Straub, Tim M.; Moe, M.; Taft, Sarah; Corley, Richard A.

    2016-09-30

    Three-dimensional computational fluid dynamics and Lagrangian particle deposition models were developed to compare the deposition of aerosolized Bacillus anthracis spores in the respiratory airways of a human with that of the rabbit, a species commonly used in the study of anthrax disease. The respiratory airway geometries for each species were derived from computed tomography (CT) or µCT images. Both models encompassed airways that extended from the external nose to the lung with a total of 272 outlets in the human model and 2878 outlets in the rabbit model. All simulations of spore deposition were conducted under transient, inhalation-exhalation breathing conditions using average species-specific minute volumes. The highest exposure concentration was modeled in the rabbit based upon prior acute inhalation studies. For comparison, human simulation was also conducted at the same concentration. Results demonstrated that regional spore deposition patterns were sensitive to airway geometry and ventilation profiles. Due to the complex airway geometries in the rabbit nose, higher spore deposition efficiency was predicted in the upper conducting airways compared to the human at the same air concentration of anthrax spores. As a result, higher particle deposition was predicted in the conducting airways and deep lung of the human compared to the rabbit lung due to differences in airway branching pattern. This information can be used to refine published and ongoing biokinetic models of inhalation anthrax spore exposures, which currently estimate deposited spore concentrations based solely upon exposure concentrations and inhaled doses that do not factor in species-specific anatomy and physiology.

  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. Draft Genome Sequence of Commercial Textile Dye-Decolorizing and -Degrading Bacillus subtilis Strain C3 Isolated in India

    Science.gov (United States)

    Kunadia, Khushbu; Nathani, Neelam M.; Kothari, Vishal; Kotadia, Rohit J.; Kothari, Charmy R.; Joshi, Anjali; Rank, Jalpa K.; Faldu, Priti R.; Shekar, M. Chandra; Viroja, Mitkumar J.; Patel, Priyank A.; Jadeja, Divyarajsinh; Reddy, Bhaskar; Pal Singh, Ravindra; Koringa, Prakash G.; Joshi, Chaitanya G.

    2016-01-01

    Bacillus subtilis C3, a commercial textile dye-decolorizing and -degrading bacterium, was isolated from the common effluent treatment plant (CEPT) of the Jetpur textile dyeing and printing industrial sector situated in the district of Rajkot, Gujarat, India. Here, we present the annotated 4.18-Mb draft genome sequence of B. subtilis C3, providing information about the metabolic pathways involved in decolorization and degradation of several commercial textile azo dyes. Thus, we confirm B. subtilis C3 as a potential candidate for bioremediation of textile effluents. PMID:26966205

  3. Draft Genome Sequence of Commercial Textile Dye-Decolorizing and -Degrading Bacillus subtilis Strain C3 Isolated in India.

    Science.gov (United States)

    Kunadia, Khushbu; Nathani, Neelam M; Kothari, Vishal; Kotadia, Rohit J; Kothari, Charmy R; Joshi, Anjali; Rank, Jalpa K; Faldu, Priti R; Shekar, M Chandra; Viroja, Mitkumar J; Patel, Priyank A; Jadeja, Divyarajsinh; Reddy, Bhaskar; Pal Singh, Ravindra; Koringa, Prakash G; Joshi, Chaitanya G; Kothari, Ramesh K

    2016-03-10

    Bacillus subtilis C3, a commercial textile dye-decolorizing and -degrading bacterium, was isolated from the common effluent treatment plant (CEPT) of the Jetpur textile dyeing and printing industrial sector situated in the district of Rajkot, Gujarat, India. Here, we present the annotated 4.18-Mb draft genome sequence of B. subtilis C3, providing information about the metabolic pathways involved in decolorization and degradation of several commercial textile azo dyes. Thus, we confirm B. subtilis C3 as a potential candidate for bioremediation of textile effluents.

  4. Characterization of dacC, which encodes a new low-molecular-weight penicillin-binding protein in Bacillus subtilis

    DEFF Research Database (Denmark)

    Pedersen, Lotte Bang; Murray, T; Popham, D L;

    1998-01-01

    The pbp gene (renamed dacC), identified by the Bacillus subtilis genome sequencing project, encodes a putative 491-residue protein with sequence homology to low-molecular-weight penicillin-binding proteins. Use of a transcriptional dacC-lacZ fusion revealed that dacC expression (i) is initiated...... at the end of stationary phase; (ii) depends strongly on transcription factor sigmaH; and (iii) appears to be initiated from a promoter located immediately upstream of yoxA, a gene of unknown function located upstream of dacC on the B. subtilis chromosome. A B. subtilis dacC insertional mutant grew...

  5. Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Applied to Quantitative Proteomics of Bacillus subtilis

    DEFF Research Database (Denmark)

    Soufi, Boumediene; Kumar, C.; Gnad, F.

    2010-01-01

    We applied stable isotope labeling by amino acids in cell culture (SILAC) to large-scale quantitative proteomics analyses of the model bacterium Bacillus subtilis in two physiological conditions: growth on succinate and growth under phosphate starvation. Using a B. subtilis strain auxotrophic...... of the most comprehensive quantitative proteomics studies in bacteria, covering more than 75% of the B. subtilis genes expressed in the log phase of growth. Furthermore, we detect and quantify dynamics of 35 Ser/Thr/Tyr phosphorylation sites under growth on succinate, and 10 phosphorylation sites under...

  6. Sterilization effect of UV light on Bacillus spores using TiO2 films depends on wavelength

    OpenAIRE

    Nhung, Le Thi Tuyet; Nagata, Hirofumi; Takahashi, Akira; Aihara, Mutsumi; Okamoto, Toshihiro; Shimohata, Takaaki; Mawatari, Kazuaki; Akutagawa, Masatake; Kinouchi, Yohsuke; Haraguchi, Masanobu

    2012-01-01

    UV light and photocatalysts such as titanium dioxide (TiO2) and silver (Ag) are useful for disinfection of water and surfaces. However, the effect of UV wavelength on photocatalytic disinfection of spores is not well understood. Inactivation of Bacillus spores has been examined using different UV wavelengths and TiO2 or TiO2/Ag composite materials. The level of UVA disinfection of Bacillus anthracis and Bacillus brevis vegetative cells increased with the presence of the TiO2 and Ag photocatal...

  7. Caracterización de cristales de calcita bioprecipitada por un aislamiento nativo de Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Carolina Montoya

    2007-02-01

    Full Text Available Bacillus subtilis es una bacteria útil en algunas aplicaciones biotecnológicas por poseer enzimas como las amilasas, las cuales desempeñan un papel importante en diferentes procesos industriales. Una de sus propiedades, poco estudiada, ha sido su capacidad de inducir bioprecipitación química de carbonato de calcio (Ca2+ + HCO3 3> CaCO3 + H+ mediante un mecanismo similar al observado en la formación de rocas, suelos y estructuras biológicas como huesos, conchas y dientes. En esta investigación se estudiaron los cristales producidos por un aislamiento nativo de B. subtilis, tomado de una mina de oro situada en Segovia (Antioquia. Se determinó su capacidad calcificante utilizando el medio de cultivo B4. La caracterización del cristal producido se realizó con lupa binocular, microscopio petrográfico de luz plana polarizada (MOLP en su modo de luz transmitida, microscopio electrónico de barrido con analizador de estado sólido (ESEM/EDX y espectroscopía infrarroja con transformada de Fourier (FTIR. A partir de los resultados obtenidos por medio de la caracterización utilizando la combinación de las técnicas analíticas que se mencionaron, fue posible determinar que el aislado nativo de B. subtilis generó y por ende es productor de cristales de carbonato de calcio (CaCO3 en su forma polimórfica de baja temperatura (calcite.Palabras clave: Bacillus subtilis, calcita, bioprecipitación, mineralogía aplicada, biomineralogía.ABSTRACTBacillus subtilis, a bacterium useful in some biotechnology applications, contains enzymes such as amylases, which play an important role in several industrial processes. One of its properties, not very well studied, is its capacity to induce the chemical bioprecipitation of CaCO3 (Ca2+ + HCO3 —> CaCO3 + H+, a similar mechanism commonly observed in the formation of rocks, soils and biological structures like bones, shells and teeth. In this work we have studied carbonate crystals produced by a B

  8. Crude glycerol from biodiesel industry as substrate for biosurfactant production by Bacillus subtilis ATCC 6633

    Directory of Open Access Journals (Sweden)

    Marylane de Sousa

    2014-04-01

    Full Text Available Glycerol, a co-product of the biodiesel industry, may be a suitable raw material for the production of high added-value compounds by the microorganisms. This study aimed to use the glycerol obtained from the biodiesel production process as the main carbon source for biosurfactant production by Bacillus subtilis ATCC 6633. Results indicated that the strain lowered the surface tension of the cell-free fermented broth to 31.5 ± 1.6 mN/m, indicating the production of biosurfactant. The critical micelle concentration (CMC = 33.6 mN/m obtained was similar to the previously reported for biossurfactants isolated from other Bacillus. The produced biosurfactant was able to emulsify n-hexadecane and soybean oil.

  9. Isothermal titration calorimetry and surface plasmon resonance allow quantifying substrate binding to different binding sites of Bacillus subtilis xylanase

    DEFF Research Database (Denmark)

    Cuyvers, Sven; Dornez, Emmie; Abou Hachem, Maher;

    2012-01-01

    Isothermal titration calorimetry and surface plasmon resonance were tested for their ability to study substrate binding to the active site (AS) and to the secondary binding site (SBS) of Bacillus subtilis xylanase A separately. To this end, three enzyme variants were compared. The first was a cat......Isothermal titration calorimetry and surface plasmon resonance were tested for their ability to study substrate binding to the active site (AS) and to the secondary binding site (SBS) of Bacillus subtilis xylanase A separately. To this end, three enzyme variants were compared. The first...

  10. Application of Bacillus subtilis 168 as a multifunctional agent for improvement of the durability of cement mortar.

    Science.gov (United States)

    Park, Sung-Jin; Park, Jong-Myong; Kim, Wha-Jung; Ghim, Sa-Youl

    2012-11-01

    Microbiological calcium carbonate precipitation (MCCP) has been investigated for its ability to improve the durability of cement mortar. However, very few strains have been applied to crack remediation and strengthening of cementitious materials. In this study, we report the biodeposition of Bacillus subtilis 168 and its ability to enhance the durability of cement material. B. subtilis 168 was applied to the surface of cement specimens. The results showed a new layer of deposited organic-inorganic composites on the surface of the cement paste. In addition, the water permeability of the cement paste treated with B. subtilis 168 was lower than that of non-treated specimens. Furthermore, artificial cracks in the cement paste were completely remediated by the biodeposition of B. subtilis 168. The compressive strength of cement mortar treated with B. subtilis 168 increased by about 19.5% when compared with samples completed with only B4 medium. Taken together, these findings suggest that the biodeposition of B. subtilis 168 could be used as a sealing and coating agent to improve the strength and water resistance of concrete. This is the first paper to report the application of Bacillus subtilis 168 for its ability to improve the durability of cement mortar through calcium carbonate precipitation.

  11. Rapid detection of Bacillus anthracis spores using a super-paramagnetic lateral-flow immunological detection system.

    Science.gov (United States)

    Wang, Dian-Bing; Tian, Bo; Zhang, Zhi-Ping; Deng, Jiao-Yu; Cui, Zong-Qiang; Yang, Rui-Fu; Wang, Xu-Ying; Wei, Hong-Ping; Zhang, Xian-En

    2013-04-15

    There is an urgent need for convenient, sensitive, and specific methods to detect the spores of Bacillus anthracis, the causative agent of anthrax, because of the bioterrorism threat posed by this bacterium. In this study, we firstly develop a super-paramagnetic lateral-flow immunological detection system for B. anthracis spores. This system involves the use of a portable magnetic assay reader, super-paramagnetic iron oxide particles, lateral-flow strips and two different monoclonal antibodies directed against B. anthracis spores. This detection system specifically recognises as few as 400 pure B. anthracis spores in 30 min. This system has a linear range of 4×10³-10⁶ CFU ml⁻¹ and reproducible detection limits of 200 spores mg⁻¹ milk powder and 130 spores mg⁻¹ soil for simulated samples. In addition, this approach shows no obvious cross-reaction with other related Bacillus spores, even at high concentrations, and has no significant dependence on the duration of the storage of the immunological strips. Therefore, this super-paramagnetic lateral-flow immunological detection system is a promising tool for the rapid and sensitive detection of Bacillus anthracis spores under field conditions.

  12. Heterologous expression, biochemical characterization, and overproduction of alkaline α-amylase from Bacillus alcalophilus in Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Li Jianghua

    2011-10-01

    Full Text Available Abstract Background Alkaline α-amylases have potential applications for hydrolyzing starch under high pH conditions in the starch and textile industries and as ingredients in detergents for automatic dishwashers and laundries. While the alkaline α-amylase gains increased industrial interest, the yield of alkaline α-amylases from wild-type microbes is low, and the combination of genetic engineering and process optimization is necessary to achieve the overproduction of alkaline α-amylase. Results The alkaline α-amylase gene from Bacillus alcalophilus JN21 (CCTCC NO. M 2011229 was cloned and expressed in Bacillus subtilis strain WB600 with vector pMA5. The recombinant alkaline α-amylase was stable at pH from 7.0 to 11.0 and temperature below 40°C. The optimum pH and temperature of alkaline α-amylase was 9.0 and 50°C, respectively. Using soluble starch as the substrate, the Km and Vmax of alkaline α-amylase were 9.64 g/L and 0.80 g/(L·min, respectively. The effects of medium compositions (starch, peptone, and soybean meal and temperature on the recombinant production of alkaline α-amylase in B. subtilis were investigated. Under the optimal conditions (starch concentration 0.6% (w/v, peptone concentration 1.45% (w/v, soybean meal concentration 1.3% (w/v, and temperature 37°C, the highest yield of alkaline α-amylase reached 415 U/mL. The yield of alkaline α-amylase in a 3-L fermentor reached 441 U/mL, which was 79 times that of native alkaline α-amylase from B. alcalophilus JN21. Conclusions This is the first report concerning the heterologous expression of alkaline α-amylase in B. subtilis, and the obtained results make it feasible to achieve the industrial production of alkaline α-amylase with the recombinant B. subtilis.

  13. On the origin of heterogeneity in (preservation) resistance of Bacillus spores: Input for a ‘systems’ analysis approach of bacterial spore outgrowth

    NARCIS (Netherlands)

    Hornstra, L.M.; ter Beek, A.; Smelt, J.P.; Kallemeijn, W.W.; Brul, S.

    2009-01-01

    Bacterial spores are the ultimate (stress) ‘survival capsules’. They allow strains from the Bacillus and Clostridium species to survive harsh environmental conditions. In addition to the decision to enter sporulation the decision to do the reverse (germinate) is also a decisive event after which the

  14. IMMUNE-RELATED GENES EXPRESSION AND PHAGOCYTOSIS AGAINST WHITE SPOT SYNDROME VIRUS AFTER ORAL DELIVERY OF VP28 USING BACILLUS SUBTILIS AS VEHICLES IN LITOPENAEUS VANNAMEI%以枯草芽孢杆菌递呈VP28对南美白对虾免疫相关基因表达和细胞特异性吞噬的影响

    Institute of Scientific and Technical Information of China (English)

    丁晶; 王彦波; 傅玲琳

    2013-01-01

    以枯草芽孢杆菌(Bacillus subtilis)为活载体口服递呈对虾白斑综合征病毒(WSSV)囊膜蛋白 VP28,评价其抗病毒感染能力、对南美白对虾免疫相关基因表达以及血淋巴细胞对病毒特异性吞噬的影响。经口服免疫枯草重组菌株B. subtilis-VP28攻毒后,对虾的相对存活率达83.3%。为探讨重组菌株的抗病机理,比较研究了免疫相关基因-proPO(酚氧化酶原)、Peroxinectin(PE)和脂多糖-β-1,3-葡聚糖结合蛋白(LGBP)基因的表达差异,并进一步分析了血淋巴细胞吞噬活性和特异性。结果表明, B. subtilis-VP28菌液能显著提高(P<0.05)对虾proPO、PE和LGBP mRNA的表达水平和血细胞对WSSV的吞噬活性, B. subtilis组对免疫相关基因也有一定的激活作用,而B. subtilis-VP28发酵上清液则能增加血细胞吞噬活性;此外, B. subtilis-VP28菌液组血细胞对WSSV具有特异性吞噬作用。研究为枯草重组菌株B. subtilis-VP28抗WSSV感染作用及其作为特殊功能水产微生态制剂的应用提供了一定的科学依据。%The regulation of immune-related genes expression and phagocytosis of White Spot Syndrome Virus (WSSV) were evaluated by oral delivery of VP28 using Bacillus subtilis as vehicles in Litopenaeus vannamei. In our initial ex-periment, by oral delivery of B. subtilis spores harboring VP28 (B. subtilis-VP28) to L. vannamei, the extremely high survival (Relative Percent Survival:83.3%) upon challenge with WSSV can be observed. The differences of genes ex-pression levels of proPO, Peroxinectin (PE) and lipopolysaccharide-and beta-1, 3-glucan-binding protein (LGBP) were demonstrated among experimental groups of B. subtilis-VP28 bacterial spores, B. subtilis-VP28 supernatants, B. subtilis and control. The result showed that immune-related genes (proPO, PE and LGBP) were significantly (P<0.05) upregu-lated in both B. subtilis-VP28 bacterial spores and B. subtilis feeding groups compared to B. subtilis-VP28

  15. Spore prevalence and toxigenicity of Bacillus cereus and Bacillus thuringiensis isolates from U.S. retail spices.

    Science.gov (United States)

    Hariram, Upasana; Labbé, Ronald

    2015-03-01

    Recent incidents of foodborne illness associated with spices as the vehicle of transmission prompted this examination of U.S. retail spices with regard to Bacillus cereus. This study focused on the levels of aerobic-mesophilic spore-forming bacteria and B cereus spores associated with 247 retail spices purchased from five states in the United States. Samples contained a wide range of aerobic-mesophilic bacterial spore counts ( 10(7) CFU/g). Using a novel chromogenic agar, B. cereus and B. thuringiensis spores were isolated from 77 (31%) and 11 (4%) samples, respectively. Levels of B. cereus were thuringiensis isolates possessed at least one type of enterotoxin gene: HBL (hemolysin BL) or nonhemolytic enterotoxin (NHE). None of the 88 isolates obtained in this study possessed the emetic toxin gene (ces). Using commercially available immunological toxin detection kits, the toxigenicity of the isolates was confirmed. The NHE enterotoxin was expressed in 98% of B. cereus and 91% of B. thuringiensis isolates that possessed the responsible gene. HBL enterotoxin was detected in 87% of B. cereus and 100% of B. thuringiensis PCR-positive isolates. Fifty-two percent of B. cereus and 54% of B. thuringiensis isolates produced both enterotoxins. Ninety-seven percent of B. cereus isolates grew at 12°C, although only two isolates grew well at 9°C. The ability of these spice isolates to form spores, produce diarrheal toxins, and grow at moderately abusive temperatures makes retail spices an important potential vehicle for foodborne illness caused by B. cereus strains, in particular those that produce diarrheal toxins.

  16. An improved system for the surface immobilisation of proteins on Bacillus thuringiensis vegetative cells and spores through a new spore cortex-lytic enzyme anchor.

    Science.gov (United States)

    Shao, Xiaohu; Ni, Hong; Lu, Ting; Jiang, Mengtian; Li, Hua; Huang, Xinfeng; Li, Lin

    2012-02-15

    An improved surface-immobilisation system was engineered to target heterologous proteins onto vegetative cells and spores of Bacillus thuringiensis plasmid-free recipient strain BMB171. The sporulation-dependent spore cortex-lytic enzyme from B. thuringiensis YBT-1520, SceA, was expressed in vegetative cells and used as the surface anchoring motif. Green fluorescent protein (GFP) and a Bacillus endo-β-1,3-1,4-glucanase (BglS) were used as the fusion partners to test the binding efficiency and the functional activities of immobilised surface proteins. The surface localisation of the SceA-GFP fusion protein on vegetative cells and spores was confirmed by Western blot, immunofluorescence microscopy and flow cytometry. The GFP fluorescence intensity from both vegetative cells and spores was measured and compared to a previously characterised surface display system using a peptidoglycan hydrolase anchor (Mbg). Results demonstrated comparable efficiency of SceA- and Mbg-mediated immobilisation on vegetative cells but a more efficient immobilisation on spores using the SceA anchor, suggesting SceA has greater potential for spore-based applications. The SceA protein was then applied to target BglS onto vegetative cells and spores, and the surface immobilisation was verified by the substantial whole-cell enzymatic activity and enhanced whole-spore enzymatic activity compared to vegetative cells. A dually active B. thuringiensis vegetative cell and spore display system could prove especially valuable for the development of regenerable and heat-stable biocatalysts that function under adverse environmental conditions, for example, an effective feed additive for improved digestion and nutrient absorption by livestock.

  17. Inactivation characteristics of ozone and electrolysis process for ballast water treatment using B. subtilis spores as a probe.

    Science.gov (United States)

    Jung, Youmi; Yoon, Yeojoon; Hong, Eunkyung; Kwon, Minhwan; Kang, Joon-Wun

    2013-07-15

    Since ballast water affects the ocean ecosystem, the International Maritime Organization (IMO) sets a standard for ballast water management and might impose much tighter regulations in the future. The aim of this study is to evaluate the inactivation efficiency of ozonation, electrolysis, and an ozonation-electrolysis combined process, using B. subtilis spores. In seawater ozonation, HOBr is the key active substance for inactivation, because of rapid reactivity of ozone with Br(-) in seawater. In seawater electrolysis, it is also HOBr, but not HOCl, because of the rapid reaction of HOCl with Br(-), which has not been recognized carefully, even though many electrolysis technologies have been approved by the IMO. Inactivation pattern was different in ozonation and electrolysis, which has some limitations with the tailing or lag-phase, respectively. However, each deficiency can be overcome with a combined process, which is most effective as a sequential application of ozonation followed by electrolysis.

  18. Enhanced and Secretory Expression of Human Granulocyte Colony Stimulating Factor by Bacillus subtilis SCK6

    Directory of Open Access Journals (Sweden)

    Shaista Bashir

    2015-01-01

    Full Text Available This study describes a simplified approach for enhanced expression and secretion of a pharmaceutically important human cytokine, that is, granulocyte colony stimulating factor (GCSF, in the culture supernatant of Bacillus subtilis SCK6 cells. Codon optimized GCSF and pNWPH vector containing SpymwC signal sequence were amplified by prolonged overlap extension PCR to generate multimeric plasmid DNA, which was used directly to transform B. subtilis SCK6 supercompetent cells. Expression of GCSF was monitored in the culture supernatant for 120 hours. The highest expression, which corresponded to 17% of the total secretory protein, was observed at 72 hours of growth. Following ammonium sulphate precipitation, GCSF was purified to near homogeneity by fast protein liquid chromatography on a QFF anion exchange column. Circular dichroism spectroscopic analysis showed that the secondary structure contents of the purified GCSF are similar to the commercially available GCSF. Biological activity, as revealed by the regeneration of neutrophils in mice treated with ifosfamine, was also similar to the commercial preparation of GCSF. This, to our knowledge, is the first study that reports secretory expression of human GCSF in B. subtilis SCK6 with final recovery of up to 96 mg/L of the culture supernatant, without involvement of any chemical inducer.

  19. Plant growth promotion by spermidine-producing Bacillus subtilis OKB105.

    Science.gov (United States)

    Xie, Shan-Shan; Wu, Hui-Jun; Zang, Hao-Yu; Wu, Li-Ming; Zhu, Qing-Qing; Gao, Xue-Wen

    2014-07-01

    The interaction between plants and plant-growth-promoting rhizobacteria (PGPR) is a complex, reciprocal process. On the one hand, plant compounds such as carbohydrates and amino acids serve as energy sources for PGPR. On the other hand, PGPR promote plant growth by synthesizing plant hormones and increasing mineral availability in the soil. Here, we evaluated the growth-promoting activity of Bacillus subtilis OKB105 and identified genes associated with this activity. The genes yecA (encoding a putative amino acid/polyamine permease) and speB (encoding agmatinase) are involved in the secretion or synthesis of polyamine in B. subtilis OKB105. Disruption of either gene abolished the growth-promoting activity of the bacterium, which was restored when polyamine synthesis was complemented. Moreover, high-performance liquid chromatography analysis of culture filtrates of OKB105 and its derivatives demonstrated that spermidine, a common polyamine, is the pivotal plant-growth-promoting compound. In addition, real-time polymerase chain reaction analysis revealed that treatment with B. subtilis OKB105 induced expansin gene (Nt-EXPA1 and Nt-EXPA2) expression and inhibited the expression of the ethylene biosynthesis gene ACO1. Furthermore, enzyme-linked immunosorbent assay analysis showed that the ethylene content in plant root cells decreased in response to spermidine produced by OKB105. Therefore, during plant interactions, OKB105 may produce and secrete spermidine, which induces expansin production and lowers ethylene levels.

  20. Stationary-Phase Mutagenesis in Stressed Bacillus subtilis Cells Operates by Mfd-Dependent Mutagenic Pathways.

    Science.gov (United States)

    Gómez-Marroquín, Martha; Martin, Holly A; Pepper, Amber; Girard, Mary E; Kidman, Amanda A; Vallin, Carmen; Yasbin, Ronald E; Pedraza-Reyes, Mario; Robleto, Eduardo A

    2016-07-05

    In replication-limited cells of Bacillus subtilis, Mfd is mutagenic at highly transcribed regions, even in the absence of bulky DNA lesions. However, the mechanism leading to increased mutagenesis through Mfd remains currently unknown. Here, we report that Mfd may promote mutagenesis in nutritionally stressed B. subtilis cells by coordinating error-prone repair events mediated by UvrA, MutY and PolI. Using a point-mutated gene conferring leucine auxotrophy as a genetic marker, it was found that the absence of UvrA reduced the Leu⁺ revertants and that a second mutation in mfd reduced mutagenesis further. Moreover, the mfd and polA mutants presented low but similar reversion frequencies compared to the parental strain. These results suggest that Mfd promotes mutagenic events that required the participation of NER pathway and PolI. Remarkably, this Mfd-dependent mutagenic pathway was found to be epistatic onto MutY; however, whereas the MutY-dependent Leu⁺ reversions required Mfd, a direct interaction between these proteins was not apparent. In summary, our results support the concept that Mfd promotes mutagenesis in starved B. subtilis cells by coordinating both known and previously unknown Mfd-associated repair pathways. These mutagenic processes bias the production of genetic diversity towards highly transcribed regions in the genome.

  1. Bacillus subtilis: from soil bacterium to super-secreting cell factory

    Directory of Open Access Journals (Sweden)

    van Dijl Jan Maarten

    2013-01-01

    Full Text Available Abstract The biotechnology industry has become a key element in modern societies. Within this industry, the production of recombinant enzymes and biopharmaceutical proteins is of major importance. The global markets for such recombinant proteins are growing rapidly and, accordingly, there is a continuous need for new production platforms that can deliver protein products in greater yields, with higher quality and at lower costs. This calls for the development of next-generation super-secreting cell factories. One of the microbial cell factories that can meet these challenges is the Gram-positive bacterium Bacillus subtilis, an inhabitant of the upper layers of the soil that has the capacity to secrete proteins in the gram per litre range. The engineering of B. subtilis into a next-generation super-secreting cell factory requires combined Systems and Synthetic Biology approaches. In this way, the bacterial protein secretion machinery can be optimized from the single molecule to the network level while, at the same time, taking into account the balanced use of cellular resources. Although highly ambitious, this is an achievable objective due to recent advances in functional genomics and Systems- and Synthetic Biological analyses of B. subtilis cells.

  2. Enhanced dipicolinic acid production during the stationary phase in Bacillus subtilis by blocking acetoin synthesis.

    Science.gov (United States)

    Toya, Yoshihiro; Hirasawa, Takashi; Ishikawa, Shu; Chumsakul, Onuma; Morimoto, Takuya; Liu, Shenghao; Masuda, Kenta; Kageyama, Yasushi; Ozaki, Katsuya; Ogasawara, Naotake; Shimizu, Hiroshi

    2015-01-01

    Bacterial bio-production during the stationary phase is expected to lead to a high target yield because the cells do not consume the substrate for growth. Bacillus subtilis is widely used for bio-production, but little is known about the metabolism during the stationary phase. In this study, we focused on the dipicolinic acid (DPA) production by B. subtilis and investigated the metabolism. We found that DPA production competes with acetoin synthesis and that acetoin synthesis genes (alsSD) deletion increases DPA productivity by 1.4-fold. The mutant showed interesting features where the glucose uptake was inhibited, whereas the cell density increased by approximately 50%, resulting in similar volumetric glucose consumption to that of the parental strain. The metabolic profiles revealed accumulation of pyruvate, acetyl-CoA, and the TCA cycle intermediates in the alsSD mutant. Our results indicate that alsSD-deleted B. subtilis has potential as an effective host for stationary-phase production of compounds synthesized from these intermediates.

  3. Molecular insights into frataxin-mediated iron supply for heme biosynthesis in Bacillus subtilis.

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    Andreas Mielcarek

    Full Text Available Iron is required as an element to sustain life in all eukaryotes and most bacteria. Although several bacterial iron acquisition strategies have been well explored, little is known about the intracellular trafficking pathways of iron and its entry into the systems for co-factor biogenesis. In this study, we investigated the iron-dependent process of heme maturation in Bacillus subtilis and present, for the first time, structural evidence for the physical interaction of a frataxin homologue (Fra, which is suggested to act as a regulatory component as well as an iron chaperone in different cellular pathways, and a ferrochelatase (HemH, which catalyses the final step of heme b biogenesis. Specific interaction between Fra and HemH was observed upon co-purification from crude cell lysates and, further, by using the recombinant proteins for analytical size-exclusion chromatography. Hydrogen-deuterium exchange experiments identified the landscape of the Fra/HemH interaction interface and revealed Fra as a specific ferrous iron donor for the ferrochelatase HemH. The functional utilisation of the in vitro-generated heme b co-factor upon Fra-mediated iron transfer was confirmed by using the B. subtilis nitric oxide synthase bsNos as a metabolic target enzyme. Complementary mutational analyses confirmed that Fra acts as an essential component for maturation and subsequent targeting of the heme b co-factor, hence representing a key player in the iron-dependent physiology of B. subtilis.

  4. Localization of Components of the RNA-Degrading Machine in Bacillus subtilis

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    Cascante-Estepa, Nora; Gunka, Katrin; Stülke, Jörg

    2016-01-01

    In bacteria, the control of mRNA stability is crucial to allow rapid adaptation to changing conditions. In most bacteria, RNA degradation is catalyzed by the RNA degradosome, a protein complex composed of endo- and exoribonucleases, RNA helicases, and accessory proteins. In the Gram-positive model organism Bacillus subtilis, the existence of a RNA degradosome assembled around the membrane-bound endoribonuclease RNase Y has been proposed. Here, we have studied the intracellular localization of the protein that have been implicated in the potential B. subtilis RNA degradosome, i.e., polynucleotide phosphorylase, the exoribonucleases J1 and J2, the DEAD-box RNA helicase CshA, and the glycolytic enzymes enolase and phosphofructokinase. Our data suggests that the bulk of these enzymes is located in the cytoplasm. The RNases J1 and J2 as well as the RNA helicase CshA were mainly localized in the peripheral regions of the cell where also the bulk of messenger RNA is localized. We were able to demonstrate active exclusion of these proteins from the transcribing nucleoid. Taken together, our findings suggest that the interactions of the enzymes involved in RNA degradation in B. subtilis are rather transient. PMID:27708634

  5. Malate metabolism in Bacillus subtilis: distinct roles for three classes of malate-oxidizing enzymes.

    Science.gov (United States)

    Meyer, Frederik M; Stülke, Jörg

    2013-02-01

    The Gram-positive soil bacterium Bacillus subtilis uses glucose and malate as the preferred carbon sources. In the presence of either glucose or malate, the expression of genes and operons for the utilization of secondary carbon sources is subject to carbon catabolite repression. While glucose is a preferred substrate in many organisms from bacteria to man, the factors that contribute to the preference for malate have so far remained elusive. In this work, we have studied the contribution of the different malate-metabolizing enzymes in B. subtilis, and we have elucidated their distinct functions. The malate dehydrogenase and the phosphoenolpyruvate carboxykinase are both essential for malate utilization; they introduce malate into gluconeogenesis. The NADPH-generating malic enzyme YtsJ is important to establish the cellular pools of NADPH for anabolic reactions. Finally, the NADH-generating malic enzymes MaeA, MalS, and MleA are involved in keeping the ATP levels high. Together, this unique array of distinct activities makes malate a preferred carbon source for B. subtilis.

  6. Characterization of ftsZ mutations that render Bacillus subtilis resistant to MinC.

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    Inês Filipa Fernandes de Oliveira

    Full Text Available BACKGROUND: Cell division in Bacillus subtilis occurs precisely at midcell. Positional control of cell division is exerted by two mechanisms: nucleoid occlusion, through Noc, which prevents division through nucleoids, and the Min system, where the combined action of the MinC, D and J proteins prevents formation of the FtsZ ring at cell poles or recently completed division sites. METHODOLOGY/PRINCIPAL FINDINGS: We used a genetic screen to identify mutations in ftsZ that confer resistance to the lethal overexpression of the MinC/MinD division inhibitor. The FtsZ mutants were purified and found to polymerize to a similar or lesser extent as wild type FtsZ, and all mutants displayed reduced GTP hydrolysis activity indicative of a reduced polymerization turnover. We found that even though the mutations conferred in vivo resistance to MinC/D, the purified FtsZ mutants did not display strong resistance to MinC in vitro. CONCLUSIONS/SIGNIFICANCE: Our results show that in B. subtilis, overproduction of MinC can be countered by mutations that alter FtsZ polymerization dynamics. Even though it would be very likely that the FtsZ mutants found depend on other Z-ring stabilizing proteins such as ZapA, FtsA or SepF, we found this not to be the case. This indicates that the cell division process in B. subtilis is extremely robust.

  7. Small regulatory RNA-induced growth rate heterogeneity of Bacillus subtilis.

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    Ruben A T Mars

    2015-03-01

    Full Text Available Isogenic bacterial populations can consist of cells displaying heterogeneous physiological traits. Small regulatory RNAs (sRNAs could affect this heterogeneity since they act by fine-tuning mRNA or protein levels to coordinate the appropriate cellular behavior. Here we show that the sRNA RnaC/S1022 from the Gram-positive bacterium Bacillus subtilis can suppress exponential growth by modulation of the transcriptional regulator AbrB. Specifically, the post-transcriptional abrB-RnaC/S1022 interaction allows B. subtilis to increase the cell-to-cell variation in AbrB protein levels, despite strong negative autoregulation of the abrB promoter. This behavior is consistent with existing mathematical models of sRNA action, thus suggesting that induction of protein expression noise could be a new general aspect of sRNA regulation. Importantly, we show that the sRNA-induced diversity in AbrB levels generates heterogeneity in growth rates during the exponential growth phase. Based on these findings, we hypothesize that the resulting subpopulations of fast- and slow-growing B. subtilis cells reflect a bet-hedging strategy for enhanced survival of unfavorable conditions.

  8. Enantioselective transesterification of glycidol catalysed by a novel lipase expressed from Bacillus subtilis.

    Science.gov (United States)

    Wang, Lei; Tai, Jian-Dong; Wang, Ren; Xun, Er-Na; Wei, Xiao-Fei; Wang, Lei; Wang, Zhi

    2010-05-10

    A novel plasmid (pBSR2) was constructed by incorporating a strong lipase promoter and a terminator into the original pBD64. The lipase gene from Bacillus subtilis strain IFFI10210 was cloned into the plasmid pBSR2 and transformed into B. subtilis A.S.1.1655 to obtain an overexpression strain. The recombinant lipase [BSL2 (B. subtilis lipase 2)] has been expressed from the novel constructed strain and used in kinetic resolution of glycidol through enantioselective transesterification. The effects of reaction conditions on the activity as well as enantioselectivity were investigated. BSL2 showed a satisfying enantioselectivity (E>30) under the optimum conditions [acyl donor: vinyl butyrate; the mole ratio of vinyl butyrate to glycidol was 3:1; organic medium: 1,2-dichloroethane with water activity (a(w))=0.33; temperature 40 degrees C]. The remaining (R)-glycidol with a high enantiomeric purity [ee (enantiomeric excess) >99%] could be obtained when the conversion was approx. 60%. The results clearly show a good potential for industrial application of BSL2 in the resolution of glycidol through enantioselective transesterification.

  9. A process for high-efficiency isoflavone deglycosylation using Bacillus subtilis natto NTU-18.

    Science.gov (United States)

    Kuo, Lun-Cheng; Wu, Ren-Yu; Lee, Kung-Ta

    2012-06-01

    In order to produce isoflavone aglycosides effectively, a process of isoflavone hydrolysis by Bacillus subtilis natto NTU-18 (BCRC 80390) was established. This process integrates the three stages for the production of isoflavone aglycosides in one single fermenter, including the growth of B. subtilis natto, production of β-glucosidase, deglycosylation of fed isoflavone glycosides. After 8 h of batch culture of B. subtilis natto NTU-18 in 2 L of soy medium, a total of 3 L of soy isoflavone glucoside solution containing 3.0 mg/mL of daidzin and 1.0 mg/mL of genistin was fed continuously over 34 h. The percentage deglycosylation of daidzin and genistin was 97.7% and 94.6%, respectively. The concentration of daidzein and genistein in the broth reached 1,066.8 μg/mL (4.2 mM) and 351 μg/mL (1.3 mM), respectively, and no residual daidzin or genistin was detected. The productivity of the bioconversion of daidzein and genistein over the 42 h of culture was 25.6 mg/L/h and 8.5 mg/L/h, respectively. This showed that this is an efficient bioconversion process for selective estrogen receptor modulator production.

  10. Three biotechnical processes using Ashbya gossypii, Candida famata, or Bacillus subtilis compete with chemical riboflavin production.

    Science.gov (United States)

    Stahmann, K P; Revuelta, J L; Seulberger, H

    2000-05-01

    Chemical riboflavin production, successfully used for decades, is in the course of being replaced by microbial processes. These promise to save half the costs, reduce waste and energy requirements, and use renewable resources like sugar or plant oil. Three microorganisms are currently in use for industrial riboflavin production. The hemiascomycetes Ashbya gossypii, a filamentous fungus, and Candida famata, a yeast, are naturally occurring overproducers of this vitamin. To obtain riboflavin production with the gram-positive bacterium Bacillus subtilis requires at least the deregulation of purine synthesis and a mutation in a flavokinase/FAD-synthetase. It is common to all three organisms that riboflavin production is recognizable by the yellow color of the colonies. This is an important tool for the screening of improved mutants. Antimetabolites like itaconate, which inhibits the isocitrate lyase in A. gossypii, tubercidin, which inhibits purine biosynthesis in C. famata, or roseoflavin, a structural analog of riboflavin used for B. subtilis, have been applied successfully for mutant selections. The production of riboflavin by the two fungi seems to be limited by precursor supply, as was concluded from feeding and gene-overexpression experiments. Although flux studies in B. subtilis revealed an increase both in maintenance metabolism and in the oxidative part of the pentose phosphate pathway, the major limitation there seems to be the riboflavin pathway. Multiple copies of the rib genes and promoter replacements are necessary to achieve competitive productivity.

  11. Effect of the derivatives of andrographolide on the morphology of Bacillus subtilis.

    Science.gov (United States)

    Aromdee, Chantana; Sriubolmas, Nongluksna; Wiyakrutta, Suthep; Suebsasna, Supawadee; Khunkitti, Watcharee

    2011-01-01

    Andrographis paniculata has been reported to have antiviral, antipyretic and anticancer activities. Andrographolide, an ent-labdane diterpene, is an active constituent in this plant. In this study, andrographolide (1) and its natural derivative 14-deoxy-11,12-didehydroandrographolide (2) and 5 other semisynthetic derivatives were tested for their activity against Gram-positive and Gram-negative bacteria and Candida albicans. Only derivatives bearing a 14-acetyl group showed activity, and this activity was only against Gram-positive bacteria. 14-Acetylandrographolide showed the highest potency against Bacillus subtilis; the other 14-acetylandrographolides with additional substitution at the 3- and 19-hydroxyl groups showed lower activity against Gram-positive bacteria. The morphology of B. subtilis after being treated with 14-acetylandrographolide was investigated with TEM. This is the first report on 14-acetylandrographolide's quantified antibacterial activity, and the crucial functional group of this ent-labdane that plays an important role in perturbing the morphogenesis of B. subtilis leading to cell death.

  12. Bacillus subtilis attachment to Aspergillus niger hyphae results in mutually altered metabolism.

    Science.gov (United States)

    Benoit, Isabelle; van den Esker, Marielle H; Patyshakuliyeva, Aleksandrina; Mattern, Derek J; Blei, Felix; Zhou, Miaomiao; Dijksterhuis, Jan; Brakhage, Axel A; Kuipers, Oscar P; de Vries, Ronald P; Kovács, Ákos T

    2015-06-01

    Interaction between microbes affects the growth, metabolism and differentiation of members of the microbial community. While direct and indirect competition, like antagonism and nutrient consumption have a negative effect on the interacting members of the population, microbes have also evolved in nature not only to fight, but in some cases to adapt to or support each other, while increasing the fitness of the community. The presence of bacteria and fungi in soil results in various interactions including mutualism. Bacilli attach to the plant root and form complex communities in the rhizosphere. Bacillus subtilis, when grown in the presence of Aspergillus niger, interacts similarly with the fungus, by attaching and growing on the hyphae. Based on data obtained in a dual transcriptome experiment, we suggest that both fungi and bacteria alter their metabolism during this interaction. Interestingly, the transcription of genes related to the antifungal and putative antibacterial defence mechanism of B. subtilis and A. niger, respectively, are decreased upon attachment of bacteria to the mycelia. Analysis of the culture supernatant suggests that surfactin production by B. subtilis was reduced when the bacterium was co-cultivated with the fungus. Our experiments provide new insights into the interaction between a bacterium and a fungus.

  13. Isolation and characterization of atrazine mineralizing Bacillus subtilis strain HB-6.

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    Jinhua Wang

    Full Text Available Atrazine is a widely used herbicide with great environmental concern due to its high potential to contaminate soil and waters. An atrazine-degrading bacterial strain HB-6 was isolated from industrial wastewater and the 16S rRNA gene sequencing identified HB-6 as a Bacillus subtilis. PCR assays indicated that HB-6 contained atrazine-degrading genes trzN, atzB and atzC. The strain HB-6 was capable of utilizing atrazine and cyanuric acid as a sole nitrogen source for growth and even cleaved the s-triazine ring and mineralized atrazine. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by cooperating with other bacteria, suggesting its huge potential for remediation of atrazine-contaminated sites. To our knowledge, there are few Bacillus subtilis strains reported that can mineralize atrazine, therefore, the present work might provide some new insights on atrazine remediation.

  14. Isolation and characterization of atrazine mineralizing Bacillus subtilis strain HB-6.

    Science.gov (United States)

    Wang, Jinhua; Zhu, Lusheng; Wang, Qi; Wang, Jun; Xie, Hui

    2014-01-01

    Atrazine is a widely used herbicide with great environmental concern due to its high potential to contaminate soil and waters. An atrazine-degrading bacterial strain HB-6 was isolated from industrial wastewater and the 16S rRNA gene sequencing identified HB-6 as a Bacillus subtilis. PCR assays indicated that HB-6 contained atrazine-degrading genes trzN, atzB and atzC. The strain HB-6 was capable of utilizing atrazine and cyanuric acid as a sole nitrogen source for growth and even cleaved the s-triazine ring and mineralized atrazine. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by cooperating with other bacteria, suggesting its huge potential for remediation of atrazine-contaminated sites. To our knowledge, there are few Bacillus subtilis strains reported that can mineralize atrazine, therefore, the present work might provide some new insights on atrazine remediation.

  15. Simulation of Bacillus subtilis biofilm growth on agar plate by diffusion-reaction based continuum model

    Science.gov (United States)

    Zhang, Xianlong; Wang, Xiaoling; Nie, Kai; Li, Mingpeng; Sun, Qingping

    2016-08-01

    Various species of bacteria form highly organized spatially-structured aggregates known as biofilms. To understand how microenvironments impact biofilm growth dynamics, we propose a diffusion-reaction continuum model to simulate the formation of Bacillus subtilis biofilm on an agar plate. The extended finite element method combined with level set method are employed to perform the simulation, numerical results show the quantitative relationship between colony morphologies and nutrient depletion over time. Considering that the production of polysaccharide in wild-type cells may enhance biofilm spreading on the agar plate, we inoculate mutant colony incapable of producing polysaccharide to verify our results. Predictions of the glutamate source biofilm’s shape parameters agree with the experimental mutant colony better than that of glycerol source biofilm, suggesting that glutamate is rate limiting nutrient for Bacillus subtilis biofilm growth on agar plate, and the diffusion-limited is a better description to the experiment. In addition, we find that the diffusion time scale is of the same magnitude as growth process, and the common-employed quasi-steady approximation is not applicable here.

  16. Induced sensitivity of Bacillus subtilis colony morphology to mechanical media compression

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    Jessica K. Polka

    2014-09-01

    Full Text Available Bacteria from several taxa, including Kurthia zopfii, Myxococcus xanthus, and Bacillus mycoides, have been reported to align growth of their colonies to small features on the surface of solid media, including anisotropies created by compression. While the function of this phenomenon is unclear, it may help organisms navigate on solid phases, such as soil. The origin of this behavior is also unknown: it may be biological (that is, dependent on components that sense the environment and regulate growth accordingly or merely physical.Here we show that B. subtilis, an organism that typically does not respond to media compression, can be induced to do so with two simple and synergistic perturbations: a mutation that maintains cells in the swarming (chained state, and the addition of EDTA to the growth media, which further increases chain length. EDTA apparently increases chain length by inducing defects in cell separation, as the treatment has only marginal effects on the length of individual cells.These results lead us to three conclusions. First, the wealth of genetic tools available to B. subtilis will provide a new, tractable chassis for engineering compression sensitive organisms. Second, the sensitivity of colony morphology to media compression in Bacillus can be modulated by altering a simple physical property of rod-shaped cells. And third, colony morphology under compression holds promise as a rapid, simple, and low-cost way to screen for changes in the length of rod-shaped cells or chains thereof.

  17. Dietary mannan oligosaccharide and Bacillus subtilis in diets for Nile tilapia (Oreochromis niloticus

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    Rafael Vieira de Azevedo

    2016-11-01

    Full Text Available A six week study was conducted to investigate the supplementation of prebiotic (Mannan oligosaccharide – MOS, from yeast Saccharomyces cerevisiae, probiotic (Bacillus subtilis – BS, C-3102 strain and their combination in diets for Nile tilapia. 192 fishes (4.03 ± 0.28 g were distributed into 16 tanks (40-L, in a completely randomized design (n=4. The following treatments were evaluated: control; prebiotic - 2 g MOS kg-1; probiotic - 2 g BS kg-1 and synbiotic - 1 g MOS kg-1 plus 1 g BS kg-1. Fishes fed diets pre-, pro- and synbiotic supplemented performed better in average daily gain, feed conversion rate, specific growth rate, protein efficiency ratio, carcass yield, total and standard length and body height than those maintained on control diets. The probiotic supplementation resulted in higher villus height and intestinal perimeter ratio than the control diet while the pre- and synbiotic supplementation in diets resulted in higher intestinal perimeter ratio. Carcass protein and ether extract were, respectively, higher and lower in fish fed synbiotic diets than other fish. The results of this study indicated that the mannan oligosaccharide and Bacillus subtilis supplementation, isolated or combined (synbiotic, could improve growth, body index, intestine morphometry and carcass composition in Nile tilapia.

  18. Point-cycle bistability and stochasticity in a regulatory circuit for Bacillus subtilis competence.

    Science.gov (United States)

    Xi, Hongguang; Duan, Lixia; Turcotte, Marc

    2013-08-01

    Bacillus subtilis is a very well-studied organism in biology. Recent results show that an evolutionary plausible alternative competence regulation circuit for this bacterium, despite presenting equivalent functionality, exhibits physiologically important differences. Thus, it is not a priori clear why Nature only selects a specific gene regulation circuit other than a plethora of equivalent others. Here, we use simulations to study this question further. Based on the wild-type Bacillus subtilis circuit, we add a positive autoregulation feedback loop to the intermediate gene comS. We use bifurcation theory to study the dynamical features of the hypothetical gene circuit versus the feedback strength of the added loop, and we rely on stochastic simulations to perform in silico experiments. We discover the existence of a bistable system: a stable limit cycle and a stable fixed point separated by an unstable limit cycle with a varying height of underlying stochastic potential. This structure is absent from the wild type. The coexistence of the unstable limit cycle with stochastic noise endows the circuit with an ability to confine, prevent or switch between its two stable attractors.

  19. Involvement of fengycin-type lipopeptides in the multifaceted biocontrol potential of Bacillus subtilis.

    Science.gov (United States)

    Ongena, Marc; Jacques, Philippe; Touré, Yacine; Destain, Jacqueline; Jabrane, Abdelhamid; Thonart, Philippe

    2005-11-01

    In this work, the potential of Bacillus subtilis strain M4 at protecting plants against fungal diseases was demonstrated in different pathosystems. We provide evidence for the role of secreted lipopeptides, and more particularly of fengycins, in the protective effect afforded by the strain against damping-off of bean seedlings caused by Pythium ultimum and against gray mold of apple in post-harvest disease. This role was demonstrated by the strong biocontrol activity of lipopeptide-enriched extracts and through the detection of inhibitory quantities of fengycins in infected tissues. Beside such a direct antagonism of the pathogen, we show that root pre-inoculation with M4 enabled the host plant to react more efficiently to subsequent pathogen infection on leaves. Fengycins could also be involved in this systemic resistance-eliciting effect of strain M4, as these molecules may induce the synthesis of plant phenolics involved in or derived from the defense-related phenylpropanoid metabolism. Much remains to be discovered about the mechanisms by which Bacillus spp suppress disease. Through this study on strain M4, we reinforce the interest in B. subtilis as a pathogen antagonist and plant defense-inducing agent. The secretion of cyclic fengycin-type lipopeptides may be tightly related to the expression of these two biocontrol traits.

  20. Growth Inhibition of Colletotrichum gloeosporioides by Trichoderma harzianum, Trichoderma koningii, Bacillus subtilis and Pseudomonas fluorescens

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    Febrilia Nur ‘Aini

    2015-11-01

    Full Text Available Colletotrichum  gloeosporioides is  a  disease  which  can  cause  significant yield  loss  of  cocoa.  The  objective  of  this  research  is  to  investigate  the  abilityof  antagonist  microbes,  Trichoderma  harzianum,  Trichoderma  koningii,  Bacillus subtilis  and Pseudomonas  fluorescens  in  controlling  gloeosporioides  biologically  in  laboratorium  condition.  The  experiment  was  carried  out  in  Crop  Protection  Laboratory,  Indonesian  Coffee  and  Cocoa  Research  Institute.  Results of  this  research  showed  that  antagonist  fungi,  T.  harzianum,  T.  koningii,  had  a stronger  ability  in  inhibiting  growth  of  C.  gloeosporioides about  83%  compared  to  the  ability  of  antagonist  bacteria,  B.  subtilis  and P.  fluorescens,  only about  49%. Key words: Growth  inhibition,  Colletotrichum  gloeosporioides,  Trichoderma  harzianum, Trichoderma koningii,  Bacillus subtilis, Pseudomonas fluorescens.

  1. Poly-γ-Glutamic Acids Contribute to Biofilm Formation and Plant Root Colonization in Selected Environmental Isolates of Bacillus subtilis.

    Science.gov (United States)

    Yu, Yiyang; Yan, Fang; Chen, Yun; Jin, Christopher; Guo, Jian-Hua; Chai, Yunrong

    2016-01-01

    Bacillus subtilis is long known to produce poly-γ-glutamic acids (γ-PGA) as one of the major secreted polymeric substances. In B. subtilis, the regulation of γ-PGA production and its physiological role are still unclear. B. subtilis is also capable of forming structurally complex multicellular communities, or biofilms, in which an extracellular matrix consisting of secreted proteins and polysaccharides holds individual cells together. Biofilms were shown to facilitate B. subtilis-plant interactions. In this study, we show that different environmental isolates of B. subtilis, all capable of forming biofilms, vary significantly in γ-PGA production. This is possibly due to differential regulation of γ-PGA biosynthesis genes. In many of those environmental isolates, γ-PGA seems to contribute to robustness and complex morphology of the colony biofilms, suggesting a role of γ-PGA in biofilm formation. Our evidence further shows that in selected B. subtilis strains, γ-PGA also plays a role in root colonization by the bacteria, pinpointing a possible function of γ-PGA in B. subtilis-plant interactions. Finally, we found that several pathways co-regulate both γ-PGA biosynthesis genes and genes for the biofilm matrix in B. subtilis, but in an opposing fashion. We discussed potential biological significance of that.

  2. Isolation and Identification of Lipopeptides Produced by Bacillus subtilis fmbJ%Bacillus subtilis fmbJ脂肽类抗菌物质的分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    别小妹; 吕凤霞; 陆兆新; 黄现青; 沈娟

    2006-01-01

    Bacillus subtilis fmbJ脂肽类抗菌物质的分离和鉴定进行了系统研究.通过HPLC层析确定Bacillus subtilis fmbJ抗菌物质由多种组分构成,其中含有保留时间与surfactin相似的成分.通过TLC层析和原位酸解确定Bacillus subtilis fmbJ抗菌物质含有两个具有闭合肽键类的物质,其中之一为迁移率Rf与标样surfactin非常相近的组分.通过ESI-MS分析检测到Bacillus subtilis fmbJ抗菌物质含有分子量与fengicin相同的m/z1449.9、m/z1463.8、m/z1477.8、m/z1491.9和m/z1505.9五种同系物,和分子量与surfactin相同的m/z1008.8、m/z1022.8和m/z1036.8三种同系物.

  3. Isolation of Bacillus subtilis as indicator in the disinfection of residual water by means of gamma radiation; Aislamiento de Bacillus subtilis como indicador en la desinfeccion de aguas residuales mediante radiacion gamma

    Energy Technology Data Exchange (ETDEWEB)

    Mata J, M.; Colin C, A. [Facultad de Quimica, UAEM, Paseo Colon esq. Tollocan s/n, Toluca, 50000 Estado de Mexico (Mexico); Lopez V, H.; Brena V, M.; Carrasco A, H.; Pavon R, S. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)

    2002-07-01

    In the attempt to get more alternatives of disinfection of residual water, the Bacillus subtilis was isolated by means of gamma radiation as a bio indicator of disinfection since it turned out to be resistant to the 5 KGy dose, comparing this one with other usual microorganisms as biondicators like E. coli and S typhimurium which turn out more sensitive to such dose. (Author)

  4. Killing of Bacillus Megaterium Spores by X-rays at the Phosphorus K-edge

    Science.gov (United States)

    Richmond, Robert C.; Frigo, Sean P.; Ehret, Charles F.; Rose, M. Franklin (Technical Monitor)

    2001-01-01

    This study continues a progression of experiments on the radiation-induced killing of bacterial spores that began at the Argonne National Laboratory in 1957. A series of aliquots of Bacillus megaterium spores were prepared onto polycarbonate filters and irradiated with photons of 2159 eV compared to 2140 eV energy on the 2-IDB beamline at the Advanced Photon Source. Flux density was approximately 10(exp 18) photons/sec/sq mm. The phosphorous K-edge absorption spectrum in these spores was determined to peak at 2159 eV, wheras 2140 eV was determined to be outside that absorption spectrum. Spores on filters were irradiated at ambient conditions, and were either immediately plated for colony formation after irradiation, or were held for postirradiation exposure to oxygen prior to plating. Slopes of survival curves from the four conditions of irradiation, i.e., two photon energies each comparing immediate plating vs postirradiation holding, were used for quantitative determination of differences in rates of spore killing over a range of radiation doses. It was found that spores irradiated at the phosphorus K-edge were killed 20% more efficiently than when irradiated with 2140 eV photons, and this was true for both immediate plating and postirradiation holding in air. Postirradiation holding in air increased killing efficiency by about 12% for both photon energies compared to plating immediately after irradiation. The increase of killing efficiency with postirradiation holding is less than expected from earlier experiments using relatively low-flux X-rays, and raises the possibility of dose-mitigation by radical-radical recombination in the case of high-flux X-rays from the synchrotron.

  5. [Participation of the antibiotics of Bac. pumilus and Bac. subtilis in the regulation of bacterial spore formation].

    Science.gov (United States)

    Lukin, A A; Korolev, V I

    1979-03-01

    Sporulation and antibiotic production, as well as the effect of exogenic antibacterial substances on bacterial sporogenesis were studied in various strains of Bac. pumilus and Bac. pumilus and Bac. subtilis. The bacteria were grown on a solid sporulation medium with and without the antibiotics. After 5-day incubation the presence of refractyl spores was determined with a phase-contrast method. It was found that in the strains of Bac. pumilus producing antibacterial substances the sporulation was normal. The loss of the capacity for synthesizing such substances resulted in asporegenicity or oligosporogenicity. This allowed a conclusion on existence of phenomenological connection between sporulation and antibiotic production. The study of the antibiotic effect on bacterial sporogenesis showed negative results which are discussed in the paper along two directions: (1) the antibiotics did not probably participate in regulation of the bacteria cell differentiation, (2) the antibiotics regulated the bacterial sporogenesis though their effect was not as yet detected because of methodical difficulties. Therefore, the problem of the antibiotic participation in regulation of sporulation in Bac. pumilus and Bac. subtilis remains open.

  6. Most Probable Number Rapid Viability PCR Method to Detect Viable Spores of Bacillus anthracis in Swab Samples

    Energy Technology Data Exchange (ETDEWEB)

    Letant, S E; Kane, S R; Murphy, G A; Alfaro, T M; Hodges, L; Rose, L; Raber, E

    2008-05-30

    This note presents a comparison of Most-Probable-Number Rapid Viability (MPN-RV) PCR and traditional culture methods for the quantification of Bacillus anthracis Sterne spores in macrofoam swabs generated by the Centers for Disease Control and Prevention (CDC) for a multi-center validation study aimed at testing environmental swab processing methods for recovery, detection, and quantification of viable B. anthracis spores from surfaces. Results show that spore numbers provided by the MPN RV-PCR method were in statistical agreement with the CDC conventional culture method for all three levels of spores tested (10{sup 4}, 10{sup 2}, and 10 spores) even in the presence of dirt. In addition to detecting low levels of spores in environmental conditions, the MPN RV-PCR method is specific, and compatible with automated high-throughput sample processing and analysis protocols.

  7. Detecting Cortex Fragments During Bacterial Spore Germination.

    Science.gov (United States)

    Francis, Michael B; Sorg, Joseph A

    2016-06-25

    The process of endospore germination in Clostridium difficile, and other Clostridia, increasingly is being found to differ from the model spore-forming bacterium, Bacillus subtilis. Germination is triggered by small molecule germinants and occurs without the need for macromolecular synthesis. Though differences exist between the mechanisms of spore germination in species of Bacillus and Clostridium, a common requirement is the hydrolysis of the peptidoglycan-like cortex which allows the spore core to swell and rehydrate. After rehydration, metabolism can begin and this, eventually, leads to outgrowth of a vegetative cell. The detection of hydrolyzed cortex fragments during spore germination can be difficult and the modifications to the previously described assays can be confusing or difficult to reproduce. Thus, based on our recent report using this assay, we detail a step-by-step protocol for the colorimetric detection of cortex fragments during bacterial spore germination.

  8. Bacillus subtilis is a Potential Degrader of Pyrene and Benzo[a]pyrene

    Directory of Open Access Journals (Sweden)

    Lynette Ekunwe

    2005-08-01

    Full Text Available Polycyclic Aromatic Hydrocarbons (PAHs are a group of compounds that pose many health threats to human and animal life. They occur in nature as a result of incomplete combustion of organic matter, as well as from many anthropogenic sources including cigarette smoke and automobile exhaust. PAHs have been reported to cause liver damage, red blood cell damage and a variety of cancers. Because of this, methods to reduce the amount of PAHs in the environment are continuously being sought. The purpose of this study was to find soil bacteria capable of degrading high molecular weight PAHs, such as pyrene (Pyr and benzo[a]pyrene (BaP, which contain more than three benzene rings and so persist in the environment. Bacillus subtilis, identified by fatty acid methyl ester (FAME analysis, was isolated from PAH contaminated soil. Because it grew in the presence of 33μg/ml each of pyrene, 1-AP and 1-HP, its biodegradation capabilities were assessed. It was found that after a four-day incubation period at 30oC in 20μg/ml pyrene or benzo[a]pyrene, B. subtilis was able to transform approximately 40% and 50% pyrene and benzo[a]pyrene, respectively. This is the first report implicating B. subtilis in PAH degradation. Whether or not the intermediates resulting from the transformation are more toxic than their parent compounds, and whether B. subtilis is capable of mineralizing pyrene or benzo[a]pyrene to carbon dioxide and water, remains to be evaluated.

  9. Effect of nanocomposite packaging containing ZnO on growth of Bacillus subtilis and Enterobacter aerogenes

    Energy Technology Data Exchange (ETDEWEB)

    Esmailzadeh, Hakimeh [National Nutrition and Food Technology Research Institute, Faculty of Nutrition and Food Technology, Shahid Beheshti University of Medical Sciences, Tehran (Iran, Islamic Republic of); Sangpour, Parvaneh, E-mail: Sangpour@merc.ac.ir [Nanotechnology and Advanced Materials Department, Materials and Energy Research Center, Karaj (Iran, Islamic Republic of); Shahraz, Farzaneh [National Nutrition and Food Technology Research Institute, Faculty of Nutrition and Food Technology, Shahid Beheshti University of Medical Sciences, Tehran (Iran, Islamic Republic of); Hejazi, Jalal [Department of Biochemistry and Nutrition, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan (Iran, Islamic Republic of); Khaksar, Ramin [National Nutrition and Food Technology Research Institute, Faculty of Nutrition and Food Technology, Shahid Beheshti University of Medical Sciences, Tehran (Iran, Islamic Republic of)

    2016-01-01

    Recent advances in nanotechnology have opened new windows in active food packaging. Nano-sized ZnO is an inexpensive material with potential antimicrobial properties. The aim of the present study is to evaluate the antibacterial effect of low density Polyethylene (LDPE) containing ZnO nanoparticles on Bacillus subtilis and Enterobacter aerogenes. ZnO nanoparticles have been synthesized by facil molten salt method and have been characterized by X-ray diffraction (XRD), and scanning electron microscopy (SEM). Nanocomposite films containing 2 and 4 wt.% ZnO nanoparticles were prepared by melt mixing in a twin-screw extruder. The growth of both microorganisms has decreased in the presence of ZnO containing nanocomposites compared with controls. Nanocomposites with 4 wt.% ZnO nanoparticles had stronger antibacterial effect against both bacteria in comparison with the 2 wt.% ZnO containing nanocomposites. B. subtilis as Gram-positive bacteria were more sensitive to ZnO containing nanocomposite films compared with E. aerogenes as Gram-negative bacteria. There were no significant differences between the migration of Zn ions from 2 and 4 wt.% ZnO containing nanocomposites and the released Zn ions were not significantly increased in both groups after 14 days compared with the first. Regarding the considerable antibacterial effects of ZnO nanoparticles, their application in active food packaging can be a suitable solution for extending the shelf life of food. - Highlights: • ZnO containing nanocomposites decreased growth of both B. subtilis and E. aerogenes. • B. subtilis was more sensitive to ZnO containing nanocomposites. • The migration of Zn ions from nanocomposites was negligible.

  10. Promoter Screening from Bacillus subtilis in Various Conditions Hunting for Synthetic Biology and Industrial Applications.

    Directory of Open Access Journals (Sweden)

    Yafeng Song

    Full Text Available The use of Bacillus subtilis in synthetic biology and metabolic engineering is highly desirable to take advantage of the unique metabolic pathways present in this organism. To do this, an evaluation of B. subtilis' intrinsic biological parts is required to determine the best strategies to accurately regulate metabolic circuits and expression of target proteins. The strengths of promoter candidates were evaluated by measuring relative fluorescence units of a green fluorescent protein reporter, integrated into B. subtilis' chromosome. A total of 84 predicted promoter sequences located upstream of different classes of proteins including heat shock proteins, cell-envelope proteins, and proteins resistant against toxic metals (based on similarity and other kinds of genes were tested. The expression levels measured ranged from 0.0023 to 4.53-fold of the activity of the well-characterized strong promoter P43. No significant shifts were observed when strains, carrying different promoter candidates, were cultured at high temperature or in media with ethanol, but some strains showed increased activity when cultured under high osmotic pressure. Randomly selected promoter candidates were tested and found to activate transcription of thermostable β-galactosidase (bgaB at a similar level, implying the ability of these sequences to function as promoter elements in multiple genetic contexts. In addition, selected promoters elevated the final production of both cytoplasmic bgaB and secreted protein α-amylase to about fourfold and twofold, respectively. The generated data allows a deeper understanding of B. subtilis' metabolism and will facilitate future work to develop this organism for synthetic biology.

  11. Rational improvement of the engineered isobutanol-producing Bacillus subtilis by elementary mode analysis

    Directory of Open Access Journals (Sweden)

    Li Shanshan

    2012-08-01

    Full Text Available Abstract Background Isobutanol is considered as a leading candidate for the replacement of current fossil fuels, and expected to be produced biotechnologically. Owing to the valuable features, Bacillus subtilis has been engineered as an isobutanol producer, whereas it needs to be further optimized for more efficient production. Since elementary mode analysis (EMA is a powerful tool for systematical analysis of metabolic network structures and cell metabolism, it might be of great importance in the rational strain improvement. Results Metabolic network of the isobutanol-producing B. subtilis BSUL03 was first constructed for EMA. Considering the actual cellular physiological state, 239 elementary modes (EMs were screened from total 11,342 EMs for potential target prediction. On this basis, lactate dehydrogenase (LDH and pyruvate dehydrogenase complex (PDHC were predicted as the most promising inactivation candidates according to flux flexibility analysis and intracellular flux distribution simulation. Then, the in silico designed mutants were experimentally constructed. The maximal isobutanol yield of the LDH- and PDHC-deficient strain BSUL05 reached 61% of the theoretical value to 0.36 ± 0.02 C-mol isobutanol/C-mol glucose, which was 2.3-fold of BSUL03. Moreover, this mutant produced approximately 70 % more isobutanol to the maximal titer of 5.5 ± 0.3 g/L in fed-batch fermentations. Conclusions EMA was employed as a guiding tool to direct rational improvement of the engineered isobutanol-producing B. subtilis. The consistency between model prediction and experimental results demonstrates the rationality and accuracy of this EMA-based approach for target identification. This network-based rational strain improvement strategy could serve as a promising concept to engineer efficient B. subtilis hosts for isobutanol, as well as other valuable products.

  12. Functional Expression of Enterobacterial O-Polysaccharide Biosynthesis Enzymes in Bacillus subtilis

    Science.gov (United States)

    Schäffer, Christina; Wugeditsch, Thomas; Messner, Paul; Whitfield, Chris

    2002-01-01

    The expression of heterologous bacterial glycosyltransferases is of interest for potential application in the emerging field of carbohydrate engineering in gram-positive organisms. To assess the feasibility of using enzymes from gram-negative bacteria, the functional expression of the genes wbaP (formerly rfbP), wecA (formerly rfe), and wbbO (formerly rfbF) from enterobacterial lipopolysaccharide O-polysaccharide biosynthesis pathways was examined in Bacillus subtilis. WbaP and WecA are initiation enzymes for O-polysaccharide formation, catalyzing the transfer of galactosyl 1-phosphate from UDP-galactose and N-acetylglucosaminyl 1-phosphate from UDP-N-acetylglucosamine, respectively, to undecaprenylphosphate. The WecA product (undecaprenylpyrophosphoryl GlcNAc) is used as an acceptor to which the bifunctional wbbO gene product sequentially adds a galactopyranose and a galactofuranose residue from the corresponding UDP sugars to form a lipid-linked trisaccharide. Genes were cloned into the shuttle vectors pRB374 and pAW10. In B. subtilis hosts, the genes were effectively transcribed under the vegII promoter control of pRB374, but the plasmids were susceptible to rearrangements and deletion. In contrast, pAW10-based constructs, in which genes were cloned downstream of the tet resistance cassette, were stable but yielded lower levels of enzyme activity. In vitro glycosyltransferase assays were performed in Escherichia coli and B. subtilis, using membrane preparations as sources of enzymes and endogenous undecaprenylphosphate as an acceptor. Incorporation of radioactivity from UDP-α-d-14C-sugar into reaction products verified the functionality of WbaP, WecA, and WbbO in either host. Enzyme activities in B. subtilis varied between 20 and 75% of those measured in E. coli. PMID:12324313

  13. Changes in the Acetylome and Succinylome of Bacillus subtilis in Response to Carbon Source.

    Directory of Open Access Journals (Sweden)

    Saori Kosono

    Full Text Available Lysine residues can be post-translationally modified by various acyl modifications in bacteria and eukarya. Here, we showed that two major acyl modifications, acetylation and succinylation, were changed in response to the carbon source in the Gram-positive model bacterium Bacillus subtilis. Acetylation was more common when the cells were grown on glucose, glycerol, or pyruvate, whereas succinylation was upregulated when the cells were grown on citrate, reflecting the metabolic states that preferentially produce acetyl-CoA and succinyl-CoA, respectively. To identify and quantify changes in acetylation and succinylation in response to the carbon source, we performed a stable isotope labeling by amino acids in cell culture (SILAC-based quantitative proteomic analysis of cells grown on glucose or citrate. We identified 629 acetylated proteins with 1355 unique acetylation sites and 204 succinylated proteins with 327 unique succinylation sites. Acetylation targeted different metabolic pathways under the two growth conditions: branched-chain amino acid biosynthesis and purine metabolism in glucose and the citrate cycle in citrate. Succinylation preferentially targeted the citrate cycle in citrate. Acetylation and succinylation mostly targeted different lysine residues and showed a preference for different residues surrounding the modification sites, suggesting that the two modifications may depend on different factors such as characteristics of acyl-group donors, molecular environment of the lysine substrate, and/or the modifying enzymes. Changes in acetylation and succinylation were observed in proteins involved in central carbon metabolism and in components of the transcription and translation machineries, such as RNA polymerase and the ribosome. Mutations that modulate protein acylation affected B. subtilis growth. A mutation in acetate kinase (ackA increased the global acetylation level, suggesting that acetyl phosphate-dependent acetylation is

  14. Direct Comparison of Physical Properties of Bacillus subtilis NCIB 3610 and B-1 Biofilms.

    Science.gov (United States)

    Kesel, Sara; Grumbein, Stefan; Gümperlein, Ina; Tallawi, Marwa; Marel, Anna-Kristina; Lieleg, Oliver; Opitz, Madeleine

    2016-04-01

    Many bacteria form surface-attached communities known as biofilms. Due to the extreme resistance of these bacterial biofilms to antibiotics and mechanical stresses, biofilms are of growing interest not only in microbiology but also in medicine and industry. Previous studies have determined the extracellular polymeric substances present in the matrix of biofilms formed by Bacillus subtilis NCIB 3610. However, studies on the physical properties of biofilms formed by this strain are just emerging. In particular, quantitative data on the contributions of biofilm matrix biopolymers to these physical properties are lacking. Here, we quantitatively investigated three physical properties of B. subtilis NCIB 3610 biofilms: the surface roughness and stiffness and the bulk viscoelasticity of these biofilms. We show how specific biomolecules constituting the biofilm matrix formed by this strain contribute to those biofilm properties. In particular, we demonstrate that the surface roughness and surface elasticity of 1-day-old NCIB 3610 biofilms are strongly affected by the surface layer protein BslA. For a second strain,B. subtilis B-1, which forms biofilms containing mainly γ-polyglutamate, we found significantly different physical biofilm properties that are also differently affected by the commonly used antibacterial agent ethanol. We show that B-1 biofilms are protected from ethanol-induced changes in the biofilm's stiffness and that this protective effect can be transferred to NCIB 3610 biofilms by the sole addition of γ-polyglutamate to growing NCIB 3610 biofilms. Together, our results demonstrate the importance of specific biofilm matrix components for the distinct physical properties of B. subtilis biofilms.

  15. Film coating of seeds with Bacillus cereus RS87 spores for early plant growth enhancement.

    Science.gov (United States)

    Jetiyanon, Kanchalee; Wittaya-Areekul, Sakchai; Plianbangchang, Pinyupa

    2008-10-01

    The plant growth-promoting rhizobacterium Bacillus cereus RS87 was previously reported to promote plant growth in various crops in both greenhouse and field trials. To apply as a plant growth promoting agent with practical use, it is essential to ease the burden of routine preparation of a fresh suspension of strain RS87 in laboratory. The objectives of this study were to investigate the feasibility of film-coating seeds with B. cereus RS87 spores for early plant growth enhancement and to reveal the indoleacetic acid (IAA) production released from strain RS87. The experiment consisted of the following 5 treatments: nontreated seeds, water-soaked seeds, film-coated seeds, seeds soaked with vegetative cells of strain RS87, and film-coated seeds with strain RS87 spores. Three experiments were conducted separately to assess seed emergence, root length, and plant height. Results showed that both vegetative cells and spores of strain RS87 significantly promoted (P seed emergence, root length and plant height over the control treatments. The strain RS87 also produced IAA. In conclusion, the film coating of seeds with spores of B. cereus RS87 demonstrated early plant growth enhancement as well as seeds using their vegetative cells. IAA released from strain RS87 would be one of the mechanisms for plant growth enhancement.

  16. Identification of the immunogenic spore and vegetative proteins of Bacillus anthracis vaccine strain A16R.

    Directory of Open Access Journals (Sweden)

    Xiankai Liu

    Full Text Available Immunoproteomics was used to screen the immunogenic spore and vegetative proteins of Bacillus anthracis vaccine strain A16R. The spore and vegetative proteins were separated by 2D gel electrophoresis and transferred to polyvinylidene difluoride membranes, and then western blotting was performed with rabbit immune serum against B.anthracis live spores. Immunogenic spots were cut and digested by trypsin. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed to identify the proteins. As a result, 11 and 45 immunogenic proteins were identified in the spores and vegetative cells, respectively; 26 of which have not been reported previously. To verify their immunogenicity, 12 of the identified proteins were selected to be expressed, and the immune sera from the mice vaccinated by the 12 expressed proteins, except BA0887, had a specific western blot band with the A16R whole cellular lytic proteins. Some of these immunogenic proteins might be used as novel vaccine candidates themselves or for enhancing the protective efficacy of a protective-antigen-based vaccine.

  17. Effects of the electrolytic treatment on Bacillus subtilis Efeito do tratamento eletrolítico em Bacillus subtis

    Directory of Open Access Journals (Sweden)

    Rodolfo Tolentino-Bisneto

    2003-11-01

    Full Text Available Conventional processes of water disinfection can generate toxic composites. It is the case of the trihalomethanes (carcinogenic formed in the contact of chlorine with organic substances present in the water. The electrolytic treatment can be a substitute for the chlorination process without the need for addition of chemical substances to the process. The effect of the electrolytic treatment using carbon cathode on the viability of the microorganism Bacillus subtilis was tested to determine the death process. By means of electronic microscopy, it was observed that the main cause of the microorganism's death was the cellular lysis due to the electroporation in the cell membrane.Processos convencionais de desinfecção de águas podem gerar compostos tóxicos. Esse é o caso dos trialometanos formados na reação do cloro com compostos orgânicos presentes na água. O tratamento eletrolítico pode ser um substituto à cloração com vantagem de não requer a adição de nenhum composto na água. O efeito do tratamento eletrolítico, utilizando eletrodos de carbono, na viabilidade de Bacillus subtilis foi testado para se determinar o mecanismo de morte. Através de microscopia eletrônica, foi possível evidenciar que a morte do microrganismo se deu pela lise celular, provavelmente provocada pela eletroporação irreversível da membrana celular.

  18. Variable Lymphocyte Receptor Recognition of the Immunodominant Glycoprotein of Bacillus anthracis Spores

    Energy Technology Data Exchange (ETDEWEB)

    Kirchdoerfer, Robert N.; Herrin, Brantley R.; Han, Byung Woo; Turnbough, Jr., Charles L.; Cooper, Max D.; Wilson, Ian A. (SNU); (Scripps); (Emory); (UAB); (Emory Vaccine)

    2012-07-25

    Variable lymphocyte receptors (VLRs) are the adaptive immune receptors of jawless fish, which evolved adaptive immunity independent of other vertebrates. In lieu of the immunoglobulin fold-based T and B cell receptors, lymphocyte-like cells of jawless fish express VLRs (VLRA, VLRB, or VLRC) composed of leucine-rich repeats and are similar to toll-like receptors (TLRs) in structure, but antibodies (VLRB) and T cell receptors (VLRA and VLRC) in function. Here, we present the structural and biochemical characterization of VLR4, a VLRB, in complex with BclA, the immunodominant glycoprotein of Bacillus anthracis spores. Using a combination of crystallography, mutagenesis, and binding studies, we delineate the mode of antigen recognition and binding between VLR4 and BclA, examine commonalities in VLRB recognition of antigens, and demonstrate the potential of VLR4 as a diagnostic tool for the identification of B. anthracis spores.

  19. Isolation, evaluation and characterization of Bacillus subtilis from cotton rhizospheric soil with biocontrol activity against Fusarium oxysporum.

    Science.gov (United States)

    Gajbhiye, Archana; Rai, Alok R; Meshram, Sudhir U; Dongre, A B

    2010-07-01

    Present investigation is based on the isolation of Bacillus subtilis from cotton rhizosphere and their evaluation as biocontrol agent against Fusarium oxysporum. The production of extracellular hydrolytic enzyme was studied for determining the antagonism. 43% of 21 isolates were identified under the B. subtilis group on the basis of biochemical characterization. 38% isolates showed competitive activity against Fusarium oxysporum and exhibit more than 50% mycelial inhibition in dual culture bioassay. The pot assay of cotton by seed treatment and soil amendment technique under green house condition showed the competent activity of the isolates in preventing the wilting of cotton seedlings due to F. oxysporum infection. SVI values of 30 day old seedlings indicated that the soil inoculation with B. subtilis BP-2 and seed treatment with B. subtilis BP-9 significantly promoted the growth of cotton seedlings. RAPD profiling revealed the diversity in the Bacillus subtilis group, ranging from 10 to 32%. The discriminative pattern among the isolates belonging to the same species was validated by 16S rDNA partial sequencing which identified them into four different strains of B. subtilis.

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

    DEFF Research Database (Denmark)

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

    2011-01-01

    Bacillus subtilis two-component system DegS/U is well known for the complexity of its regulation. The cytosolic sensory kinase DegS does not receive a single predominant input signal like most two-component kinases, instead it integrates a wide array of metabolic inputs that modulate its activity...

  1. The ability of the biological control agent Bacillus subtilis, strain BB, to colonise vegetable brassicas endophytically following seed inoculation

    NARCIS (Netherlands)

    Wulff, E.G.; Vuurde, van J.W.L.; Hockenhull, J.

    2003-01-01

    The ability of Bacillus subtilis, strain BB, to colonise cabbage seedlings endophytically was examined following seed inoculation. Strain BB was recovered from different plant parts including leaves (cotyledons), stem (hypocotyl) and roots. While high bacterial populations persisted in the roots and

  2. The extraordinary specificity of xanthine phosphoribosyltransferase from Bacillus subtilis elucidated by reaction kinetics, ligand binding, and crystallography

    DEFF Research Database (Denmark)

    Arent, Susan; Kadziola, Anders; Larsen, Sine;

    2006-01-01

    Xanthine phosphoribosyltransferase (XPRTase) from Bacillus subtilis is a representative of the highly xanthine specific XPRTases found in Gram-positive bacteria. These XPRTases constitute a distinct subclass of 6-oxopurine PRTases, which deviate strongly from the major class of H(X)GPRTases with ...

  3. The Bacillus subtilis transition state regulator AbrB binds to the-35 promoter region of comK

    NARCIS (Netherlands)

    Hamoen, LW; Kausche, D; Marahiel, MA; van Sinderen, D; Venema, G; Serror, P

    2003-01-01

    Genetic competence is a differentiation process initiated by Bacillus subtilis as a result of nutritional deprivation, and is controlled by a complex signal transduction cascade. The promoter of comK, encoding the competence transcription factor, is regulated by at least four different transcription

  4. Heterologous Gene Expression in Lactococcus lactis subsp. lactis : Synthesis, Secretion, and Processing of the Bacillus subtilis Neutral Protease

    NARCIS (Netherlands)

    Guchte, Maarten van de; Kodde, Jan; Vossen, Jos M.B.M. van der; Kok, Jan; Venema, Gerard

    1990-01-01

    The Bacillus subtilis nprE gene lacking its own promoter sequence was inserted in the lactococcal expression vector pMG36e. Upon introduction of the recombinant plasmid into Lactococcus lactis subsp. lactis strain MG1363, neutral protease activity could be visualized by the appearance of large clear

  5. Influence of ad Libitum Feeding of Piglets With Bacillus Subtilis Fermented Liquid Feed on Gut Flora, Luminal Contents and Health

    Science.gov (United States)

    He, Yuyong; Mao, Chunxia; Wen, Hong; Chen, Zhiyu; Lai, Tao; Li, Lingyu; Lu, Wei; Wu, Huadong

    2017-01-01

    Some scholars caution that long-term ad libitum feeding with probiotic fermented food poses potential health risks to baby animals. We conducted a feeding experiment to investigate the influence of ad libitum feeding of pre-and post-weaned piglets with a Bacillus subtilis fermented diet on the gut microbiome, gut metabolomic profiles, bile acid metabolism, proinflammatory cytokines and faecal consistency. Compared with piglets fed a Bacillus subtilis-supplemented pellet diet, piglets fed the Bacillus subtilis fermented liquid diet had lower intestinal bacterial diversity (P > 0.05), higher intestinal fungal diversity (P > 0.05), more Firmicutes (P > 0.05), fewer Bacteroidetes, Actinobacteria and Proteobacteria (P > 0.05), higher concentrations of 3-hydroxypropionic acid (P acid (P lactic acid (P acid (P > 0.05) and lithocholic acid (P  0.05). The data show that ad libitum feeding of piglets with a Bacillus subtilis fermented liquid diet during the suckling and early post-weaning periods promotes the growth of lactic acid bacteria, bile salt hydrolase-active bacteria and 7a-dehydroxylase-active bacteria in the intestinal lumen; disturbs the normal production of lactic acid, orotic acid and unconjugated bile acids; and increases circulating interleukin-6 levels and diarrhoea incidence. PMID:28291252

  6. Autoregulation of subtilin biosynthesis in Bacillus subtilis: the role of the spa-box in subtilin-responsive promoters

    NARCIS (Netherlands)

    Kleerebezem, M.; Bongers, R.; Rutten, G.; Vos, de W.M.; Kuipers, O.P.

    2004-01-01

    The production of the type 1 antimicrobial peptide (AMP) subtilin by Bacillus subtilis is regulated in a cell-density-dependent manner [Kleerebezem M, de Vos WM, Kuipers OP. The lantibiotics nisin and subtilin act as extracellular regulators of their own biosynthesis. In: Dunny GM, Winans SC, editor

  7. Characterization of the replication region of the Bacillus subtilis plasmid pLS20 : a novel type of replicon

    NARCIS (Netherlands)

    Meijer, WJJ; De Boer, AJ; van Tongeren, S; Venema, G; Bron, S

    1995-01-01

    A 3.1 kb fragment of the large (~55 kb) Bacillus subtilis plasmid pLS20 containing all the information for autonomous replication was cloned and sequenced. In contrast to the parental plasmid, derived minireplicons were unstably maintained. Using deletion analysis the fragment essential and sufficie

  8. Transcriptome analysis of sorbic acid-stressed Bacillus subtilis reveals a nutrient limitation response and indicates plasma membrane remodeling

    NARCIS (Netherlands)

    A. ter Beek; B.J.F. Keijser; A. Boorsma; A. Zakrzewska; R. Orij; G.J. Smits; S. Brul

    2008-01-01

    The weak organic acid sorbic acid is a commonly used food preservative, as it inhibits the growth of bacteria, yeasts, and molds. We have used genome-wide transcriptional profiling of Bacillus subtilis cells during mild sorbic acid stress to reveal the growth-inhibitory activity of this preservative

  9. Towards the entire proteome of the model bacterium Bacillus subtilis by gel-based and gel-free approaches

    NARCIS (Netherlands)

    Wolff, Susanne; Antelmann, Haike; Albrecht, Dirk; Becher, Doerte; Bernhardt, Joerg; Bron, Sierd; Buettner, Knut; van Dijl, Jan Maarten; Eymann, Christine; Otto, Andreas; Tam, Le Thi; Hecker, Michael

    2007-01-01

    With the emergence of mass spectrometry in protein science and the availability of complete genome sequences, proteomics has gone through a rapid development. The soil bacterium Bacillus subtilis, as one of the first DNA sequenced species, represents a model for Gram-positive bacteria and its proteo

  10. Defining the structure of the general stress regulon of Bacillus subtilis using targeted microarray analysis and random forest classification.

    NARCIS (Netherlands)

    Nannapaneni, P.; Hertwig, F.; Depke, M.; Hecker, M.; Mader, U.; Volker, U.; Steil, L.; Hijum, S.A.F.T. van

    2012-01-01

    The structure of the SigB-dependent general stress regulon of Bacillus subtilis has previously been characterized by proteomics approaches as well as DNA array-based expression studies. However, comparing the SigB targets published in three previous major transcriptional profiling studies it is obvi

  11. The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca

    NARCIS (Netherlands)

    Romero, Diego; de Vicente, Antonio; Rakotoaly, Rivo H.; Dufour, Samuel E.; Veening, Jan-Willem; Arrebola, Eva; Cazorla, Francisco M.; Kuipers, Oscar P.; Paquot, Michel; Perez-Garcia, Alejandro; Stacey, Gary

    2007-01-01

    Podosphaera fusca is the main causal agent of cucurbit powdery mildew in Spain. Four Bacillus subtilis strains, UMAF6614, UMAF6619, UMAF6639, and UMAF8561, with proven ability to suppress the disease on melon in detached leaf and seedling assays, were subjected to further analyses to elucidate the m

  12. Kinetics of Mn(II) oxidation by spores of the marine Bacillus sp. SG-1

    Science.gov (United States)

    Toyoda, Kazuhiro; Tebo, Bradley M.

    2016-09-01

    The kinetics of Mn(II) oxidation by spores of the marine Bacillus sp. SG-1 was measured under controlled conditions of the initial Mn(II) concentration, spore concentration, chemical speciation, pH, O2, and temperature. Mn(II) oxidation experiments were performed with spore concentrations ranging from 0.7 to 11 × 109 spores/L, a pH range from 5.8 to 8.1, temperatures between 4 and 58 °C, a range of dissolved oxygen from 2 to 270 μM, and initial Mn(II) concentrations from 1 to 200 μM. The Mn(II) oxidation rates were directly proportional to the spore concentrations over these ranges of concentration. The Mn(II) oxidation rate increased with increasing initial Mn(II) concentration to a critical concentration, as described by the Michaelis-Menten model (Km = ca. 3 μM). Whereas with starting Mn(II) concentrations above the critical concentration, the rate was almost constant in low ionic solution (I = 0.05, 0.08). At high ionic solution (I = 0.53, 0.68), the rate was inversely correlated with Mn(II) concentration. Increase in the Mn(II) oxidation rate with the dissolved oxygen concentration followed the Michaelis-Menten model (Km = 12-19 μM DO) in both a HEPES-buffered commercial drinking (soft) water and in artificial and natural seawater. Overall, our results suggest that the mass transport limitations of Mn(II) ions due to secondary Mn oxide products accumulating on the spores cause a significant decrease of the oxidation rate at higher initial Mn(II) concentration on a spore basis, as well as in more concentrated ionic solutions. The optimum pH for Mn(II) oxidation was approximately 7.0 in low ionic solutions (I = 0.08). The high rates at the alkaline side (pH > 7.5) may suggest a contribution by heterogeneous reactions on manganese bio-oxides. The effect of temperature on the Mn(II) oxidation rate was studied in three solutions (500 mM NaCl, ASW, NSW solutions). Thermal denaturation occurred at 58 °C and spore germination was evident at 40 °C in all three

  13. Research advances in preventing animal diseases by utilising Bacillus subtilis%枯草芽孢杆菌在防御动物疾病中的研究进展

    Institute of Scientific and Technical Information of China (English)

    叶小兰; 杨倩

    2011-01-01

    This article reviews the research advances in current use of Bacillus subtilis in antagonizing pathogens,immunostimulatory,spore vaccine as well as intestinal breeding to provide references for further studies in preventing animal diseases by utilizing B.subtilis.%结合国内外对枯草芽孢杆菌的研究结果,对枯草芽孢杆菌在拮抗病原菌、免疫刺激、芽孢疫苗及肠道内繁殖等方面的研究进展进行了综述,旨在为枯草芽孢杆菌在防御动物疾病中的深入研究提供参考。

  14. A genetic algorithm-Bayesian network approach for the analysis of metabolomics and spectroscopic data: application to the rapid identification of Bacillus spores and classification of Bacillus species

    OpenAIRE

    Goodacre Royston; Correa Elon

    2011-01-01

    Abstract Background The rapid identification of Bacillus spores and bacterial identification are paramount because of their implications in food poisoning, pathogenesis and their use as potential biowarfare agents. Many automated analytical techniques such as Curie-point pyrolysis mass spectrometry (Py-MS) have been used to identify bacterial spores giving use to large amounts of analytical data. This high number of features makes interpretation of the data extremely difficult We analysed Py-...

  15. Inhibition of the Aspergillus flavus Growth and Aflatoxin B1 Contamination on Pistachio Nut by Fengycin and Surfactin-Producing Bacillus subtilis UTBSP1

    Science.gov (United States)

    Farzaneh, Mohsen; Shi, Zhi-Qi; Ahmadzadeh, Masoud; Hu, Liang-Bin; Ghassempour, Alireza

    2016-01-01

    In this study, the treatment of pistachio nuts by Bacillus subtilis UTBSP1, a promising isolate to degrade aflatoxin B1 (AFB1), caused to reduce the growth of Aspergillus flavus R5 and AFB1 content on pistachio nuts. Fluorescence probes revealed that the cell free supernatant fluid from UTBSP1 affects spore viability considerably. Using high-performance liquid chromatographic (HPLC) method, 10 fractions were separated and collected from methanol extract of cell free supernatant fluid. Two fractions showed inhibition zones against A. flavus. Mass spectrometric analysis of the both antifungal fractions revealed a high similarity between these anti-A. flavus compounds and cyclic-lipopeptides of surfactin, and fengycin families. Coproduction of surfactin and fengycin acted in a synergistic manner and consequently caused a strong antifungal activity against A. flavus R5. There was a positive significant correlation between the reduction of A. flavus growth and the reduction of AFB1 contamination on pistachio nut by UTBSP1. The results indicated that fengycin and surfactin-producing B. subtilis UTBSP1 can potentially reduce A. flavus growth and AFB1 content in pistachio nut. PMID:27298596

  16. Inhibition of the Aspergillus flavus Growth and Aflatoxin B1 Contamination on Pistachio Nut by Fengycin and Surfactin-Producing Bacillus subtilis UTBSP1

    Directory of Open Access Journals (Sweden)

    Mohsen Farzaneh

    2016-06-01

    Full Text Available In this study, the treatment of pistachio nuts by Bacillus subtilis UTBSP1, a promising isolate to degrade aflatoxin B1 (AFB1, caused to reduce the growth of Aspergillus flavus R5 and AFB1 content on pistachio nuts. Fluorescence probes revealed that the cell free supernatant fluid from UTBSP1 affects spore viability considerably. Using high-performance liquid chromatographic (HPLC method, 10 fractions were separated and collected from methanol extract of cell free supernatant fluid. Two fractions showed inhibition zones against A. flavus. Mass spectrometric analysis of the both antifungal fractions revealed a high similarity between these anti-A. flavus compounds and cyclic-lipopeptides of surfactin, and fengycin families. Coproduction of surfactin and fengycin acted in a synergistic manner and consequently caused a strong antifungal activity against A. flavus R5. There was a positive significant correlation between the reduction of A. flavus growth and the reduction of AFB1 contamination on pistachio nut by UTBSP1. The results indicated that fengycin and surfactin-producing B. subtilis UTBSP1 can potentially reduce A. flavus growth and AFB1 content in pistachio nut.

  17. Isolation and molecular characterization of thermostable phytase from Bacillus subtilis (BSPhyARRMK33).

    Science.gov (United States)

    Reddy, Chinreddy Subramanyam; Achary, V Mohan Murali; Manna, Mrinalini; Singh, Jitender; Kaul, Tanushri; Reddy, Malireddy K

    2015-03-01

    The thermostable phytase gene was isolated from Bacillus subtilis ARRMK33 (BsPhyARRMK33). The gene has an ORF of 1152 bp and that encodes a protein of 383 amino acids. Sequence analysis showed high homology with Bacillus sp. phytase proteins, but no similarity was found with other phytases. SDS-PAGE analysis exhibited a predicted molecular mass of 42 kDa. Homology modeling of BsPhyARRMK33 protein based on Bacillus amyloliquefaciens crystal structure disclosed its β-propeller structure. BsPhyARRMK33 recombinant plasmid in pET-28a(+) was expressed in Rosetta gami B DE3 cells and the maximum phytase activity 15.3 U mg(-1) obtained. The enzyme exhibits high thermostability at various temperatures and broad pH ranges. The recombinant protein retained 74% of its original activity after incubation at 95 °C for 10 min. In the presence of Ca(2+), the recombinant phytase activity was maximal where as it was inhibited by EDTA. The optimal pH and temperature for the recombinant phytase activity is achieved at 7.0 and 55 °C, respectively. Thermostable nature and wide range of pH are promising features of recombinant BsPhyARRMK33 protein that may be employed as an efficient alternative to commercially known phytases and thereby alleviate environmental eutrophication.

  18. Online monitoring of Escherichia coli and Bacillus thuringiensis spore inactivation after advanced oxidation treatment.

    Science.gov (United States)

    Sherchan, Samendra P; Snyder, Shane A; Gerba, Charles P; Pepper, Ian L

    2014-01-01

    Various studies have shown that advanced oxidation processes (AOPs) such as UV light in combination with hydrogen peroxide is an efficient process for the removal of a large variety of emerging contaminants including microorganisms. The mechanism of destruction in the presence of hydrogen peroxide (H2O2) is the enhanced formation of hydroxyl (·OH) radicals, which have a high oxidation potential. The goal of this study was to utilize in-line advanced oxidation to inactivate microbes, and document the inactivation via an in-line, real-time sensor. Escherichia coli cells and Bacillus thuringiensis spores were exposed to UV/H2O2 treatment in DI water, and the online sensor BioSentry(®) was evaluated for its potential to monitor inactivation in real-time. B. thuringiensis was selected as a non-pathogenic surrogate for B. anthracis, the causative agent of anthrax and a proven biological weapon. UV radiation and UV/H2O2 exposure resulted in a >6 log10 reduction of the viable culturable counts of E. coli vegetative cells, and a 3 log10 reduction of B. thuringiensis spores. Scanning electron microscopy of the treated samples revealed severe damage on the surface of most E. coli cells, yet there was no significant change observed in the morphology of the B. thuringiensis spores. Following AOP exposure, the BioSentry sensor showed an increase in the categories of unknown, rod and spores counts, but overall, did not correspond well with viable count assays. Data from this study show that advanced oxidation processes effectively inactivate E. coli vegetative cells, but not B. thuringiensis spores, which were more resistant to AOP. Further, the BioSentry in-line sensor was not successful in documenting destruction of the microbial cells in real-time.

  19. Rapid Detection of Viable Bacillus anthracis Spores in Environmental Samples by Using Engineered Reporter Phages

    Science.gov (United States)

    Sharp, Natasha J.; Molineux, Ian J.; Page, Martin A.

    2016-01-01

    Bacillus anthracis, the causative agent of anthrax, was utilized as a bioterrorism agent in 2001 when spores were distributed via the U.S. postal system. In responding to this event, the Federal Bureau of Investigation used traditional bacterial culture viability assays to ascertain the extent of contamination of the postal facilities within 24 to 48 h of environmental sample acquisition. Here, we describe a low-complexity, second-generation reporter phage assay for the rapid detection of viable B. anthracis spores in environmental samples. The assay uses an engineered B. anthracis reporter phage (Wβ::luxAB-2) which transduces bioluminescence to infected cells. To facilitate low-level environmental detection and maximize the signal response, expression of luxAB in an earlier version of the reporter phage (Wβ::luxAB-1) was optimized. These alterations prolonged signal kinetics, increased light output, and improved assay sensitivity. Using Wβ::luxAB-2, detection of B. anthracis spores was 1 CFU in 8 h from pure cultures and as low as 10 CFU/g in sterile soil but increased to 105 CFU/g in unprocessed soil due to an unstable signal and the presence of competing bacteria. Inclusion of semiselective medium, mediated by a phage-expressed antibiotic resistance gene, maintained signal stability and enabled the detection of 104 CFU/g in 6 h. The assay does not require spore extraction and relies on the phage infecting germinating cells directly in the soil sample. This reporter phage displays promise for the rapid detection of low levels of spores on clean surfaces and also in grossly contaminated environmental samples from complex matrices such as soils. PMID:26873316

  20. Rapid Detection of Viable Bacillus anthracis Spores in Environmental Samples by Using Engineered Reporter Phages.

    Science.gov (United States)

    Sharp, Natasha J; Molineux, Ian J; Page, Martin A; Schofield, David A

    2016-04-01

    Bacillus anthracis, the causative agent of anthrax, was utilized as a bioterrorism agent in 2001 when spores were distributed via the U.S. postal system. In responding to this event, the Federal Bureau of Investigation used traditional bacterial culture viability assays to ascertain the extent of contamination of the postal facilities within 24 to 48 h of environmental sample acquisition. Here, we describe a low-complexity, second-generation reporter phage assay for the rapid detection of viableB. anthracis spores in environmental samples. The assay uses an engineered B. anthracis reporter phage (Wβ::luxAB-2) which transduces bioluminescence to infected cells. To facilitate low-level environmental detection and maximize the signal response, expression of luxABin an earlier version of the reporter phage (Wβ::luxAB-1) was optimized. These alterations prolonged signal kinetics, increased light output, and improved assay sensitivity. Using Wβ::luxAB-2, detection of B. anthracis spores was 1 CFU in 8 h from pure cultures and as low as 10 CFU/g in sterile soil but increased to 10(5)CFU/g in unprocessed soil due to an unstable signal and the presence of competing bacteria. Inclusion of semiselective medium, mediated by a phage-expressed antibiotic resistance gene, maintained signal stability and enabled the detection of 10(4)CFU/g in 6 h. The assay does not require spore extraction and relies on the phage infecting germinating cells directly in the soil sample. This reporter phage displays promise for the rapid detection of low levels of spores on clean surfaces and also in grossly contaminated environmental samples from complex matrices such as soils.