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Sample records for bacillus licheniformis

  1. Isolation and characterization of cellulolytic Bacillus licheniformis ...

    African Journals Online (AJOL)

    Eight cellulose degrading bacteria were isolated from compost and were identified as Bacillus licheniformis by 16S rRNA sequencing. Among the eight isolates, Bacillus licheniformis B4, B7 and B8 showed the highest cellulase activity. B. licheniformis B4 and B8 showed the maximum cellulase activity during the stationary ...

  2. Heavy metals and their radionuclides uptake by Bacillus Licheniformis

    International Nuclear Information System (INIS)

    Ramadan, A.A.; Ahmed, M.M.; Abo-state, M.A.M.; Sarhan, M.; Faroqe, M.

    2007-01-01

    Bacillus licheniformis is a gram positive spore forming bacterium. Different concentrations of cobalt affected the ability of Co uptake and growth of Bacillus licheniformis. As the concentration increased, both the uptake and growth were decreased. Maximum Co uptake was found at ph 7.0, while for growth was ph 8.0. The optimum temperature for uptake and growth was 40 degree C and 20% inoculum size represents the maximum cobalt uptake by Bacillus licheniformis. Also, maximum uptake was recorded after 72 hours, incubation period. As the concentration of cesium was increased till 400 mg/l, the uptake was also increased. The optimum cesium uptake and growth was at ph 8.0. The optimum growth was at 45 degree C while Cs uptake was found at 35 degree C and 15% inoculum size represented the maximum Cs uptake. After 72 hour incubation period, maximum Cs uptake was recorded. Generally, Bacillus licheniformis removed more than 80% of Co and 50% of Cs from the broth medium. Addition of clay to Bacillus licheniformis increased both Co or Cs uptake. Bacillus licheniformis was gamma resistant and 10 KGy reduced the viability by 5.3 log cycles. The irradiated and non-irradiated cultures can grow on 500 or 700 mg Co or Cs. Bacillus licheniformis removed 99.32% of the Co radionuclides and 99.28% of Cs radionuclides

  3. Cell Physiology and Protein Secretion of Bacillus licheniformis Compared to Bacillus subtilis

    NARCIS (Netherlands)

    Voigt, Birgit; Antelmann, Haike; Albrecht, Dirk; Ehrenreich, Armin; Maurer, Karl-Heinz; Evers, Stefan; Gottschalk, Gerhard; van Dijl, Jan Maarten; Schweder, Thomas; Hecker, Michael

    2009-01-01

    The genome sequence of Bacillus subtilis was published in 1997 and since then many other bacterial genomes have been sequenced, among them Bacillus licheniformis in 2004. B. subtilis and B. licheniformis are closely related and feature similar saprophytic lifestyles in the soil. Both species can

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

    Science.gov (United States)

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

    2015-01-01

    The aim of the present study was to investigate the characteristic diversity and stability of antimicrobial compounds produced by two probiotic strains of Bacillus licheniformis (MCC2514 and MCC2512). Antimicrobial compounds from the two strains notably varied, related to stability and potency. The inhibitory spectrum of B. licheniformis MCC2512 was higher than MCC2514, but, related to the effect on Micrococcus luteus ATCC9341, MCC2514 (LD50 = 450 AU ml(-1)) was more potent than MCC2512 (LD50 = 750 AU ml(-1)). The compounds were thermo-resistant and stable at a wide range of pH and exhibited considerable resistance to digestive enzymes and bile salts (anionic biological detergents), contributing to their appropriate application in various food systems. The isolate B. licheniformis MCC2512 gave a positive response to Bacillus subtilis-based biosensors BSF2470 and BS168.BS2, confirming the mode of action on the cell wall and subtilin-type, respectively. For B. licheniformis MCC2514, the mode of action was characterized by constructing B. subtilis reporters that interfered in five major biosynthetic pathways, i.e., biosynthesis of DNA, RNA, protein, the cell wall and fatty acids. B. licheniformis MCC2514 responded to the yvgS reporter, indicating it as an RNA synthesis inhibitor. Overall, the investigation reveals variability of the antimicrobial compounds from B. licheniformis of different origins and for their possible application as biopreservative agents. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  5. EXPERIMENTAL-INFECTION IN MICE WITH BACILLUS-LICHENIFORMIS

    DEFF Research Database (Denmark)

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

    1995-01-01

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

  6. Accumulation of some metal ions on Bacillus licheniformis

    International Nuclear Information System (INIS)

    Hafez, M.B.; El-Desouky, W.; Fouad, A.

    2001-01-01

    Pure species of Bacillus licheniformis was used to remove ions from aqueous and simulated waste solutions. Metal ion accumulation on B. licheniformis was fast. Maximum uptake occurred at pH 4± 0.5 and at 25 ± 3 deg C. One gram of dry B. licheniformis was found to accumulate 115 mg cerium, 34 mg copper and 11 mg cobalt from aqueous solutions. The presence of certain foreign ions such as calcium, sodium and potassium decreased the uptake of ions by B. licheniformis, while citrate and EDTA prevent the uptake. Electron microscopic investigations showed that cerium (III), copper (II) and cobalt (II) accumulated extracellulary around the surface wall of B. licheniformis cells. A bio-adsorption mechanism between the metal ions and B. licheniformis cell wall was proposed. (author)

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

    Science.gov (United States)

    Gomaa, Eman Zakaria

    2012-02-01

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

  8. Aroma characteristics of Moutai-flavour liquor produced with Bacillus licheniformis by solid-state fermentation.

    Science.gov (United States)

    Zhang, R; Wu, Q; Xu, Y

    2013-07-01

    The potential of Bacillus licheniformis as a starter culture for aroma concentration improvement in the fermentation of Chinese Moutai-flavour liquor was elucidated. The volatile compounds produced by B. licheniformis were identified by GC-MS, in which C4 compounds, pyrazines, volatile acids, aromatic and phenolic compounds were the main ingredients. The strains B. licheniformis (MT-6 and MT-15) produced more volatile compound concentrations, mainly C4 compounds, than the type strain of B. licheniformis (ATCC 14580) at the fermentation temperature of 55°C. Meanwhile, more volatile compound concentrations were produced by B. licheniformis in solid-state fermentation than in submerged state fermentation. Thus, the strains MT-6 and MT-15 were used as the Bacillus starter culture for investigating Moutai-flavour liquor production. The distilled liquor inoculated with Bacillus starter culture was significantly different from the liquor without inoculum. This was particularly evident in the fore-run part of the distilled sample which was inoculated with Bacillus starter culture, where volatile compounds greatly increased compared to the control. Furthermore, the distilled liquor with Bacillus starter culture showed improved results in sensory appraisals. These results indicated that B. licheniformis was one of the main species influencing the aroma characteristics of Moutai-flavour liquor. This is the first report of an investigation into the effect of Bacillus starter cultures on the flavour features of Moutai-flavour liquor, which verified that Bacillus licheniformis can enhance aroma concentration in Moutai-flavour liquor. Bacillus starter culture brought C4 compounds, pyrazines, volatile acids, aromatic and phenolic compounds to the liquor, which gave a better result in sensory appraisals. © 2013 The Society for Applied Microbiology.

  9. 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.

  10. Complete genome sequence of the industrial bacterium Bacillus licheniformis and comparisons with closely related Bacillus species

    Science.gov (United States)

    Rey, Michael W; Ramaiya, Preethi; Nelson, Beth A; Brody-Karpin, Shari D; Zaretsky, Elizabeth J; Tang, Maria; de Leon, Alfredo Lopez; Xiang, Henry; Gusti, Veronica; Clausen, Ib Groth; Olsen, Peter B; Rasmussen, Michael D; Andersen, Jens T; Jørgensen, Per L; Larsen, Thomas S; Sorokin, Alexei; Bolotin, Alexander; Lapidus, Alla; Galleron, Nathalie; Ehrlich, S Dusko; Berka, Randy M

    2004-01-01

    Background Bacillus licheniformis is a Gram-positive, spore-forming soil bacterium that is used in the biotechnology industry to manufacture enzymes, antibiotics, biochemicals and consumer products. This species is closely related to the well studied model organism Bacillus subtilis, and produces an assortment of extracellular enzymes that may contribute to nutrient cycling in nature. Results We determined the complete nucleotide sequence of the B. licheniformis ATCC 14580 genome which comprises a circular chromosome of 4,222,336 base-pairs (bp) containing 4,208 predicted protein-coding genes with an average size of 873 bp, seven rRNA operons, and 72 tRNA genes. The B. licheniformis chromosome contains large regions that are colinear with the genomes of B. subtilis and Bacillus halodurans, and approximately 80% of the predicted B. licheniformis coding sequences have B. subtilis orthologs. Conclusions Despite the unmistakable organizational similarities between the B. licheniformis and B. subtilis genomes, there are notable differences in the numbers and locations of prophages, transposable elements and a number of extracellular enzymes and secondary metabolic pathway operons that distinguish these species. Differences include a region of more than 80 kilobases (kb) that comprises a cluster of polyketide synthase genes and a second operon of 38 kb encoding plipastatin synthase enzymes that are absent in the B. licheniformis genome. The availability of a completed genome sequence for B. licheniformis should facilitate the design and construction of improved industrial strains and allow for comparative genomics and evolutionary studies within this group of Bacillaceae. PMID:15461803

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

    Digital Repository Service at National Institute of Oceanography (India)

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

    investigation. Analysis of the nucleotide sequence of the 16S rDNA gene of Bacillus sp. SAB1 showed a strong similarity (100%) with the 16S rDNA gene of Bacillus licheniformis HNL09. The bioactive compounds produced by B. licheniformis SAB1 (GenBank accession...

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

    Directory of Open Access Journals (Sweden)

    VINTILĂ T.

    2007-01-01

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

  13. Thermostable Bacteriocin BL8 from Bacillus licheniformis isolated from marine sediment.

    Science.gov (United States)

    Smitha, S; Bhat, S G

    2013-03-01

    To isolate and characterize bacteriocin, BL8, from the bacteria identified as Bacillus licheniformis from marine environment. One-hundred and twelve bacterial isolates from sediment and water samples collected off the coast of Cochin, India, were screened for antibacterial activity. Strain BTHT8, identified as Bacillus licheniformis, inhibited the growth of Gram-positive test organisms. The active component labelled as bacteriocin BL8 was partially purified by ammonium sulphate fractionation and was subjected to glycine SDS-PAGE. The band exhibiting antimicrobial activity was electroeluted and analysed using MALDI-TOF mass spectrometry, and the molecular mass was determined as 1.4 kDa. N-terminal amino acid sequencing of BL8 gave a 13 amino acid sequence stretch. Bacteriocin BL8 was stable even after boiling at 100 °C for 30 min and over a wide pH range of 1-12. A novel, pH-tolerant and thermostable bacteriocin BL8, active against the tested Gram-positive bacteria, was isolated from Bacillus licheniformis. This study reports a stable, low molecular weight bacteriocin from Bacillus licheniformis. This bacteriocin can be used to address two important applications: as a therapeutic agent and as a biopreservative in food processing industry. © 2012 The Society for Applied Microbiology.

  14. A Phosphate Starvation-Inducible Ribonuclease of Bacillus licheniformis.

    Science.gov (United States)

    Nguyen, Thanh Trung; Nguyen, Minh Hung; Nguyen, Huy Thuan; Nguyen, Hoang Anh; Le, Thi Hoi; Schweder, Thomas; Jürgen, Britta

    2016-08-28

    The BLi03719 protein of Bacillus licheniformis DSM13 belongs to the most abundant extracellular proteins under phosphate starvation conditions. In this study, the function of this phosphate starvation inducible protein was determined. An amino-acid sequence analysis of the BLi03719-encoding gene showed a high similarity with genes encoding the barnase of Bacillus amyloliquefaciens FZB42 and binase-like RNase of Bacillus pumilus SARF-032. The comparison of the control strain and a BLi03719-deficient strain revealed a strongly reduced extracellular ribonuclease activity of the mutant. Furthermore, this knockout mutant exhibited delayed growth with yeast RNA as an alternative phosphate and carbon source. These results suggest that BLi03719 is an extracellular ribonuclease expressed in B. licheniformis under phosphate starvation conditions. Finally, a BLi03719 mutant showed an advantageous effect on the overexpression of the heterologous amyE gene under phosphate-limited growth conditions.

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

    DEFF Research Database (Denmark)

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

    1995-01-01

    A retrospective study of bovine abortions associated with Bacillus licheniformis is described. The material consisted of 2445 bovine abortions submitted for diagnostics from 1986 through 1993. Initially, B, licheniformis had been isolated from 81 cases. Sections of these cases were reexamined...... isolations, especially from the placenta, lungs, and abomasal contents, combined with the histological findings points to B, licheniformis abortions as being of haematogenous origin with subsequent transplacental spread to the fetus....

  16. Genetic evidence for a novel competence inhibitor in the industrially important Bacillus licheniformis.

    Science.gov (United States)

    Muth, Christine; Buchholz, Meike; Schmidt, Christina; Volland, Sonja; Meinhardt, Friedhelm

    2017-12-01

    Natural genetic competence renders bacteria able to take up and, in case there is sufficient homology to the recipient's chromosome, integrate exogenously supplied DNA. Well studied in Bacillus subtilis, genetic competence is-in several aspects-known to be differently regulated in Bacillus licheniformis. We now report on the identification of a novel, chromosomally encoded homolog of a competence inhibitor in B. licheniformis (ComI) that has hitherto only been described as a plasmid borne trait in the ancestral B. subtilis NCIB3610. Bioinformatical analysis that included 80 Bacillus strains covering 20 different species revealed a ComI encoding gene in all of the examined B. licheniformis representatives, and was identified in few among the other species investigated. The predicted ComI of B. licheniformis is a highly conserved peptide consisting of 28 amino acids. Since deletion of comI in B. licheniformis DSM13 resulted in twofold increased transformation efficiency by genetic competence and overexpression resulted in threefold decreased transformability, the function as a competence inhibitor became evident.

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

    Directory of Open Access Journals (Sweden)

    T. VINTILĂ

    2007-05-01

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

  18. TRANSDUCTION OF BACILLUS LICHENIFORMIS AND BACILLUS SUBTILIS BY EACH OF TWO PHAGES1

    Science.gov (United States)

    Taylor, Martha J.; Thorne, Curtis B.

    1963-01-01

    Taylor, Martha J. (U.S. Army Biological Laboratories, Fort Detrick, Frederick, Md.) and Curtis B. Thorne. Transduction of Bacillus licheniformis and Bacillus subtilis by each of two phages. J. Bacteriol. 86:452–461. 1963.—A second transducing bacteriophage, designated SP-15, was isolated from the same soil-sample culture filtrate that supplied the Bacillus subtilis transducing phage, SP-10, reported earlier from this laboratory. SP-10 and SP-15 differ serologically and in several other respects, but share the ability to propagate on B. subtilis W-23-Sr (streptomycin-resistant) and B. licheniformis ATCC 9945a, and to mediate general transduction in either species when propagated homologously. Attempts to transduce between the species have failed. SP-10 forms plaques readily on both W-23-Sr and 9945a; SP-15 forms minute plaques on W-23-Sr and has shown no evidence of any lytic activity on 9945a. Maximal recoveries of prototrophic colonies from mixtures of SP-10 with auxotrophs of either W-23-Sr or 9945a were obtained only when excess phage was neutralized by post-transduction treatment with specific phage antiserum. Such treatment was not necessary for maximal recovery of transductants effected by SP-15. Unlike SP-10, SP-15 propagated on W-23-Sr did not transduce B. subtilis 168 (indole−). SP-15 transduced B. licheniformis more efficiently than did SP-10. Neither phage was able to transduce B. licheniformis as efficiently as it transduced B. subtilis. The differing influences of multiplicity of infection were compared for the two phages in both species. PMID:14066421

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

    Directory of Open Access Journals (Sweden)

    Shi Gui-Yang

    2009-10-01

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

  20. Central carbon metabolism influences cellulase production in Bacillus licheniformis.

    Science.gov (United States)

    Wang, J; Liu, S; Li, Y; Wang, H; Xiao, S; Li, C; Liu, B

    2018-01-01

    Bacillus licheniformis that can produce cellulase including endo glucanase and glucosidase is an important industrial microbe for cellulose degradation. The purpose of this research was to assess the effect of endo glucanase gene bglC and glucosidase gene bglH on the central metabolic flux in B. licheniformis. bglC and bglH were knocked out using homologous recombination method, respectively, and the corresponding knockout strains were obtained for 13 C metabolic flux analysis. A significant change was observed in metabolic fluxes after 13 C metabolic flux ratio analysis. In both of the knockout strains, the increased fluxes of the pentose phosphate pathway and malic enzyme reaction enabled an elevated supply of NADPH which provided enough reducing power for the in vivo synthesis reactions. The fluxes through tricarboxylic acid cycle and anaplerotic reactions increased fast in the two knockout strains, which meant more energy generated. The changed fluxes in central carbon metabolism provided a holistic view of the physiological status in B. licheniformis and possible targets for further strain engineering. Cellulase is very important in the field of agriculture and bioenergy because of its degrading effect on cellulosic biomass. This study presented the effect of central carbon metabolism on cellulase production in Bacillus licheniformis. The study also provided a holistic view of the physiological status in B. licheniformis. The shifted metabolism provided a quantitative evaluation of the biosynthesis of cellulase and a priority ranked target list for further strain engineering. © 2017 The Society for Applied Microbiology.

  1. Exploring the Metabolomic Responses of Bacillus licheniformis to Temperature Stress by Gas Chromatography/Mass Spectrometry.

    Science.gov (United States)

    Dong, Zixing; Chen, Xiaoling; Cai, Ke; Chen, Zhixin; Wang, Hongbin; Jin, Peng; Liu, Xiaoguang; Permaul, Kugenthiren; Singh, Suren; Wang, Zhengxiang

    2018-03-28

    Owing to its high protein secretion capacity, simple nutritional requirements, and GRAS (generally regarded as safe) status, Bacillus licheniformis is widely used as a host for the industrial production of enzymes, antibiotics, and peptides. However, as compared with its close relative Bacillus subtilis , little is known about the physiology and stress responses of B. licheniformis . To explore its temperature-stress metabolome, B. licheniformis strains ATCC 14580 and B186, with respective optimal growth temperatures of 42°C and 50°C, were cultured at 42°C, 50°C, and 60°C and their corresponding metabolic profiles were determined by gas chromatography/mass spectrometry and multivariate statistical analyses. It was found that with increased growth temperatures, the two B. licheniformis strains displayed elevated cellular levels of proline, glutamate, lysine, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, and octadecanoic acid, and decreased levels of glutamine and octadecenoic acid. Regulation of amino acid and fatty acid metabolism is likely to be associated with the evolution of protective biochemical mechanisms of B. licheniformis . Our results will help to optimize the industrial use of B. licheniformis and other important Bacillus species.

  2. Expression of alpha-amylase in Bacillus licheniformis.

    OpenAIRE

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

    1986-01-01

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

  3. Effect of Ultrasonic Waves on the Heat Resistance of Bacillus cereus and Bacillus licheniformis Spores

    Science.gov (United States)

    Burgos, J.; Ordóñez, J. A.; Sala, F.

    1972-01-01

    Heat resistance of Bacillus cereus and Bacillus licheniformis spores in quarter-strength Ringer solution decreases markedly after ultrasonic treatments which are unable to kill a significant proportion of the spore population. This effect does not seem to be caused by a loss of Ca2+ or dipicolinic acid. The use of ultrasonics to eliminate vegetative cells or to break aggregates in Bacillus spore suspensions to be used subsequently in heat resistance experiments appears to be unadvisable. PMID:4627969

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

    Directory of Open Access Journals (Sweden)

    Khan Sara

    2016-01-01

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

  5. Lichenysin is produced by most Bacillus licheniformis strains.

    Science.gov (United States)

    Madslien, E H; Rønning, H T; Lindbäck, T; Hassel, B; Andersson, M A; Granum, P E

    2013-10-01

    The aim of this study was to elucidate the prevalence of lichenysin production in Bacillus licheniformis and to see whether this feature was restricted to certain genotypes. Secondly, we wanted to see whether cytotoxicity reflected the measured levels of lichenysin. Fifty-three genotyped strains of B. licheniformis, representing a wide variety of sources, were included. lchAA gene fragments were detected in all strains by polymerase chain reaction (PCR). All 53 strains produced lichenysins with four molecular masses as confirmed by LC-MS/MS (liquid chromatography-tandem mass spectrometry) analysis. The amounts of lichenysin varied more than two orders of magnitude between strains and were irrespective of genotype. Finally, there was a strong association between lichenysin concentrations and toxicity towards boar spermatozoa, erythrocytes and Vero cells. Lichenysin synthesis was universal among the 53 B. licheniformis strains examined. The quantities varied considerably between strains, but were not specifically associated with genotype. Cytotoxicity was evident at lichenysin concentrations above 10 μg ml(-1) , which is in accordance with previous studies. This study might be of interest to those working on B. licheniformis for commercial use as well as for authorities who make risk assessments of B. licheniformis when used as a food and feed additive. © 2013 The Society for Applied Microbiology.

  6. Bacillus licheniformis in geogenic dust induces inflammation in respiratory epithelium.

    Science.gov (United States)

    Pickering, Janessa; Teo, Teck Hui; Thornton, Ruth B; Kirkham, Lea-Ann; Zosky, Graeme R; Clifford, Holly D

    2018-07-01

    Exposure to environmental geogenic (or earth-derived) dust can lead to more frequent and severe infections in the human airway. Particulate matter respiratory diseases. We have previously demonstrated that mice exposed to geogenic dust PM 10 experienced an exacerbation of inflammatory responses to influenza A virus. Whether geogenic dust PM 10 also exacerbates respiratory bacterial infection is not yet known, nor are the components of the dust that drive these responses. We treated airway bronchial epithelial cells (NuLi-1) with UV-irradiated geogenic dust PM 10 from six remote Western Australian towns. High levels of IL-6 and IL-8 production were observed, as well as persistent microbial growth. 16 S rRNA sequencing of the growth identified the microbe as Bacillus licheniformis, a spore-forming, environmentally abundant bacterium. We next investigated the interaction of B. licheniformis with respiratory epithelium in vitro to determine whether this exacerbated infection with a bacterial respiratory pathogen (non-typeable Haemophilus influenzae, NTHi). Heat treatment (100 °C) of all PM 10 samples eliminated B. licheniformis contamination and reduced epithelial inflammatory responses, suggesting that heat-labile and/or microbial factors were involved in the host response to geogenic dust PM 10 . We then exposed NuLi-1 epithelium to increasing doses of the isolated Bacillus licheniformis (multiplicity of infection of 10:1, 1:1 or 0.1:1 bacteria: cells) for 1, 3, and 24 h. B. licheniformis and NTHi infection (association and invasion) was assessed using a standard gentamicin survival assay, and epithelial release of IL-6 and IL-8 was measured using a bead based immunoassay. B. licheniformis was cytotoxic to NuLi-1 cells at 24 h. At 3 h post-challenge, B. licheniformis elicited high IL-6 and IL-8 inflammatory responses from NuLi-1 cells compared with cells treated with heat-treated geogenic dust PM 10 (p respiratory epithelium. The impact on respiratory

  7. Determining the source of Bacillus cereus and Bacillus licheniformis isolated from raw milk, pasteurized milk and yoghurt.

    Science.gov (United States)

    Banykó, J; Vyletelová, M

    2009-03-01

    Strain-specific detection of Bacillus cereus and Bacillus licheniformis in raw and pasteurized milk, and yoghurt during processing. Randomly selected isolates of Bacillus spp. were subjected to PCR analysis, where single primer targeting to the repetitive sequence Box elements was used to fingerprint the species. The isolates were separated into six different fingerprint patterns. The results show that isolates clustered together at about the 57% similarity level with two main groups at the 82% and 83% similarity levels, respectively. Contamination with identical strains both of B. cereus and B. licheniformis in raw and pasteurized milk was found as well as contaminated with different strains (in the case of raw milk and yoghurt/pasteurized milk and yoghurt). Several BOX types traced in processed milk samples were not discovered in the original raw milk. BOX-PCR fingerprinting is useful for characterizing Bacillus populations in a dairy environment. It can be used to confirm environmental contamination, eventually clonal transfer of Bacillus strains during the technological processing of milk. Despite the limited number of strains analysed, the two Bacillus species yielded adequately detectable banding profiles, permitting differentiation of bacteria at the strain level and showing their diversity throughout dairy processing.

  8. Amobilisasi Sel Bacillus licheniformis KA-08 dalam Menghasilkan Keratinase Termostabil

    Directory of Open Access Journals (Sweden)

    Anthoni Agustien

    2012-10-01

    Full Text Available Isolate local Bacillus licheniformis KA-08 known extracellular thermostable keratinase producers. Scale up of thermostable keratinase production can be with cells immobilized. The objective of the research is to thermostable keratinase production of B. licheniformis KA-08 cells immobilization. Thermostable keratinase activities were determined with modification of Brandelli and Riffel method. Protein concentration of enzyme determined with Lowry method. Immobilization of cells by Ca-alginate matrix with Adinarayana method, alginate concentration and amount of alginate bead effects with Beshay method. The result extracellular thermostable keratinase of B. licheniformis KA-08 cells immobilized was maximum produced at 12 times incubation with activity as 9.25 U/mg. Three percent alginate has optimum activity. Three hundred alginate beads has optimum activity. Cells immobilized ofB. licheniformis KA-08 has scale up of thermostable keratinase activity at 2 times than free cells. Thermostable keratinase produced by cell immobilized was nine cycles.

  9. Identification of strong promoters based on the transcriptome of Bacillus licheniformis.

    Science.gov (United States)

    Liu, Xin; Yang, Haiyan; Zheng, Junwei; Ye, Yanrui; Pan, Li

    2017-06-01

    To expand the repertoire of strong promoters for high level expression of proteins based on the transcriptome of Bacillus licheniformis. The transcriptome of B. licheniformis ATCC14580 grown to the early stationary phase was analyzed and the top 10 highly expressed genes/operons out of the 3959 genes and 1249 operons identified were chosen for study promoter activity. Using beta-galactosidase gene as a reporter, the candidate promoter pBL9 exhibited the strongest activity which was comparable to that of the widely used strong promoter p43. Furthermore, the pro-transglutaminase from Streptomyces mobaraensis (pro-MTG) was expressed under the control of promoter pBL9 and the activity of pro-MTG reached 82 U/ml after 36 h, which is 23% higher than that of promoter p43 (66.8 U/ml). In our analyses of the transcriptome of B. licheniformis, we have identified a strong promoter pBL9, which could be adapted for high level expression of proteins in the host Bacillus subtilis.

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

    DEFF Research Database (Denmark)

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

    1997-01-01

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

  11. Inhibition of Bacillus licheniformis LMG 19409 from ropy cider by enterocin AS-48.

    Science.gov (United States)

    Grande, M J; Lucas, R; Abriouel, H; Valdivia, E; Ben Omar, N; Maqueda, M; Martínez-Cañamero, M; Gálvez, A

    2006-08-01

    To determine the activity of enterocin AS-48 against ropy-forming Bacillus licheniformis from cider. Enterocin AS-48 was tested on B. licheniformis LMG 19409 from ropy cider in MRS-G broth, fresh-made apple juice and in two commercial apple ciders (A and B). Bacillus licheniformis was rapidly inactivated in MRS-G by 0.5 microg ml(-1)AS-48 and in fresh-made apple juice by 3 microg ml(-1). Concentration-dependent inactivation of this bacterium in two commercial apple ciders (A and B) stored at 4, 15 and 30 degrees C for 15 days was also demonstrated. Counts from heat-activated endospores in cider A plus AS-48 decreased very slowly. Application of combined treatments of heat (95 degrees C) and enterocin AS-48 reduced the time required to achieved complete inactivation of intact spores in cider A to 4 min for 6 microg ml(-1) and to 1 min for 12 microg ml(-1). D and z values also decreased as the bacteriocin concentration increased. Enterocin AS-48 can inhibit ropy-forming B. licheniformis in apple cider and increase the heat sensitivity of spores. Results from this study support the potential use of enterocin AS-48 to control B. licheniformis in apple cider.

  12. Role of the gerA operon in L-alanine germination of Bacillus licheniformis spores

    Directory of Open Access Journals (Sweden)

    Løvdal Irene S

    2012-03-01

    Full Text Available Abstract Background The genome of Bacillus licheniformis DSM 13 harbours three neighbouring open reading frames showing protein sequence similarities to the proteins encoded from the Bacillus subtilis subsp. subtilis 168 gerA operon, GerAA, GerAB and GerAC. In B. subtilis, these proteins are assumed to form a germinant receptor involved in spore germination induced by the amino acid L-alanine. Results In this study we show that disruption of the gerAA gene in B. licheniformis MW3 hamper L-alanine and casein hydrolysate-triggered spore germination, measured by absorbance at 600 nm and confirmed by phase contrast microscopy. This ability was restored by complementation with a plasmid-borne copy of the gerA locus. Addition of D-alanine in the casein hydrolysate germination assay abolished germination of both B. licheniformis MW3 and the complementation mutant. Germination of both B. licheniformis MW3 and the gerA disruption mutant was induced by the non-nutrient germinant Ca2+-Dipicolinic acid. Conclusions These results demonstrate that the B. licheniformis MW3 gerA locus is involved in germination induced by L-alanine and potentially other components present in casein hydrolysate.

  13. Role of the gerA operon in L-alanine germination of Bacillus licheniformis spores

    Science.gov (United States)

    2012-01-01

    Background The genome of Bacillus licheniformis DSM 13 harbours three neighbouring open reading frames showing protein sequence similarities to the proteins encoded from the Bacillus subtilis subsp. subtilis 168 gerA operon, GerAA, GerAB and GerAC. In B. subtilis, these proteins are assumed to form a germinant receptor involved in spore germination induced by the amino acid L-alanine. Results In this study we show that disruption of the gerAA gene in B. licheniformis MW3 hamper L-alanine and casein hydrolysate-triggered spore germination, measured by absorbance at 600 nm and confirmed by phase contrast microscopy. This ability was restored by complementation with a plasmid-borne copy of the gerA locus. Addition of D-alanine in the casein hydrolysate germination assay abolished germination of both B. licheniformis MW3 and the complementation mutant. Germination of both B. licheniformis MW3 and the gerA disruption mutant was induced by the non-nutrient germinant Ca2+-Dipicolinic acid. Conclusions These results demonstrate that the B. licheniformis MW3 gerA locus is involved in germination induced by L-alanine and potentially other components present in casein hydrolysate. PMID:22420404

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

    NARCIS (Netherlands)

    Creusot, N.P.

    2006-01-01

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

  15. Enzyme activities and antibiotic susceptibility of colonial variants of Bacillus subtilis and Bacillus licheniformis.

    OpenAIRE

    Carlisle, G E; Falkinham, J O

    1989-01-01

    A nonmucoid colonial variant of a mucoid Bacillus subtilis strain produced less amylase activity and a transparent colonial variant of a B. licheniformis strain produced less protease activity compared with their parents. Antibiotic susceptibility patterns of the colonial variants differed, and increased resistance to beta-lactam antibiotics was correlated with increased production of extracellular beta-lactamase.

  16. Nitrogen removal and water microbiota in grass carp culture following supplementation with Bacillus licheniformis BSK-4.

    Science.gov (United States)

    Liang, Quan; Zhang, Xiaoping; Lee, Khui Hung; Wang, Yibing; Yu, Kan; Shen, Wenying; Fu, Luoqin; Shu, Miaoan; Li, Weifen

    2015-11-01

    This experiment was designed to study the effects of Bacillus licheniformis BSK-4 on nitrogen removal and microbial community structure in a grass carp (Ctenopharyngodon idellus) culture. The selected strain Bacillus licheniformis BSK-4 significantly decreased nitrite, nitrate and total nitrogen levels in water over an extended, whereas increased ammonia level. Pyrosequencing showed that Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes were dominant in grass carp culture water. Compared with the control group, the number of Proteobacteria and Firmicutes were increased, while Actinobacteria and Bacteroidetes decreased in treatment group. At the genus level, some genera, such as Bacillus, Prosthecobacter, Enterococcus, etc., appear only in the treatment, while many other genera exist only in the control group; Lactobacillus, Luteolibacter, Phenylobacterium, etc. were increased in treatment group compared to those in control group. As above, the results suggested that supplementation with B. licheniformis BSK-4 could remove some nitrogen and cause alterations of the microbial composition in grass carp water.

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

    Science.gov (United States)

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

    2017-02-01

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

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

    Science.gov (United States)

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

    2013-09-20

    Conjugative shuttle vectors of the pKVM series, based on an IncP transfer origin and the pMAD vector with a temperature sensitive replication were constructed to establish a markerless gene deletion protocol for Bacilli without natural competence such as the exoenzyme producer Bacillus licheniformis. The pKVM plasmids can be conjugated to strains of B. licheniformis and B. subtilis. For chromosomal gene deletion, regions flanking the target gene are fused and cloned in a pKVM vector prior to conjugative transfer from Escherichia coli to B. licheniformis. Appropriate markers on the vector backbone allow for the identification of the integration at the target locus and thereafter the vector excision, both events taking place via homologous recombination. The functionality of the deletion system was demonstrated with B. licheniformis by a markerless 939 bp in-frame deletion of the yqfD gene and the deletion of a 31 kbp genomic segment carrying a PBSX-like prophage. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Proteomics analysis of Bacillus licheniformis in response to oligosaccharides elicitors.

    Science.gov (United States)

    Reffatti, Patricia Fernanda; Roy, Ipsita; Odell, Mark; Keshavarz, Tajalli

    2014-01-01

    The role of oligosaccharides as biotic elicitors has been recognised in the enhanced production of antibiotics from fungal and bacterial cultures. The yield of bacitracin A in cultures of Bacillus licheniformis was increased after supplementation with oligoguluronate (OG), and mannan oligosaccharides (MO) and its mechanism at transcription level been established already. However, the elicitation mechanism at post transcriptional level has not been reported so far. In this paper we investigate changes in proteomics of B. licheniformis in presence of the oligosaccharide elicitors OG and MO. Differentially expressed proteins were examined using 2D-PAGE stained with colloidal Coomassie and were further identified by LC-MS/MS. We identified 19 differentially expressed proteins including those involved in carbon metabolism, energy generation, amino acid biosynthesis, oxidative and general stress response. The novel findings of this work, together with previous reports, contribute to the unravelling of the overall mechanism of elicitation in B. licheniformis cultures and reliability of the use of these elicitors for potential industrial application. Copyright © 2014 Elsevier Inc. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Somen Acharya

    2012-08-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  2. A proteomic view of cell physiology of the industrial workhorse Bacillus licheniformis

    NARCIS (Netherlands)

    Voigt, Birgit; Schroeter, Rebecca; Schweder, Thomas; Juergen, Britta; Albrecht, Dirk; van Dijl, Jan Maarten; Maurer, Karl-Heinz; Hecker, Michael

    2014-01-01

    Bacillus licheniformis is known for its high protein secretion capacity and is being applied extensively as a host for the industrial production of enzymes such as proteases and amylases. In its natural environment as well as in fermentation processes the bacterium is often facing adverse conditions

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

    NARCIS (Netherlands)

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

    2014-01-01

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

  4. Effects of High Pressure on Bacillus licheniformis Spore Germination and Inactivation.

    Science.gov (United States)

    Borch-Pedersen, Kristina; Mellegård, Hilde; Reineke, Kai; Boysen, Preben; Sevenich, Robert; Lindbäck, Toril; Aspholm, Marina

    2017-07-15

    Bacillus and Clostridium species form spores, which pose a challenge to the food industry due to their ubiquitous nature and extreme resistance. Pressurization at 300 MPa likely triggers germination by opening dipicolinic acid (DPA) channels present in the inner membrane of the spores. In this work, we expose spores of Bacillus licheniformis , a species associated with food spoilage and occasionally with food poisoning, to high pressure (HP) for holding times of up to 2 h. By using mutant spores lacking one or several GRs, we dissect the roles of the GerA, Ynd, and GerK GRs in moderately HP (mHP; 150 MPa)-induced spore germination. We show that Ynd alone is sufficient for efficient mHP-induced spore germination. GerK also triggers germination with mHP, although at a reduced germination rate compared to that of Ynd. GerA stimulates mHP-induced germination but only in the presence of either the intact GerK or Ynd GR. These results suggests that the effectiveness of the individual GRs in mHP-induced germination differs from their effectiveness in nutrient-induced germination, where GerA plays an essential role. In contrast to Bacillus subtilis spores, treatment with very HP (vHP) of 550 MPa at 37°C did not promote effective germination of B. licheniformis spores. However, treatment with vHP in combination with elevated temperatures (60°C) gave a synergistic effect on spore germination and inactivation. Together, these results provide novel insights into how HP affects B. licheniformis spore germination and inactivation and the role of individual GRs in this process. IMPORTANCE Bacterial spores are inherently resistant to food-processing regimes, such as high-temperature short-time pasteurization, and may therefore compromise food durability and safety. The induction of spore germination facilitates subsequent inactivation by gentler processing conditions that maintain the sensory and nutritional qualities of the food. High-pressure (HP) processing is a nonthermal

  5. Markerless deletion of putative alanine dehydrogenase genes in Bacillus licheniformis using a codBA-based counterselection technique.

    Science.gov (United States)

    Kostner, David; Rachinger, Michael; Liebl, Wolfgang; Ehrenreich, Armin

    2017-11-01

    Bacillus licheniformis strains are used for the large-scale production of industrial exoenzymes from proteinaceous substrates, but details of the amino acid metabolism involved are largely unknown. In this study, two chromosomal genes putatively involved in amino acid metabolism of B. licheniformis were deleted to clarify their role. For this, a convenient counterselection system for markerless in-frame deletions was developed for B. licheniformis. A deletion plasmid containing up- and downstream DNA segments of the chromosomal deletion target was conjugated to B. licheniformis and integrated into the genome by homologous recombination. Thereafter, the counterselection was done by using a codBA cassette. The presence of cytosine deaminase and cytosine permease exerted a conditionally lethal phenotype on B. licheniformis cells in the presence of the cytosine analogue 5-fluorocytosine. Thereby clones were selected that lost the integrated vector sequence and the anticipated deletion target after a second recombination step. This method allows the construction of markerless mutants in Bacillus strains in iterative cycles. B. licheniformis MW3 derivatives lacking either one of the ORFs BL03009 or BL00190, encoding a putative alanine dehydrogenase and a similar putative enzyme, respectively, retained the ability to grow in minimal medium supplemented with alanine as the carbon source. In the double deletion mutant MW3 ΔBL03009 ΔBL00190, however, growth on alanine was completely abolished. These data indicate that the two encoded enzymes are paralogues fulfilling mutually replaceable functions in alanine utilization, and suggest that in B. licheniformis MW3 alanine utilization is initiated by direct oxidative transamination to pyruvate and ammonium.

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

    Directory of Open Access Journals (Sweden)

    Rebecca Schroeter

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

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

    Science.gov (United States)

    Schroeter, Rebecca; Hoffmann, Tamara; Voigt, Birgit; Meyer, Hanna; Bleisteiner, Monika; Muntel, Jan; Jürgen, Britta; Albrecht, Dirk; Becher, Dörte; Lalk, Michael; Evers, Stefan; Bongaerts, Johannes; Maurer, Karl-Heinz; Putzer, Harald; Hecker, Michael; Schweder, Thomas; Bremer, Erhard

    2013-01-01

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

  8. pH modulates arsenic toxicity in Bacillus licheniformis DAS-2.

    Science.gov (United States)

    Tripti, K; Shardendu

    2016-08-01

    The toxic characteristics of arsenic species, As(V) and As(III) result in ecological risks. Arsenic tolerant bacterium was isolated and identified as the Bacillus licheniformis DAS-2 through 16SrDNA sequencing. B. licheniformis DAS-2 was efficient to tolerate and remove both the As(V)[MIC 8mM] and As(III)[MIC 6mM] from the growth medium. The potential for the removal/uptake of arsenic from the 3, 5 and 7mM As(V) enriched growth media was 100%, 60% and 35% respectively and from the 1, 3 and 5mM As(III) enrichment it was 100%, 99% and 58% respectively at neutral pH. 80% of uptake As(V) was reduced to As(III) in 3mM As(V) enrichment which was gradually decreased to only 17% at 7mM As(V) enrichment at neutral pH. The arsenic toxicity in B. licheniformis DAS-2 was found modulated by pH and was examined through alteration in growth, uptake/removal, reduction and measurement of chemical toxicity. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. L-alanine-induced germination in Bacillus licheniformis -the impact of native gerA sequences.

    Science.gov (United States)

    Madslien, Elisabeth H; Granum, Per Einar; Blatny, Janet M; Lindbäck, Toril

    2014-04-22

    L-alanine, acting through the GerA receptor, was recently found to be an efficient germinant in Bacillus licheniformis ATCC14580/DSM13. In this study, we show that several of 46 examined B. licheniformis strains germinate remarkably slower than the type strain when exposed to L-alanine. These strains are not necessarily closely related, as determined by MLST (multi-locus sequence typing). Three of the slow-germinating strains were further examined in order to see whether nucleotide substitutions in the gerA sequences were responsible for the slow L-alanine germination. This was performed by complementing the transformable type strain derivate MW3ΔgerAA with gerA variants from the three slow-germinating strains; NVH1032, NVH1112 and NVH800. A wide selection of B. licheniformis strains was evaluated for L-alanine-induced germination efficiency. Our results show that gerA substitutions could only partially explain why spores of some B. licheniformis strains responded slower than others in the presence of L-alanine.

  10. Enhanced production of bacitracin by a mutant strain bacillus licheniformis UV-MN-HN-8 (enhanced bacitracin production by mutagenesis)

    International Nuclear Information System (INIS)

    Aftab, M.N.; Ikram-ul-Haq; Baig, S.

    2010-01-01

    The present study is focused on the improvement of Bacillus licheniformis through random mutagenesis to obtain mutant having enhanced production of bacitracin. Many isolates of Bacillus licheniformis were isolated and the isolate GP-40 produced maximum bacitracin production (16 +- 0.72 IU/mL). Treatment of Bacillus licheniformis GP-40 with ultraviolet (UV) radiations increased bacitracin production to 29 +- 0.69 IU/mL. Similarly, treatment of vegetative cells of GP-40 with chemicals like N-methyl N'-nitro N-nitroso guanidine (MNNG) and Nitrous acid (HNO/sub 2/) increased bacitracin production to 35 +- 1.35 IU/mL and 29 +- 0.89 IU/mL respectively. Studies regarding the combined effect of UV and chemical treatment on parental cells exhibited significantly higher titers of bacitracin with maximum bacitracin production reached to 47.6 +- 0.92 IU/mL. An increase of 2.97 fold production of bacitracin in comparison to wild type was observed. Mutant strain was highly stable and produced consistent yield of bacitracin even after 15 generations. On the basis of kinetic variables, notably mu (h-/sup 1/)max, Yp/x, qp, Qp and Qx mutant strain B. licheniformis UV-MN-HN-8 was found to be a hyper producer of bacitracin. (author)

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

    Science.gov (United States)

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

    2014-03-01

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

  12. Development of an Efficient Genome Editing Tool in Bacillus licheniformis Using CRISPR-Cas9 Nickase.

    Science.gov (United States)

    Li, Kaifeng; Cai, Dongbo; Wang, Zhangqian; He, Zhili; Chen, Shouwen

    2018-03-15

    Bacillus strains are important industrial bacteria that can produce various biochemical products. However, low transformation efficiencies and a lack of effective genome editing tools have hindered its widespread application. Recently, clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 techniques have been utilized in many organisms as genome editing tools because of their high efficiency and easy manipulation. In this study, an efficient genome editing method was developed for Bacillus licheniformis using a CRISPR-Cas9 nickase integrated into the genome of B. licheniformis DW2 with overexpression driven by the P43 promoter. The yvmC gene was deleted using the CRISPR-Cas9n technique with homology arms of 1.0 kb as a representative example, and an efficiency of 100% was achieved. In addition, two genes were simultaneously disrupted with an efficiency of 11.6%, and the large DNA fragment bacABC (42.7 kb) was deleted with an efficiency of 79.0%. Furthermore, the heterologous reporter gene aprN , which codes for nattokinase in Bacillus subtilis , was inserted into the chromosome of B. licheniformis with an efficiency of 76.5%. The activity of nattokinase in the DWc9nΔ7/pP43SNT-S sacC strain reached 59.7 fibrinolytic units (FU)/ml, which was 25.7% higher than that of DWc9n/pP43SNT-S sacC Finally, the engineered strain DWc9nΔ7 (Δ epr Δ wprA Δ mpr Δ aprE Δ vpr Δ bprA Δ bacABC ), with multiple disrupted genes, was constructed using the CRISPR-Cas9n technique. Taken together, we have developed an efficient genome editing tool based on CRISPR-Cas9n in B. licheniformis This tool could be applied to strain improvement for future research. IMPORTANCE As important industrial bacteria, Bacillus strains have attracted significant attention due to their production of biological products. However, genetic manipulation of these bacteria is difficult. The CRISPR-Cas9 system has been applied to genome editing in some bacteria, and CRISPR-Cas9n was proven to

  13. Nitrous oxide emission by the non-denitrifying, nitrate ammonifier Bacillus licheniformis

    OpenAIRE

    Sun, Yihua; De Vos, Paul; Heylen, Kim

    2016-01-01

    Background Firmicutes have the capacity to remove excess nitrate from the environment via either denitrification, dissimilatory nitrate reduction to ammonium or both. The recent renewed interest in their nitrogen metabolism has revealed many interesting features, the most striking being their wide variety of dissimilatory nitrate reduction pathways. In the present study, nitrous oxide production from Bacillus licheniformis, a ubiquitous Gram-positive, spore-forming species with many industria...

  14. Evaluation of antifungal metabolites activity from bacillus licheniformis OE-04 against Colletotrichum gossypii.

    Science.gov (United States)

    Nawaz, Hafiz Husnain; Nelly Rajaofera, M J; He, Qiguang; Anam, Usmani; Lin, Chunhua; Miao, Weiguo

    2018-04-01

    Anthracnose disease in the cotton plant caused by fungal pathogen Colletotrichum gossypii. It is supposed to be most critical diseases in the cotton crop as it causes infection and leads to complete damaging of the cotton crop by infecting the leaves, stems, and bolls in the field. The disease control is challenging due to the absence of an effective fungicide without damaging the farmer health and environment. So the series of experiments were designed to assess the antagonistic activity of biosurfactant released by strain Bacillus licheniformis OE-04 against the anthracnose causing agent in cotton and this strain was screened out from forty eight strain of rhizobacteria. We also estimated the heat stability and pH range and toxicity of biosurfactant produced by strain 0E-04. The results showed that biosurfactant has maximum antifungal activity against C. gossypii. In vitro study concluded that the biosurfactant can reduce fungal activity by inhibiting the spore germination of C. gossypii. Moreover, the biosurfactant also has wide pH and temperature range. We observed Antifungal activity of biosurfactant at 5 to 10 pH range and temperature range was also wide from room temperature to 100 °C. We also observed the toxicity of biosurfactant produced by Bacillus licheniformis against zebra fish (Danio rerio). We were noticed that biosurfactant have least harmful effect with maximum concentration. The study confirmed that biosurfactant of Bacillus licheniformis have high pH and heat stability range with least harmful effects so it can be a good replacement of chemical pesticides for cotton anthracnose control. Copyright © 2018 Elsevier Inc. All rights reserved.

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

    International Nuclear Information System (INIS)

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

    2015-01-01

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

  16. RNA-Seq of Bacillus licheniformis: active regulatory RNA features expressed within a productive fermentation

    Science.gov (United States)

    2013-01-01

    Background The production of enzymes by an industrial strain requires a complex adaption of the bacterial metabolism to the conditions within the fermenter. Regulatory events within the process result in a dynamic change of the transcriptional activity of the genome. This complex network of genes is orchestrated by proteins as well as regulatory RNA elements. Here we present an RNA-Seq based study considering selected phases of an industry-oriented fermentation of Bacillus licheniformis. Results A detailed analysis of 20 strand-specific RNA-Seq datasets revealed a multitude of transcriptionally active genomic regions. 3314 RNA features encoded by such active loci have been identified and sorted into ten functional classes. The identified sequences include the expected RNA features like housekeeping sRNAs, metabolic riboswitches and RNA switches well known from studies on Bacillus subtilis as well as a multitude of completely new candidates for regulatory RNAs. An unexpectedly high number of 855 RNA features are encoded antisense to annotated protein and RNA genes, in addition to 461 independently transcribed small RNAs. These antisense transcripts contain molecules with a remarkable size range variation from 38 to 6348 base pairs in length. The genome of the type strain B. licheniformis DSM13 was completely reannotated using data obtained from RNA-Seq analyses and from public databases. Conclusion The hereby generated data-sets represent a solid amount of knowledge on the dynamic transcriptional activities during the investigated fermentation stages. The identified regulatory elements enable research on the understanding and the optimization of crucial metabolic activities during a productive fermentation of Bacillus licheniformis strains. PMID:24079885

  17. RNA-Seq of Bacillus licheniformis: active regulatory RNA features expressed within a productive fermentation.

    Science.gov (United States)

    Wiegand, Sandra; Dietrich, Sascha; Hertel, Robert; Bongaerts, Johannes; Evers, Stefan; Volland, Sonja; Daniel, Rolf; Liesegang, Heiko

    2013-10-01

    The production of enzymes by an industrial strain requires a complex adaption of the bacterial metabolism to the conditions within the fermenter. Regulatory events within the process result in a dynamic change of the transcriptional activity of the genome. This complex network of genes is orchestrated by proteins as well as regulatory RNA elements. Here we present an RNA-Seq based study considering selected phases of an industry-oriented fermentation of Bacillus licheniformis. A detailed analysis of 20 strand-specific RNA-Seq datasets revealed a multitude of transcriptionally active genomic regions. 3314 RNA features encoded by such active loci have been identified and sorted into ten functional classes. The identified sequences include the expected RNA features like housekeeping sRNAs, metabolic riboswitches and RNA switches well known from studies on Bacillus subtilis as well as a multitude of completely new candidates for regulatory RNAs. An unexpectedly high number of 855 RNA features are encoded antisense to annotated protein and RNA genes, in addition to 461 independently transcribed small RNAs. These antisense transcripts contain molecules with a remarkable size range variation from 38 to 6348 base pairs in length. The genome of the type strain B. licheniformis DSM13 was completely reannotated using data obtained from RNA-Seq analyses and from public databases. The hereby generated data-sets represent a solid amount of knowledge on the dynamic transcriptional activities during the investigated fermentation stages. The identified regulatory elements enable research on the understanding and the optimization of crucial metabolic activities during a productive fermentation of Bacillus licheniformis strains.

  18. Lack of specific hybridization between the lep genes of Salmonella typhimurium and Bacillus licheniformis

    NARCIS (Netherlands)

    van Dijl, J M; Jong, de Anne; Smith, H; Bron, Sierd; Venema, G

    1991-01-01

    This paper describes an attempt to clone the Bacillus licheniformis lep gene, encoding signal peptidase, using the Salmonella typhimurium lep gene as a hybridization probe. Although a hybridizing fragment was obtained, DNA sequence analysis indicated that it did not contain the lep gene. Instead,

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

    Directory of Open Access Journals (Sweden)

    Prabha Devi

    2010-04-01

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

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

    NARCIS (Netherlands)

    Creusot, N.P.; Gruppen, H.

    2007-01-01

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

  1. Comprehensive transcriptome and improved genome annotation of Bacillus licheniformis WX-02.

    Science.gov (United States)

    Guo, Jing; Cheng, Gang; Gou, Xiang-Yong; Xing, Feng; Li, Sen; Han, Yi-Chao; Wang, Long; Song, Jia-Ming; Shu, Cheng-Cheng; Chen, Shou-Wen; Chen, Ling-Ling

    2015-08-19

    The updated genome of Bacillus licheniformis WX-02 comprises a circular chromosome of 4286821 base-pairs containing 4512 protein-coding genes. We applied strand-specific RNA-sequencing to explore the transcriptome profiles of B. licheniformis WX-02 under normal and high-salt conditions (NaCl 6%). We identified 2381 co-expressed gene pairs constituting 871 operon structures. In addition, 1169 antisense transcripts and 90 small RNAs were detected. Systematic comparison of differentially expressed genes under different conditions revealed that genes involved in multiple functions were significantly repressed in long-term high salt adaptation process. Genes related to promotion of glutamic acid synthesis were activated by 6% NaCl, potentially explaining the high yield of γ-PGA under salt condition. This study will be useful for the optimization of crucial metabolic activities in this bacterium. Copyright © 2015. Published by Elsevier B.V.

  2. Isolation and characterization of an antifungal protein from Bacillus licheniformis HS10.

    Science.gov (United States)

    Wang, Zhixin; Wang, Yunpeng; Zheng, Li; Yang, Xiaona; Liu, Hongxia; Guo, Jianhua

    2014-11-07

    Bacillus licheniformis HS10 is a good biocontrol agent against Pseudoperonospora cubensis which caused cucumber downy disease. To identify and characterize the antifungal proteins produced by B.licheniformis HS10, the proteins from HS10 were isolated by using 30-60% ammonium sulfate precipitation, and purified with column chromatography on DEAE Sepharose Fast Flow, RESOURCE Q and Sephadex G-75. And the SDS-PAGE and MALDI-TOF/TOF-MS analysis results demonstrated that the antifungal protein was a monomer with molecular weight of about 55 kDa, identified as carboxypeptidase. Our experiments also showed that the antifungal protein from B. licheniformis HS10 had significantly inhibition on eight different kinds of plant pathogenic fungi, and it was stable with good biological activity at as high as 100°C for 30 min and in pH value ranged from 6 to 10. The biological activity was negatively affected by protease K and 10mM metal cations except Ca(2+). Copyright © 2014 Elsevier Inc. All rights reserved.

  3. Effects of Dietary Bacillus licheniformis on Gut Physical Barrier, Immunity, and Reproductive Hormones of Laying Hens.

    Science.gov (United States)

    Wang, Yang; Du, Wei; Lei, Kai; Wang, Baikui; Wang, Yuanyuan; Zhou, Yingshan; Li, Weifen

    2017-09-01

    Previous study showed that dietary Bacillus licheniformis (B. licheniformis) administration contributes to the improvement of laying performance and egg quality in laying hens. In this study, we aimed to further evaluate its underlying mechanisms. Three hundred sixty Hy-Line Variety W-36 hens (28 weeks of age) were randomized into four groups, each group with six replications (n = 15). The control group received the basal diet and the treatment groups received the same basal diets supplemented with 0.01, 0.03, and 0.06% B. licheniformis powder (2 × 10 10  cfu/g) for an 8-week trial. The results demonstrate that B. licheniformis significantly enhance the intestinal barrier functions via decreasing gut permeability, promoting mucin-2 transcription, and regulating inflammatory cytokines. The systemic immunity of layers in B. licheniformis treatment groups is improved through modulating the specific and non-specific immunity. In addition, gene expressions of hormone receptors, including estrogen receptor α, estrogen receptor β, and follicle-stimulating hormone receptor, are also regulated by B. licheniformis. Meanwhile, compared with the control, B. licheniformis significantly increase gonadotropin-releasing hormone level, but markedly reduce ghrelin and inhibin secretions. Overall, our data suggest that dietary inclusion of B. licheniformis can improve the intestinal barrier function and systemic immunity and regulate reproductive hormone secretions, which contribute to better laying performance and egg quality of hens.

  4. The Cooperative and Interdependent Roles of GerA, GerK, and Ynd in Germination of Bacillus licheniformis Spores.

    Science.gov (United States)

    Borch-Pedersen, Kristina; Lindbäck, Toril; Madslien, Elisabeth H; Kidd, Shani W; O'Sullivan, Kristin; Granum, Per Einar; Aspholm, Marina

    2016-07-15

    When nutrients are scarce, Bacillus species form metabolically dormant and extremely resistant spores that enable survival over long periods of time under conditions not permitting growth. The presence of specific nutrients triggers spore germination through interaction with germinant receptors located in the spore's inner membrane. Bacillus licheniformis is a biotechnologically important species, but it is also associated with food spoilage and food-borne disease. The B. licheniformis ATCC 14580/DSM13 genome exhibits three gerA family operons (gerA, gerK, and ynd) encoding germinant receptors. We show that spores of B. licheniformis germinate efficiently in response to a range of different single l-amino acid germinants, in addition to a weak germination response seen with d-glucose. Mutational analyses revealed that the GerA and Ynd germination receptors function cooperatively in triggering an efficient germination response with single l-amino acid germinants, whereas the GerK germination receptor is essential for germination with d-glucose. Mutant spores expressing only GerA and GerK or only Ynd and GerK show reduced or severely impaired germination responses, respectively, with single l-amino acid germinants. Neither GerA nor Ynd could function alone in stimulating spore germination. Together, these results functionally characterize the germination receptor operons present in B. licheniformis We demonstrate the overlapping germinant recognition patterns of the GerA and Ynd germination receptors and the cooperative functionalities between GerA, Ynd, and GerK in inducing germination. To ensure safe food production and durable foods, there is an obvious need for more knowledge on spore-forming bacteria. It is the process of spore germination that ultimately leads to food spoilage and food poisoning. Bacillus licheniformis is a biotechnologically important species that is also associated with food spoilage and food-borne disease. Despite its importance, the

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

    NARCIS (Netherlands)

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

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

  6. High-Level Heat Resistance of Spores of Bacillus amyloliquefaciens and Bacillus licheniformis Results from the Presence of a spoVA Operon in a Tn1546 Transposon

    Science.gov (United States)

    Berendsen, Erwin M.; Koning, Rosella A.; Boekhorst, Jos; de Jong, Anne; Kuipers, Oscar P.; Wells-Bennik, Marjon H. J.

    2016-01-01

    Bacterial endospore formers can produce spores that are resistant to many food processing conditions, including heat. Some spores may survive heating processes aimed at production of commercially sterile foods. Recently, it was shown that a spoVA operon, designated spoVA2mob, present on a Tn1546 transposon in Bacillus subtilis, leads to profoundly increased wet heat resistance of B. subtilis spores. Such Tn1546 transposon elements including the spoVA2mob operon were also found in several strains of Bacillus amyloliquefaciens and Bacillus licheniformis, and these strains were shown to produce spores with significantly higher resistances to wet heat than their counterparts lacking this transposon. In this study, the locations and compositions of Tn1546 transposons encompassing the spoVA2mob operons in B. amyloliquefaciens and B. licheniformis were analyzed. Introduction of these spoVA2mob operons into B. subtilis 168 (producing spores that are not highly heat resistant) rendered mutant 168 strains that produced high-level heat resistant spores, demonstrating that these elements in B. amyloliquefaciens and B. licheniformis are responsible for high level heat resistance of spores. Assessment of growth of the nine strains of each species between 5.2°C and 57.7°C showed some differences between strains, especially at lower temperatures, but all strains were able to grow at 57.7°C. Strains of B. amyloliquefaciens and B. licheniformis that contain the Tn1546 elements (and produce high-level heat resistant spores) grew at temperatures similar to those of their Tn1546-negative counterparts that produce low-level heat resistant spores. The findings presented in this study allow for detection of B. amyloliquefaciens and B. licheniformis strains that produce highly heat resistant spores in the food chain. PMID:27994575

  7. Identification of key genes involved in polysaccharide bioflocculant synthesis in Bacillus licheniformis.

    Science.gov (United States)

    Chen, Zhen; Liu, Peize; Li, Zhipeng; Yu, Wencheng; Wang, Zhi; Yao, Haosheng; Wang, Yuanpeng; Li, Qingbiao; Deng, Xu; He, Ning

    2017-03-01

    The present study reports the sequenced genome of Bacillus licheniformis CGMCC 2876, which is composed of a 4,284,461 bp chromosome that contains 4,188 protein-coding genes, 72 tRNA genes, and 21 rRNA genes. Additional analysis revealed an eps gene cluster with 16 open reading frames. Conserved Domains Database analysis combined with qPCR experiments indicated that all genes in this cluster were involved in polysaccharide bioflocculant synthesis. Phosphoglucomutase and UDP-glucose pyrophosphorylase were supposed to be key enzymes in polysaccharide secretion in B. licheniformis. A biosynthesis pathway for the production of polysaccharide bioflocculant involving the integration of individual genes was proposed based on functional analysis. Overexpression of epsDEF from the eps gene cluster in B. licheniformis CGMCC 2876 increased the flocculating activity of the recombinant strain by 90% compared to the original strain. Similarly, the crude yield of polysaccharide bioflocculant was enhanced by 27.8%. Overexpression of the UDP-glucose pyrophosphorylase gene not only increased the flocculating activity by 71% but also increased bioflocculant yield by 13.3%. Independent of UDP-N-acetyl-D-mannosamine dehydrogenase gene, flocculating activity, and polysaccharide yield were negatively impacted by overexpression of the UDP-N-acetylglucosamine 2-epimerase gene. Overall, epsDEF and gtaB2 were identified as key genes for polysaccharide bioflocculant synthesis in B. licheniformis. These results will be useful for further engineering of B. licheniformis for industrial bioflocculant production. Biotechnol. Bioeng. 2017;114: 645-655. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  8. High-level expression of nattokinase in Bacillus licheniformis by manipulating signal peptide and signal peptidase.

    Science.gov (United States)

    Cai, D; Wei, X; Qiu, Y; Chen, Y; Chen, J; Wen, Z; Chen, S

    2016-09-01

    Nattokinase is an enzyme produced by Bacillus licheniformis and has potential to be used as a drug for treating cardiovascular disease due to its beneficial effects of preventing fibrin clots etc. However, the low activity and titre of this protein produced by B. licheniformis often hinders its application of commercial production. The aim of this work is to improve the nattokinase production by manipulating signal peptides and signal peptidases in B. licheniformis. The P43 promoter, amyL terminator and AprN target gene were used to form the nattokinase expression vector, pHY-SP-NK, which was transformed into B. licheniformis and nattokinase was expressed successfully. A library containing 81 predicted signal peptides was constructed for nattokinase expression in B. licheniformis, with the maximum activity being obtained under the signal peptide of AprE. Among four type I signal peptidases genes (sipS, sipT, sipV, sipW) in B. licheniformis, the deletion of sipV resulted in a highest decrease in nattokinase activity. Overexpression of sipV in B. licheniformis led to a nattokinase activity of 35·60 FU ml(-1) , a 4·68-fold improvement over activity produced by the initial strain. This work demonstrates the potential of B. licheniformis for industrial production of nattokinase through manipulation of signal peptides and signal peptidases expression. This study has screened the signal peptides of extracellular proteins of B. licheniformis for nattokinase production. Four kinds of Type I signal peptidases genes have been detected respectively in B. licheniformis to identify which one played the vital role for nattokinase production. This study provided a promising strain for industry production of nattokinase. © 2016 The Society for Applied Microbiology.

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

    NARCIS (Netherlands)

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

    2015-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Muhammad Nauman Aftab

    2012-03-01

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

  11. The two putative comS homologs of the biotechnologically important Bacillus licheniformis do not contribute to competence development.

    Science.gov (United States)

    Jakobs, Mareike; Hoffmann, Kerstin; Liesegang, Heiko; Volland, Sonja; Meinhardt, Friedhelm

    2015-03-01

    In Bacillus subtilis, natural genetic competence is subject to complex genetic regulation and quorum sensing dependent. Upon extracellular accumulation of the peptide-pheromone ComX, the membrane-bound sensor histidine kinase ComP initiates diverse signaling pathways by activating-among others-DegQ and ComS. While DegQ favors the expression of extracellular enzymes rather than competence development, ComS is crucial for competence development as it prevents proteolytic degradation of ComK, the key transcriptional activator of all genes required for the uptake and integration of DNA. In Bacillus licheniformis, ComX/ComP sensed cell density negatively influences competence development, suggesting differences from the quorum-sensing-dependent control mechanism in Bacillus subtilis. Here, we show that each of six investigated strains possesses both of two different, recently identified putative comS genes. When expressed from an inducible promoter, none of the comS candidate genes displayed an impact on competence development neither in B. subtilis nor in B. licheniformis. Moreover, disruption of the genes did not reduce transformation efficiency. While the putative comS homologs do not contribute to competence development, we provide evidence that the degQ gene as for B. subtilis negatively influences genetic competency in B. licheniformis.

  12. Nitrous oxide emission by the non-denitrifying, nitrate ammonifier Bacillus licheniformis.

    Science.gov (United States)

    Sun, Yihua; De Vos, Paul; Heylen, Kim

    2016-01-19

    Firmicutes have the capacity to remove excess nitrate from the environment via either denitrification, dissimilatory nitrate reduction to ammonium or both. The recent renewed interest in their nitrogen metabolism has revealed many interesting features, the most striking being their wide variety of dissimilatory nitrate reduction pathways. In the present study, nitrous oxide production from Bacillus licheniformis, a ubiquitous Gram-positive, spore-forming species with many industrial applications, is investigated. B. licheniformis has long been considered a denitrifier but physiological experiments on three different strains demonstrated that nitrous oxide is not produced from nitrate in stoichiometric amounts, rather ammonium is the most important end-product, produced during fermentation. Significant strain dependency in end-product ratios, attributed to nitrite and ammonium, and medium dependency in nitrous oxide production were also observed. Genome analyses confirmed the lack of a nitrite reductase to nitric oxide, the key enzyme of denitrification. Based on the gene inventory and building on knowledge from other non-denitrifying nitrous oxide emitters, hypothetical pathways for nitrous oxide production, involving NarG, NirB, qNor and Hmp, are proposed. In addition, all publically available genomes of B. licheniformis demonstrated similar gene inventories, with specific duplications of the nar operon, narK and hmp genes as well as NarG phylogeny supporting the evolutionary separation of previously described distinct BALI1 and BALI2 lineages. Using physiological and genomic data we have demonstrated that the common soil bacterium B. licheniformis does not denitrify but is capable of fermentative dissimilatory nitrate/nitrite reduction to ammonium (DNRA) with concomitant production of N2O. Considering its ubiquitous nature and non-fastidious growth in the lab, B. licheniformis is a suitable candidate for further exploration of the actual mechanism of N2O

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

    Directory of Open Access Journals (Sweden)

    Zare Mirakabadi, A.

    2012-11-01

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

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

    Science.gov (United States)

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

    2015-12-02

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

  15. A proteomic view of cell physiology of the industrial workhorse Bacillus licheniformis.

    Science.gov (United States)

    Voigt, Birgit; Schroeter, Rebecca; Schweder, Thomas; Jürgen, Britta; Albrecht, Dirk; van Dijl, Jan Maarten; Maurer, Karl-Heinz; Hecker, Michael

    2014-12-10

    Bacillus licheniformis is known for its high protein secretion capacity and is being applied extensively as a host for the industrial production of enzymes such as proteases and amylases. In its natural environment as well as in fermentation processes the bacterium is often facing adverse conditions such as oxidative or osmotic stress or starvation for nutrients. During the last years detailed proteome and transcriptome analyses have been performed to study the adaptation of B. licheniformis cells to various stresses (heat, ethanol, oxidative or salt stress) and starvation conditions (glucose, nitrogen or phosphate starvation). A common feature of the response to all tested conditions is the downregulation of many genes encoding house-keeping proteins and, consequently, a reduced synthesis of the corresponding proteins. Induction of the general stress response (σ(B) regulon) is only observed in cells subjected to heat, ethanol or salt stress. This paper summarizes our current knowledge on general and specific stress and starvation responses of this important industrial bacterium. The importance of selected marker genes and proteins for the monitoring and optimization of B. licheniformis based fermentation processes is discussed. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2015-03-01

    γ-poly glutamic acid (γ-PGA) has received considerable attention for pharmaceutical and biomedical applications. γ-PGA from the newly isolate Bacillus licheniformis NRC20 was purified and characterized using diffusion distance agar plate, mass spectrometry and thin layer chromatography. All analysis indicated that γ-PGA is a homopolymer composed of glutamic acid. Its molecular weight was determined to be 1266 kDa. It was composed of L- and D-glutamic acid residues. An amplicon of 3050 represents the γ-PGA-coding genes was obtained, sequenced and submitted in genbank database. Its amino acid sequence showed high similarity with that obtained from B. licheniformis strains. The bacterium NRC 20 was independent of L-glutamic acid but the polymer production enhanced when cultivated in medium containing L-glutamic acid as the sole nitrogen source. Finally we can conclude that γ-PGA production from B. licheniformis NRC20 has many promised applications in medicine, industry and nanotechnology. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2015-02-11

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

  18. Degradation of aldrin by bacillus licheniformis, isolated from water and sediment from the Cienaga Grande, Santa Marta, Colombia

    International Nuclear Information System (INIS)

    Sanchez Diazgranados, Jose Gregorio; Henry Lopez, Carlos Andres

    2012-01-01

    The bacterium bacillus licheniformis was isolated from sediment and water samples from estuary lagoon Cienaga Grande de Santa Marta (CGSM), Colombian Caribbean. The aim of the work was to use this microorganism as an alternative in the degradation of organic persistent pollutants. b. licheniformis was able to tolerate aerobic conditions and concentrations of the pesticide organochlorine, aldrin. The test was made during 30 days with 60 ng/l of aldrin in order to evaluate the degradation capacity of this bacterium. Identification and isolation of b. licheniformis was made through morphological (gram test), as well as biochemical characterization (bbl crystal system). Aldrin concentration was determined by gas chromatography. Results show that b. licheniformis had a degradation capacity of 24% from total concentration. Factors like solar light exposition and volatilization had an extra influence of 31% on aldrin degradation.

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

    NARCIS (Netherlands)

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

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

  20. The response of Bacillus licheniformis to heat and ethanol stress and the role of the SigB regulon.

    Science.gov (United States)

    Voigt, Birgit; Schroeter, Rebecca; Jürgen, Britta; Albrecht, Dirk; Evers, Stefan; Bongaerts, Johannes; Maurer, Karl-Heinz; Schweder, Thomas; Hecker, Michael

    2013-07-01

    The heat and ethanol stress response of Bacillus licheniformis DSM13 was analyzed at the transcriptional and/or translational level. During heat shock, regulons known to be heat-induced in Bacillus subtilis 168 are upregulated in B. licheniformis, such as the HrcA, SigB, CtsR, and CssRS regulon. Upregulation of the SigY regulon and of genes controlled by other extracytoplasmic function (ECF) sigma factors indicates a cell-wall stress triggered by the heat shock. Furthermore, tryptophan synthesis enzymes were upregulated in heat stressed cells as well as regulons involved in usage of alternative carbon and nitrogen sources. Ethanol stress led to an induction of the SigB, HrcA, and CtsR regulons. As indicated by the upregulation of a SigM-dependent protein, ethanol also triggered a cell wall stress. To characterize the SigB regulon of B. licheniformis, we analyzed the heat stress response of a sigB mutant. It is shown that the B. licheniformis SigB regulon comprises additional genes, some of which do not exist in B. subtilis, such as BLi03885, encoding a hypothetical protein, the Na/solute symporter gene BLi02212, the arginase homolog-encoding gene BLi00198 and mcrA, encoding a protein with endonuclease activity. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Proteomic analysis of enzyme production by Bacillus licheniformis using different feather wastes as the sole fermentation media.

    Science.gov (United States)

    Parrado, J; Rodriguez-Morgado, B; Tejada, M; Hernandez, T; Garcia, C

    2014-04-10

    This study evaluates the use of different types of feathers as fermentation media for enzyme production. Bacillus licheniformis was grown on the feathers, which lead to total biodegradation due to bacterial enzymatic hydrolytic excretion. B. licheniformis excretes protease and lipase activity, with feather concentration being the main parameter controlling their generation. Using a proteomic approach, the proteins excreted during fermentation were identified, and the influence of the chemical composition of the feathers on protein secretion was tested. The identified proteins are hydrolytic enzymes such as keratinase, gamma-glutamyltranspeptidase, chitosanases, and glicosidases. The diversity of proteins is related to the chemical complexity of the feathers. Understanding the composition of a hydrolytic system, when B. licheniformis is cultured on different feathers, may assist in utilizing such a system for producing different hydrolytic enzymes. The data indicate that proteomics can be a valuable tool for describing the physiological state of B. licheniformis cell populations growing on different wastes. Copyright © 2014 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

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

    2015-03-01

    The alkaline protease gene, Apr, from Bacillus licheniformis 2709 was cloned into an expression vector pET - 28b (+), to yield the recombinant plasmid pET-28b (+) - Apr. The pET-28b (+) - Apr was expressed in a high expression strain E. coli BL21. The amino acid sequence deduced from the DNA sequence analysis revealed a 98% identity to that of Bacillus licheniformis 2709. Sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE) was used to access the protein expression. SDS-PAGE analysis indicated a protein of Mr of 38.8 kDa. The medium components and condition of incubation were optimized for the growth state of a recombinant strain. The optimal composition of production medium was composed of glucose 8 g/L, peptone 8 g/L and salt solution 10 mL. The samples were incubated on a rotary shaker of 180 r/min at 37°C for 24 h. Copyright © 2014. Published by Elsevier B.V.

  3. Fermentation stage-dependent adaptations of Bacillus licheniformis during enzyme production

    Science.gov (United States)

    2013-01-01

    Background Industrial fermentations can generally be described as dynamic biotransformation processes in which microorganisms convert energy rich substrates into a desired product. The knowledge of active physiological pathways, reflected by corresponding gene activities, allows the identification of beneficial or disadvantageous performances of the microbial host. Whole transcriptome RNA-Seq is a powerful tool to accomplish in-depth quantification of these gene activities, since the low background noise and the absence of an upper limit of quantification allow the detection of transcripts with high dynamic ranges. Such data enable the identification of potential bottlenecks and futile energetic cycles, which in turn can lead to targets for rational approaches to productivity improvement. Here we present an overview of the dynamics of gene activity during an industrial-oriented fermentation process with Bacillus licheniformis, an important industrial enzyme producer. Thereby, valuable insights which help to understand the complex interactions during such processes are provided. Results Whole transcriptome RNA-Seq has been performed to study the gene expression at five selected growth stages of an industrial-oriented protease production process employing a germination deficient derivative of B. licheniformis DSM13. Since a significant amount of genes in Bacillus strains are regulated posttranscriptionally, the generated data have been confirmed by 2D gel-based proteomics. Regulatory events affecting the coordinated activity of hundreds of genes have been analyzed. The data enabled the identification of genes involved in the adaptations to changing environmental conditions during the fermentation process. A special focus of the analyses was on genes contributing to central carbon metabolism, amino acid transport and metabolism, starvation and stress responses and protein secretion. Genes contributing to lantibiotics production and Tat-dependent protein secretion have

  4. High-resolution proteome maps of Bacillus licheniformis cells growing in minimal medium.

    Science.gov (United States)

    Voigt, Birgit; Albrecht, Dirk; Sievers, Susanne; Becher, Dörte; Bongaerts, Johannes; Evers, Stefan; Schweder, Thomas; Maurer, Karl-Heinz; Hecker, Michael

    2015-08-01

    Bacillus licheniformis is an important host for the industrial production of enzymes mainly because of its ability to secrete large amounts of protein. We analyzed the proteome of B. licheniformis cells growing in a minimal medium. Beside the cytosolic proteome, the membrane and the extracellular proteome were studied. We could identify 1470 proteins; 1168 proteins were classified as cytosolic proteins, 195 proteins with membrane-spanning domains were classified as membrane proteins, and 107 proteins, with either putative signals peptides or flagellin-like sequences, were classified as secreted proteins. The identified proteins were grouped into functional categories and used to reconstruct cellular functions and metabolic pathways of growing B. licheniformis cells. The largest group was proteins with functions in basic metabolic pathways such as carbon metabolism, amino acid and nucleotide synthesis and synthesis of fatty acids and cofactors. Many proteins detected were involved in DNA replication, transcription, and translation. Furthermore, a high number of proteins employed in the transport of a wide variety of compounds were found to be expressed in the cells. All MS data have been deposited in the ProteomeXchange with identifier PXD000791 (http://proteomecentral.proteomexchange.org/dataset/PXD000791). © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Transcriptional characteristics associated with lichenysin biosynthesis in Bacillus licheniformis from Chinese Maotai-flavor liquor making.

    Science.gov (United States)

    Wu, Qun; Zhang, Rong; Peng, Suqin; Xu, Yan

    2015-01-28

    This work investigated the biosynthetic mechanism of lichenysin, the newly identified nonvolatile matrix component in Chinese liquors. Transcriptomes were analyzed in three producers, Bacillus licheniformis CGMCC 3961, 3962, and 3963, which were isolated from Maotai-flavor liquor-making process and produced 386.3, 553.5, and 795.2 μg/L lichenysin in a simulative liquor fermentation process. Lichenysin synthetase genes lchAA-AD in these three producers were expressed much more highly than those of the nonproducer B. licheniformis ATCC 14580 (>18.4-fold). In addition, ABC transporters were the most significant responsive metabolic pathway, and the expression levels of peptide transporter genes dppABCDE all increased more than 19.2-fold. When B. licheniformis CGMCC 3963 was cultured in synthetic medium, the expression of dppABCDE and lichenysin both increased with the addition of casein hydrolysate (containing various peptides). This indicated that peptide would act as a substrate for lichenysin synthesis. This work sheds new light on the mechanism for lichenysin biosynthesis.

  6. High-level production of α-amylase by manipulating the expression of alanine racamase in Bacillus licheniformis.

    Science.gov (United States)

    He, Penghui; Zhang, Zeying; Cai, Dongbo; Chen, Yaozhong; Wang, Hao; Wei, Xuetuan; Li, Shunyi; Chen, Shouwen

    2017-09-01

    To improve target protein production by manipulating expression levels of alanine racemase in Bacillus licheniformis. The gene of dal was identified to be responsible for alanine racemase function. Based on the selection marker of dal, a food-grade expression system was constructed in B. licheniformis, and effects of different dal expression levels mediated by promoters on α-amylase production were investigated. The highest α-amylase activity (155 U/ml) was obtained in BL10D/pP43SAT-PtetDal, increased by 27% compared with that of the control strain BL10/pP43SAT in tetracycline-based system (123 U/ml). Moreover, the dal transcriptional level was not correlated positively with that of amyL. A food-grade system for high-level production of α-amylase was constructed in B. licheniformis, revealing that expression levels of selection marker significantly affected target protein production.

  7. In vitro Ca(2+)-dependent maturation of milk-clotting recombinant Epr: minor extracellular protease: from Bacillus licheniformis.

    Science.gov (United States)

    Ageitos, José Manuel; Vallejo, Juan Andrés; Serrat, Manuel; Sánchez-Pérez, Angeles; Villa, Tomás G

    2013-06-01

    The minor extracellular protease (Epr) is secreted into the culture medium during Bacillus licheniformis, strain USC13, stationary phase of growth. Whereas, B. subtilis Epr has been reported to be involved in swarming; the B. licheniformis protease is also involved in milk-clotting as shown by the curd forming ability of culture broths expressing this protein. The objectives of this study are the characterization of recombinant B. licheniformis Epr (minor extracellular protease) and the determination of its calcium-dependent activation process. In this work, we have cloned and expressed B. licheniformis Epr in Escherichia coli. We were also able to construct a tridimensional model for Epr based on its homology to Thermococcus kodakarensis pro-tk-subtilisin 2e1p, fervidolysin from Fervidobacterium pennivorans 1rv6, and B. lentus 1GCI subtilisin. Recombinant Epr was accumulated into inclusion bodies; after protein renaturation, Epr undergoes an in vitro calcium-dependent activation, similar to that described for tk protease. The recombinant Epr is capable of producing milk curds with the same clotting activity previously described for the native B. licheniformis Epr enzyme although further rheological and industrial studies should be carried out to confirm its real applicability. This work represents for the first time that Epr may be successfully expressed in a non-bacilli microorganism.

  8. Influence of dietary inclusion of Bacillus licheniformis on laying performance, egg quality, antioxidant enzyme activities, and intestinal barrier function of laying hens.

    Science.gov (United States)

    Lei, K; Li, Y L; Yu, D Y; Rajput, I R; Li, W F

    2013-09-01

    This experiment was conducted to evaluate the effects of dietary inclusion of Bacillus licheniformis on laying performance, egg quality, antioxidant enzyme activities, and intestinal barrier function of laying hens. Hy-Line Variety W-36 hens (n = 540; 28 wk of age) were randomized into 6 groups, each group with 6 replications (n = 15). The control group received the basal diet formulated with maize and soybean meal. The treatment groups received the same basal diets supplemented with 0.01, 0.02, 0.03, 0.06, and 0.09% Bacillus licheniformis powder (2 × 10(10) cfu/g) for an 8-wk trial. The results showed that dietary supplementation with 0.01 and 0.03% B. licheniformis significantly increased egg production and egg mass. However, no significant differences were observed in egg weight, feed consumption, and feed conversion efficiency among the 6 groups. Supplementation with different levels of B. licheniformis was found to be effective in improvement of egg quality by increasing egg shell thickness and strength. Compared with control, d-lactate content, diamine oxidase activity, and adrenocorticotropic hormone level in serum decreased significantly, and the level of estradiol and follicle-stimulating hormone increased significantly in plasma of all the experimental groups. Dietary supplementation with B. licheniformis increased the intestinal villus height and reduced the crypt depth. In conclusion, dietary inclusion of B. licheniformis could improve laying performance and egg quality significantly in a dose-dependent manner by decreasing the stress response, upregulating the growth hormone, and improving intestinal health.

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

    Directory of Open Access Journals (Sweden)

    Devendra H Dusane

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

  10. Disruption of Microbial Biofilms by an Extracellular Protein Isolated from Epibiotic Tropical Marine Strain of Bacillus licheniformis

    Science.gov (United States)

    Dusane, Devendra H.; Damare, Samir R.; Nancharaiah, Yarlagadda V.; Ramaiah, N.; Venugopalan, Vayalam P.; Kumar, Ameeta Ravi; Zinjarde, Smita S.

    2013-01-01

    Background Marine epibiotic bacteria produce bioactive compounds effective against microbial biofilms. The study examines antibiofilm ability of a protein obtained from a tropical marine strain of Bacillus licheniformis D1. Methodology/Principal Findings B. licheniformis strain D1 isolated from the surface of green mussel, Perna viridis showed antimicrobial activity against pathogenic Candida albicans BH, Pseudomonas aeruginosa PAO1 and biofouling Bacillus pumilus TiO1 cultures. The antimicrobial activity was lost after treatment with trypsin and proteinase K. The protein was purified by ultrafiltration and size-exclusion chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis revealed the antimicrobial agent to be a 14 kDa protein designated as BL-DZ1. The protein was stable at 75°C for 30 min and over a pH range of 3.0 to 11.0. The sequence alignment of the MALDI-fingerprint showed homology with the NCBI entry for a hypothetical protein (BL00275) derived from B. licheniformis ATCC 14580 with the accession number gi52082584. The protein showed minimum inhibitory concentration (MIC) value of 1.6 µg/ml against C. albicans. Against both P. aeruginosa and B. pumilus the MIC was 3.12 µg/ml. The protein inhibited microbial growth, decreased biofilm formation and dispersed pre-formed biofilms of the representative cultures in polystyrene microtiter plates and on glass surfaces. Conclusion/Significance We isolated a protein from a tropical marine strain of B. licheniformis, assigned a function to the hypothetical protein entry in the NCBI database and described its application as a potential antibiofilm agent. PMID:23691235

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

    DEFF Research Database (Denmark)

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

    2014-01-01

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

  12. Modeling the recovery of heat-treated Bacillus licheniformis Ad978 and Bacillus weihenstephanensis KBAB4 spores at suboptimal temperature and pH using growth limits.

    Science.gov (United States)

    Trunet, C; Mtimet, N; Mathot, A-G; Postollec, F; Leguerinel, I; Sohier, D; Couvert, O; Carlin, F; Coroller, L

    2015-01-01

    The apparent heat resistance of spores of Bacillus weihenstephanensis and Bacillus licheniformis was measured and expressed as the time to first decimal reduction (δ value) at a given recovery temperature and pH. Spores of B. weihenstephanensis were produced at 30°C and 12°C, and spores of B. licheniformis were produced at 45°C and 20°C. B. weihenstephanensis spores were then heat treated at 85°C, 90°C, and 95°C, and B. licheniformis spores were heat treated at 95°C, 100°C, and 105°C. Heat-treated spores were grown on nutrient agar at a range of temperatures (4°C to 40°C for B. weihenstephanensis and 15°C to 60°C for B. licheniformis) or a range of pHs (between pH 4.5 and pH 9.5 for both strains). The recovery temperature had a slight effect on the apparent heat resistance, except very near recovery boundaries. In contrast, a decrease in the recovery pH had a progressive impact on apparent heat resistance. A model describing the heat resistance and the ability to recover according to the sporulation temperature, temperature of treatment, and recovery temperature and pH was proposed. This model derived from secondary mathematical models for growth prediction. Previously published cardinal temperature and pH values were used as input parameters. The fitting of the model with apparent heat resistance data obtained for a wide range of spore treatment and recovery conditions was highly satisfactory. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  13. Cloning, purification and characterization of a thermostable β-galactosidase from Bacillus licheniformis strain KG9.

    Science.gov (United States)

    Matpan Bekler, F; Stougaard, P; Güven, K; Gül Güven, R; Acer, Ö

    2015-06-28

    A thermo— and alkalitolerant Bacillus licheniformis KG9 isolated from Taşlıdere hot water spring in Batman/Turkey was found to produce a thermostable β—galactosidase. Phylogenetic analysis showed that the 16S rRNA gene from B. licheniformis strain KG9 was 99.9% identical to that of the genome sequenced B. licheniformis strain DSM 13. Analysis of the B. licheniformis DSM 13 genomic sequence revealed four putative β—galactosidase genes. PCR primers based on the genome sequence of strain DSM 13 were used to isolate the corresponding β—galactosidase genes from B. licheniformis strain KG9. The calculated molecular weights of the β—galactosidases I, II, III, and IV using sequencing data were 30, 79, 74, and 79 kDa, respectively. The genes were inserted into an expression vector and recombinant β—galactosidase was produced in Escherichia coli. Of the four β—galactosidase genes identified in strain KG9, three of them were expressed as active, intracellular enzymes in E. coli. One of the recombinant enzymes, β—galactosidase III, was purified and characterized. Optimal temperature and pH was determined to be at 60 ºC and pH 6.0, respectively. Km was determined to be 1.3 mM and 13.3 mM with oNPG (ortho—nitrophenyl—β—D—galactopyranoside) and lactose as substrates, respectively, and Vmax was measured to 1.96 μmol/min and 1.55 μmol/min with oNPG and lactose, respectively.

  14. Draft genome comparison of representatives of the three dominant genotype groups of dairy Bacillus licheniformis strains.

    Science.gov (United States)

    Dhakal, Rajat; Seale, R Brent; Deeth, Hilton C; Craven, Heather; Turner, Mark S

    2014-06-01

    The spore-forming bacterium Bacillus licheniformis is a common contaminant of milk and milk products. Strains of this species isolated from dairy products can be differentiated into three major groups, namely, G, F1, and F2, using random amplification of polymorphic DNA (RAPD) analysis; however, little is known about the genomic differences between these groups and the identity of the fragments that make up their RAPD profiles. In this work we obtained high-quality draft genomes of representative strains from each of the three RAPD groups (designated strain G-1, strain F1-1, and strain F2-1) and compared them to each other and to B. licheniformis ATCC 14580 and Bacillus subtilis 168. Whole-genome comparison and multilocus sequence typing revealed that strain G-1 contains significant sequence variability and belongs to a lineage distinct from the group F strains. Strain G-1 was found to contain genes coding for a type I restriction modification system, urease production, and bacitracin synthesis, as well as the 8-kbp plasmid pFL7, and these genes were not present in strains F1-1 and F2-1. In agreement with this, all isolates of group G, but no group F isolates, were found to possess urease activity and antimicrobial activity against Micrococcus. Identification of RAPD band sequences revealed that differences in the RAPD profiles were due to differences in gene lengths, 3' ends of predicted primer binding sites, or gene presence or absence. This work provides a greater understanding of the phylogenetic and phenotypic differences observed within the B. licheniformis species.

  15. Bioreactor studies on the production of bacitracin by mutant strain of bacillus licheniformis BLS-NTTG 4

    International Nuclear Information System (INIS)

    Sabeen, M.; Arif, R.; Saleem, M.; Ghouri, S.M.; Baig, S.; Syed, Q.

    2005-01-01

    The present study deals with the production of antibiotic bacitracin by Bacillus licheniformis using submerged fermentation technique in stirred fermenter. Altogether 15 samples were isolated from local habitat such as Government College Lahore. Of all the culture tested BLS 13 gave maximum titer (81 plus minus li. Micro /ml) in the fermentation broth. To develop the process for the production of antibiotic bacitracin agro industrial wastes used as a sole source of carbon in different fermentation medias (M1-M10). The culture of Bacillus licheniformis BLS 13 was improved by UV irradiation by exposing the cells of BLS-UV 16 mutated for 60 minutes gave the maximum production i.e. 129.1 plus minus 0.7 i. Micro/ml. The parental strain (i.e. BLS 13 was also improved by using chemical mutagen NTTG which enhance the antibiotic activity, and the mutated strain number i.e. BLS-NTTG 4 gave the maximum production 143.0 plus minus 1.0 i. Micro/ml. In stirred fermenter chemically mutated strain was used to check the productivity of bacitracin in stirred fermenter was increased up to 156 plus minus 1 i. micro /ml. (author)

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

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    Jasmin Dischinger

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

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

    Science.gov (United States)

    Dischinger, Jasmin; Josten, Michaele; Szekat, Christiane; Sahl, Hans-Georg; Bierbaum, Gabriele

    2009-01-01

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

  18. Construction and analysis of a genome-scale metabolic network for Bacillus licheniformis WX-02.

    Science.gov (United States)

    Guo, Jing; Zhang, Hong; Wang, Cheng; Chang, Ji-Wei; Chen, Ling-Ling

    2016-05-01

    We constructed the genome-scale metabolic network of Bacillus licheniformis (B. licheniformis) WX-02 by combining genomic annotation, high-throughput phenotype microarray (PM) experiments and literature-based metabolic information. The accuracy of the metabolic network was assessed by an OmniLog PM experiment. The final metabolic model iWX1009 contains 1009 genes, 1141 metabolites and 1762 reactions, and the predicted metabolic phenotypes showed an agreement rate of 76.8% with experimental PM data. In addition, key metabolic features such as growth yield, utilization of different substrates and essential genes were identified by flux balance analysis. A total of 195 essential genes were predicted from LB medium, among which 149 were verified with the experimental essential gene set of B. subtilis 168. With the removal of 5 reactions from the network, pathways for poly-γ-glutamic acid (γ-PGA) synthesis were optimized and the γ-PGA yield reached 83.8 mmol/h. Furthermore, the important metabolites and pathways related to γ-PGA synthesis and bacterium growth were comprehensively analyzed. The present study provides valuable clues for exploring the metabolisms and metabolic regulation of γ-PGA synthesis in B. licheniformis WX-02. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  19. Chitosanase purified from bacterial isolate Bacillus licheniformis of ruined vegetables displays broad spectrum biofilm inhibition.

    Science.gov (United States)

    Muslim, Sahira Nsayef; Al-Kadmy, Israa M S; Hussein, Nadheema Hammood; Mohammed Ali, Alaa Naseer; Taha, Buthainah Mohammed; Aziz, Sarah Naji; Kheraif, Abdulaziz Abdullah Al; Divakar, Darshan Devang; Ramakrishnaiah, Ravikumar

    2016-11-01

    A number of bacterial species produces chitosanases which has variety of applications because of its high biodegradability, non-toxicity and antimicrobial assets. In the present study chitosanase is purified from new bacterial species Bacillus licheniformis from spoiled vegetable. This novel strain of Bacillus licheniformis isolated from spoilt cucumber and pepper samples has the ability to produce the chitosanase enzyme when grown on chitosan substrate. Study also examined its antibiofilm properties against diverse bacterial species with biofilm forming ability. The purified chitosanase inhibited the biofilm formation ability for all Gram-negative and Gram-positive biofilm-forming bacteria [biofilm producers] tested in this study in congo red agar and microtiter plate's methods. Highly antibiofilm activity of chitosanase was recorded against Pseudomonas aeruginosa followed by Klebsiella pneumoniae with reduction of biofilm formation upto 22 and 29%, respectively compared with [100] % of control. Biofilm formation has multiple role including ability to enhance resistance and self-protection from external stress. This chitosanase has promising benefit as antibiofilm agent against biofilm forming pathogenic bacteria and has promising application as alternative antibiofilm agents to combat the growing number of multidrug resistant pathogen-associated infections, especially in situation where biofilms are involved. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Structural characterization of Bacillus licheniformis Dahb1 exopolysaccharide-antimicrobial potential and larvicidal activity on malaria and Zika virus mosquito vectors.

    Science.gov (United States)

    Abinaya, Muthukumar; Vaseeharan, Baskaralingam; Divya, Mani; Vijayakumar, Sekar; Govindarajan, Marimuthu; Alharbi, Naiyf S; Khaled, Jamal M; Al-Anbr, Mohammed N; Benelli, Giovanni

    2018-04-27

    Microbial polysaccharides produced by marine species play a key role in food and cosmetic industry, as they are nontoxic and biodegradable polymers. This investigation reports the isolation of exopolysaccharide from Bacillus licheniformis Dahb1 and its biomedical applications. Bacillus licheniformis Dahb1 exopolysaccharide (Bl-EPS) was extracted using the ethanol precipitation method and structurally characterized. FTIR and 1 H-NMR pointed out the presence of various functional groups and primary aromatic compounds, respectively. Bl-EPS exhibited strong antioxidant potential confirmed via DPPH radical, reducing power and superoxide anion scavenging assays. Microscopic analysis revealed that the antibiofilm activity of Bl-EPS (75 μg/ml) was higher against Gram-negative (Pseudomonas aeruginosa and Proteus vulgaris) bacteria over Gram-positive species (Bacillus subtilis and Bacillus pumilus). Bl-EPS led to biofilm inhibition against Candida albicans when tested at 75 μg/ml. The hemolytic assay showed low cytotoxicity of Bl-EPS at 5 mg/ml. Besides, Bl-EPS achieved LC 50 values < 80 μg/ml against larvae of mosquito vectors Anopheles stephensi and Aedes aegypti. Overall, our findings pointed out the multipurpose bioactivity of Bl-EPS, which deserves further consideration for pharmaceutical, environmental and entomological applications.

  1. The difference in in vivo sensitivity between Bacillus licheniformis PerR and Bacillus subtilis PerR is due to the different cellular environments.

    Science.gov (United States)

    Kim, Jung-Hoon; Won, Young-Bin; Ji, Chang-Jun; Yang, Yoon-Mo; Ryu, Su-Hyun; Ju, Shin-Yeong; Kwon, Yumi; Lee, Yeh-Eun; Lee, Jin-Won

    2017-02-26

    PerR, a member of Fur family of metal-dependent regulators, is a major peroxide sensor in many Gram positive bacteria, and controls the expression of genes involved in peroxide resistance. Bacillus licheniformis, a close relative to the well-studied model organism Bacillus subtilis, contains three PerR-like proteins (PerR BL , PerR2 and PerR3) in addition to Fur and Zur. In the present study, we characterized the role of PerR BL in B. licheniformis. In vitro and in vivo studies indicate that PerR BL , like PerR BS , uses either Fe 2+ or Mn 2+ as a corepressor and only the Fe 2+ -bound form of PerR BL senses low levels of H 2 O 2 by iron-mediated histidine oxidation. Interestingly, regardless of the difference in H 2 O 2 sensitivity, if any, between PerR BL and PerR BS , B. licheniformis expressing PerR BL or PerR BS could sense lower levels of H 2 O 2 and was more sensitive to H 2 O 2 than B. subtilis expressing PerR BL or PerR BS . This result suggests that the differences in cellular milieu between B. subtilis and B. licheniformis, rather than the intrinsic differences in PerR BS and PerR BL per se, affect the H 2 O 2 sensing ability of PerR inside the cell and the H 2 O 2 resistance of cell. In contrast, B. licheniformis and B. subtilis expressing Staphylococcus aureus PerR (PerR SA ), which is more sensitive to H 2 O 2 than PerR BL and PerR BS , were more resistant to H 2 O 2 than those expressing either PerR BL or PerR BS . This result indicates that the sufficient difference in H 2 O 2 susceptibility of PerR proteins can override the difference in cellular environment and affect the resistance of cell to H 2 O 2 . Copyright © 2017 Elsevier Inc. All rights reserved.

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

    OpenAIRE

    Bravo Rodríguez,Vicente; Jurado Alameda,Encarnación; Martínez Gallegos,Juan Francisco; Reyes Requena,Antonia; García López,Ana Isabel; Sampaio Cabral,Joaquim Manuel; Fernandes,Pedro; Pina da Fonseca,Luis Joaquim

    2006-01-01

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

  3. Proteomic profiling of Bacillus licheniformis reveals a stress response mechanism in the synthesis of extracellular polymeric flocculants.

    Science.gov (United States)

    Yu, Wencheng; Chen, Zhen; Shen, Liang; Wang, Yuanpeng; Li, Qingbiao; Yan, Shan; Zhong, Chuan-Jian; He, Ning

    2016-04-01

    Some bioflocculants composed of extracellular polymeric substances are produced under peculiar conditions. Bacillus licheniformis CGMCC2876 is a microorganism that secretes both extracellular polysaccharides (EPS) and poly-gamma-glutamic acid (γ-PGA) under stress conditions. In this work, SWATH acquisition LC-MS/MS method was adopted for differential proteomic analysis of B. licheniformis, aiming at determining the bacterial stress mechanism. Compared with LB culture, 190 differentially expressed proteins were identified in B. licheniformis CGMCC2876 cultivated in EPS culture, including 117 up-regulated and 73 down-regulated proteins. In γ-PGA culture, 151 differentially expressed proteins, 89 up-regulated and 62 down-regulated, were found in the cells. Up-regulated proteins involved in amino acid biosynthesis were found to account for 43% and 41% of the proteomes in EPS and γ-PGA cultivated cells, respectively. Additionally, a series of proteins associated with amino acid degradation were found to be repressed under EPS and γ-PGA culture conditions. Transcriptional profiling via the qPCR detection of selected genes verified the proteomic analysis. Analysis of free amino acids in the bacterial cells further suggested the presence of amino acid starvation conditions. EPS or γ-PGA was synthesized to alleviate the effect of amino acid limitation in B. licheniformis. This study identified a stress response mechanism in the synthesis of macromolecules in B. licheniformis, providing potential culture strategies to improve the production of two promising bioflocculants. © 2015 Wiley Periodicals, Inc.

  4. Kinetic and equilibrium study of uranium(VI) adsorption by Bacillus licheniformis

    International Nuclear Information System (INIS)

    Zheng-ji Yi; University of Science and Technology Beijing, Beijing; Jun Yao

    2012-01-01

    Uranium pollution is a severe problem worldwide. Biosorption has been proposed as one of the most promising technologies for the removal of uranyl cations. Here we report on the adsorption behavior of uranium(VI) [U(VI)] on Bacillus licheniformis biomass to explore the potentiality of its application in uranium contamination control. The adsorption equilibrium, adsorption kinetics, and effects of temperature, pH and initial biosorbent dosage on the adsorption equilibrium were investigated in detail through batch experiments. The adsorption process is pronouncedly affected by the solution pH and the optimum pH range should be 4.5-5.0.Temperature range from 25 to 45 deg C has a certain effect on the rate of biosorption, but little effect on the equilibrium adsorption capacity. The U(VI) percentage removal increased concurrently with increasing biomass dosage, whereas the adsorption capacity decreased. The process follows the Langmuir isotherm model. The adsorption kinetics data were fitted very well by the pseudo-first-order rate model. Finally, the calculation results of thermodynamic constant (ΔG a = 9.98 kJ/mol) reveal that the adsorption process can be identified as a spontaneous chemical process. The present results suggest that B. licheniformis has considerable potential for the removal of uranyl from aqueous solution. (author)

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

    Science.gov (United States)

    Gong, Jin-Song; Li, Wei; Zhang, Dan-Dan; Xie, Min-Feng; Yang, Biao; Zhang, Rong-Xian; Li, Heng; Lu, Zhen-Ming; Xu, Zheng-Hong; Shi, Jin-Song

    2015-12-17

    In the present study, we isolated a trypsin-producing strain DMN6 from the leather waste and identified it as Bacillus licheniformis through a two-step screening strategy. The trypsin activity was increased up to 140 from 20 U/mL through culture optimization. The enzyme was purified to electrophoretic homogeneity with a molecular mass of 44 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the specific activity of purified enzyme is 350 U/mg with Nα-Benzoyl-L-arginine ethylester as the substrate. The optimum temperature and pH for the trypsin are 65 °C and pH 9.0, respectively. Also, the enzyme can be significantly activated by Ba(2+). This enzyme is relatively stable in alkaline environment and displays excellent activity at low temperatures. It could retain over 95% of enzyme activity after 180 min of incubation at 45 °C. The distinguished activity under low temperature and prominent stability enhance its catalytic potential. In the current work, the open reading frame was obtained with a length of 1371 nucleotides that encoded a protein of 456 amino acids. These data would warrant the B. licheniformis trypsin as a promising candidate for catalytic application in collagen preparation and leather bating through further protein engineering.

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

    Directory of Open Access Journals (Sweden)

    Jin-Song Gong

    2015-12-01

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

  7. Effect of amino acids on tannase biosynthesis by Bacillus licheniformis KBR6.

    Science.gov (United States)

    Mohapatra, Pradeep K Das; Pati, Bikas R; Mondal, Keshab C

    2009-04-01

    Microbial tannase (tannin acyl hydrolase, EC 3.1.1.20), a hydrolysable tannin-degrading enzyme, has gained importance in various industrial processes, and is used extensively in the manufacture of instant tea, beer, wine, and gallic acid. Tannase is an inducible enzyme, and hydrolysable tannin, especially tannic acid, is the sole inducer. This study is of the effect of various amino acids and their analogues on tannase biosynthesis by Bacillus licheniformis KBR6 to ascertain the mode of action of these growth factors on tannase biosynthesis from microbial origin. Enzyme production was carried out in enriched tannic acid medium through submerged fermentation for 20 h at 35 degrees C. Different amino acids at a concentration of 0.05 g% (w/v) were added to the culture medium immediately after sterilization. Culture supernatant was used as the source of the enzyme and the quantity of tannase was estimated by the colorimetric assay method. Growth of the organism was estimated according to biomass dry weight. Maximum tannase (2.87-fold that of the control) was synthesized by B. licheniformis KBR6 when alanine was added to the culture medium. Other amino acids, such as DL-serine, L-cystine, glycine, L-ornithine, aspartic acid, L-glutamic acid, DL-valine, L-leucine and L-lysine, also induced tannase synthesis. L-Cysteine monohydrochloride and DL-threonine were the most potent inhibitors. Regulation of tannase biosynthesis by B. licheniformis in the presence of various amino acids is shown. This information will be helpful for formulating an enriched culture medium for industrial-scale tannase production.

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

    Science.gov (United States)

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

    2014-10-01

    In this study, a thermophilic Bacillus licheniformis strain X10 was newly isolated for 2,3-butanediol (2,3-BD) production from lignocellulosic hydrolysate. Strain X10 could utilize glucose and xylose simultaneously without carbon catabolite repression. In addition, strain X10 possesses high tolerance to fermentation inhibitors including furfural, vanillin, formic acid, and acetic acid. In a fed-batch fermentation, 74.0g/L of 2,3-BD was obtained from corn stover hydrolysate, with a productivity of 2.1g/Lh and a yield of 94.6%. Thus, this thermophilic B. licheniformis strain is a candidate for the development of efficient industrial production of 2,3-BD from corn stover hydrolysate. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Effects of Bacillus licheniformis on the growth performance and expression of lipid metabolism-related genes in broiler chickens challenged with Clostridium perfringens-induced necrotic enteritis.

    Science.gov (United States)

    Zhou, Mengjia; Zeng, Dong; Ni, Xueqin; Tu, Teng; Yin, Zhongqiong; Pan, Kangcheng; Jing, Bo

    2016-03-08

    Necrotic enteritis (NE), caused by Clostridium perfringens, has cost the poultry industry $2 billion in losses. This study aimed to investigate the effect of Bacillus licheniformis as dietary supplement on the growth, serum antioxidant status, and expression of lipid-metabolism genes of broiler chickens with C. perfringens-induced NE. A total of 240 one-day-old broilers were randomly grouped into four: a negative control, an NE experimental model (PC), chickens fed a diet supplemented with 30 % of fishmeal from day 14 onwards and challenged with coccidiosis vaccine (FC), and NE group supplied with feed containing 1.0 × 10(6) CFU/g B. licheniformis (BL). Body weight gain, feed conversion ratio, serum antioxidant status, and lipid-metabolism-gene expression were analyzed. In the PC group, FCR increased significantly whereas serum catalase and glutathione peroxidase activity decreased compared with NC group. Dietary B. licheniformis supplementation improved FCR and oxidative stress in experimental avian NE. Using Bacillus licheniformis as a direct-fed microbial (DFM) could also significantly upregulate catabolism-related genes, namely, peroxisome proliferator-activated receptor-α and carnitine palmitoyltransferase-1, in livers and changed the expression of lipid-anabolism genes. These results suggested that dietary B. licheniformis supplementation can enhance growth and antioxidant ability, as well as change the expression of genes related to fatty-acid synthesis and oxidation in the livers of NE-infected broilers.

  10. Effect of glucose on poly-γ-glutamic acid metabolism in Bacillus licheniformis.

    Science.gov (United States)

    Yu, Wencheng; Chen, Zhen; Ye, Hong; Liu, Peize; Li, Zhipeng; Wang, Yuanpeng; Li, Qingbiao; Yan, Shan; Zhong, Chuan-Jian; He, Ning

    2017-02-08

    Poly-gamma-glutamic acid (γ-PGA) is a promising macromolecule with potential as a replacement for chemosynthetic polymers. γ-PGA can be produced by many microorganisms, including Bacillus species. Bacillus licheniformis CGMCC2876 secretes γ-PGA when using glycerol and trisodium citrate as its optimal carbon sources and secretes polysaccharides when using glucose as the sole carbon source. To better understand the metabolic mechanism underlying the secretion of polymeric substances, SWATH was applied to investigate the effect of glucose on the production of polysaccharides and γ-PGA at the proteome level. The addition of glucose at 5 or 10 g/L of glucose decreased the γ-PGA concentration by 31.54 or 61.62%, respectively, whereas the polysaccharide concentration increased from 5.2 to 43.47%. Several proteins playing related roles in γ-PGA and polysaccharide synthesis were identified using the SWATH acquisition LC-MS/MS method. CcpA and CcpN co-enhanced glycolysis and suppressed carbon flux into the TCA cycle, consequently slowing glutamic acid synthesis. On the other hand, CcpN cut off the carbon flux from glycerol metabolism and further reduced γ-PGA production. CcpA activated a series of operons (glm and epsA-O) to reallocate the carbon flux to polysaccharide synthesis when glucose was present. The production of γ-PGA was influenced by NrgB, which converted the major nitrogen metabolic flux between NH 4 + and glutamate. The mechanism by which B. licheniformis regulates two macromolecules was proposed for the first time in this paper. This genetic information will facilitate the engineering of bacteria for practicable strategies for the fermentation of γ-PGA and polysaccharides for diverse applications.

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

    Science.gov (United States)

    Lu, Tao

    2009-07-01

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

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

    Directory of Open Access Journals (Sweden)

    Jian-Rong Wang

    2015-01-01

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

  13. Efficient Simultaneous Saccharification and Fermentation of Inulin to 2,3-Butanediol by Thermophilic Bacillus licheniformis ATCC 14580

    Science.gov (United States)

    Li, Lixiang; Chen, Chao; Li, Kun; Wang, Yu; Gao, Chao; Ma, Cuiqing

    2014-01-01

    2,3-Butanediol (2,3-BD) is an important starting material for the manufacture of bulk chemicals. For efficient and large-scale production of 2,3-BD through fermentation, low-cost substrates are required. One such substrate, inulin, is a polydisperse fructan found in a wide variety of plants. In this study, a levanase with high inulinase activity and high pH and temperature stability was identified in Bacillus licheniformis strain ATCC 14580. B. licheniformis strain ATCC 14580 was found to efficiently produce 2,3-BD from fructose at 50°C. Then, the levanase was used for simultaneous saccharification and fermentation (SSF) of inulin to 2,3-BD. A fed-batch SSF yielded 103.0 g/liter 2,3-BD in 30 h, with a high productivity of 3.4 g/liter · h. The results suggest that the SSF process developed with the thermophilic B. licheniformis strain used might be a promising alternative for efficient 2,3-BD production from the favorable substrate inulin. PMID:25107977

  14. Bacillus licheniformis affects the microbial community and metabolic profile in the spontaneous fermentation of Daqu starter for Chinese liquor making.

    Science.gov (United States)

    Wang, Peng; Wu, Qun; Jiang, Xuejian; Wang, Zhiqiang; Tang, Jingli; Xu, Yan

    2017-06-05

    Chinese liquor is produced from spontaneous fermentation starter (Daqu) that provides the microbes, enzymes and flavors for liquor fermentation. To improve the flavor character of Daqu, we inoculated Bacillus licheniformis and studied the effect of this strain on the community structure and metabolic profile in Daqu fermentation. The microbial relative abundance changed after the inoculation, including the increase in Bacillus, Clavispora and Aspergillus, and the decrease in Pichia, Saccharomycopsis and some other genera. This variation was also confirmed by pure culture and coculture experiments. Seventy-three metabolites were identified during Daqu fermentation process. After inoculation, the average content of aromatic compounds were significantly enriched from 0.37mg/kg to 0.90mg/kg, and the average content of pyrazines significantly increased from 0.35mg/kg to 5.71mg/kg. The increase in pyrazines was positively associated with the metabolism of the inoculated Bacillus and the native genus Clavispora, because they produced much more pyrazines in their cocultures. Whereas the increase in aromatic compounds might be related to the change of in situ metabolic activity of several native genera, in particular, Aspergillus produced more aromatic compounds in cocultures with B. licheniformis. It indicated that the inoculation of B. licheniformis altered the flavor character of Daqu by both its own metabolic activity and the variation of in situ metabolic activity. Moreover, B. licheniformis inoculation influenced the enzyme activity of Daqu, including the significant increase in amylase activity (from 1.3gstarch/g/h to 1.7gstarch/g/h), and the significant decrease in glucoamylase activity (from 627.6mgglucose/g/h to 445.6mgglucose/g/h) and esterase activity (from 28.1mgethylcaproate/g/100h to 17.2mgethylcaproate/g/100h). These effects of inoculation were important factors for regulating the metabolism of microbial communities, hence for improving the flavor profile

  15. Improving the reversibility of thermal denaturation and catalytic efficiency of Bacillus licheniformis α-amylase through stabilizing a long loop in domain B.

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    Zhu Li

    Full Text Available The reversibility of thermal denaturation and catalytic efficiency of Bacillus licheniformis α-amylase were improved through site-directed mutagenesis. By using multiple sequence alignment and PoPMuSiC algorithm, Ser187 and Asn188, which located within a long loop in Domain B of Bacillus licheniformis α-amylase, were selected for mutation. In addition, Ala269, which is adjacent to Ser187 and Asn188, was also investigated. Seven mutants carrying the mutations S187D, N188T, N188S, A269K, A269K/S187D, S187D/N188T, and A269K/S187D/N188T were generated and characterized. The most thermostable mutant, A269K/S187D/N188T, exhibited a 9-fold improvement in half-life at 95°C and pH 5.5, compared with that of the wild-type enzyme. Mutant A269K/S187D/N188T also exhibited improved catalytic efficiency. The catalytic efficiency of mutant A269K/S187D/N188T reached 5.87×103±0.17 g·L-1·s-1 at pH 5.5, which is 1.84-fold larger than the corresponding value determined for the wild-type enzyme. Furthermore, the structure analysis showed that immobilization of the loop containing Ser187 and Asn188 plays a significant role in developing the properties of Bacillus licheniformis α-amylase.

  16. Improving the reversibility of thermal denaturation and catalytic efficiency of Bacillus licheniformis α-amylase through stabilizing a long loop in domain B.

    Science.gov (United States)

    Li, Zhu; Duan, Xuguo; Chen, Sheng; Wu, Jing

    2017-01-01

    The reversibility of thermal denaturation and catalytic efficiency of Bacillus licheniformis α-amylase were improved through site-directed mutagenesis. By using multiple sequence alignment and PoPMuSiC algorithm, Ser187 and Asn188, which located within a long loop in Domain B of Bacillus licheniformis α-amylase, were selected for mutation. In addition, Ala269, which is adjacent to Ser187 and Asn188, was also investigated. Seven mutants carrying the mutations S187D, N188T, N188S, A269K, A269K/S187D, S187D/N188T, and A269K/S187D/N188T were generated and characterized. The most thermostable mutant, A269K/S187D/N188T, exhibited a 9-fold improvement in half-life at 95°C and pH 5.5, compared with that of the wild-type enzyme. Mutant A269K/S187D/N188T also exhibited improved catalytic efficiency. The catalytic efficiency of mutant A269K/S187D/N188T reached 5.87×103±0.17 g·L-1·s-1 at pH 5.5, which is 1.84-fold larger than the corresponding value determined for the wild-type enzyme. Furthermore, the structure analysis showed that immobilization of the loop containing Ser187 and Asn188 plays a significant role in developing the properties of Bacillus licheniformis α-amylase.

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

    Science.gov (United States)

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

    2013-11-01

    This study was conducted to investigate the effects of fructooligosaccharide (FOS) and Bacillus licheniformis (B. licheniformis) and their interaction on innate immunity, antioxidant capability and disease resistance of triangular bream Megalobrama terminalis (average initial weight 30.5 ± 0.5 g). Nine experimental diets were formulated to contain three FOS levels (0, 0.3% and 0.6%) and three B. licheniformis levels (0, 1 × 10(7), 5 × 10(7) CFU g(-1)) according to a 3 × 3 factorial design. At the end of the 8-week feeding trial, fish were challenged by Aeromonas hydrophila (A. hydrophila) and survival rate was recorded for the next 7 days. The results showed that leucocyte counts, alternative complement activity as well as total serum protein and globulin contents all increased significantly (P licheniformis levels increased from 0 to 1 × 10(7) CFU g(-1), while little difference (P > 0.05) was observed in these parameters in terms of dietary FOS levels. Both plasma alkaline phosphatase and phenoloxidase activities were significantly (P 0.05) by both FOS and B. licheniformis. Liver catalase, glutathione peroxidase as well as plasma SOD activities of fish fed 1 × 10(7) CFU g(-1)B. licheniformis were all significantly (P 0.05) by either FOS levels or B. licheniformis contents, whereas a significant (P licheniformis. The results of this study indicated that dietary FOS and B. licheniformis could significantly enhance the innate immunity and antioxidant capability of triangular bream, as well as improve its disease resistance. The best combination of these two prebiotics and/or probiotics was 0.3% FOS and 1 × 10(7) CFU g(-1)B. licheniformis. Copyright © 2013 Elsevier Ltd. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Sreethar Swaathy

    2014-12-01

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

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

    International Nuclear Information System (INIS)

    Ikram-ul-Haq, M.; Javed, M.M.; Hameed, U.; Adnan, F.

    2010-01-01

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

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

    Science.gov (United States)

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

    2016-04-01

    Poly(γ-glutamic acid) (PGA) is a polymer composed of L- and/or D-glutamic acids that is produced by Bacillus sp. Because the polymer has various features as water soluble, edible, non-toxic and so on, it has attracted attention as a candidate for many applications such as foods, cosmetics and so on. However, although it is well known that the intracellular metabolism of Bacillus sp. is mainly regulated by catabolite control, the effect of the catabolite control on the PGA producing Bacillus sp. is largely unknown. This study is the first report of metabolome analysis on the PGA producing Bacillus sp. that reveals the effect of carbon catabolite control on the metabolism of PGA producing Bacillus licheniformis ATCC 9945. Results showed that the cells cultivated in glycerol-containing medium showed higher PGA production than the cells in glucose-containing medium. Furthermore, metabolome analysis revealed that the activators of CcpA and CodY, global regulatory proteins of the intracellular metabolism, accumulated in the cells cultivated in glycerol-containing and glucose-containing medium, respectively, with CodY apparently inhibiting PGA production. Moreover, the cells seemed to produce glutamate from citrate and ammonium using glutamine synthetase/glutamate synthase. Pulsed addition of di-ammonium hydrogen citrate, as suggested by the metabolome result, was able to achieve the highest value so far for PGA production in B. licheniformis. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. Growth-Rate Dependent Regulation of tRNA Level and Charging in Bacillus licheniformis.

    Science.gov (United States)

    Ferro, Iolanda; Liebeton, Klaus; Ignatova, Zoya

    2017-10-13

    Cellular growth crucially depends on protein synthesis and the abundance of translational components. Among them, aminoacyl-tRNAs play a central role in biosynthesis and shape the kinetics of mRNA translation, thus influencing protein production. Here, we used microarray-based approaches to determine the charging levels and tRNA abundance of Bacillus licheniformis. We observed an interesting cross-talk among tRNA expression, charging pattern, and growth rate. For a large subset of tRNAs, we found a co-regulated and augmented expression at high growth rate. Their tRNA aminoacylation level is kept relatively constant through riboswitch-regulated expression of the cognate aminoacyl-tRNA-synthetase (AARS). We show that AARSs with putative riboswitch-controlled expression are those charging tRNAs with amino acids which disfavor cell growth when individually added to the nutrient medium. Our results suggest that the riboswitch-regulated AARS expression in B. licheniformis is a powerful mechanism not only to maintain a constant ratio of aminoacyl-tRNA independent of the growth rate but concomitantly to control the intracellular level of free amino acids. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Modeling heat resistance of Bacillus weihenstephanensis and Bacillus licheniformis spores as function of sporulation temperature and pH.

    Science.gov (United States)

    Baril, Eugénie; Coroller, Louis; Couvert, Olivier; Leguérinel, Ivan; Postollec, Florence; Boulais, Christophe; Carlin, Frédéric; Mafart, Pierre

    2012-05-01

    Although sporulation environmental factors are known to impact on Bacillus spore heat resistance, they are not integrated into predictive models used to calculate the efficiency of heating processes. This work reports the influence of temperature and pH encountered during sporulation on heat resistance of Bacillus weihenstephanensis KBAB4 and Bacillus licheniformis AD978 spores. A decrease in heat resistance (δ) was observed for spores produced either at low temperature, at high temperature or at acidic pH. Sporulation temperature and pH maximizing the spore heat resistance were identified. Heat sensitivity (z) was not modified whatever the sporulation environmental factors were. A resistance secondary model inspired by the Rosso model was proposed. Sporulation temperatures and pHs minimizing or maximizing the spore heat resistance (T(min(R)), T(opt(R)), T(max(R)), pH(min(R)) and pH(opt(R))) were estimated. The goodness of the model fit was assessed for both studied strains and literature data. The estimation of the sporulation temperature and pH maximizing the spore heat resistance is of great interest to produce spores assessing the spore inactivation in the heating processes applied by the food industry. Copyright © 2011 Elsevier Ltd. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Ratu SAFITRI

    2015-10-01

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

  4. Improving flavor metabolism of Saccharomyces cerevisiae by mixed culture with Bacillus licheniformis for Chinese Maotai-flavor liquor making.

    Science.gov (United States)

    Meng, Xing; Wu, Qun; Wang, Li; Wang, Diqiang; Chen, Liangqiang; Xu, Yan

    2015-12-01

    Microbial interactions could impact the metabolic behavior of microbes involved in food fermentation, and therefore they are important for improving food quality. This study investigated the effect of Bacillus licheniformis, the dominant bacteria in the fermentation process of Chinese Maotai-flavor liquor, on the metabolic activity of Saccharomyces cerevisiae. Results indicated that S. cerevisiae inhibited the growth of B. licheniformis in all mixed culture systems and final viable cell count was lower than 20 cfu/mL. Although growth of S. cerevisiae was barely influenced by B. licheniformis, its metabolism was changed as initial inoculation ratio varied. The maximum ethanol productions were observed in S. cerevisiae and B. licheniformis at 10(6):10(7) and 10(6):10(8) ratios and have increased by 16.8 % compared with single culture of S. cerevisiae. According to flavor compounds, the culture ratio 10(6):10(6) showed the highest level of total concentrations of all different kinds of flavor compounds. Correlation analyses showed that 12 flavor compounds, including 4 fatty acids and their 2 corresponding esters, 1 terpene, and 5 aromatic compounds, that could only be produced by S. cerevisiae were significantly correlated with the initial inoculation amount of B. licheniformis. These metabolic changes in S. cerevisiae were not only a benefit for liquor aroma, but may also be related to its inhibition effect in mixed culture. This study could help to reveal the microbial interactions in Chinese liquor fermentation and provide guidance for optimal arrangement of mixed culture fermentation systems.

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

    Science.gov (United States)

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

    2015-01-01

    Naturally stressed habitats are known to be repositories for novel microorganisms with potential bioremediation applications. In this study, we isolated a [Co(III)-EDTA](-) reducing bacterium Bacillus licheniformis SPB-2 from a solar salt pan that is exposed to constant cycles of hydration and desiccation in nature. [Co(III)-EDTA](-) generated during nuclear waste management process is difficult to remove from the waste due to its high stability and solubility. It is reduced form i.e. [Co(II)-EDTA](2-) is less stable though it is toxic. This study showed that B. licheniformis SPB-2 reduced 1mM [Co(III)-EDTA](-) in 14 days when grown in a batch mode. However, subsequent cycles showed an increase in the reduction activity, which was observed up to four cycles. Interestingly, the present study also showed that [Co(III)-EDTA](-) acted as an inducer for B. licheniformis SPB-2 spore germination. Vegetative cells germinated from the spores were found to be involved in [Co(III)-EDTA](-) reduction. More detailed investigations showed that after [Co(III)-EDTA](-) reduction, i.e. [Co(II)-EDTA](2-) complex was removed by B. licheniformis SPB-2 from the bulk liquid by adsorption phenomenon. The bacterium showed a D10 value (radiation dose required to kill 90% cells) of ∼250 Gray (Gy), which signifies the potential use of B. licheniformis SPB-2 for bioremediation of moderately active nuclear waste. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. A pathway closely related to the (D)-tagatose pathway of gram-negative enterobacteria identified in the gram-positive bacterium Bacillus licheniformis.

    Science.gov (United States)

    Van der Heiden, Edwige; Delmarcelle, Michaël; Lebrun, Sarah; Freichels, Régine; Brans, Alain; Vastenavond, Christian M; Galleni, Moreno; Joris, Bernard

    2013-06-01

    We report the first identification of a gene cluster involved in d-tagatose catabolism in Bacillus licheniformis. The pathway is closely related to the d-tagatose pathway of the Gram-negative bacterium Klebsiella oxytoca, in contrast to the d-tagatose 6-phosphate pathway described in the Gram-positive bacterium Staphylococcus aureus.

  7. A Pathway Closely Related to the d-Tagatose Pathway of Gram-Negative Enterobacteria Identified in the Gram-Positive Bacterium Bacillus licheniformis

    OpenAIRE

    Van der Heiden, Edwige; Delmarcelle, Michaël; Lebrun, Sarah; Freichels, Régine; Brans, Alain; Vastenavond, Christian M.; Galleni, Moreno; Joris, Bernard

    2013-01-01

    We report the first identification of a gene cluster involved in d-tagatose catabolism in Bacillus licheniformis. The pathway is closely related to the d-tagatose pathway of the Gram-negative bacterium Klebsiella oxytoca, in contrast to the d-tagatose 6-phosphate pathway described in the Gram-positive bacterium Staphylococcus aureus.

  8. Heterogeneous nucleation helps the search for initial crystallization conditions of γ-glutamyl transpeptidase from Bacillus licheniformis

    International Nuclear Information System (INIS)

    Lin, Long-Liu; Merlino, Antonello

    2013-01-01

    An additional example in which heterogeneous nucleation has helped in the search for crystallization conditions of a protein is reported. Optimization of the crystallization conditions led to the formation of single crystals of γ-glutamyl transpeptidase from B. licheniformis that diffracted to about 3.0 Å resolution. Here, the crystallization and preliminary X-ray diffraction studies of Bacillus licheniformis γ-glutamyl transpeptidase (BlGT) are reported. The serendipitous finding of heterogeneous nucleants in the initial experiments provided the first crystallization conditions for the protein. Crystals were grown by hanging-drop vapour diffusion using a precipitant solution consisting of 20%(w/v) PEG 3350, 0.2 M magnesium chloride hexahydrate, 0.1 M Tris–HCl pH 8.2. The protein crystallized in the orthorhombic space group P2 1 2 1 2 1 , with one heterodimer per asymmetric unit and unit-cell parameters a = 60.90, b = 61.97, c = 148.24 Å. The BlGT crystals diffracted to 2.95 Å resolution

  9. Subcellular localization of alkaline phosphatase in Bacillus licheniformis 749/C by immunoelectron microscopy with colloidal gold

    International Nuclear Information System (INIS)

    Tinglu, G.; Ghosh, A.; Ghosh, B.K.

    1984-01-01

    Subcellular distribution of the alkaline phosphatase of Bacillus licheniformis 749/C was determined by an immunoelectron microscopy method. Anti-alkaline phosphatase antibody labeled with 15- to 18-nm colloidal gold particles (gold-immunoglobulin G [IgG] complex) were used for the study. Both the plasma membrane and cytoplasmic material were labeled with the gold-IgG particles. These particles formed clusters in association with the plasma membrane; in contrast, in the cytoplasm the particles were largely dispersed, and only a few clusters were found. The gold-IgG binding was quantitatively estimated by stereological analysis of labeled, frozen thin sections. This estimation of a variety of control samples showed that the labeling was specific for the alkaline phosphatase. Cluster formation of the gold -IgG particles in association with the plasma membrane suggests that existence of specific alkaline phosphatase binding sites (receptors) in the plasma membrane of B. licheniformis 749/C. 27 references, 6 figures, 1 table

  10. A Pathway Closely Related to the d-Tagatose Pathway of Gram-Negative Enterobacteria Identified in the Gram-Positive Bacterium Bacillus licheniformis

    Science.gov (United States)

    Van der Heiden, Edwige; Lebrun, Sarah; Freichels, Régine; Brans, Alain; Vastenavond, Christian M.; Galleni, Moreno; Joris, Bernard

    2013-01-01

    We report the first identification of a gene cluster involved in d-tagatose catabolism in Bacillus licheniformis. The pathway is closely related to the d-tagatose pathway of the Gram-negative bacterium Klebsiella oxytoca, in contrast to the d-tagatose 6-phosphate pathway described in the Gram-positive bacterium Staphylococcus aureus. PMID:23524682

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

    Directory of Open Access Journals (Sweden)

    Paulo Roberto Martins Bonilha

    2006-09-01

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

  12. Effect of Degradation of Zearalenone-Contaminated Feed by Bacillus licheniformis CK1 on Postweaning Female Piglets

    Directory of Open Access Journals (Sweden)

    Guanhua Fu

    2016-10-01

    Full Text Available Zearalenone (ZEA, an estrogenic mycotoxin, is mainly produced by Fusarium fungi. In this study, Bacillus licheniformis CK1 isolated from soil with the capability of degrading ZEA was evaluated for its efficacy in reducing the adverse effects of ZEA in piglets. The gilts were fed one of the following three diets for 14 days: a basic diet for the control group; the basic diet supplemented with ZEA-contaminated basic diet for the treatment 1 (T1 group; and the basic diet supplemented with fermented ZEA-contaminated basic diet by CK1 for the treatment 2 (T2 group. The actual ZEA contents (analyzed were 0, 1.20 ± 0.11, 0.47 ± 0.22 mg/kg for the control, T1, and T2 diets, respectively. The results showed that the T1 group had significantly increased the size of vulva and the relative weight of reproductive organs compared to the control group at the end of the trial. The T1 group significantly decreased the concentration of the luteinizing hormone (LH compared with the control and T2 groups. Expression of ERβ was significantly up-regulated in the T2 group compared with the control. In addition, expression of ERβ was not different between the control and the T1 group. In summary, our results suggest that Bacillus licheniformis CK1 could detoxify ZEA in feed and reduce the adverse effects of ZEA in the gilts.

  13. Effect of Degradation of Zearalenone-Contaminated Feed by Bacillus licheniformis CK1 on Postweaning Female Piglets.

    Science.gov (United States)

    Fu, Guanhua; Ma, Junfei; Wang, Lihong; Yang, Xin; Liu, Jeruei; Zhao, Xin

    2016-10-17

    Zearalenone (ZEA), an estrogenic mycotoxin, is mainly produced by Fusarium fungi. In this study, Bacillus licheniformis CK1 isolated from soil with the capability of degrading ZEA was evaluated for its efficacy in reducing the adverse effects of ZEA in piglets. The gilts were fed one of the following three diets for 14 days: a basic diet for the control group; the basic diet supplemented with ZEA-contaminated basic diet for the treatment 1 (T1) group; and the basic diet supplemented with fermented ZEA-contaminated basic diet by CK1 for the treatment 2 (T2) group. The actual ZEA contents (analyzed) were 0, 1.20 ± 0.11, 0.47 ± 0.22 mg/kg for the control, T1, and T2 diets, respectively. The results showed that the T1 group had significantly increased the size of vulva and the relative weight of reproductive organs compared to the control group at the end of the trial. The T1 group significantly decreased the concentration of the luteinizing hormone (LH) compared with the control and T2 groups. Expression of ERβ was significantly up-regulated in the T2 group compared with the control. In addition, expression of ERβ was not different between the control and the T1 group. In summary, our results suggest that Bacillus licheniformis CK1 could detoxify ZEA in feed and reduce the adverse effects of ZEA in the gilts.

  14. The inability of Bacillus licheniformis perR mutant to grow is mainly due to the lack of PerR-mediated fur repression.

    Science.gov (United States)

    Kim, Jung-Hoon; Yang, Yoon-Mo; Ji, Chang-Jun; Ryu, Su-Hyun; Won, Young-Bin; Ju, Shin-Yeong; Kwon, Yumi; Lee, Yeh-Eun; Youn, Hwan; Lee, Jin-Won

    2017-06-01

    PerR, a member of Fur family protein, is a metal-dependent H 2 O 2 sensing transcription factor that regulates genes involved in peroxide stress response. Industrially important bacterium Bacillus licheniformis contains three PerR-like proteins (PerR BL , PerR2, and PerR3) compared to its close relative Bacillus subtilis. Interestingly, unlike other bacteria including B. subtilis, no authentic perR BL null mutant could be established for B. licheniformis. Thus, we constructed a conditional perR BL mutant using a xylose-inducible promoter, and investigated the genes under the control of PerR BL . PerR BL regulon genes include katA, mrgA, ahpC, pfeT, hemA, fur, and perR as observed for PerR BS . However, there is some variation in the expression levels of fur and hemA genes between B. subtilis and B. licheniformis in the derepressed state. Furthermore, katA, mrgA, and ahpC are strongly induced, whereas the others are only weakly or not induced by H 2 O 2 treatment. In contrast to the B. subtilis perR null mutant which frequently gives rise to large colony phenotype mainly due to the loss of katA, the suppressors of B. licheniformis perR mutant, which can form colonies on LB agar, were all catalase-positive. Instead, many of the suppressors showed increased levels of siderophore production, suggesting that the suppressor mutation is linked to the fur gene. Consistent with this, perR fur double mutant could grow on LB agar without Fe supplementation, whereas perR katA double mutant could only grow on LB agar with Fe supplementation. Taken together, our data suggest that in B. licheniformis, despite the similarity in PerR BL and PerR BS regulon genes, perR is an essential gene required for growth and that the inability of perR null mutant to grow is mainly due to elevated expression of Fur.

  15. Inducible expression of trehalose synthase in Bacillus licheniformis.

    Science.gov (United States)

    Li, Youran; Gu, Zhenghua; Zhang, Liang; Ding, Zhongyang; Shi, Guiyang

    2017-02-01

    Trehalose synthase (TreS) could transform maltose into trehalose via isomerization. It is a crucial enzyme in the process of trehalose enzymatical transformation. In this study, plasmid-based inducible expression systems were constructed to produce Thermomonospora curvata TreS in B. licheniformis. Xylose operons from B. subtilis, B. licheniformis and B. megaterium were introduced to regulate the expression of the gene encoding TreS. It was functionally expressed, and the BlsTs construct yielded the highest enzyme activity (12.1 U/mL). Furthermore, the effect of different cultural conditions on the inducible expression of BlsTs was investigated, and the optimal condition was as follows: 4% maltodextrin, 0.4% soybean powder, 1% xylose added after 10 h of growth and an induction time of 12 h at 37 °C. As a result, the maximal yield reached 24.7 U/mL. This study contributes to the industrial application of B. licheniformis, a GRAS workhorse for enzyme production. Copyright © 2016 Elsevier Inc. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Diana Sossa-Urrego

    2008-09-01

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

  17. Purification and characterization of a novel lichenase from Bacillus licheniformis GZ-2.

    Science.gov (United States)

    Gao, Zhen

    2016-01-01

    A novel lichenase from Bacillus licheniformis GZ-2 was purified to homogeneity by two steps ion-exchange chromatography with a specific activity of 8231.3 U/mg. The purified enzyme showed as a single protein band with a molecular mass of 25 kDa. The optimum pH and temperature for the enzyme activity were 6.5 and 60 °C, respectively. The enzyme exhibited strict specificity for β-1,3-1,4-d-glucans. The kinetic parameters Km and Vmax were 5.11 mg/mL and 2097 µmol/Min/mg for lichenan and 7.42 mg/mL and 1440 µmol/Min/mg for barley β-glucan. Compared to most of the reported β-1,3-1,4-glucanases (lichenase), the activity of the purified enzyme for lichenan was much higher than that for barley β-glucan. The main products of β-glucan hydrolyzed by the lichenase were cellubiosyltriose (DP3) and cellutriosyltraose (DP4). The lichenase gene from B. licheniformis GZ-2 was cloned and sequenced. The open reading frame of gene gz-2 contained 642 bp coding for a 214 amino acid mature protein. The gene was cloned into an expression vector pET 28a and expressed in Escherichia coli BL21. The activity in cell lysate supernatant was 137.9 U/mg. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

  18. The Poly-γ-D-Glutamic Acid Capsule of Bacillus licheniformis, a Surrogate of Bacillus anthracis Capsule Induces Interferon-Gamma Production in NK Cells through Interactions with Macrophages.

    Science.gov (United States)

    Lee, Hae-Ri; Jeon, Jun Ho; Rhie, Gi-Eun

    2017-05-28

    The poly-γ- D -glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis , provides protection of the bacterium from phagocytosis and allows its unimpeded growth in the host. We investigated crosstalk between murine natural killer (NK) cells and macrophages stimulated with the PGA capsule of Bacillus licheniformis , a surrogate of the B. anthracis capsule. PGA induced interferon-gamma production from NK cells cultured with macrophages. This effect was dependent on macrophage-derived IL-12 and cell-cell contact interaction with macrophages through NK cell receptor NKG2D and its ligand RAE-1. The results showed that PGA could enhance NK cell activation by inducing IL-12 production in macrophages and a contact-dependent crosstalk with macrophages.

  19. Heat and desiccation are the predominant factors affecting inactivation of Bacillus licheniformis and Bacillus thuringiensis spores during simulated composting.

    Science.gov (United States)

    Stanford, K; Harvey, A; Barbieri, R; Xu, S; Reuter, T; Amoako, K K; Selinger, L B; McAllister, T A

    2016-01-01

    The suitability of composting for disposal of livestock mortalities due to Bacillus anthracis was assessed by measuring viability of surrogate spores from two strains each of Bacillus licheniformis and Bacillus thuringiensis after a heating cycle modelled on a cattle composting study. Sporulation was attempted from 10 to 37°C, but poor yields at lower temperatures resulted in 25, 30 and 37°C being selected to generate sufficient spores (8 log10  CFU ml(-1) ) for experiments. Spores were inoculated into 3 g autoclaved dried-ground compost rehydrated with 6 ml water or silica beads in a factorial design for each strain, sporulation temperature, matrix and sampling day (0, 25, 50, 100, 150). Maximum incubation temperature was 62°C, but spores were maintained at ≥55°C for 78 of 150 days. Although significant differences existed among Bacillus strains and sporulation temperatures, numbers of viable spores after 150 days averaged 1·3 log10  CFU g(-1) , a 5·2 log10 reduction from day 0. Spore inactivation was likely due to heat and desiccation as matrices were autoclaved prior to incubation, negating impacts of microflora. Results support composting for disposal of anthrax mortalities, provided long-term thermophillic heating is achieved. Due to limited sporulation at 10°C, livestock mortalities from anthrax at this or lower ambient temperatures would likely be of lower risk for disease transmission. © 2015 The Society for Applied Microbiology.

  20. Deletion of meso-2,3-butanediol dehydrogenase gene budC for enhanced D-2,3-butanediol production in Bacillus licheniformis

    Science.gov (United States)

    2014-01-01

    Background D-2,3-butanediol has many industrial applications such as chiral reagents, solvents, anti-freeze agents, and low freezing point fuels. Traditional D-2,3-butanediol producing microorganisms, such as Klebsiella pneumonia and K. xoytoca, are pathogenic and not capable of producing D-2,3-butanediol at high optical purity. Bacillus licheniformis is a potential 2,3-butanediol producer but the wild type strain (WX-02) produces a mix of D- and meso-type isomers. BudC in B. licheniformis is annotated as 2,3-butanediol dehydrogenase or acetoin reductase, but no pervious experiment was performed to verify this hypothesis. Results We developed a genetically modified strain of B. licheniformis (WX-02 ΔbudC) as a D-2,3-butanediol producer with high optimal purity. A marker-less gene deletion protocol based on a temperature sensitive knock-out plasmid T2-Ori was used to knock out the budC gene in B. licheniformis WX-02. The budC knock-out strain successfully abolished meso-2,3-butanediol production with enhanced D-2,3-butanediol production. No meso-BDH activity was detectable in cells of this strain. On the other hand, the complementary strain restored the characteristics of wild strain, and produced meso-2,3-butanediol and possessed meso-BDH activity. All of these data suggested that budC encoded the major meso-BDH catalyzing the reversible reaction from acetoin to meso-2,3-butanediol in B. licheniformis. The budC knock-out strain produced D-2,3-butanediol isomer only with a high yield of 30.76 g/L and a productivity of 1.28 g/L-h. Conclusions We confirmed the hypothesis that budC gene is responsible to reversibly transfer acetoin to meso-2,3-butanediol in B. licheniformis. A mutant strain of B. licheniformis with depleted budC gene was successfully developed and produced high level of the D-2,3-butanediol with high optimal purity. PMID:24475980

  1. Production of tannase by the immobilized cells of Bacillus licheniformis KBR6 in Ca-alginate beads.

    Science.gov (United States)

    Mohapatra, P K D; Mondal, K C; Pati, B R

    2007-06-01

    The present study was aimed at finding the optimal conditions for immobilization of Bacillus licheniformis KBR6 cells in calcium-alginate (Ca-alginate) beads and determining the operational stability during the production of tannin-acyl-hydrolase (tannase) under semicontinous cultivation. The active cells of B. licheniformis KBR6 were immobilized in Ca-alginate and used for the production of tannase. The influence of alginate concentration (5, 10, 20 and 30 g l(-1)) and initial cell loading on enzyme production were studied. The production of tannase increased significantly with increasing alginate concentration and reached a maximum enzyme yield of 0.56 +/- 0.03 U ml(-1) at 20 g l(-1). This was about 1.70-fold higher than that obtained by free cells. The immobilized cells produced tannase consistently over 13 repeated cycles and reached a maximum level at the third cycle. Scanning electron microscope study indicated that the cells in Ca-alginate beads remain in normal shape. The Ca-alginate entrapment is a promising immobilization method of B. licheniformis KBR6 for repeated tannase production. Tannase production by immobilized cells is superior to that of free cells because it leads to higher volumetric activities within the same period of fermentation. This is the first report of tannase production from immobilized bacterial cells. The bacterium under study can produce higher amounts of tannase with respect to other fungal strains within a short cultivation period.

  2. In vitro antagonistic activity and the protective effect of probiotic Bacillus licheniformis Dahb1 in zebrafish challenged with GFP tagged Vibrio parahaemolyticus Dahv2.

    Science.gov (United States)

    Girija, Vairavan; Malaikozhundan, Balasubramanian; Vaseeharan, Baskaralingam; Vijayakumar, Sekar; Gobi, Narayanan; Del Valle Herrera, Marian; Chen, Jiann-Chu; Santhanam, Perumal

    2018-01-01

    In vitro antagonistic activity and the protective effect of probiotic Bacillus licheniformis Dahb1 in zebrafish (Danio rerio) challenged with GFP tagged Vibrio parahaemolyticus Dahv2 was studied. The cell free extract of probiotic B. licheniformis Dahb1 at 100 μg mL -1 showed growth inhibition of V. parahaemolyticus Dahv2 in vitro. B. licheniformis Dahb1 also inhibited the biofilm growth of GFP tagged V. parahaemolyticus Dahv2 at 100 μg mL -1 in vitro. The growth and survival of zebrafish was tested using probiotic B. licheniformis Dahb1. Weight (1.28 g) of zebrafish that received the cell free extract was much higher than in control (1.04 g). The mortality of zebrafish infected with GFP tagged V. parahaemolyticus Dahv2 at 10 7 Cfu mL -1 (Group IV) was 100%, whereas a complete survival of zebrafish that received the cell free extract of B. licheniformis Dahb1 at 10 7 Cfu mL -1 (Group VII) was observed after 30 days. The number of GFP tagged V. parahaemolyticus Dahv2 colonies in the intestine and gills significantly reduced after treatment with the cell free extract of B. licheniformis Dahb1. Furthermore, a significant decrease in the fluorescent colonies of GFP tagged V. parahaemolyticus Dahv2 was observed after treatment with the cell free extract of B. licheniformis Dahb1 under confocal laser scanning microscopy (CLSM). In conclusion, the cell free extract of B. licheniformis Dahb1 could prevent Vibrio infection by enhancing the growth and survival of zebrafish. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Genome-Based Identification of Active Prophage Regions by Next Generation Sequencing in Bacillus licheniformis DSM13

    Science.gov (United States)

    Hertel, Robert; Rodríguez, David Pintor; Hollensteiner, Jacqueline; Dietrich, Sascha; Leimbach, Andreas; Hoppert, Michael; Liesegang, Heiko; Volland, Sonja

    2015-01-01

    Prophages are viruses, which have integrated their genomes into the genome of a bacterial host. The status of the prophage genome can vary from fully intact with the potential to form infective particles to a remnant state where only a few phage genes persist. Prophages have impact on the properties of their host and are therefore of great interest for genomic research and strain design. Here we present a genome- and next generation sequencing (NGS)-based approach for identification and activity evaluation of prophage regions. Seven prophage or prophage-like regions were identified in the genome of Bacillus licheniformis DSM13. Six of these regions show similarity to members of the Siphoviridae phage family. The remaining region encodes the B. licheniformis orthologue of the PBSX prophage from Bacillus subtilis. Analysis of isolated phage particles (induced by mitomycin C) from the wild-type strain and prophage deletion mutant strains revealed activity of the prophage regions BLi_Pp2 (PBSX-like), BLi_Pp3 and BLi_Pp6. In contrast to BLi_Pp2 and BLi_Pp3, neither phage DNA nor phage particles of BLi_Pp6 could be visualized. However, the ability of prophage BLi_Pp6 to generate particles could be confirmed by sequencing of particle-protected DNA mapping to prophage locus BLi_Pp6. The introduced NGS-based approach allows the investigation of prophage regions and their ability to form particles. Our results show that this approach increases the sensitivity of prophage activity analysis and can complement more conventional approaches such as transmission electron microscopy (TEM). PMID:25811873

  4. Effects of dietary Bacillus licheniformis on growth performance, immunological parameters, intestinal morphology and resistance of juvenile Nile tilapia (Oreochromis niloticus) to challenge infections.

    Science.gov (United States)

    Han, Biao; Long, Wei-Qing; He, Ju-Yun; Liu, Yong-Jian; Si, Yu-Qi; Tian, Li-Xia

    2015-10-01

    The effects of oral administration of Bacillus licheniformis on growth performance, immunity, intestinal morphology and disease resistance of juvenile tilapia were investigated. Six experimental diets supplemented with different concentrations of B. licheniformis (0%, 0.02%, 0.04%, 0.06%, 0.08% and 0.1% of AlCare(®), containing live germ 2 × 10(10) CFU/g) were formulated, viz. control, T1, T2, T3, T4 and T5. Each diet was randomly assigned to triplicate groups of 30 fishes (3.83 ± 0.03 g). After 10 weeks of feeding trial, weight gain (WG), final body wet weight (FBW) and specific growth rate (SGR) increased significantly in groups T2, T3, T4 and T5 compared with control and T1 (p 0.05). Compared with control, dietary B. licheniformis supplementation increased the content of complement C3 in serum significantly (P licheniformis supplementation (P > 0.05). When tilapia were challenged against Streptococcus iniae, survival rate improved significantly when tilapia fed with T2, T3, T4 and T5 (P licheniformis (T2, T3, T4, T5) tended to be regularly arranged and exhibited less exfoliation, twist and fusion. These results indicated that dietary supplementation of B. licheniformis not only increased the growth, immune response and disease resistance of juvenile tilapia, but also influenced anterior intestinal development and integrity. Furthermore, in our study, the optimal concentration of B. licheniformis in diets for tilapia was greater than or equal to 4.4 × 10(6) CFU/g. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. High production of succinyl isoflavone glycosides by Bacillus licheniformis ZSP01 resting cells in aqueous miscible organic medium.

    Science.gov (United States)

    Zhang, Sen; Chen, Guoguang; Chu, Jianlin; Wu, Bin; He, Bingfang

    2015-01-01

    To achieve efficient production of succinyldaidzin and succinylgenistin, resting cells of a solvent-stable strain Bacillus licheniformis ZSP01 were used to react with pure isoflavones or soybean flour extract in a reaction medium with 10% dimethyl sulfoxide. Strikingly, 0.8 mM daidzein, 0.8 mM genistein, 2.0 mM daidzin, and 2.0 mM genistin were transformed to succinyl isoflavone glycosides in 27 H (yield >90%). The soybean flour extract (6.1%, w/v) contained 0.32 mM daidzein, 0.84 mM daidzin, 0.38 mM genistein, and 1.04 mM genistin. Over 95% of total isoflavones (daidzein, daidzin, genistein, and genistin) in the soybean flour extract were converted to succinyl isoflavone glycosides after 27 H. Strain ZSP01 shows both high glycosylation and succinylation activities. These results suggest that B. licheniformis ZSP01 could be useful for the efficient production of succinyl soybean isoflavone glycosides. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

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

    Science.gov (United States)

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

    2017-04-01

    Poly-γ-glutamic acid (γ-PGA), a natural biopolymer, is widely used in cosmetics, medicine, food, water treatment, and agriculture owing to its features of moisture sequestration, cation chelation, non-toxicity and biodegradability. Intracellular glutamic acid, the substrate of γ-PGA, is a limiting factor for high yield in γ-PGA production. Bacillus subtilis and Bacillus licheniformis are both important γ-PGA producing strains, and B. subtilis synthesizes glutamic acid in vivo using the unique GOGAT/GS pathway. However, little is known about the glutamate synthesis pathway in B. licheniformis. The aim of this work was to characterize the glutamate dehydrogenase (RocG) in glutamic acid synthesis from B. licheniformis with both in vivo and in vitro experiments. By re-directing the carbon flux distribution, the rocG gene deletion mutant WX-02ΔrocG produced intracellular glutamic acid with a concentration of 90ng/log(CFU), which was only 23.7% that of the wild-type WX-02 (380ng/log(CFU)). Furthermore, the γ-PGA yield of mutant WX-02ΔrocG was 5.37g/L, a decrease of 45.3% compared to the wild type (9.82g/L). In vitro enzymatic assays of RocG showed that RocG has higher affinity for 2-oxoglutarate than glutamate, and the glutamate synthesis rate was far above degradation. This is probably the first study to reveal the glutamic acid synthesis pathway and the specific functions of RocG in B. licheniformis. The results indicate that γ-PGA production can be enhanced through improving intracellular glutamic acid synthesis. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Hyper production of cellulose degrading endo (1,4 β-d-glucanase from Bacillus licheniformis KIBGE-IB2

    Directory of Open Access Journals (Sweden)

    Asad Karim

    2015-04-01

    Full Text Available Cellulase hydrolyzes β (1,4 glycosidic linkages of cellulose polymer to soluble sugar. An extracellular enzyme production by Bacillus licheniformis KIBGE-IB2 (GenBank accession No. GU216259 was studied under various environmental conditions. Maximum enzyme production was measured in the liquid fermentation medium after 48 h, containing (gL−1, CMC, 5.0; peptone, 15.0; yeast extract, 15.0; CaCl2·2H2O, 0.001; FeSO4·7H2O, 0.001; K2HPO4, 5.0; NaH2PO4, 5.0 and MgSO4·7H2O, 1.0. The optimal pH and temperature for enzyme production was found to be 6.0 and 37°C, respectively. It was also found that beside soluble sugars, a significant amount of enzyme production was obtained when biomass (wheat bran and orange peel were examined as a sole carbon source. The current findings indicate that endo (1,4 β-d-glucanase from B. licheniformis KIBGE-IB2 can be beneficial for commercial purpose.

  8. Optimization of novel and greener approach for the coproduction of uricase and alkaline protease in Bacillus licheniformis by Box-Behnken model.

    Science.gov (United States)

    Pawar, Shweta V; Rathod, Virendra K

    2018-01-02

    This study explores a novel concept of coproduction of uricase and alkaline protease by Bacillus licheniformis using single substrate in single step. Seven local bacterial strains were screened for uricase production, amongst which B. licheniformis is found to produce highest uricase along with alkaline protease. Optimization of various factors influencing maximum enzyme coproduction by B. licheniformis is performed. Maximum enzyme productivity of 0.386 U/mL uricase and 0.507 U/mL alkaline protease is obtained at 8 hr of incubation period, 1% (v/v) inoculum, and at 0.2% (w/v) uric acid when the organism is cultivated at 25°C, 180 rpm, in a media containing xylose as a carbon source, urea as a nitrogen source, and initial pH of 9.5. The statistical experimental design method of Box-Behnken was further applied to obtain optimal concentration of significant parameters such as pH (9.5), uric acid concentration (0.1%), and urea concentration (0.05%). The maximum uricase and alkaline protease production by B. licheniformis using Box-Behnken design was 0.616 and 0.582 U/mL, respectively, with 1.6- and 1.13-fold increase as compared to one factor at a time optimized media. This study will be useful to develop an economic, commercially viable, and scalable process for simultaneous production of uricase and protease enzymes.

  9. The effect of ionic strength on the adsorption of H{sup +}, Cd{sup 2+}, Pb{sup 2+}, and Cu{sup 2+} by Bacillus subtilis and Bacillus licheniformis: A surface complexation model

    Energy Technology Data Exchange (ETDEWEB)

    Daughney, C.J. [McGill Univ., Montreal, Quebec (Canada). Earth and Planetary Sciences; Fein, J.B. [Univ. of Notre Dame, IN (United States)

    1998-02-01

    To quantify metal adsorption onto bacterial surfaces, recent studies have applied surface complexation theory to model the specific chemical and electrostatic interactions occurring at the solution-cell wall interface. However, to date, the effect of ionic strength on these interactions has not been investigated. In this study, the authors perform acid-base titrations of suspensions containing Bacillus subtilis or Bacillus licheniformis in 0.01 or 0.1 M NaNO{sub 3}, and they evaluate the constant capacitance and basic Stern double-layer models for their ability to describe ionic-strength-dependent behavior. The constant capacitance model provides the best description of the experimental data. The constant capacitance model parameters vary between independently grown bacterial cultures, possibly due to cell wall variation arising from genetic exchange during reproduction. The authors perform metal-B. subtilis and metal-B. licheniformis adsorption experiments using Cd, Pb, and Cu, and they solve for stability constants describing metal adsorption onto distinct functional groups on the bacterial cell walls. They find that these stability constants vary substantially but systematically between the two bacterial species at the two different ionic strengths.

  10. Analyzing AbrB-Knockout Effects through Genome and Transcriptome Sequencing of Bacillus licheniformis DW2

    Science.gov (United States)

    Shu, Cheng-Cheng; Wang, Dong; Guo, Jing; Song, Jia-Ming; Chen, Shou-Wen; Chen, Ling-Ling; Gao, Jun-Xiang

    2018-01-01

    As an industrial bacterium, Bacillus licheniformis DW2 produces bacitracin which is an important antibiotic for many pathogenic microorganisms. Our previous study showed AbrB-knockout could significantly increase the production of bacitracin. Accordingly, it was meaningful to understand its genome features, expression differences between wild and AbrB-knockout (ΔAbrB) strains, and the regulation of bacitracin biosynthesis. Here, we sequenced, de novo assembled and annotated its genome, and also sequenced the transcriptomes in three growth phases. The genome of DW2 contained a DNA molecule of 4,468,952 bp with 45.93% GC content and 4,717 protein coding genes. The transcriptome reads were mapped to the assembled genome, and obtained 4,102∼4,536 expressed genes from different samples. We investigated transcription changes in B. licheniformis DW2 and showed that ΔAbrB caused hundreds of genes up-regulation and down-regulation in different growth phases. We identified a complete bacitracin synthetase gene cluster, including the location and length of bacABC, bcrABC, and bacT, as well as their arrangement. The gene cluster bcrABC were significantly up-regulated in ΔAbrB strain, which supported the hypothesis in previous study of bcrABC transporting bacitracin out of the cell to avoid self-intoxication, and was consistent with the previous experimental result that ΔAbrB could yield more bacitracin. This study provided a high quality reference genome for B. licheniformis DW2, and the transcriptome data depicted global alterations across two strains and three phases offered an understanding of AbrB regulation and bacitracin biosynthesis through gene expression. PMID:29599755

  11. Hyperproduction of γ-glutamyl transpeptidase from Bacillus licheniformis ER15 in the presence of high salt concentration.

    Science.gov (United States)

    Bindal, Shruti; Gupta, Rani

    2017-02-07

    Microbial γ-glutamyl transpeptidases (GGTs) have been exploited in biotechnological, pharmaceutical, and food sectors for the synthesis of various γ-glutamyl compounds. But, till date, no bacterial GGTs are commercially available in the market because of lower levels of production from various sources. In the current study, production of GGT from Bacillus licheniformis ER15 was investigated to achieve high GGT titers. Hyperproduction of GGT from B. licheniformis ER15 was achieved with 6.4-fold enhancement (7921.2 ± 198.7 U/L) by optimization of culture medium following one-variable-at-a-time strategy and statistical approaches. Medium consisting of Na 2 HPO 4 : 0.32% (w/v); KH 2 PO 4 : 0.15% (w/v); starch: 0.1% (w/v); soybean meal: 0.5% (w/v); NaCl: 4.0% (w/v), and MgCl 2 : 5 mM was found to be optimal for maximum GGT titers. Maximum GGT titers were obtained, in the optimized medium at 37°C and 200 rpm, after 40 h. It was noteworthy that GGT production was a linear function of sodium chloride concentration, as observed during response surface methodology. While investigating the role of NaCl on GGT production, it was found that NaCl drastically decreased subtilisin concentration and indirectly increasing GGT recovery. B. licheniformis ER15 is proved to be a potential candidate for large-scale production of GGT enzyme and its commercialization.

  12. Studies on the production of glucose isomerase by Bacillus licheniformis

    Directory of Open Access Journals (Sweden)

    Nwokoro Ogbonnaya

    2015-09-01

    Full Text Available This work reports the effects of some culture conditions on the production of glucose isomerase by Bacillus licheniformis. The bacterium was selected based on the release of 3.62 mg/mL fructose from the fermentation of glucose. Enzyme was produced using a variety of carbon substrates but the highest enzyme activity was detected in a medium containing 0.5% xylose and 1% glycerol (specific activity = 6.88 U/mg protein. Media containing only xylose or glucose gave lower enzyme productivies (specific activities= 4.60 and 2.35 U/mg protein respectively. The effects of nitrogen substrates on glucose isomerase production showed that yeast extract supported maximum enzyme activity (specific activity = 5.24 U/mg protein. Lowest enzyme activity was observed with sodium trioxonitrate (specific activity = 2.44 U/mg protein. In general, organic nitrogen substrates supported higher enzyme productivity than inorganic nitrogen substrates. Best enzyme activity was observed in the presence of Mg2+ (specific activity = 6.85 U/mg protein while Hg2+ was inhibitory (specific activity = 1.02 U/mg protein. The optimum pH for best enzyme activity was 6.0 while optimum temperature for enzyme production was 50ºC.

  13. GFP tagged Vibrio parahaemolyticus Dahv2 infection and the protective effects of the probiotic Bacillus licheniformis Dahb1 on the growth, immune and antioxidant responses in Pangasius hypophthalmus.

    Science.gov (United States)

    Gobi, Narayanan; Malaikozhundan, Balasubramanian; Sekar, Vijayakumar; Shanthi, Sathappan; Vaseeharan, Baskaralingam; Jayakumar, Rengarajan; Khudus Nazar, Abdul

    2016-05-01

    In this study, the pathogenicity of GFP tagged Vibrio parahaemolyticus Dahv2 and the protective effect of the probiotic strain, Bacillus licheniformis Dahb1 was studied on the Asian catfish, Pangasius hypophthalmus. The experiment was carried out for 24 days with three groups and one group served as the control (without treatment). In the first group, P. hypophthalmus was orally infected with 1 mL of GFP tagged V. parahaemolyticus Dahv2 at two different doses (10(5) and 10(7) cfu mL(-1)). In the second group, P. hypophthalmus was orally administrated with 1 ml of the probiotic B. licheniformis Dahb1 at two different doses (10(5) and 10(7) cfu mL(-1)). In the third group, P. hypophthalmus was orally infected first with 1 mL of GFP tagged V. parahaemolyticus Dahv2 followed by the administration of 1 mL of B. licheniformis Dahb1 (combined treatment) at two different doses (10(5) and 10(7) cfu mL(-1)). The growth, immune (myeloperoxidase, respiratory burst, natural complement haemolytic and lysozyme activity) and antioxidant (glutathione-S-transferase, reduced glutathione and total glutathione) responses of P. hypophthalmus were reduced after post infection of GFP tagged V. parahaemolyticus Dahv2 compared to control. However, after administration with the probiotic B. licheniformis Dahb1 at 10(5) cfu mL(-1), P. hypophthalmus showed significant increase in the growth, immune and antioxidant responses compared to 10(7) cfu mL(-1). On the otherhand, the growth, immune and antioxidant responses of P. hypophthalmus infected and administrated with combined GFP tagged Vibrio + Bacillus at 10(5) cfu mL(-1) were relatively higher than that of GFP tagged V. parahaemolyticus Dahv2 and control groups but lower than that of probiotic B. licheniformis Dahb1 groups. The results of the present study conclude that the probiotic B. licheniformis Dahb1 at 10(5) cfu mL(-1) has the potential to protect the P. hypophthalmus against V. parahaemolyticus Dahv2 infection by enhancing the growth

  14. Removal of SO{sub 2} at low temperature using dead Bacillus licheniformis

    Energy Technology Data Exchange (ETDEWEB)

    Lishan Jia; Hao Song; Weiping Fang; Qingbiao Li; Jing Gao; Juanjuan Li; Qian Zhang [Xiamen University, Xiamen (China). China Department of Chemical Engineering and Biochemical Engineering

    2010-03-15

    In this paper we studied the adsorption and desorption behavior of SO{sub 2} by the dead Bacillus licheniformis R08 biomass. The effects of water vapor, temperature and O{sub 2} on the removal of SO{sub 2} by the biomass were studied. FTIR and XPS were used to characterize the mechanism of the SO{sub 2} adsorption on the biomass. The experimental results showed that water vapor and temperature deeply influenced the adsorption of SO{sub 2} by the biomass. However, O{sub 2} cannot oxidize SO{sub 2} to SO{sub 3} on the biomass. FTIR and XPS results showed that oxygenous and nitrogenous functional groups on the cell walls of biomass may be related to the SO{sub 2} adsorption and three sulfur species were formed on the biomass in adsorption process. In the desorption process, weakly adsorbed SO{sub 2} could be desorbed by increasing temperature and the biomass can be reused for 10 cycles. 27 refs., 6 figs., 2 tabs.

  15. Optimization of endoglucanase production from thermophilic strain of Bacillus licheniformis RT-17 and its application for saccharification of sugarcane bagasse

    International Nuclear Information System (INIS)

    Tariq, R.; Qadir, F.; Ahmed, A.; Shariq, M.; Zafar, U.; Khan, S.A.

    2018-01-01

    Thermostable cellulases are required for a variety of commercial processes. Bacillus is a house of thermostable proteins. Screening of indigenously isolated strains of bacteria revealed the promising production of cellulase by a strain, RT-17, at 50 degree C. The strain was identified on the basis of biochemical and molecular characteristics as B. licheniformis. The factors affecting cellulase production from B. licheniformis RT-17 were evaluated for their significant effect using Plackett Burman Design and were optimized by employing Box-Behnken Design. The model predicted 9.808 IU/ml of endoglucanase (EG) under optimum conditions of 50 degree C; 10% inoculum size; pH 5; and 1% peptone in fermentation medium. Practically, a titer of 9.128 IU/ml was obtained, showed the validity of the model. The enzyme preparation from B. licheniformis RT-17 was applied in combination with xylanase and pectinase preparations from indigenous yeasts for the hydrolysis of sugarcane bagasse (SCB). A higher degree of synergy (7.1 folds) was observed when yeast pectinase was used with bacterial cellulase for the hydrolysis of alkali treated SCB. Whereas, the degree of synergy was lower when bacterial cellulase was mixed with yeast xylanase. The study revealed the possibility of utilization of combination of yeast and bacterial enzymes for biomass saccharification. (author)

  16. An organic solvent-, detergent-, and thermo-stable alkaline protease from the mesophilic, organic solvent-tolerant Bacillus licheniformis 3C5.

    Science.gov (United States)

    Rachadech, W; Navacharoen, A; Ruangsit, W; Pongtharangkul, T; Vangnai, A S

    2010-01-01

    Bacillus licheniformis 3C5, isolated as mesophilic bacterium, exhibited tolerance towards a wide range of non-polar and polar organic solvents at 45 degrees C. It produced an extracellular organic solvent-stable protease with an apparent molecular mass of approximately 32 kDa. The inhibitory effect of PMSF and EDTA suggested it is likely to be an alkaline serine protease. The protease was active over abroad range of temperatures (45-70 degrees C) and pH (8-10) range with an optimum activity at pH 10 and 65 degrees C. It was comparatively stable in the presence ofa relatively high concentration (35% (v/v)) of organic solvents and various types of detergents even at a relatively high temperature (45 degrees C). The protease production by B. licheniformis 3C5 was growth-dependent. The optimization of carbon and nitrogen sources for cell growth and protease production revealed that yeast extract was an important medium component to support both cell growth and the protease production. The overall properties of the protease produced by B. licheniformis 3C5 suggested that this thermo-stable, solvent-stable, detergent-stable alkaline protease is a promising potential biocatalyst for industrial and environmental applications.

  17. A novel strategy to improve protein secretion via overexpression of the SppA signal peptide peptidase in Bacillus licheniformis.

    Science.gov (United States)

    Cai, Dongbo; Wang, Hao; He, Penghui; Zhu, Chengjun; Wang, Qin; Wei, Xuetuan; Nomura, Christopher T; Chen, Shouwen

    2017-04-24

    Signal peptide peptidases play an important role in the removal of remnant signal peptides in the cell membrane, a critical step for extracellular protein production. Although these proteins are likely a central component for extracellular protein production, there has been a lack of research on whether protein secretion could be enhanced via overexpression of signal peptide peptidases. In this study, both nattokinase and α-amylase were employed as prototypical secreted target proteins to evaluate the function of putative signal peptide peptidases (SppA and TepA) in Bacillus licheniformis. We observed dramatic decreases in the concentrations of both target proteins (45 and 49%, respectively) in a sppA deficient strain, while the extracellular protein yields of nattokinase and α-amylase were increased by 30 and 67% respectively in a strain overexpressing SppA. In addition, biomass, specific enzyme activities and the relative gene transcriptional levels were also enhanced due to the overexpression of sppA, while altering the expression levels of tepA had no effect on the concentrations of the secreted target proteins. Our results confirm that SppA, but not TepA, plays an important functional role for protein secretion in B. licheniformis. Our results indicate that the sppA overexpression strain, B. licheniformis BL10GS, could be used as a promising host strain for the industrial production of heterologous secreted proteins.

  18. Enhancement of Exochitinase Production by Bacillus licheniformis AT6 Strain and Improvement of N-Acetylglucosamine Production.

    Science.gov (United States)

    Aounallah, Mohamed Amine; Slimene-Debez, Imen Ben; Djebali, Kais; Gharbi, Dorra; Hammami, Majdi; Azaiez, Sana; Limam, Ferid; Tabbene, Olfa

    2017-02-01

    A strain producing chitinase, isolated from potato stem tissue, was identified as Bacillus licheniformis by biochemical properties and 16S RNA sequence analysis. Statistical experimental designs were used to optimize nine independent variables for chitinase production by B. licheniformis AT6 strain in submerged fermentation. Using Plackett-Burman design, (NH 4 ) 2 SO 4 , MgSO 4 .7H 2 O, colloidal chitin, MnCl 2 2H 2 O, and temperature were found to influence chitinase production significantly. According to Box-Behnken response surface methodology, the optimal fermentation conditions allowing maximum chitinase production were (in gram per liter): (NH 4 ) 2 SO 4 , 7; K 2 HPO 4 , 1; NaCl, 1; MgSO 4 .7H 2 O, 0.1; yeast extract, 0.5; colloidal chitin, 7.5; MnCl 2 .2H 2 O, 0.2; temperature 35 °C; pH medium 7. The optimization strategy led to a 10-fold increase in chitinase activity (505.26 ± 22.223 mU/mL versus 50.35 ± 19.62 mU/mL for control basal medium). A major protein band with a molecular weight of 61.9 kDa corresponding to chitinase activity was clearly detected under optimized conditions. Chitinase activity produced in optimized medium mainly releases N-acetyl glucosamine (GlcNAc) monomer from colloidal chitin. This enzyme also acts as an exochitinase with β-N-acetylglucosaminidase. These results suggest that B. licheniformis AT6 secreting exochitinase is highly efficient in GlcNAc production which could in turn be envisaged as a therapeutic agent or as a conservator against the alteration of several ailments.

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

    Directory of Open Access Journals (Sweden)

    Jung-Hoon Kim

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

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

    Science.gov (United States)

    Ju, Shin-Yeong; Yang, Yoon-Mo; Ryu, Su-Hyun; Kwon, Yumi; Won, Young-Bin; Lee, Yeh-Eun; Youn, Hwan; Lee, Jin-Won

    2016-01-01

    The ferric uptake regulator (Fur) family proteins include sensors of Fe (Fur), Zn (Zur), and peroxide (PerR). Among Fur family proteins, Fur and Zur are ubiquitous in most prokaryotic organisms, whereas PerR exists mainly in Gram positive bacteria as a functional homologue of OxyR. Gram positive bacteria such as Bacillus subtilis, Listeria monocytogenes and Staphylococcus aureus encode three Fur family proteins: Fur, Zur, and PerR. In this study, we identified five Fur family proteins from B. licheniformis: two novel PerR-like proteins (BL00690 and BL00950) in addition to Fur (BL05249), Zur (BL03703), and PerR (BL00075) homologues. Our data indicate that all of the five B. licheniformis Fur homologues contain a structural Zn2+ site composed of four cysteine residues like many other Fur family proteins. Furthermore, we provide evidence that the PerR-like proteins (BL00690 and BL00950) as well as PerRBL (BL00075), but not FurBL (BL05249) and ZurBL (BL03703), can sense H2O2 by histidine oxidation with different sensitivity. We also show that PerR2 (BL00690) has a PerR-like repressor activity for PerR-regulated genes in vivo. Taken together, our results suggest that B. licheniformis contains three PerR subfamily proteins which can sense H2O2 by histidine oxidation not by cysteine oxidation, in addition to Fur and Zur. PMID:27176811

  1. Poly-γ-glutamic acid produced from Bacillus licheniformis CGMCC 2876 as a potential substitute for polyacrylamide in the sugarcane industry.

    Science.gov (United States)

    Yan, Shan; Yao, Haosheng; Chen, Zhen; Zeng, Shengquan; Xi, Xi; Wang, Yuanpeng; He, Ning; Li, Qingbiao

    2015-01-01

    As an environmentally friendly and industrially useful biopolymer, poly-γ-glutamic acid (γ-PGA) from Bacillus licheniformis CGMCC 2876 was characterized by the high-resolution mass spectrometry and (1)H NMR. A flocculating activity of 11,474.47 U mL(-1) obtained with γ-PGA, and the effects of carbon sources, ions, and chemical properties (D-/L-composition and molecular weight) on the production and flocculating activity of γ-PGA were discussed. Being a bioflocculant in the sugar refinery process, the color and turbidity of the sugarcane juice was IU 1,877.36 and IU 341.41 with 0.8 ppm of γ-PGA, respectively, which was as good as the most widely used chemically synthesized flocculant in the sugarcane industry--polyacrylamide with 1 ppm. The γ-PGA produced from B. licheniformis CGMCC 2876 could be a promising alternate of chemically synthesized flocculants in the sugarcane industry. © 2015 American Institute of Chemical Engineers.

  2. Selection of Suitable Carbon, Nitrogen and Sulphate Source for the Production of Alkaline Protease by Bacillus licheniformis NCIM-2042

    Directory of Open Access Journals (Sweden)

    Biswanath BHUNIA

    2010-06-01

    Full Text Available In this study, selection of suitable carbon, nitrogen and sulphate sources were carried out by one-variable-at-time approach for the production of alkaline protease enzyme by Bacillus licheniformis NCIM-2042. Maximum levels of alkaline protease were found in culture media supplemented with magnesium sulphate, starch and soybean meal as a good sulphate, carbon and nitrogen sources which influenced the maximum yield of this enzyme (137.694.57, 135.231.73 and 134.741.77, respectively in comparison with the other sulphate, carbon and nitrogen sources.

  3. Gene expression and molecular characterization of a chaperone protein HtpG from Bacillus licheniformis.

    Science.gov (United States)

    Lo, Hui-Fen; Chen, Bo-En; Lin, Min-Guan; Chi, Meng-Chun; Wang, Tzu-Fan; Lin, Long-Liu

    2016-04-01

    Heat shock protein 90 (Hsp90/HtpG) is a highly abundant and ubiquitous ATP-dependent molecular chaperone consisting of three flexibly linked regions, an N-terminal nucleotide-binding domain, middle domain, and a C-terminal domain. Here the putative htpG gene of Bacillus licheniformis was cloned and heterologously expressed in Escherichia coli M15 cells. Native-gel electrophoresis, size exclusion chromatography, and cross-linking analysis revealed that the recombinant protein probably exists as a mixture of monomer, dimer and other oligomers in solution. The optimal conditions for the ATPase activity of B. licheniformis HtpG (BlHtpG) were 45°C and pH 7.0 in the presence of 0.5mM Mg(2+) ions. The molecular architecture of this protein was stable at higher temperatures with a transition point (Tm) of 45°C at neutral pH, whereas the Tm value was reduced to 40.8°C at pH 10.5. Acrylamide quenching experiment further indicated that the dynamic quenching constant (Ksv) of BlHtpG became larger at higher pH values. BlHtpG also experienced a significant change in the protein conformation upon the addition of ATP and organic solvents. Collectively, our experiment data may provide insights into the molecular properties of BlHtpG and identify the alteration of protein structure to forfeit the ATPase activity at alkaline conditions. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Efficient expression of nattokinase in Bacillus licheniformis: host strain construction and signal peptide optimization.

    Science.gov (United States)

    Wei, Xuetuan; Zhou, Yinhua; Chen, Jingbang; Cai, Dongbo; Wang, Dan; Qi, Gaofu; Chen, Shouwen

    2015-02-01

    Nattokinase (NK) possesses the potential for prevention and treatment of thrombus-related diseases. In this study, high-level expression of nattokinase was achieved in Bacillus licheniformis WX-02 via host strain construction and signal peptides optimization. First, ten genes (mpr, vpr, aprX, epr, bpr, wprA, aprE, bprA, hag, amyl) encoding for eight extracellular proteases, a flagellin and an amylase were deleted to obtain B. licheniformis BL10, which showed no extracellular proteases activity in gelatin zymography. Second, the gene fragments of P43 promoter, Svpr, nattokinase and TamyL were combined into pHY300PLK to form the expression vector pP43SNT. In BL10 (pP43SNT), the fermentation activity and product activity per unit of biomass of nattokinase reached 14.33 FU/mL and 2,187.71 FU/g respectively, which increased by 39 and 156 % compared to WX-02 (pP43SNT). Last, Svpr was replaced with SsacC and SbprA, and the maximum fermentation activity (33.83 FU/mL) was achieved using SsacC, which was 229 % higher than that of WX-02 (pP43SNT). The maximum NK fermentation activity in this study reaches the commercial production level of solid state fermentation, and this study provides a promising engineered strain for industrial production of nattokinase, as well as a potential platform host for expression of other target proteins.

  5. Laboratory investigation of microbiologically influenced corrosion of C1018 carbon steel by nitrate reducing bacterium Bacillus licheniformis

    International Nuclear Information System (INIS)

    Xu, Dake; Li, Yingchao; Song, Fengmei; Gu, Tingyue

    2013-01-01

    Nitrate injection is used to suppress reservoir souring in oil and gas fields caused by Sulfate Reducing Bacteria (SRB) through promotion of nitrate respiration by Nitrate Reducing Bacteria (NRB). However, it is not well publicized that nitrate reduction by NRB can cause Microbiologically Influenced Corrosion (MIC) because nitrate reduction coupled with iron oxidation is thermodynamically favorable. NRB benefits bioenergetically from this redox reaction under biocatalysis. This work showed that the Bacillus licheniformis biofilm, when grown as an NRB biofilm, caused a 14.5 μm maximum pit depth and 0.89 mg/cm 2 normalized weight loss against C1018 carbon steel in one-week lab tests

  6. Heat activation and stability of amylases from Bacillus species

    African Journals Online (AJOL)

    Administrator

    2007-05-16

    May 16, 2007 ... as Bacillus macerans, Bacillus coagulans Bacillus licheniformis, Bacillus circulans, Bacillus megaterium, Bacillus polymyxa and Bacillus subtilis. Heat treatment at 70oC denatured the β-amylase component of the amylase source while α-amylase retained its potency at this temperature. Calcium.

  7. Characterization of lipopeptides produced by Bacillus licheniformis using liquid chromatography with accurate tandem mass spectrometry.

    Science.gov (United States)

    Favaro, Gabriella; Bogialli, Sara; Di Gangi, Iole Maria; Nigris, Sebastiano; Baldan, Enrico; Squartini, Andrea; Pastore, Paolo; Baldan, Barbara

    2016-10-30

    The plant endophyte Bacillus licheniformis, isolated from leaves of Vitis vinifera, was studied to individuate and characterize the presence of bioactive lipopeptides having amino acidic structures. Crude extracts of liquid cultures were analyzed by ultra-high-performance liquid chromatography (UHPLC) coupled to a quadrupole time-of-flight (QTOF) mass analyzer. Chromatographic conditions were optimized in order to obtain an efficient separation of the different isobaric lipopeptides, avoiding merged fragmentations of co-eluted isomeric compounds and reducing possible cross-talk phenomena. Composition of the amino acids was outlined through the interpretation of the fragmentation behavior in tandem high-resolution mass spectrometry (HRMS/MS) mode, which showed both common-class and peculiar fragment ions. Both [M + H](+) and [M + Na](+) precursor ions were fragmented in order to differentiate some isobaric amino acids, i.e. Leu/Ile. Neutral losses characteristic of the iso acyl chain were also evidenced. More than 90 compounds belonging to the classes of surfactins and lichenysins, known as biosurfactant molecules, were detected. Sequential LC/HRMS/MS analysis was used to identify linear and cyclic lipopeptides, and to single out the presence of a large number of isomers not previously reported. Some critical issues related to the simultaneous selection of different compounds by the quadrupole filter were highlighted and partially solved, leading to tentative assignments of several structures. Linear lichenysins are described here for the first time. The approach was proved to be useful for the characterization of non-target lipopeptides, and proposes a rationale MS experimental scheme aimed to investigate the difference in amino acid sequence and/or in the acyl chain of the various congeners, when standards are not available. Results expanded the knowledge about production of linear and cyclic bioactive compounds from Bacillus licheniformis, clarifying the

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

    Directory of Open Access Journals (Sweden)

    González, Elisa

    1998-02-01

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

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

  9. Production of amylolytic enzymes by bacillus spp

    International Nuclear Information System (INIS)

    Dawood, Elham Shareif

    1997-12-01

    Sixty six bacteria and twenty fungi were isolated from various sources. These varied from rotten fruites to local drinks and soil samples from different parts of Sudan. On the basis of index of amylolytic activity, forty one bacteria and twelve fungi were found to hydrolyse strach. The best ten strach hydrolysing isolates were identified all as bacilli (Bacillus licheniformis SUD-K 1 , SUD-K 2 , SUD-K 4 , SUD-O, SUD-SRW, SUD-BRW, SUD-By, Bacillus subtilis SUD-K 3 , and Bacillus circulans SUD-D and SUD-K 7 ). Their amylase productivity was studied with respect to temperature and time. Amylolytic activity was measured by spectrophotometer, the highest activity was produced in around 24 hours of growth in all; six of which gave the highest amylase activity at 50 deg C and the rest at 45C. Based on the thermal production six isolates were chosen for further investigation. These were Bacillus licheniformis SUD-K 1 , SUD-K 2 , SUD-K 4 , SUD-O, Bacillus subtilis SUD-K 3 and Bacillus circulans SUD-K 7 . The inclusion of strach and Mg ++ ions in the culture medium gave the highest enzyme yield. The Ph 9.0 was found to be the optimum for amylase production for all isolates except Bacillus subtilis SUD-K 3 which had an optimum at pH 7.0. Three isolates (Bacillus licheniformis SUD-K 1 , SUD-K 4 and SUD-O recorded highestamylase production in a medium supplemented with peptone while the rest (Bacillus licheniformis SUD-K 2 , Bacillus subtilis SUD-K 3 and Bacillus circulans SUD-K 7 ) gave highest amylase productivity in a medium supplemented with malt extract. Four isolates (Bacillus licheniformis SUD-K 1 and Bacillus subtilis SUD-K 3 gave maximum amylase production in a medium containing 0.5% soluble strach while the rest (gave maximum amylase production at 2%. Soluble strach was found to be best substrate among the different carbon sources tested. The maximum temperature for amylase activity ranged from 60-70 deg C and 1% strach concentration was optimum for all isolates

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

    DEFF Research Database (Denmark)

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

    2007-01-01

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

  11. Identification and High-level Production of Pulcherrimin in Bacillus licheniformis DW2.

    Science.gov (United States)

    Li, Xiaoyun; Wang, Dong; Cai, Dongbo; Zhan, Yangyang; Wang, Qin; Chen, Shouwen

    2017-12-01

    Pulcherrimin, a potential biocontrol agent produced by microorganisms, has the promising applications in the agricultural, medical, and food areas, and the low yield of pulcherrimin has hindered its applications. In this study, the red pigment produced by Bacillus licheniformis DW2 was identified as pulcherrimin through the spectrometry analysis and genetic manipulation, and the component of the medium used for pulcherrimin production was optimized. Based on our results, the addition of 1.0 g L -1 Tween 80 could improve the yield of pulcherrimin, and glucose and (NH 4 ) 2 SO 4 were served as the optimal carbon and nitrogen sources for pulcherrimin synthesis, respectively. Furthermore, an orthogonal array design was applied for optimization of the medium. Under optimized condition, the maximum yield of pulcherrimin was 331.17 mg L -1 , 5.30-fold higher than that of the initial condition, which was the maximum yield reported for pulcherrimin production. Collectively, this study provided a promising strain and a feasible approach to achieve the high-level production of antimicrobial pulcherrimin.

  12. Mechanistic aspects of biogenic synthesis of CdS nanoparticles using Bacillus licheniformis

    Science.gov (United States)

    Tripathi, R. M.; Singh Bhadwal, Akhshay; Singh, Priti; Shrivastav, Archana; Singh, M. P.; Shrivastav, B. R.

    2014-06-01

    A novel eco-friendly effort has been made for the synthesis of cadmium sulfide (CdS) nanoparticles using bacterial biomass. Although some articles have been reported on CdS nanoparticles synthesis by bacteria, here we have synthesized CdS nanoparticles using non-pathogenic bacteria Bacillus licheniformis MTCC 9555. UV-Vis spectroscopy was carried out to confirm the formation of CdS nanoparticles; the peak occurring at 368 nm gives the indication of synthesis of CdS nanoparticles. The size and morphology of the synthesized CdS nanoparticles were analyzed by transmission electron microscopy (TEM) and the nanoparticles are found to have a narrow size of 5.1 ± 0.5 nm with spherical morphology. Further, the nanoparticles were examined by energy dispersive x-ray (EDX) spectroscopy to identify the presence of elements and confirmed the existence of Cd and S in single nanoparticles. X-ray diffraction (XRD) analysis exhibited 2θ values corresponding to CdS nanocrystals. Fourier transform infrared spectroscopy (FTIR) provides the evidence for the presence of proteins as possible biomolecules responsible for the stabilization of the synthesized CdS nanoparticles.

  13. Selection of Suitable Carbon, Nitrogen and Sulphate Source for the Production of Alkaline Protease by Bacillus licheniformis NCIM-2042

    Directory of Open Access Journals (Sweden)

    Biswanath BHUNIA

    2010-06-01

    Full Text Available In this study, selection of suitable carbon, nitrogen and sulphate sources were carried out by one-variable-at-time approach for the production of alkaline protease enzyme by Bacillus licheniformis NCIM-2042. Maximum levels of alkaline protease were found in culture media supplemented with magnesium sulphate, starch and soybean meal as a good sulphate, carbon and nitrogen sources which influenced the maximum yield of this enzyme (137.69�4.57, 135.23�1.73 and 134.74�1.77, respectively in comparison with the other sulphate, carbon and nitrogen sources.

  14. Inhibitory effects of Bacillus probionts on growth and toxin production of Vibrio harveyi pathogens of shrimp.

    Science.gov (United States)

    Nakayama, T; Lu, H; Nomura, N

    2009-12-01

    To investigate the effects of Bacillus subtilis, Bacillus licheniformis and Bacillus megaterium in terms of toxin and growth of pathogenic Vibrio harveyi. Three Bacillus probionts were isolated from probiotic BZT aquaculture and identified using a 16S rDNA sequence. Growth inhibition assay showed that supernatants from the 24-h culture of three Bacillus species were able to inhibit the growth of V. harveyi (LMG 4044); B. subtilis was the most effective based on the well diffusion method. Results of a liquid culture model showed that B. subtilis was also widely effective in inhibiting three strains of V. harveyi (isolated from Thailand, the Philippines and LMG 4044), and that both B. licheniformis and B. megaterium inhibit the growth of V. harveyi isolated from the Philippines. Moreover, a haemolytic activity assay demonstrated that V. harveyi (IFO 15634) was significantly decreased by the addition of B. licheniformis or B. megaterium supernatant. Bacillus subtilis inhibited Vibrio growth, and both B. licheniformis and B. megaterium suppressed haemolytic activity in Vibrio. The cell-free supernatants produced by Bacillus probionts inhibit Vibrio disease, and Bacillus probionts might have an influence on Vibrio cell-to-cell communications.

  15. Untangling the transcription regulatory network of the bacitracin synthase operon in Bacillus licheniformis DW2.

    Science.gov (United States)

    Wang, Dong; Wang, Qin; Qiu, Yimin; Nomura, Christopher T; Li, Junhui; Chen, Shouwen

    The bacitracin synthetase gene cluster in Bacillus licheniformis DW2 is composed of the bacABC operon encoding a non-ribosomal peptide synthetase and bacT encoding a thioesterase. Although the bacitracin gene cluster has been well studied, little is known about how this gene cluster is regulated. This study provides insight into how the transcription factors Spo0A and AbrB regulate bacitracin biosynthesis. Deletion of spo0A resulted in drastically reduced expression of bacA and bacT, and subsequently bacitracin production. On the other hand, the expression of bacA and bacT increased significantly in B. licheniformis DW2ΔabrB and DW2Δ0AΔabrB compared to the wild-type strain DW2. The bacitracin yields on cell numbers (U/CFU) in DW2ΔabrB and DW2Δ0A/pHY300-0A-sad67 were 17.5% and 14.9% higher than that of the wild-type strain. An electrophoretic mobility shift assay (EMSA) further confirmed that AbrB could directly bind to the promoter regions of bacA and bacT. These results indicate that AbrB acts as a repressor of bacitracin biosynthesis by inhibiting bacA and bacT expression, while Spo0A indirectly promotes bacitracin biosynthesis by repressing abrB expression. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  16. L-Glutamic acid production by Bacillus spp. isolated from vegetable ...

    African Journals Online (AJOL)

    Ogiri” (fermented vegetable proteins) in Nigeria. The isolates were identified as Bacillus subtilis (6), (27.3%), Bacillus pumilus (5), (22.7%), Bacillus licheniformis (5), (27.3%) and Bacillus polymyxa (6), (22.7%). Four species of the Bacillus isolates ...

  17. Disruption in the cecal microbiota of chickens challenged with Clostridium perfringens and other factors was alleviated by Bacillus licheniformis supplementation.

    Science.gov (United States)

    Lin, Yicen; Xu, Shuai; Zeng, Dong; Ni, Xueqin; Zhou, Mengjia; Zeng, Yan; Wang, Hesong; Zhou, Yi; Zhu, Hui; Pan, Kangcheng; Li, Guangyao

    2017-01-01

    Clostridium perfringens can induce necrotic enteritis of chickens, which causes large economic losses every year. Bacillus licheniformis, a probiotic, can inhibit the growth of pathogenic bacteria such as Clostridium perfringens, thereby improving the health status of chickens. However, from a microbial ecology perspective, the mechanisms by which alterations to the gut microbiota improve health remain unknown. In this study, we used Illumina MiSeq sequencing to investigate the cecal microbiota of a negative control group (NC), a C. perfringens and Eimeria challenge group with fishmeal supplementation (PC), a group supplemented with fishmeal and infected with coccidia (FC), and group PC with B. licheniformis supplementation (BL). We found that the health status of C. perfringens-challenged chickens was compromised, and that B. licheniformis improved the growth of the chickens challenged with pathogens. Microbial diversity analysis and taxonomic profiling of groups NC, PC, and FC revealed a disturbed cecal microflora of the birds with C. perfringens. We also characterized the microbiota of the chickens in the BL group using several methods. Principal coordinate analysis demonstrated that, compared with group PC, the bacterial community structure of group BL was more similar to that of group NC. Linear discriminant analysis with effect size revealed less differentially represented bacterial taxa between groups BL and NC than between groups PC and NC. In addition, groups BL and NC appeared to have similar overrepresented microbial taxa (such as Bacteroides, Helicobacter, Megamonas, and Akkermansia) compared with group PC. Finally, a phylogenetic investigation of communities by reconstruction of unobserved states analysis indicated that large differences existed between group PC and groups NC and BL. In conclusion, pre-treatment with B. licheniformis reduced the disturbance of the cecal microbiome induced by challenge with C. perfringens and other factors in broiler

  18. In vitro antimicrobial effect of Satureja wiedemanniana against Bacillus species isolated from raw meat samples.

    Science.gov (United States)

    Yucel, Nihal; Aslim, Belma; Ozdoğan, Hakan

    2009-08-01

    In this study a total of 30 raw meat samples obtained from Ankara, Turkey were screened for the presence of Bacillus species. Among the meat samples analyzed, the predominant species isolated was Bacillus circulans; other Bacillus species were identified as Bacillus firmus, Bacillus lentus, Bacillus megaterium, Bacillus licheniformis, Bacillus mycoides, Bacillus sphaericus, and Bacillus cereus. Minced meat samples were more contaminated with Bacillus species than sliced beef sample. From these samples, 242 Bacillus species isolates were obtained, which were investigated for proteolytic and lipolytic activity, associated with meat spoilage. Interestingly, some Bacillus strains produced the highest values of proteolytic/lipolytic activities. Nineteen Bacillus strains were selected among the 242 isolates according to their proteolytic/lipolytic activity with a clear zone diameter of > or =6 mm. The essential oil of Satureja wiedemanniana (Lalem) Velen was also tested against these 19 Bacillus species that had proteolytic and lipolytic activity. The essential oil yield obtained from the aerial parts of the plant was 0.35% (vol/wt). The inhibition zones of the essential oil obtained against all the Bacillus species were in the range of 5.0-12.0 mm. The oil showed high antimicrobial activities against B. licheniformis M 6(26), M 11(16), and M 12(1) strains. B. licheniformis 12(1) showed high lipolytic activity (18.0 mm). Also, B. licheniformis M 6(26) and M 11(16) showed high proteolytic activity (16.0 and 14.0 mm). These results may suggest that an essential oil of S. wiedemanniana can be used as a natural preservative in meat against spoilage bacteria.

  19. Production of amylolytic enzymes by bacillus spp

    Energy Technology Data Exchange (ETDEWEB)

    Dawood, Elham Shareif [Department of Botany, Faculty of Science, University of Khartoum, Khartoum (Sudan)

    1997-12-01

    Sixty six bacteria and twenty fungi were isolated from various sources. These varied from rotten fruites to local drinks and soil samples from different parts of Sudan. On the basis of index of amylolytic activity, forty one bacteria and twelve fungi were found to hydrolyse strach. The best ten strach hydrolysing isolates were identified all as bacilli (Bacillus licheniformis SUD-K{sub 1}, SUD-K{sub 2}, SUD-K{sub 4}, SUD-O, SUD-SRW, SUD-BRW, SUD-By, Bacillus subtilis SUD-K{sub 3}, and Bacillus circulans SUD-D and SUD-K{sub 7}). Their amylase productivity was studied with respect to temperature and time. Amylolytic activity was measured by spectrophotometer, the highest activity was produced in around 24 hours of growth in all; six of which gave the highest amylase activity at 50 deg C and the rest at 45C. Based on the thermal production six isolates were chosen for further investigation. These were Bacillus licheniformis SUD-K{sub 1}, SUD-K{sub 2}, SUD-K{sub 4}, SUD-O, Bacillus subtilis SUD-K{sub 3} and Bacillus circulans SUD-K{sub 7}. The inclusion of strach and Mg{sup ++} ions in the culture medium gave the highest enzyme yield. The Ph 9.0 was found to be the optimum for amylase production for all isolates except Bacillus subtilis SUD-K{sub 3} which had an optimum at pH 7.0. Three isolates (Bacillus licheniformis SUD-K{sub 1}, SUD-K{sub 4} and SUD-O recorded highestamylase production in a medium supplemented with peptone while the rest (Bacillus licheniformis SUD-K{sub 2}, Bacillus subtilis SUD-K{sub 3} and Bacillus circulans SUD-K{sub 7}) gave highest amylase productivity in a medium supplemented with malt extract. Four isolates (Bacillus licheniformis SUD-K{sub 1} and Bacillus subtilis SUD-K{sub 3} gave maximum amylase production in a medium containing 0.5% soluble strach while the rest (gave maximum amylase production at 2%. Soluble strach was found to be best substrate among the different carbon sources tested. The maximum temperature for amylase activity

  20. Induction of Osmoadaptive Mechanisms and Modulation of Cellular Physiology Help Bacillus licheniformis Strain SSA 61 Adapt to Salt Stress

    Energy Technology Data Exchange (ETDEWEB)

    Paul, Sangeeta; Aggarwal, Chetana; Thakur, Jyoti Kumar; Bandeppa, G. S.; Khan, Md. Aslam; Pearson, Lauren M.; Babnigg, Gyorgy; Giometti, Carol S.; Joachimiak, Andrzej

    2015-01-06

    Bacillus licheniformis strain SSA 61, originally isolated from Sambhar salt lake, was observed to grow even in the presence of 25 % salt stress. Osmoadaptive mechanisms of this halotolerant B. licheniformis strain SSA 61, for long-term survival and growth under salt stress, were determined. Proline was the preferentially accumulated compatible osmolyte. There was also increased accumulation of antioxidants ascorbic acid and glutathione. Among the different antioxidative enzymes assayed, superoxide dismutase played the most crucial role in defense against salt-induced stress in the organism. Adaptation to stress by the organism involved modulation of cellular physiology at various levels. There was enhanced expression of known proteins playing essential roles in stress adaptation, such as chaperones DnaK and GroEL, and general stress protein YfkM and polynucleotide phosphorylase/polyadenylase. Proteins involved in amino acid biosynthetic pathway, ribosome structure, and peptide elongation were also overexpressed. Salt stress-induced modulation of expression of enzymes involved in carbon metabolism was observed. There was up-regulation of a number of enzymes involved in generation of NADH and NADPH, indicating increased cellular demand for both energy and reducing power.

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

    Science.gov (United States)

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

    2015-04-01

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

  2. Mechanistic aspects of biogenic synthesis of CdS nanoparticles using Bacillus licheniformis

    International Nuclear Information System (INIS)

    Tripathi, R M; Shrivastav, Archana; Bhadwal, Akhshay Singh; Singh, Priti; Singh, M P; Shrivastav, B R

    2014-01-01

    A novel eco-friendly effort has been made for the synthesis of cadmium sulfide (CdS) nanoparticles using bacterial biomass. Although some articles have been reported on CdS nanoparticles synthesis by bacteria, here we have synthesized CdS nanoparticles using non-pathogenic bacteria Bacillus licheniformis MTCC 9555. UV-Vis spectroscopy was carried out to confirm the formation of CdS nanoparticles; the peak occurring at 368 nm gives the indication of synthesis of CdS nanoparticles. The size and morphology of the synthesized CdS nanoparticles were analyzed by transmission electron microscopy (TEM) and the nanoparticles are found to have a narrow size of 5.1 ± 0.5 nm with spherical morphology. Further, the nanoparticles were examined by energy dispersive x-ray (EDX) spectroscopy to identify the presence of elements and confirmed the existence of Cd and S in single nanoparticles. X-ray diffraction (XRD) analysis exhibited 2θ values corresponding to CdS nanocrystals. Fourier transform infrared spectroscopy (FTIR) provides the evidence for the presence of proteins as possible biomolecules responsible for the stabilization of the synthesized CdS nanoparticles. (papers)

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

    Science.gov (United States)

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

    2017-07-01

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

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

    Science.gov (United States)

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

    2016-10-01

    Poly(γ-glutamic acid) mainly produced by Bacillus spp. is an industrially important compound due to several useful features. Among them, molecular weight is an important characteristic affecting on the physical properties such as viscosities and negative charge densities. However, it is difficult to control the molecular size of PGA since it decreases during fermentation. Previous study reported that PGA produced in the media containing different carbon sources such as glucose and glycerol showed differences in molecular weight. Therefore in this study, the effect of carbon source on the PGA molecular weight was examined; with the aim of developing a strategy to maintain the high molecular weight of PGA during fermentation. Our result showed that the weight average molecular weight (Mw) of PGA of Bacillus licheniformis ATCC 9945 cultivated in the media containing PTS-sugars were higher than the medium containing glycerol (non-PTS). The result of metabolome analysis indicated the possibility of CodY (a global regulator protein) activation in the cells cultivated in the media containing PTS-sugars. To mimic this effect, branched-chain amino acids (BCAAs), which are activators of CodY, were added to a medium containing glycerol. As the result, the Mw of PGA in the BCAAs-supplemented media were maintained and high during the early production phase compared to the non BCAAs-supplemented medium. These results indicate that BCAAs can repress the PGA molecular weight reduction during fermentation in B. licheniformis ATCC 9945. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  5. Production of tannase through submerged fermentation of tannin-containing plant extracts by Bacillus licheniformis KBR6.

    Science.gov (United States)

    Das Mohapatra, Pradeep K; Mondal, Keshab C; Pati, Bikas R

    2006-01-01

    Tannins are water-soluble polyphenolic compounds found in plants as secondary metabolites. The presence of these substances in the barks of eight different plants was initially examined and their crude extracts were used separately as a substrate for production of tannase through submerged fermentation by Bacillus licheniformis KBR6. Tannase production as well as biodegradation of the substrate reached a maximum within 15 to 18 h against crude tannin extract obtained from Anacardium occidentale. Among different concentrations of the crude tannin tested, 0.5% (w/v) induced maximum synthesis of enzyme. Tannase production was higher by almost two-fold in the presence of crude tannin compared to pure tannic acid used as a substrate. It seems that industrial production of tannase, using bark extract of A. occidentale can be a very simple and suitable alternative to presently used procedures.

  6. THERMOPHILIC BACILLUS LICHENIFORMIS RBS 5 ISOLATED FROM HOT TUNISIAN SPRING CO-PRODUCING ALKALINE AND THERMOSTABLE α-AMYLASE AND PROTEASE ENZYMES

    Directory of Open Access Journals (Sweden)

    Rakia Ben Salem

    2016-06-01

    Full Text Available Bacillus licheniformis RBS 5 was isolated from thermal spring in Tunisia. The isolate coproduce α-amylase and protease enzymes. The α-amylase activity showed an optimal activity at approximately 65°C and in wide pH interval ranging from 4 to 9. This enzyme was stable over the range of 45 to 70°C after 30 min of incubation and in the pH range of 8 to 10. Protease activity was optimal; at 80°C, pH 12. This enzyme was stable until 60°C over the pH range of 10 to 12. EDTA at concentration of 5 mM reduces slightly both activities evoking the serine alkaline protease. Cationic ions (Ca2+, Cu2+, Zn2+, and Mg 2+ have an inhibition effect on α-amylase. However, protease activity was enhanced by Ca2+, Cu2+ and Mg 2+; the other cations reduce slightly the proteolytic activity. SDS and H2O2 were found as inhibitors for both activities whereas Triton X-100 and perfume have no effect. Taken together, these traits make protease activity of B. licheniformis RBS 5 as efficient for use in detergent industry.

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

    Directory of Open Access Journals (Sweden)

    De Felice Maurilio

    2002-04-01

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

  8. Cloning, Expression, and Characterization of budC Gene Encoding meso-2,3-Butanediol Dehydrogenase from Bacillus licheniformis.

    Science.gov (United States)

    Xu, Guo-Chao; Bian, Ya-Qian; Han, Rui-Zhi; Dong, Jin-Jun; Ni, Ye

    2016-02-01

    The budC gene encoding a meso-2,3-butanediol dehydrogenase (BlBDH) from Bacillus licheniformis was cloned and overexpressed in Escherichia coli BL21(DE3). Sequence analysis reveals that this BlBDH belongs to short-chain dehydrogenase/reductase (SDR) superfamily. In the presence of NADH, BlBDH catalyzes the reduction of diacetyl to (3S)-acetoin (97.3% ee), and further to (2S,3S)-2,3-butanediol (97.3% ee and 96.5% de). Similar to other meso-2,3-BDHs, it shows oxidative activity to racemic 2,3-butanediol whereas no activity toward racemic acetoin in the presence of NAD(+). For diacetyl reduction and 2,3-butanediol oxidation, the pH optimum of BlBDH is 5.0 and 10.0, respectively. Unusually, it shows relatively high activity over a wide pH range from 5.0 to 8.0 for racemic acetoin reduction. BlBDH shows lower K m and higher catalytic efficiency toward racemic acetoin (K m = 0.47 mM, k cat /K m = 432 s(-1)·mM(-1)) when compared with 2,3-butanediol (K m = 7.25 mM, k cat /K m = 81.5 s(-1)·mM(-1)), indicating its physiological role in favor of reducing racemic acetoin into 2,3-butanediol. The enzymatic characterization of BlBDH provides evidence for the directed engineering of B. licheniformis for producing enantiopure 2,3-butanediol.

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

    Directory of Open Access Journals (Sweden)

    M.S. Hassaan

    2014-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Shamba Chatterjee

    2015-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Adyani Azizah Abd Halim

    2017-01-01

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

  12. In situ analysis of Bacillus licheniformis biofilms: amyloid-like polymers and eDNA are involved in the adherence and aggregation of the extracellular matrix.

    Science.gov (United States)

    Randrianjatovo-Gbalou, I; Rouquette, P; Lefebvre, D; Girbal-Neuhauser, E; Marcato-Romain, C-E

    2017-05-01

    This study attempts to determine which of the exopolymeric substances are involved in the adherence and aggregation of a Bacillus licheniformis biofilm. The involvement of extracellular proteins and eDNA were particularly investigated using DNase and proteinase K treatment. The permeability of the biofilms increased fivefold after DNase I treatment. The quantification of the matrix components showed that, irrespective to the enzyme tested, eDNA and amyloid-like polymers were removed simultaneously. Size-exclusion chromatography analyses supported these observations and revealed the presence of associated nucleic acid and protein complexes in the biofilm lysates. These data suggest that some extracellular DNA and amyloid-like proteins were closely interlaced within the matrix. Finally, confocal laser scanning microscopy imaging gave supplementary clues about the 3D organization of the biofilms, confirming that eDNA and exoproteins were essentially layered under and around the bacterial cells, whereas the amyloid-like fractions were homogeneously distributed within the matrix. These results confirm that some DNA-amyloid complexes play a key role in the modulation of the mechanical resistance of B. licheniformis biofilms. The study highlights the need to consider the whole structure of biofilms and to target the interactions between matrix components. A better understanding of B. licheniformis biofilm physiology and the structural organization of the matrix will strengthen strategies of biofilm control. © 2017 The Society for Applied Microbiology.

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

    Science.gov (United States)

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

    2010-10-01

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

  14. A New Bacillus licheniformis Mutant Strain Producing Serine Protease Efficient for Hvdrolvqis of Sov Meal Proteins.

    Science.gov (United States)

    Kostyleva, E V; Sereda, A S; Velikoretskaya, I A; Nefedova, L I; Sharikov, A Yu; Tsurikova, N V; Lobanov, N S; Semenova, M V; Sinitsyn, A P

    2016-07-01

    Induced mutagenesis with y-irradiation of the industrial strain Bacillus licheniformis-60 VKM B-2366,D was used to obtain a new highly active producer of an extracellular serine protease, Bacillus licheni- formis7 145. Samples of dry.concentrated preparations of serine protease produced by the original and mutant strains were obtained, and identity of their protein composition was'established. Alkaline serine protease sub- tilisin DY was the main component of the preparations. The biochemical and physicochemical properties of the Protolkheterm-145 enzyme preparation obtained from the mutant strain were studied. It exhibited pro- teolytic activity (1.5 times higher than the preparation from the initial strain) within broad ranges of pH (5- 11) and temperature (30-70'C).-Efficient hydrolysis of extruded soy meal protein at high concentrations (2 to 50%) in-the reaction mixture was.the main advantage of the Protolikheterm 145 preparation. Compared to,. the preparation obtained using the initial strain, the new preparation with increased proteolytic-activity pro- vided for more complete hydrolysis of the main non-nutritious soy,proteins.(glycinin and 0-conglycinin) with the yield of soluble protein increased by 19-28%, which decreased the cost of bioconversion of the protein- aceous material and indicated promise of the new preparation in resource-saving technologies for processing soy meals and cakes.

  15. Utilization of corn starch as sustrate for ß-Amylase by Bacillus SPP

    African Journals Online (AJOL)

    Corn starch was used as substrate for ß -amylase production from ten(10) amylolytic species of the genus Bacillus isolated locally from soil, waste water and food sources. Ten bacillus strains was made up of two strains each of Bacillus macerans, Bacillus licheniformis and Bacillus circulans. Also included are B. coagulans, ...

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

    Directory of Open Access Journals (Sweden)

    Vengadaramana, A.

    2012-01-01

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

  17. Functional and molecular characterization of a lipopeptide surfactant from the marine sponge-associated eubacteria Bacillus licheniformis NIOT-AMKV06 of Andaman and Nicobar Islands, India.

    Science.gov (United States)

    Lawrance, Anburajan; Balakrishnan, Meena; Joseph, Toms Cheriath; Sukumaran, Dheenan Palaiya; Valsalan, Vinithkumar Nambali; Gopal, Dharani; Ramalingam, Kirubagaran

    2014-05-15

    The production of a lipopeptide surfactant from the sponge-associated eubacteria Bacillus licheniformis NIOT-AMKV06 from the Andaman and Nicobar Islands was investigated. The highest production was attained with glucose and yeast extracts as the carbon and nitrogen sources (1.789 mg mL(-1)), respectively. The surfactant was highly stable over a pH range of 5.0-10 and a temperature range of 20-70°C with high NaCl concentrations. Excellent emulsification activity was exhibited by the purified surfactant with crude oil, kerosene, and diesel. A two-fold increase in surfactant production (3.0 mg mL(-1)) was observed using the newly formulated medium in this study. The surfactant biosynthesis gene cluster (sfp, sfpO, and srfA) from B. licheniformis NIOT-AMKV06 was heterologously expressed in Escherichia coli, and the production was increased three-fold (11.78 g L(-1)) over the original strain. The results confirm the potential of the surfactant for use in bioremediation of hydrocarbons in a marine environment and for enhanced oil recovery. To our knowledge, this is the first report on the ability of a hydrocarbon degrading B. licheniformis from marine sponges for the biosynthesis of a potent lipopeptide surfactant possessing characteristics of maximum stability, outstanding surfactant activity, and exceptional emulsifying capability. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Increasing the bioflocculant production and identifying the effect of overexpressing epsB on the synthesis of polysaccharide and γ-PGA in Bacillus licheniformis.

    Science.gov (United States)

    Liu, Peize; Chen, Zhen; Yang, Lijie; Li, Qingbiao; He, Ning

    2017-09-26

    Polysaccharides and poly-γ-glutamic acid (γ-PGA) are biomacromolecules that have been reported as bioflocculants, and they exhibit high flocculating activity in many industrial applications. Bacillus licheniformis CGMCC 2876 can produce polysaccharide and γ-PGA bioflocculants under different culture conditions. Several key genes are involved in the metabolic pathway of polysaccharides in B. licheniformis, but the impacts of the regulation of these genes on the production of polysaccharide bioflocculants have not been illustrated completely. To increase the bioflocculant production and identify the correlation between the synthesis of polysaccharides and γ-PGA in B. licheniformis, a few key genes were investigated to explore their influence on the synthesis of the bioflocculants. Overexpressing epsB from the eps gene cluster not only improved the bioflocculant crude yield by 13.98% but also enhanced the flocculating activity by 117.92%. The composition of the bioflocculant from the epsB recombinant strain was 28.95% total sugar, 3.464% protein and 44.03% γ-PGA, while in the original strain, these components represented 53.67%, 3.246% and 34.13%, respectively. In combination with an analysis of the transcriptional levels of several key genes involved in γ-PGA synthesis in B. licheniformis, we inferred that epsB played a key role in the synthesis of both polysaccharide and γ-PGA. The bioflocculant production of the epsB recombinant strain was further evaluated during batch fermentation in a 2 L fermenter; the flocculating activity reached 9612.75 U/mL, and the bioflocculant yield reached 10.26 g/L after 72 h, representing increases of 224% and 36.62%, respectively, compared with the original strain. Moreover, we found that the tandem expression of phosphoglucomutase (pgcA) and UTP-glucose-1-phosphate uridylyltransferase (gtaB1) could enhance the crude yield of the bioflocculant by 20.77% and that the overexpression of epsA could enhance the bioflocculant

  19. The potential of Bacillus licheniformis strains for in situ enhanced oil recovery

    Energy Technology Data Exchange (ETDEWEB)

    Yakimov, Michail M.; Timmis, Kenneth N. [Microbial Ecology Group, Division of Microbiology, GBF-National Research Centre for Biotechnology, Braunschweig (Germany); Amro, Mohammed M.; Kessel, Dagobert G. [German Petroleum Institute, Clausthal-Zellerfeld (Germany); Bock, Michael; Boseker, Klaus [BGR, Federal Institute for Geoscience and Natural Resources, Hannover (Germany); Fredrickson, Herbert L. [Environmental Laboratory, Waterways Experimental Station, USAGE, Vicksburg, MS (United States)

    1997-07-15

    The ability of microorganisms isolated from oil reservoirs to increase oil recovery by in situ growth and metabolism following the injection of laboratory grown microbial cells and nutrients were studied. Four strains isolated from Northern German oil reservoirs at depths of 866 to 1520 m, and identified as Bacillus licheniformis, were characterized taxonomically and physiologically. All strains grew on a variety of substrates at temperatures of up to 55C and at salinities of up to 12% NaCl. Extracellular polymer production occurred both aerobically and anaerobically over a wide range of temperatures, pressures and salinities, though it was optimal at temperatures around 50C and at salinities between 5 and 10% NaCl. Strain BNP29 was able to produce significant amounts of biomass, polymer, fermentation alcohols and acids in batch culture experiments under simulated reservoir conditions. Oil recovery (core flooding) experiments with strain BNP29 and a sucrose-based nutrient were performed with lime-free and lime-containing, oil-bearing sandstone cores. Oil recovery efficiencies varied from 9.3 to 22.1% of the water flood residual oil saturation. Biogenic acid production that accompanied oil production, along with selective plugging, are important mechanisms leading to increased oil recovery, presumably through resulting changes in rock porosity and alteration of wettability. These data show that strain BNP29 exhibits potential for the development of enhanced oil recovery processes

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

    Directory of Open Access Journals (Sweden)

    Madslien Elisabeth H

    2012-10-01

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

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

    Science.gov (United States)

    Suthar, Harish; Nerurkar, Anuradha

    2016-09-01

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

  2. Utilization of Industrial Waste for the Production of Bio-Preservative from Bacillus licheniformis Me1 and Its Application in Milk and Milk-Based Food Products.

    Science.gov (United States)

    Nithya, Vadakedath; Prakash, Maya; Halami, Prakash M

    2018-06-01

    The bio-preservative efficacy of a partially purified antibacterial peptide (ppABP) produced by Bacillus licheniformis Me1 in an economical medium developed using agro-industry waste was evaluated by direct application in milk and milk-based food products. The addition of ppABP in milk samples stored at 4 ± 2 °C and 28 ± 2 °C resulted in the growth inhibition of pathogens Listeria monocytogenes Scott A, Micrococcus luteus ATCC 9341, and Staphylococcus aureus FRI 722. The shelf life of milk samples with added ppABP increased to 4 days at 28 ± 2 °C, whereas curdling and off-odor were noticed in samples without ppABP. Furthermore, the milk samples with ppABP were sensorily acceptable. Antilisterial effect was also observed in cheese and paneer samples treated with ppABP. These results clearly indicate that the ppABP of B. licheniformis Me1 can be utilized as a bio-preservative to control the growth of spoilage and pathogenic bacteria, thereby reducing the risk of food-borne diseases.

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Biswanath Bhunia

    2012-01-01

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

  5. Molecular cloning, overexpression, purification and crystallographic analysis of a GH43 β-xylosidase from Bacillus licheniformis.

    Science.gov (United States)

    Diogo, José Alberto; Zanphorlin, Leticia Maria; Sato, Hélia Harumi; Murakami, Mario Tyago; Ruller, Roberto

    2015-08-01

    β-Xylosidases (EC 3.2.1.37) catalyze the hydrolysis of short xylooligosaccharides into xylose, which is an essential step in the complete depolymerization of xylan, the major hemicellulosic polysaccharide of plant cell walls, and has great biotechnological relevance for the production of lignocellulose-based biofuels and the paper industry. In this study, a GH43 β-xylosidase identified from the bacterium Bacillus licheniformis (BlXylA) was cloned into the the pET-28a bacterial expression vector, recombinantly overexpressed in Escherichia coli BL21(DE3) cells and purified to homogeneity by metal-affinity and size-exclusion chromatography. The protein was crystallized in the presence of the organic solvent 2-methyl-2,4-pentanediol and a single crystal diffracted to 2.49 Å resolution. The X-ray diffraction data were indexed in the monoclinic space group C2, with unit-cell parameters a = 152.82, b = 41.9, c = 71.79 Å, β = 91.7°. Structural characterization of this enzyme will contribute to a better understanding of the structural requirements for xylooligosaccharide specificity within the GH43 family.

  6. Purification, characterization and end product analysis of dextran degrading endodextranase from Bacillus licheniformis KIBGE-IB25.

    Science.gov (United States)

    Zohra, Rashida Rahmat; Aman, Afsheen; Ansari, Asma; Haider, Muhammad Samee; Qader, Shah Ali Ul

    2015-07-01

    Degradation of high molecular weight dextran for obtaining low molecular weight dextran is based on the hydrolysis using chemical and enzymatic methods. Current research study focused on production, purification and characterization of dextranase from a newly isolated strain of Bacillus licheniformis KIBGE-IB25. Dextranase was purified up to 36 folds with specific activity of 1405 U/mg and molecular weight of 158 kDa. It was found that enzyme performs optimum cleavage of dextran (5000 Da, 0.5%) at 35 °C in 15 min at pH 4.5 with a Km and Vmax of 0.374 mg/ml and 182 μmol/min, respectively. Relative amino acid composition analysis of purified enzyme suggested the presence of higher number of hydrophobic, acidic and glycosylation promoting amino acids. The N-terminal sequence of dextranase KIBGE-IB25 was AYTVTLYLQG. It exhibited distinct amino acid sequence yet shared some inherent characteristics with glycosyl hydrolases (GH) family 49 and also testified the presence of O-glycosylation at N-terminal end. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Muhammad Nadeem

    2010-10-01

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

  8. Enthalpies of proton adsorption onto Bacillus licheniformis at 25, 37, 50, and 75 °C

    Science.gov (United States)

    Gorman-Lewis, Drew

    2011-03-01

    Understanding bacterial surface reactivity requires many different lines of investigation. Toward this end, we used isothermal titration calorimetry to measure heats of proton adsorption onto a Gram positive thermophile Bacillus licheniformis at 25, 37, 50, and 75 °C. Proton adsorption under all conditions exhibited exothermic heat production. Below pH 4.5, exothermic heats decreased as temperature increased above 37 °C; above pH 4.5, there was no significant difference in heats evolved at the temperatures investigated. Total proton uptake did not vary significantly with temperature. Site-specific enthalpies and entropies were calculated by applying a 4-site, non-electrostatic surface complexation model to the calorimetric data. Interpretation of site-specific enthalpies and entropies of proton adsorption for site L1, L2, and L4 are consistent with previous interpretations of phosphoryl, carboxyl, and hydroxyl/amine site-identities, respectively, and with previous calorimetric measurements of proton adsorption onto mesophilic species. Enthalpies and entropies for surface site L3 are not consistent with the commonly inferred phosphoryl site-identity and are more consistent with sulfhydryl functional groups. These results reveal intricacies of surface reactivity that are not detectable by other methods.

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Sanket J. Joshi

    2016-11-01

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

  11. Production of antimicrobial substances by Bacillus subtilis LFE-1, B. firmus HO-1 and B. licheniformis T6-5 isolated from an oil reservoir in Brazil.

    Science.gov (United States)

    Korenblum, E; der Weid, I; Santos, A L S; Rosado, A S; Sebastián, G V; Coutinho, C M L M; Magalhães, F C M; Paiva, M M; Seldin, L

    2005-01-01

    Forty Bacillus strains isolated from a Brazilian oil reservoir were tested against each other to select strains producing antimicrobial substances (AMS). Three strains, Bacillus subtilis (LFE-1), Bacillus firmus (H2O-1) and Bacillus licheniformis (T6-5), were selected due to their ability to inhibit more than 65% of the Bacillus strains tested. These three strains were also investigated for their capability to inhibit sulphate-reducing bacteria (SRB). Furthermore, physiological and biochemical characteristics of the antimicrobial compounds produced by the selected strains were determined. Among the forty strains tested, 36 (90%) strains were able to inhibit at least one Bacillus strain used as indicator in plate assays and three of them (LFE-1, T6-5 and H2O-1) were able to inhibit 65, 70 and 97.5% of the 40 strains studied here respectively. Clear zones of inhibition were observed when H2O-1 was tested against SRB-containing consortium T6-lab and Desulfovibrio alaskensis strain NCIMB 13491, while strain T6-5 was able to inhibit only the D. alaskensis strain. The three substances showed to be insensitive to different enzymes and chemicals, were heat stable and the substances produced by strains T6-5 and H2O-1 were active over a wide pH range. Three different AMS produced by Bacillus strains from an oil reservoir, two of them with activity against SRB, are presented here. The preliminary characterization of these AMS points to their potential use as biocides in the petroleum industry for controlling problems associated with SRB.

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

    Science.gov (United States)

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

    2008-01-23

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

  13. Residues Phe103 and Phe149 are critical for the co-chaperone activity of Bacillus licheniformis GrpE.

    Science.gov (United States)

    Lin, Min-Guan; Chi, Meng-Chun; Chen, Bo-En; Wang, Tzu-Fan; Lo, Huei-Fen; Lin, Long-Liu

    2015-01-01

    A tryptophan-free Bacillus licheniformis nucleotide exchange factor (BlGrpE) and its Trp mutants (F70W, F103W, F149W, F70/103W, F70/149W, F103/149W and F70/103/149W) were over-expressed and purified to near homogeneity. Simultaneous addition of B. licheniformis DnaJ, NR-peptide and individual variants synergistically stimulated the ATPase activity of a recombinant DnaK (BlDnaK) from the same bacterium by 3.1-14.7-fold, which are significantly lower than the synergistic stimulation (18.9-fold) of BlGrpE. Protein-protein interaction analysis revealed that Trp mutants relevant to amino acid positions 103 and 149 lost the ability to bind BlDnaK. Circular dichroism measurements indicate that F70W displayed a comparable level of secondary structure to that of BlGrpE, and the wild-type protein and the Trp mutants as well all experienced a reversible behavior of thermal denaturation. Guanidine hydrochloride (GdnHCl)-induced unfolding transition for BlGrpE was calculated to be 1.25 M corresponding to ΔG(N-U) of 4.29 kcal/mol, whereas the unfolding transitions of mutant proteins were in the range of 0.77-1.31 M equivalent to ΔG(N-U) of 2.41-4.14 kcal/mol. Taken together, the introduction of tryptophan residue, especially at positions 103 and 149, into the primary structure of BlGrpE has been proven to be detrimental to structural integrity and proper function of the protein. Copyright © 2014 Elsevier B.V. All rights reserved.

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

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    S. A. Ahmed

    2012-03-01

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

  15. Isolation and Characterization of Thermophilic Bacteria from Jordanian Hot Springs: Bacillus licheniformis and Thermomonas hydrothermalis Isolates as Potential Producers of Thermostable Enzymes.

    Science.gov (United States)

    Mohammad, Balsam T; Al Daghistani, Hala I; Jaouani, Atef; Abdel-Latif, Saleh; Kennes, Christian

    2017-01-01

    The aim of this study was the isolation and characterization of thermophilic bacteria from hot springs in Jordan. Ten isolates were characterized by morphological, microscopic, biochemical, molecular, and physiological characteristics. Sequencing of the 16S rDNA of the isolates followed by BLAST search revealed that nine strains could be identified as Bacillus licheniformis and one isolate as Thermomonas hydrothermalis . This is the first report on the isolation of Thermomonas species from Jordanian hot springs. The isolates showed an ability to produce some thermostable enzymes such as amylase, protease, cellulose, gelatins, and lecithin. Moreover, the UPGMA dendrogram of the enzymatic characteristics of the ten isolates was constructed; results indicated a high phenotypic diversity, which encourages future studies to explore further industrial and environmental applications.

  16. Isolation and Characterization of Thermophilic Bacteria from Jordanian Hot Springs: Bacillus licheniformis and Thermomonas hydrothermalis Isolates as Potential Producers of Thermostable Enzymes

    Directory of Open Access Journals (Sweden)

    Balsam T. Mohammad

    2017-01-01

    Full Text Available The aim of this study was the isolation and characterization of thermophilic bacteria from hot springs in Jordan. Ten isolates were characterized by morphological, microscopic, biochemical, molecular, and physiological characteristics. Sequencing of the 16S rDNA of the isolates followed by BLAST search revealed that nine strains could be identified as Bacillus licheniformis and one isolate as Thermomonas hydrothermalis. This is the first report on the isolation of Thermomonas species from Jordanian hot springs. The isolates showed an ability to produce some thermostable enzymes such as amylase, protease, cellulose, gelatins, and lecithin. Moreover, the UPGMA dendrogram of the enzymatic characteristics of the ten isolates was constructed; results indicated a high phenotypic diversity, which encourages future studies to explore further industrial and environmental applications.

  17. Stimulatory effect of medium ingredients on alkaline protease production by bacillus licheniformis N-2 and compatibility studies with commercial detergents

    International Nuclear Information System (INIS)

    Nadeem, M.; Baig, S.; Qazi, J.I.

    2008-01-01

    Suitable concentration of ingredients of the growth medium played a vital role in production of alkaline protease by Bacillus licheniformis. Maximum enzyme activity (875.05 PU/ml) was achieved when the bacterium was grown in the medium containing glucose (1%), soybean meal (1%), K/sub 2/ HPO/sub 4/ (0.5%), MgSO/sub 4/ 7H/sub 2/O (0.05%), NaCI (0.05%), CaCI/sub 2/ 2H/sub 2/O (0.05%) at 37 degree C on 24 h incubation period with agitation of 140 rpm in shake flask cultures. More than 1% glucose decreased the enzyme production. The protease had excellent stability with wide range of Commercial detergents such as Ariel, Bonus, Bright Total, Surf Excel, Wheel and non-branded detergents, recommending its use as an effective additive in detergent formulation. (author)

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

    Science.gov (United States)

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

    2016-03-01

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

  19. Evolving transpeptidase and hydrolytic variants of γ-glutamyl transpeptidase from Bacillus licheniformis by targeted mutations of conserved residue Arg109 and their biotechnological relevance.

    Science.gov (United States)

    Bindal, Shruti; Sharma, Sujata; Singh, Tej P; Gupta, Rani

    2017-05-10

    γ-Glutamyl transpeptidase (GGT) catalyzes the transfer of the γ-glutamyl moiety from donor compounds such as l-glutamine (Gln) and glutathione (GSH) to an acceptor. During the biosynthesis of various γ-glutamyl-containing compounds using GGT enzyme, auto-transpeptidation reaction leads to the formation of unwanted byproducts. Therefore, in order to alter the auto-transpeptidase activity of the GGT enzyme, the binding affinity of Gln should be modified. Structural studies of the Bacillus licheniformis GGT (BlGT) complexed with the glutamic acid has shown that glutamic acid has strong ionic interactions through its α-carboxlic group with the guanidine moiety of Arg109. This interaction appears to be an important contributor for the binding affinity of Gln. In view of this, six mutants of Bacillus licheniformis ER15 GGT (BlGGT) viz. Arg109Lys, Arg109Ser, Arg109Met, Arg109Leu, Arg109Glu and Arg109Phe were prepared. As seen from the structure of BlGT, the mutation of Arg109 to Lys109 may reduce the affinity for Gln to some extent, whereas the other mutations are expected to lower the affinity much more. Biophysical characterization and functional studies revealed that Arg109Lys mutant has increased transpeptidation activity and catalytic efficiency than the other mutants. The Arg109Lys mutant showed high conversion rates for l-theanine synthesis as well. Moreover, the Arg109Met mutant showed increased hydrolytic activity as it completely altered the binding of Gln at the active site. Also, the salt stability of the enzyme was significantly improved on replacing Arg109 by Met109 which is required for hydrolytic applications of GGTs in food industries. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Bacillus licheniformis BlaR1 L3 Loop Is a Zinc Metalloprotease Activated by Self-Proteolysis

    Science.gov (United States)

    Sépulchre, Jérémy; Amoroso, Ana; Joris, Bernard

    2012-01-01

    In Bacillus licheniformis 749/I, BlaP β-lactamase is induced by the presence of a β-lactam antibiotic outside the cell. The first step in the induction mechanism is the detection of the antibiotic by the membrane-bound penicillin receptor BlaR1 that is composed of two functional domains: a carboxy-terminal domain exposed outside the cell, which acts as a penicillin sensor, and an amino-terminal domain anchored to the cytoplasmic membrane, which works as a transducer-transmitter. The acylation of BlaR1 sensor domain by the antibiotic generates an intramolecular signal that leads to the activation of the L3 cytoplasmic loop of the transmitter by a single-point cleavage. The exact mechanism of L3 activation and the nature of the secondary cytoplasmic signal launched by the activated transmitter remain unknown. However, these two events seem to be linked to the presence of a HEXXH zinc binding motif of neutral zinc metallopeptidases. By different experimental approaches, we demonstrated that the L3 loop binds zinc ion, belongs to Gluzincin metallopeptidase superfamily and is activated by self-proteolysis. PMID:22623956

  1. The clinical impact of antimicrobial resistance genomics in competition with she-camels recurrent mastitis metabolomics due to heterogeneous Bacillus licheniformis field isolates

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    Nesreen Allam Tantawy Allam

    2017-11-01

    Full Text Available Background and Aim: Recently, cases of mastitis refractory to treatment have been reported frequently. There are limited routine laboratory investigations on Camelidae infections. Mastitis has been estimated to affect more than 25% of lactating she-camel with up to 70% milk loss. The details of Bacillus spp. pathogenesis in mastitis are not yet fully described. The present study is the first detailed phenotypic and genotypic characterization of Bacillus licheniformis isolates from recurrent mastitic she-camels with sepsis in Egypt. Materials and Methods: The udders of 100 she-camels were investigated, samples collected from smallholders' farmers in 10 localities within three governorates in Egypt: Marsa Matrouh, Giza, and Sharkia governorates. The pathogens ascend from udder inducing abortion at different trimesters of pregnancy. Polymerase chain reactions-mediated proofs of identity were applied for diagnostic and taxonomic purposes, where the 16S rRNA gene sequence and the β subunit of RNA polymerase encoding gene rpoB are the molecular targets. Results: The genetic elements classified the subspecies to B. licheniformis 61.4%, in addition to, Corynebacterium bovis 29.8%. The somatic cell count (≤1x107 cells/ml and California mastitis test reactivity (+3 or +4 of milk clinically classified the she-camels population (n=100 under investigation into 50, 20, and 30 as healthy, subclinical, and clinical mastitic she-camels, respectively. During bacterial isolation, 80 species were noticed, of which 71.25% (57/80 and 28.75% (23/80 were Gram-positive and negative, respectively, in two clinical forms: Single (40%, n=16/40 and mixed (60%, n=34/40 bacterial infections. In vitro, 100% sensitivity for gentamycin (10 μg and ofloxacin (5 μg was noted; however, it was reduced to 50%. Moreover, during in vivo treatments cloxacillin (5 μg upraised as the most effective alternative with 90% sensitivity. Conclusion: Neither recurrent mastitis nor Bacillus

  2. The clinical impact of antimicrobial resistance genomics in competition with she-camels recurrent mastitis metabolomics due to heterogeneous Bacillus licheniformis field isolates

    Science.gov (United States)

    Allam, Nesreen Allam Tantawy; Sedky, Doaa; Mira, Enshrah Khalil

    2017-01-01

    Background and Aim:: Recently, cases of mastitis refractory to treatment have been reported frequently. There are limited routine laboratory investigations on Camelidae infections. Mastitis has been estimated to affect more than 25% of lactating she-camel with up to 70% milk loss. The details of Bacillus spp. pathogenesis in mastitis are not yet fully described. The present study is the first detailed phenotypic and genotypic characterization of Bacillus licheniformis isolates from recurrent mastitic she-camels with sepsis in Egypt. Materials and Methods: The udders of 100 she-camels were investigated, samples collected from smallholders’ farmers in 10 localities within three governorates in Egypt: Marsa Matrouh, Giza, and Sharkia governorates. The pathogens ascend from udder inducing abortion at different trimesters of pregnancy. Polymerase chain reactions-mediated proofs of identity were applied for diagnostic and taxonomic purposes, where the 16S rRNA gene sequence and the β subunit of RNA polymerase encoding gene rpoB are the molecular targets. Results:: The genetic elements classified the subspecies to B. licheniformis 61.4%, in addition to, Corynebacterium bovis 29.8%. The somatic cell count (≤1×107 cells/ml) and California mastitis test reactivity (+3 or +4) of milk clinically classified the she-camels population (n=100) under investigation into 50, 20, and 30 as healthy, subclinical, and clinical mastitic she-camels, respectively. During bacterial isolation, 80 species were noticed, of which 71.25% (57/80) and 28.75% (23/80) were Gram-positive and negative, respectively, in two clinical forms: Single (40%, n=16/40) and mixed (60%, n=34/40) bacterial infections. In vitro, 100% sensitivity for gentamycin (10 µg) and ofloxacin (5 µg) was noted; however, it was reduced to 50%. Moreover, during in vivo treatments, cloxacillin (5 µg) upraised as the most effective alternative with 90% sensitivity. Conclusion: Neither recurrent mastitis nor Bacillus

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

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    Meng-Chun Chi

    2013-02-01

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

  4. Characterization of the glutamate-specific endopeptidase from Bacillus licheniformis expressed in Escherichia coli.

    Science.gov (United States)

    Ye, Wei; Wang, Haiying; Ma, Yi; Luo, Xiaochun; Zhang, Weimin; Wang, Jufang; Wang, Xiaoning

    2013-10-10

    Glutamate-specific endopeptidase from Bacillus licheniformis (GSE-BL) is widely used in peptide recovery and synthesis because of its unique substrate specificity. However, the mechanism underlying its specificity is still not thoroughly understood. In this study, the roles of the prosegment and key amino acids involved in the proteolytic activity of GSE-BL were investigated. Loss of the GSE-BL prosegment severely restricted enzymatic activity toward Z-Phe-Leu-Glu-pNA. A homologous model of GSE-BL revealed that it contains the catalytic triad "His47, Asp96 and Ser 167", which was further confirmed by site-directed mutagenesis. In vitro mutagenesis further indicated that Val2, Arg89 and His190 are essential for enzymatic activity toward Z-Phe-Leu-Glu-pNA. Moreover, the catalytic efficiency of Phe57Ala GSE-BL toward Z-Phe-Leu-Glu-pNA was 50% higher than that of the native mature GSE-BL. This is the first study to fully elucidate the key amino acids for proteolytic activity of GSE-BL. Mature GSE-BL could be obtained through self-cleavage alone when Lys at -1 position was replaced by Glu, providing a new strategy for the preparation of mature GSE-BL. This study yielded some valuable insights into the substrate specificity of glutamate-specific endopeptidase, establishing a foundation for broadening the applications of GSE-BL. Copyright © 2013 Elsevier B.V. All rights reserved.

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

    NARCIS (Netherlands)

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

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

  6. High Resolution X-ray Diffraction Dataset for Bacillus licheniformis Gamma Glutamyl Transpeptidase-acivicin complex: SUMO-Tag Renders High Expression and Solubility.

    Science.gov (United States)

    Kumari, Shobha; Pal, Ravi Kant; Gupta, Rani; Goel, Manisha

    2017-02-01

    Gamma glutamyl transpeptidase, (GGT) is a ubiquitous protein which plays a central role in glutathione metabolism and has myriad clinical implications. It has been shown to be a virulence factor for pathogenic bacteria, inhibition of which results in reduced colonization potential. However, existing inhibitors are effective but toxic and therefore search is on for novel inhibitors, which makes it imperative to understand the interactions of various inhibitors with the protein in substantial detail. High resolution structures of protein bound to different inhibitors can serve this purpose. Gamma glutamyl transpeptidase from Bacillus licheniformis is one of the model systems that have been used to understand the structure-function correlation of the protein. The structures of the native protein (PDB code 4OTT), of its complex with glutamate (PDB code 4OTU) and that of its precursor mimic (PDB code 4Y23) are available, although at moderate/low resolution. In the present study, we are reporting the preliminary analysis of, high resolution X-ray diffraction data collected for the co-crystals of B. licheniformis, Gamma glutamyl transpeptidase, with its inhibitor, Acivicin. Crystals belong to the orthorhombic space group P2 1 2 1 2 1 and diffract X-ray to 1.45 Å resolution. This is the highest resolution data reported for all GGT structures available till now. The use of SUMO fused expression system enhanced yield of the target protein in the soluble fraction, facilitating recovery of protein with high purity. The preliminary analysis of this data set shows clear density for the inhibitor, acivicin, in the protein active site.

  7. Lichenysin production is improved in codY null Bacillus licheniformis by addition of precursor amino acids.

    Science.gov (United States)

    Zhu, Chengjun; Xiao, Fang; Qiu, Yimin; Wang, Qin; He, Zhili; Chen, Shouwen

    2017-08-01

    Lichenysin is categorized into the family of lipopeptide biosurfactants and has a variety of applications in the petroleum industry, bioremediation, pharmaceuticals, and the food industry. Currently, large-scale production is limited due to the low yield. This study found that lichenysin production was repressed by supplementation of extracellular amino acids. The global transcriptional factor CodY was hypothesized to prevent lichenysin biosynthesis under an amino acid-rich condition in Bacillus licheniformis. Thus, the codY null strain was constructed, and lichenysin production was increased by 31.0% to 2356 mg/L with the addition of precursor amino acids, and the lichenysin production efficiency was improved by 42.8% to 98.2 mg/L• h. Correspondingly, the transcription levels of the lichenysin synthetase gene lchAA, and its corresponding regulator genes comA, degQ, and degU, were upregulated. Also, the codY deletion enhanced biosynthesis of lichenysin precursor amino acids (Gln, Ile, Leu, and Val) and reduced the formation of byproducts, acetate, acetoin, and 2,3-butanediol. This study firstly reported that lichenysin biosynthesis was negatively regulated by CodY and lichenysin production could be further improved with the precursor amino acid amendment in the codY null strain.

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

    Science.gov (United States)

    Tabassum, Romana; Khaliq, Shazia; Rajoka, Muhammad Ibrahim; Agblevor, Foster

    2014-01-01

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

  9. Enhanced production of tetramethylpyrazine in Bacillus licheniformis BL1 by bdhA disruption and 2,3-butanediol supplementation.

    Science.gov (United States)

    Meng, Wu; Xiao, Dongguang; Wang, Ruiming

    2016-03-01

    The 2,3-butanediol (2,3-BD) dehydrogenase gene (bdhA) of Bacillus licheniformis BL1 was disrupted to construct the tetramethylpyrazine (TMP)-producing BLA strain. During microaerobic fermentation, the bdhA-disrupted BLA strain produced 46.98 g TMP/l, and this yield was 23.99% higher than that produced by the parent BL1 strain. In addition, the yield of acetoin, which is a TMP precursor, also increased by 28.98% in BLA. The TMP production by BL1 was enhanced by supplementing the fermentation medium with 2,3-BD. The yield of TMP improved from 37.89 to 44.77 g/l as the concentration of 2,3-BD increased from 0 to 2 g/l. The maximum TMP and acetoin yields increased by 18.16 and 17.87%, respectively with the increase in 2,3-BD concentration from 0 to 2 g/l. However, no increase was observed when the concentration of 2,3-BD in the matrix was ≥3 g/l. This study provides a valuable strategy to enhance TMP and acetoin productivity of mutagenic strains by gene manipulation and optimizing fermentation conditions.

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

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    Surawut Sangmanee

    2016-06-01

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

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

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    Yulia Sokurenko

    2016-01-01

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

  12. Purification and characterization of a thermostable alkaline cellulase produced by Bacillus licheniformis 380 isolated from compost

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    ÉVILIN G. DE MARCO

    2017-10-01

    Full Text Available ABSTRACT During composting processes, the degradation of organic waste is accomplished and driven by a succession of microbial populations exhibiting a broad range of functional competencies. A total of 183 bacteria, isolated from a composting process, were evaluated for cellulase activity at different temperatures (37, 50, 60, and 70°C and pH values. Out of the 22 isolates that showed activity, isolate 380 showed the highest cellulase activity. Its ability to produce cellulase was evaluated in culture medium supplemented with carboxymethyl cellulose, microcrystalline cellulose, wheat straw, and rice husk. The culture medium supplemented with carboxymethyl cellulose induced higher enzyme activity after 6 hours of incubation (0.12 UEA mL-1 min-1. For wheat straw and rice husk, the results were 0.08 UEA mL-1 min-1 for both, while for microcrystalline cellulose, 0.04 UEA mL-1 min-1 were observed. The highest carboxymethyl cellulase activity was observed at 60°C (0.14 UEA mL-1 min-1 for both crude and partially purified enzyme after 30 and 120 min of incubation, respectively. Alkalinization of the medium was observed during cultivation in all substrates. The cellulase had a molecular mass of 20 kDa determined by SDS-Page. Isolate 380 was identified as Bacillus licheniformis. This work provides a basis for further studies on composting optimization.

  13. Identification of Bacitracin Produced by Local Isolate of Bacillus ...

    African Journals Online (AJOL)

    Bacillus licheniformis was isolated from soil of different house gardens. Diagnosis was performed according to Gram stain, motility, shape forming, aerobic condition and other tests. Bacitracin was primary identified after its activity was tested against some species of Gram positive and Gram negative bacteria. Identification ...

  14. Growth measurement of some amylolytic bacillus species in three media

    International Nuclear Information System (INIS)

    Ajayi, A.O.

    2009-01-01

    Study of the growth pattern of some Bacillus species on starchy substrates showed that the metabolic activity affected the enzymatic activity. B. subtilis (WBS), B. licheniformis (WBL) and B. coagulans (MBC) generally had higher growth rate. B. circulans (SBC) and B. coagulans (WBC) had higher growth on cornstarch medium with corresponding higher beta-amylase production as compared to other strains such as B. polymyxa. Ten of the 13 Bacillus species studied had better performance on cornstarch than on soluble starch except B. macerans (MBM), B. macerans (SMB2) and B. subtilis (WBS). The enzyme production ranged from 0.022 unit/cfu x 102 to 0.912 unit/cfu x 102 on cornstarch and 0.01 unit/cfu x 102 to 0.693 unit/cfu x 102 on soluble starch. Relatively higher a-amylase activity was observed in B. subtilis, B. licheniformis, B. macerans and B. circulans (WBC1). (author)

  15. Diversity of Protease-Producing Bacillus spp. From Fresh Indonesian Tempeh Based on 16S rRNA Gene Sequence

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    Tati Barus

    2017-01-01

    Full Text Available Tempeh is a type of traditional fermented food in Indonesia. The fermentation can be performed by Rhizopus microsporus as a main microorganism. However, Bacillus spp. is found in abundance in tempeh production. Nevertheless, information regarding the diversity of Bacillus spp. in tempeh production has not been reported yet. Therefore, the aim of this investigation was to study the genetic diversity of Bacillus spp. in tempeh production based on the 16S ribosomal RNA sequence. In this study, about 22 of 24 fresh tempeh from Jakarta, Bogor, and Tangerang were used. A total of 52 protease-producing Bacillus spp. isolates were obtained. Based on 16S ribosomal RNA results, all 52 isolates were identified to be similar to B. pumilus, B. subtilis, B. megaterium, B. licheniformis, B. cereus, B. thuringiensis, B. amyloliquefaciens, Brevibacillus brevis, and Bacillus sp. All the identified isolates were divided into two large clusters: 1 a cluster of B. cereus, B. thuringiensis, Bacillus sp., and B. brevis and 2 a cluster of B. pumilus, B. subtilis, B. megaterium, B. licheniformis, and B. amyloliquefaciens. Information about the Bacillus spp. role in determining the quality of tempeh has not been reported and this is a preliminary study of Bacillus spp. from tempeh.

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

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    H.S. El-Sheshtawy

    2015-06-01

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

  17. Reclassification of Bacillus axarquiensis Ruiz-Garcia et al. 2005 and Bacillus malacitensis Ruiz-Garcia et al. 2005 as later heterotypic synonyms of Bacillus mojavensis Roberts et al. 1994.

    Science.gov (United States)

    Wang, Li-Ting; Lee, Fwu-Ling; Tai, Chun-Ju; Yokota, Akira; Kuo, Hsiao-Ping

    2007-07-01

    The Bacillus subtilis group encompasses the taxa Bacillus subtilis subsp. subtilis, B. licheniformis, B. amyloliquefaciens, B. atrophaeus, B. mojavensis, B. vallismortis, B. subtilis subsp. spizizenii, B. sonorensis, B. velezensis, B. axarquiensis and B. malacitensis. In this study, the taxonomic relatedness between the species B. axarquiensis, B. malacitensis and B. mojavensis was investigated. Sequence analysis of the 16S rRNA gene and the gene for DNA gyrase subunit B (gyrB) confirmed the very high similarities between these three type strains and a reference strain of B. mojavensis (>99 and >97 %, respectively). DNA-DNA hybridization experiments revealed high relatedness values between the type strains of B. axarquiensis, B. malacitensis and B. mojavensis and between these strains and a reference strain of B. mojavensis (83-98 %). Based on these molecular taxonomic data and the lack of phenotypic distinctive characteristics, Bacillus axarquiensis and Bacillus malacitensis should be reclassified as later heterotypic synonyms of Bacillus mojavensis.

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

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    Gillian E. Gardiner

    2012-10-01

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

  19. Extracellular facile biosynthesis, characterization and stability of gold nanoparticles by Bacillus licheniformis.

    Science.gov (United States)

    Singh, Sneha; Vidyarthi, Ambarish Sharan; Nigam, Vinod Kumar; Dev, Abhimanyu

    2014-02-01

    The development of a reliable, eco-friendly process for synthesis of gold nanoparticles (AuNPs) has gained impetus in recent years to counter the drawbacks of chemical and physical methods. This study illustrates simple, green synthesis of AuNPs in vitro using cell lysate supernatant (CLS) of non-pathogenic bacteria and to investigate its potential antimicrobial activity. Gold nanoparticles were synthesized by the reduction of precursor AuCl4- ions using the CLS of Bacillus licheniformis at 37°C upon 24 h of incubation. The nanoparticles were characterized for their morphology, particle size, optical absorption, zeta potential, and stability. Further the antimicrobial activity was assayed using cup-plate method. The process of biosynthesis was extracellular and the gold ions were reduced to stable nanogold of average size 38 nm. However, upon storage of AuNPs for longer duration at room temperature stability was influenced in terms of increase in particle size and decrease in zeta potential with respect to as synthesized nanoparticles. SEM micrographs revealed the spherical shape of AuNPs and EDX analysis confirmed the presence of gold in the sample. Also clear zone of inhibition was observed against Bacilllus subtilis MTCC 8364, Pseudomonas aeruginosa MTCC 7925, and Escherichia coli MTCC 1698 confirming the antimicrobial activity of AuNPs. The bioprocess under study was simple and less time consuming as compared to other methods as the need for harvesting AuNPs from within the microbial cells via downstream process will be eliminated. Nanoparticles exhibited good stability even in absence of external stabilizing agents. AuNPs showed good antimicrobial activity against several Gram-negative and Gram-positive pathogenic bacteria. The extracellular biosynthesis from CLS may serve as a suitable alternative for large scale synthesis of gold nanoparticles in vitro. The synthesis from lysed bacterial cell strongly suggests that exposure of microbial whole cells to the

  20. Characterization of thermostable cellulase produced by Bacillus strains isolated from solid waste of carrageenan

    Science.gov (United States)

    Listyaningrum, N. P.; Sutrisno, A.; Wardani, A. K.

    2018-03-01

    Cellulase-producing bacteria was isolated from solid waste of carrageenan and identified as Bacillus licheniformis C55 by 16S rRNA sequencing. The optimum condition for cellulase production was obtained at pH and temperature of 8.0 and 50°C, respectively in a medium containing glucose as carbon source and 1.0% carboxymethyl cellulose (CMC) to stimulate the cellulase production. Most remarkably, the enzyme retained its relative activity over 50% after incubation at 50°C for 90 minutes. Substrate specificity suggested that the enzyme is an endoglucanase. The molecular mass of Bacillus licheniformis C55 crude cellulase was found about 18 kDa by SDS-PAGE analysis. This thermostable enzyme would facilitate development of more efficient and cost-effective forms of the process to convert lignocellulosic biomass into high-value products.

  1. Effects of Two Surfactants and Beta-Cyclodextrin on Beta-Cypermethrin Degradation by Bacillus licheniformis B-1.

    Science.gov (United States)

    Zhao, Jiayuan; Chi, Yuanlong; Liu, Fangfang; Jia, Dongying; Yao, Kai

    2015-12-23

    The biodegradation efficiency of beta-cypermethrin (β-CY) is low especially at high concentrations mainly due to poor contact between this hydrophobic pesticide and microbial cells. In this study, the effects of two biodegradable surfactants (Tween-80 and Brij-35) and β-cyclodextrin (β-CD) on the growth and cell surface hydrophobicity (CSH) of Bacillus licheniformis B-1 were studied. Furthermore, their effects on the solubility, biosorption, and degradation of β-CY were investigated. The results showed that Tween-80 could slightly promote the growth of the strain while Brij-35 and β-CD exhibited little effect on its growth. The CSH of strain B-1 and the solubility of β-CY were obviously changed by using Tween-80 and Brij-35. The surfactants and β-CD could enhance β-CY biosorption and degradation by the strain, and the highest degradation was obtained in the presence of Brij-35. When the surfactant or β-CD concentration was 2.4 g/L, the degradation rate of β-CY in Brij-35, Tween-80, and β-CD treatments was 89.4%, 50.5%, and 48.1%, respectively. The half-life of β-CY by using Brij-35 was shortened by 69.1 h. Beta-CY content in the soil with both strain B-1 and Brij-35 decreased from 22.29 mg/kg to 4.41 mg/kg after incubation for 22 d. This work can provide a promising approach for the efficient degradation of pyrethroid pesticides by microorganisms.

  2. Improved keratinase production for feather degradation by Bacillus ...

    African Journals Online (AJOL)

    Optimal medium was used to improve the production of keratinase by Bacillus licheniformis ZJUEL31410, which has a promising application in the transformation of feather into soluble protein. The results of single factor design revealed that the concentration of feather at 20 g/l and the initial pH at value 8 was the best for ...

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

    Directory of Open Access Journals (Sweden)

    Jason Gioia

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

  4. Effect of temperature, pH and metal lons on the activity and stability of alkaline protease from novel bacillus licheniformis mzk03

    International Nuclear Information System (INIS)

    Sayem, S.M.A.; Hoq, M.M.; Alam, M.J.

    2006-01-01

    The effect of temperature, pH and metal ions on the activity and stability of crude protease from Bacillus licheniformis MZK03 was studied. The fermentation in shake culture revealed that maximum level of enzyme was produced at 37 degree C and pH 8.5 after 39 hr at 120 rpm. It lost its activity rapidly above 50 degree C and half-life of the protease at this temperature was 50 min with optimum activity at 40 degree C. It was most stable at pH 8.5 and lost its activity rapidly above pH 10.0, and at pH 11.0 reached 30% of the activity obtained at pH 9.0. The enzyme lost its activity completely at pH 13.0. Optimum proteolytic activity was found at 40 degree C and pH 9.5. The enzyme activity was accelerated by the addition of Mg/sup 2+/, Ca/sup 2+/ and Mn/sup 2+/, whereas it was inhibited by Hg/sup 2+/. (author)

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

    Directory of Open Access Journals (Sweden)

    Zahra Ghobadi Nejad

    2014-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Leon Rabinovitch

    1976-01-01

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

  7. Synthesis and Physicochemical Characterization of D-Tagatose-1-Phosphate: The Substrate of the Tagatose-1-Phosphate Kinase in the Phosphotransferase System-Mediated D-Tagatose Catabolic Pathway of Bacillus licheniformis.

    Science.gov (United States)

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

    2015-01-01

    We report the first enzymatic synthesis of D-tagatose-1-phosphate (Tag-1P) by the multicomponent phosphoenolpyruvate:sugar phosphotransferase system (PEP-PTS) present in tagatose-grown cells of Klebsiella pneumoniae. Physicochemical characterization by (31)P and (1)H nuclear magnetic resonance spectroscopy reveals that, in solution, this derivative is primarily in the pyranose form. Tag-1P was used to characterize the putative tagatose-1-phosphate kinase (TagK) of the Bacillus licheniformis PTS-mediated D-tagatose catabolic pathway (Bli-TagP). For this purpose, a soluble protein fusion was obtained with the 6 His-tagged trigger factor (TF(His6)) of Escherichia coli. The active fusion enzyme was named TagK-TF(His6). Tag-1P and D-fructose-1-phosphate are substrates for the TagK-TF(His6) enzyme, whereas the isomeric derivatives D-tagatose-6-phosphate and D-fructose-6-phosphate are inhibitors. Studies of catalytic efficiency (kcat/Km) reveal that the enzyme specificity is markedly in favor of Tag-1P as the substrate. Importantly, we show in vivo that the transfer of the phosphate moiety from PEP to the B. licheniformis tagatose-specific Enzyme II in E. coli is inefficient. The capability of the PTS general cytoplasmic components of B. subtilis, HPr and Enzyme I to restore the phosphate transfer is demonstrated. © 2015 S. Karger AG, Basel.

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

    Lifescience Database Archive (English)

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

  9. Antimicrobial Susceptibility of Bacillus Strains Isolated from Primary Starters for African Traditional Bread Production and Characterization of the Bacitracin Operon and Bacitracin Biosynthesis

    Science.gov (United States)

    Sørensen, Kim I.; Thorsen, Line; Stuer-Lauridsen, Birgitte; Abdelgadir, Warda S.; Nielsen, Dennis S.; Derkx, Patrick M. F.; Jespersen, Lene

    2012-01-01

    Bacillus spp. are widely used as feed additives and probiotics. However, there is limited information on their resistance to various antibiotics, and there is a growing concern over the transfer of antibiotic resistance genes. The MIC for 8 antibiotics was determined for 85 Bacillus species strains, Bacillus subtilis subsp. subtilis (n = 29), Bacillus licheniformis (n = 38), and Bacillus sonorensis (n = 18), all of which were isolated from starters for Sudanese bread production. All the strains were sensitive to tetracycline (8.0 mg/liter), vancomycin (4.0 mg/liter), and gentamicin (4.0 mg/liter) but resistant to streptomycin. Sensitivity to clindamycin, chloramphenicol, and kanamycin was species specific. The erythromycin resistance genes ermD and ermK were detected by PCR in all of the erythromycin-resistant (MIC, ≥16.0 mg/liter) B. licheniformis strains and one erythromycin-sensitive (MIC, 4.0 mg/liter) B. licheniformis strain. Several amino acid changes were present in the translated ermD and ermK nucleotide sequences of the erythromycin-sensitive strain, which could indicate ErmD and ErmK protein functionalities different from those of the resistance strains. The ermD and ermK genes were localized on an 11.4-kbp plasmid. All of the B. sonorensis strains harbored the bacitracin synthetase gene, bacA, and the transporter gene bcrA, which correlated with their observed resistance to bacitracin. Bacitracin was produced by all the investigated species strains (28%), as determined by ultra-high-definition quadrupole time-of-flight liquid chromatography-mass spectrometry (UHD-QTOF LC/MS). The present study has revealed species-specific variations in the antimicrobial susceptibilities of Bacillus spp. and provides new information on MIC values, as well as the occurrence of resistance genes in Bacillus spp., including the newly described species B. sonorensis. PMID:22941078

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Lifescience Database Archive (English)

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

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

    Science.gov (United States)

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

    2017-01-01

    Multifarious applications of Bacillus licheniformis VS16-derived biosurfactant were explored. Labeo rohita fingerlings were injected intraperitoneally with 0.1 mL of phosphate-buffered saline (PBS) containing purified biosurfactant at 0 (control), 55 (S55), 110 (S110), 220 (S220), or 330 (S330) μg mL-1 concentrations. Various immunological parameters and the expression of immune-related genes were measured at 7, 14, and 21 days post-administration (dpa). At 21 dpa, fish were challenged with Aeromonas hydrophila and mortality was recorded for 14 days. Immune parameters such as lysozyme levels (39.29 ± 2.14 U mL-1), alternative complement pathway (61.21 ± 2.38 U mL-1), and phagocytic activities (33.37 ± 1.2%) were maximum (P Biosurfactant was effective in inhibiting biofilm formation up to 54.71 ± 1.27%. Moreover, it efficiently removed cadmium (Cd) from tested vegetables such as carrot, radish, ginger, and potato, with the highest removal efficiency (60.98 ± 1.29%) recorded in ginger contaminated with Cd. Collectively, these results suggest that isolated biosurfactant could be used in the aquaculture industry, in addition to its potential application to the food industry. PMID:28400765

  13. Evaluation of the use of amplified 16S rRNA gene-restriction fragment length polymorphism analysis to detect enterobacter cloacae and bacillus licheniformis for microbial enhanced oil recovery field pilot

    Energy Technology Data Exchange (ETDEWEB)

    Fujiwara, Kazuhiro; Tanaka, Shinji; Otsuka, Makiko; Ichimura, Naoya [Lansai Research Institute, Kyoto (Japan); Yonebayashi, Hideharu [Japan National Oil Corp., Chiba (Japan); Hong, Chengxie; Enomoto, Heiji [Tohoku University, Miyagi (Japan)

    1999-09-01

    Evaluation of effectiveness of restriction fragment length polymorphism (RFLP) analysis of the 16S rRNA gene of microorganisms injected into an oil reservoir, for monitoring their levels over time, was conducted. Two microorganisms, enterobacter cloacae TRC-322 and Bacillus licheniformis TRC-18-2-a, were focused in this paper among the microorganisms selected for injection, and gene fragments of the 16S rRNA gene of these microorganisms were amplified by polymerase chain reaction (PCP), using one set of universal primers. Samples of the reservoir brine and reservoir rock were obtained; the microorganisms inhabiting in the reservoir were isolated from these samples, and the 16S rRNA gene of these microorganisms was amplified, condition remaining the same. RFLP analysis was performed on the 16S rRNA gene of each of these microorganisms, using restriction endonucleases HhaI, MspI, AluI and TaqI as necessary. Comparison of the resultant rRNA gene fragments, demonstrated that closely-related species displaying RFLP profile similar to that of E. cloacae TRC-322 or B. licheniformis TRC-18-2-a were not among the microorganisms isolated from the reservoir. PCR-RFLP analysis of the 16S rRNA gene, using the protocol; presented in this paper, is effective to detect the presence appropriate injecting microorganisms. This method was also effective for studying microorganisms isolated from the reservoir, which have the ability to grow on a molasses. (author)

  14. 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. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

    Lifescience Database Archive (English)

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

  16. Molten Globule-Like Partially Folded State of Bacillus licheniformis α-Amylase at Low pH Induced by 1,1,1,3,3,3-Hexafluoroisopropanol

    Directory of Open Access Journals (Sweden)

    Adyani Azizah Abd Halim

    2014-01-01

    Full Text Available Effect of 1,1,1,3,3,3-hexafluoroisopropanol (HFIP on acid-denatured Bacillus licheniformis α-amylase (BLA at pH 2.0 was investigated by far-UV CD, intrinsic fluorescence, and ANS fluorescence measurements. Addition of increasing HFIP concentrations led to an increase in the mean residue ellipticity at 222 nm (MRE222 nm up to 1.5 M HFIP concentration beyond which it sloped off. A small increase in the intrinsic fluorescence and a marked increase in the ANS fluorescence were also observed up to 0.4 M HFIP concentration, both of which decreased thereafter. Far- and near-UV CD spectra of the HFIP-induced state observed at 0.4 M HFIP showed significant retention of the secondary structures closer to native BLA but a disordered tertiary structure. Increase in the ANS fluorescence intensity was also observed with the HFIP-induced state, suggesting exposure of the hydrophobic clusters to the solvent. Furthermore, thermal denaturation of HFIP-induced state showed a non-cooperative transition. Taken together, all these results suggested that HFIP-induced state of BLA represented a molten globule-like state at pH 2.0.

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

    Science.gov (United States)

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

    2016-04-01

    In the present study, we synthesized and characterized a probiotic Bacillus licheniformis cell free extract (BLCFE) coated silver nanoparticles (BLCFE-AgNPs). These BLCFE-AgNPs were characterized by UV-visible spectrophotometer, XRD, EDX, FTIR, TEM and AFM. A strong surface plasmon resonance centered at 422 nm in UV-visible spectrum indicates the formation of AgNPs. The XRD spectrum of silver nanoparticles exhibited 2θ values corresponding to the silver nanocrystal. TEM and AFM showed the AgNPs were spherical in shape within the range of 18.69-63.42 nm and the presence of silver was confirmed by EDX analysis. Light and Confocal Laser Scanning Microscope (CLSM) images showed a weak adherence and disintegrated biofilm formation of Vibrio parahaemolyticus Dav1 treated with BLCFE-AgNPs compared to control. This result suggests that BLCFE-AgNps may be used for the control of biofilm forming bacterial populations in the biomedical field. In addition, acute toxicity results concluded that BLCFE-AgNPs were less toxic to the fresh water crustacean Ceriodaphnia cornuta (50 μg/ml) when compared to AgNO3 (22 μg/ml). This study also reports a short term analysis (24 h) of uptake and depuration of BLCFE-AgNPs in C. cornuta. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. 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.

  19. Introduction of a unique tryptophan residue into various positions of Bacillus licheniformis DnaK.

    Science.gov (United States)

    Chen, Bo-En; Lin, Min-Guan; Lo, Huei-Fen; Wang, Tzu-Fan; Chi, Meng-Chun; Lin, Long-Liu

    2013-01-01

    Site-directed mutagenesis together with biochemical and biophysical techniques were used to probe effects of single-tryptophan-incorporated mutations on a bacterial molecular chaperone, Bacillus licheniformis DnaK (BlDnaK). Specifically, five phenylalanine residues (Phe(120), Phe(174), Phe(186), Phe(378) and Phe(396)) of BlDnaK were individually replaced by single tryptophans, thus creating site-specific probes for the fluorescence analysis of the protein. The steady-state ATPase activity for BlDnaK, F120W, F174W, F186W, F378W, and F396W was determined to be 76.01, 52.82, 25.32, 53.31, 58.84, and 47.53 nmol Pi/min/mg, respectively. Complementation test revealed that the single mutation at codons 120, 186, and 378 of the dnaK gene still allowed an Escherichia coli dnaK756-Ts strain to grow at a stringent temperature of 44°C. Simultaneous addition of co-chaperones and NR-peptide did not synergistically stimulate the ATPase activity of F174W and F396W, and these two proteins were unable to assist the refolding of GdnHCl-denatured luciferase. The heat-induced denaturation of all variants could be fitted adequately to a three-state model, in agreement with the observation for the wild-type protein. By CD spectral analysis, GdnHCl-induced unfolding transition for BlDnaK was 1.51 M corresponding to ΔG(N-U) of 1.69 kcal/mol; however, the transitions for mutant proteins were 1.07-1.55 M equivalent to ΔG(N-U) of 0.94-2.93 kcal/mol. The emission maximum of single-tryptophan-incorporated variants was in the range of 333.2-335.8 nm. Acrylamide quenching analysis showed that the mutant proteins had a dynamic quenching constant of 3.0-4.2 M(-1). Taken together, these results suggest that the molecular properties of BlDnaK have been significantly changed upon the individual replacement of selected phenylalanine residues by tryptophan. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Optimization of Levan Production by Cold-Active Bacillus licheniformis ANT 179 and Fructooligosaccharide Synthesis by Its Levansucrase.

    Science.gov (United States)

    Xavier, Janifer Raj; Ramana, Karna Venkata

    2017-03-01

    Fructooligosaccharides (FOS) and levan attract much attention due to a wide range of applications in food technology and pharmaceutical and cosmetic industry. Bacillus licheniformis ANT 179, isolated from Antarctica soil, produced levansucrase and levan in a medium containing sucrose as carbon substrate. In this study, characterization of levansucrase and production of short-chain FOS and levan were investigated. Temperature and pH optimum of the enzyme were found to be 60 °C and pH 6.0, respectively. The optimization of fermentation conditions for levan production using sugarcane juice by response surface methodology (RSM) was carried out. Central composite rotatable design was used to study the main and the interactive effects of medium components: sugarcane juice and casein peptone concentration on levan production by the bacterium. The optimized medium with sugarcane juice at 20 % (v/v) and casein peptone at 2 % (w/v) was found to be optimal at an initial pH of 7.0 and incubation temperature of 35 °C for 48 h. Under these conditions, the maximum levan concentration was 50.25 g/L on wet weight basis and 16.35 g/L on dry weight basis. The produced inulin type FOS (kestose and neokestose) and levan were characterized by Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR) analysis. The study revealed that the levansucrase could form FOS from sucrose. The locally available low-cost substrate such as sugarcane juice in the form of a renewable substrate is proposed to be suitable even for scale-up production of enzyme and FOS for industrial applications. The levan and FOS synthesized by the bacterium are suitable for food applications and biomedical uses as the bacterium has GRAS status and devoid of endotoxin as compared to other Gram-negative bacteria.

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

    Science.gov (United States)

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

    2002-04-01

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

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

    Directory of Open Access Journals (Sweden)

    Muhammad Nadeem

    2013-04-01

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

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

    Science.gov (United States)

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

    2014-02-15

    Waxy maize starch was subjected to α-amylase (Bacillus licheniformis) hydrolysis in buffered medium to determine the evolution of reaction in quantitative terms and also in terms of the morphology and crystallinity of the partially hydrolyzed starch granules. Gathered data allowed studying the pattern of action of this α-amylase over waxy maize starch granules, with particular focus on a preferential hydrolysis of the amorphous regions of starch. Results showed that waxy maize starch hydrolysis followed a two-stage kinetic profile with an initial stage characterized by high reaction rate, followed by a slower second stage. The change of hydrolysis rate occurred at approximately 6h of reaction, a time for which X-ray diffraction data quantitatively analyzed by three different techniques showed a maximum of crystallinity in partially hydrolyzed granules. Scanning electron microscopy images illustrated the action of α-amylases which implied the exoerosion of the granules surface, the entry of α-amylases into the granules through radial channels, their endoerosion towards the granule exterior, and their fragmentation. Fragmentation of waxy maize starch granules revealed internal layered structures of starch which were interpreted as hydrolyzed/non-hydrolyzed growth rings. Under the conditions chosen, kinetic, electron microscopy and X-ray data all gave evidence of a preferential action of α-amylase from Bacillus licheniformis towards the less ordered regions of waxy maize starch. Results showed that, provided the proper hydrolysis time is chosen, starch granules with increased crystallinity can be obtained by a pure enzymatic treatment. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Detoxification and anti-nutrients reduction of Jatropha curcas seed cake by Bacillus fermentation.

    Science.gov (United States)

    Phengnuam, Thanyarat; Suntornsuk, Worapot

    2013-02-01

    Jatropha curcas seed cake is a by-product generated from oil extraction of J. curcas seed. Although it contains a high amount of protein, it has phorbol esters and anti-nutritional factors such as phytate, trypsin inhibitor, lectin and saponin. It cannot be applied directly in the food or animal feed industries. This investigation was aimed at detoxifying the toxic and anti-nutritional compounds in J. curcas seed cake by fermentation with Bacillus spp. Two GRAS (generally recognized as safe) Bacillus strains used in the study were Bacillus subtilis and Bacillus licheniformis with solid-state and submerged fermentations. Solid-state fermentation was done on 10 g of seed cake with a moisture content of 70% for 7 days, while submerged fermentation was carried out on 10 g of seed cake in 100 ml distilled water for 5 days. The fermentations were incubated at the optimum condition of each strain. After fermentation, bacterial growth, pH, toxic and anti-nutritional compounds were determined. Results showed that B. licheniformis with submerged fermentation were the most effective method to degrade toxic and anti-nutritional compounds in the seed cake. After fermentation, phorbol esters, phytate and trypsin inhibitor were reduced by 62%, 42% and 75%, respectively, while lectin could not be eliminated. The reduction of phorbol esters, phytate and trypsin inhibitor was related to esterase, phytase and protease activities, respectively. J. curcas seed cake could be mainly detoxified by bacterial fermentation and the high-protein fermented seed cake could be potentially applied to animal feed. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  5. Rational designed mutagenesis of levansucrase from Bacillus licheniformis 8-37-0-1 for product specificity study.

    Science.gov (United States)

    He, Chunjuan; Yang, Yirui; Zhao, Renfei; Qu, Jingyao; Jin, Lan; Lu, Lili; Xu, Li; Xiao, Min

    2018-04-01

    Levansucrases, which belong to the glycoside hydrolase family 68 (GH68), synthesize β (2-6)-linked fructan levan with sucrose as substrate. We described the use of a levansucrase (Bl_SacB) from Bacillus licheniformis 8-37-0-1 for catalysis of fructosyl transfer to obtain high levan yield previously. In the present study, six variants (Y246A, N251A, K372A, R369A, R369S, and R369K) were constructed through sequence alignment and structural analysis to explore the synthesis mechanism of Bl_SacB. The selected residues were predicted to localize to the substrate-entering channel of the active cavity and close to or remote from the catalytic triad. The products of these variants ranged from homopolymers levan to fructo-oligosaccharides (FOSs). The primary FOSs were identified through MS and NMR analyses as neolevan-type neokestose [β-D-Fru-(2-6)-α-D-Glc-(1-2)-β-D-Fru], levan-type 6-kestose [β-D-Fru-(2-6)-β-D-Fru-(2-1)-α-D-Glc], and inulin-type 1-kestose [β-D-Fru-(2-1)-β-D-Fru-(2-1)-α-D-Glc]. The mutation at Tyr 246 located remote from the catalytic triad led to the production of short-chain oligosaccharides with degree of polymerization (DP) of up to 25. The replaced Arg 369 located close to the catalytic triad resulted in either elimination of polysaccharide synthesis or complete change in the dominant linkage of the products. The Michaelis constants (K m ) of Y246A, N251A, K372A, and R369K were found to be similar to that of the wild type (WT). However, the turnover number (k cat ) and the value of transfructosylation versus hydrolysis activity of the six variants decreased compared with those of the WT. Hence, the residues located on the surface of the substrate-entering channel of Bl_SacB can be critical in product linkage type and/or elongation mechanism.

  6. Developing a new production host from a blueprint: Bacillus pumilus as an industrial enzyme producer.

    Science.gov (United States)

    Küppers, Tobias; Steffen, Victoria; Hellmuth, Hendrik; O'Connell, Timothy; Bongaerts, Johannes; Maurer, Karl-Heinz; Wiechert, Wolfgang

    2014-03-24

    Since volatile and rising cost factors such as energy, raw materials and market competitiveness have a significant impact on the economic efficiency of biotechnological bulk productions, industrial processes need to be steadily improved and optimized. Thereby the current production hosts can undergo various limitations. To overcome those limitations and in addition increase the diversity of available production hosts for future applications, we suggest a Production Strain Blueprinting (PSB) strategy to develop new production systems in a reduced time lapse in contrast to a development from scratch.To demonstrate this approach, Bacillus pumilus has been developed as an alternative expression platform for the production of alkaline enzymes in reference to the established industrial production host Bacillus licheniformis. To develop the selected B. pumilus as an alternative production host the suggested PSB strategy was applied proceeding in the following steps (dedicated product titers are scaled to the protease titer of Henkel's industrial production strain B. licheniformis at lab scale): Introduction of a protease production plasmid, adaptation of a protease production process (44%), process optimization (92%) and expression optimization (114%). To further evaluate the production capability of the developed B. pumilus platform, the target protease was substituted by an α-amylase. The expression performance was tested under the previously optimized protease process conditions and under subsequently adapted process conditions resulting in a maximum product titer of 65% in reference to B. licheniformis protease titer. In this contribution the applied PSB strategy performed very well for the development of B. pumilus as an alternative production strain. Thereby the engineered B. pumilus expression platform even exceeded the protease titer of the industrial production host B. licheniformis by 14%. This result exhibits a remarkable potential of B. pumilus to be the

  7. Isolation and characterization of a novel analyte from Bacillus subtilis SC-8 antagonistic to Bacillus cereus.

    Science.gov (United States)

    Lee, Nam Keun; Yeo, In-Cheol; Park, Joung Whan; Kang, Byung-Sun; Hahm, Young Tae

    2010-09-01

    In this study, an effective substance was isolated from Bacillus subtilis SC-8, which was obtained from traditionally fermented soybean paste, cheonggukjang. The substance was purified by HPLC, and its properties were analyzed. It had an adequate antagonistic effect on Bacilluscereus, and its spectrum of activity was narrow. When tested on several gram-negative and gram-positive foodborne pathogenic bacteria such as Salmonella enterica, Salmonella enteritidis, Staphylococcus aureus, and Listeria monocytogenes, no antagonistic effect was observed. Applying the derivative from B. subtilis SC-8 within the same genus did not inhibit the growth of major soybean-fermenting bacteria such as Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloquefaciens. The range of pH stability of the purified antagonistic substance was wide (from 4.0 to >10.0), and the substance was thermally stable up to 60 degrees C. In the various enzyme treatments, the antagonistic activity of the purified substance was reduced with proteinase K, protease, and lipase; its activity was partially destroyed with esterase. Spores of B. cereus did not grow at all in the presence of 5mug/mL of the purified antagonistic substance. The isolated antagonistic substance was thought to be an antibiotic-like lipopeptidal compound and was tentatively named BSAP-254 because it absorbed to UV radiation at 254nm. Copyright 2010 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  8. In silico exploration of Red Sea Bacillus genomes for natural product biosynthetic gene clusters

    KAUST Repository

    Othoum, Ghofran K

    2018-05-22

    BackgroundThe increasing spectrum of multidrug-resistant bacteria is a major global public health concern, necessitating discovery of novel antimicrobial agents. Here, members of the genus Bacillus are investigated as a potentially attractive source of novel antibiotics due to their broad spectrum of antimicrobial activities. We specifically focus on a computational analysis of the distinctive biosynthetic potential of Bacillus paralicheniformis strains isolated from the Red Sea, an ecosystem exposed to adverse, highly saline and hot conditions.ResultsWe report the complete circular and annotated genomes of two Red Sea strains, B. paralicheniformis Bac48 isolated from mangrove mud and B. paralicheniformis Bac84 isolated from microbial mat collected from Rabigh Harbor Lagoon in Saudi Arabia. Comparing the genomes of B. paralicheniformis Bac48 and B. paralicheniformis Bac84 with nine publicly available complete genomes of B. licheniformis and three genomes of B. paralicheniformis, revealed that all of the B. paralicheniformis strains in this study are more enriched in nonribosomal peptides (NRPs). We further report the first computationally identified trans-acyltransferase (trans-AT) nonribosomal peptide synthetase/polyketide synthase (PKS/ NRPS) cluster in strains of this species.ConclusionsB. paralicheniformis species have more genes associated with biosynthesis of antimicrobial bioactive compounds than other previously characterized species of B. licheniformis, which suggests that these species are better potential sources for novel antibiotics. Moreover, the genome of the Red Sea strain B. paralicheniformis Bac48 is more enriched in modular PKS genes compared to B. licheniformis strains and other B. paralicheniformis strains. This may be linked to adaptations that strains surviving in the Red Sea underwent to survive in the relatively hot and saline ecosystems.

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

    Directory of Open Access Journals (Sweden)

    Maria Teresa Fernández

    2011-07-01

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

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

    Science.gov (United States)

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

    2016-06-01

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

  11. Co-Metabolic Degradation of β-Cypermethrin and 3-Phenoxybenzoic Acid by Co-Culture of Bacillus licheniformis B-1 and Aspergillus oryzae M-4.

    Science.gov (United States)

    Zhao, Jiayuan; Chi, Yuanlong; Xu, Yingchao; Jia, Dongying; Yao, Kai

    2016-01-01

    The degradation efficiency of organic contaminants and their associated metabolites by co-culture of microbes is mainly limited by toxic intermediates from co-metabolic degradation. In this study, we investigated the degradation of β-cypermethrin (β-CY) and 3-phenoxybenzoic acid (3-PBA) by co-culture of Bacillus licheniformis B-1 and Aspergillus oryzae M-4, as well as the influences of β-CY and 3-PBA metabolites on their degradation and the growth of strains B-1 and M-4. Our results indicated that 100 mg/L β-CY was degraded by 78.85%, and 3-PBA concentration was 0.05 mg/L after 72 h. Compared with using only strain B-1, the half-life (t1/2) of β-CY by using the two strains together was shortened from 84.53 h to 38.54 h, and the yield coefficient of 3-PBA was decreased from 0.846 to 0.001. At 100 mg/L of 3-PBA and gallic acid, β-CY and 3-PBA degradation were only 17.68% and 40.45%, respectively. As the toxic intermediate derived from co-metabolic degradation of β-CY by strain B-1, 3-PBA was efficiently degraded by strain M-4, and gallic acid, as the toxic intermediate from co-metabolic degradation of 3-PBA by strain M-4, was efficiently degraded by strain B-1. These results provided a promising approach for efficient biodegradation of β-CY and 3-PBA.

  12. Co-Metabolic Degradation of β-Cypermethrin and 3-Phenoxybenzoic Acid by Co-Culture of Bacillus licheniformis B-1 and Aspergillus oryzae M-4.

    Directory of Open Access Journals (Sweden)

    Jiayuan Zhao

    Full Text Available The degradation efficiency of organic contaminants and their associated metabolites by co-culture of microbes is mainly limited by toxic intermediates from co-metabolic degradation. In this study, we investigated the degradation of β-cypermethrin (β-CY and 3-phenoxybenzoic acid (3-PBA by co-culture of Bacillus licheniformis B-1 and Aspergillus oryzae M-4, as well as the influences of β-CY and 3-PBA metabolites on their degradation and the growth of strains B-1 and M-4. Our results indicated that 100 mg/L β-CY was degraded by 78.85%, and 3-PBA concentration was 0.05 mg/L after 72 h. Compared with using only strain B-1, the half-life (t1/2 of β-CY by using the two strains together was shortened from 84.53 h to 38.54 h, and the yield coefficient of 3-PBA was decreased from 0.846 to 0.001. At 100 mg/L of 3-PBA and gallic acid, β-CY and 3-PBA degradation were only 17.68% and 40.45%, respectively. As the toxic intermediate derived from co-metabolic degradation of β-CY by strain B-1, 3-PBA was efficiently degraded by strain M-4, and gallic acid, as the toxic intermediate from co-metabolic degradation of 3-PBA by strain M-4, was efficiently degraded by strain B-1. These results provided a promising approach for efficient biodegradation of β-CY and 3-PBA.

  13. Enzymatic synthesis of γ-L-glutamyl-S-allyl-L-cysteine, a naturally occurring organosulfur compound from garlic, by Bacillus licheniformis γ-glutamyltranspeptidase.

    Science.gov (United States)

    Chen, Yi-Yu; Lo, Huei-Fen; Wang, Tzu-Fan; Lin, Min-Guan; Lin, Long-Liu; Chi, Meng-Chun

    2015-01-01

    In the practical application of Bacillus licheniformis γ-glutamyltranspeptidase (BlGGT), we describe a straightforward enzymatic synthesis of γ-L-glutamyl-S-allyl-L-cysteine (GSAC), a naturally occurring organosulfur compound found in garlic, based on a transpeptidation reaction involving glutamine as the γ-glutamyl donor and S-allyl-L-cysteine as the acceptor. With the help of thin layer chromatography technique and computer-assisted image analysis, we performed the quantitative determination of GSAC. The optimum conditions for a biocatalyzed synthesis of GSAC were 200 mM glutamine, 200 mM S-allyl-L-cysteine, 50 mM Tris-HCl buffer (pH 9.0), and BlGGT at a final concentration of 1.0 U/mL. After a 15-h incubation of the reaction mixture at 60 °C, the GSAC yield for the free and immobilized enzymes was 19.3% and 18.3%, respectively. The enzymatic synthesis of GSAC was repeated under optimal conditions at 1-mmol preparative level. The reaction products together with the commercially available GSAC were further subjected to an ESI-MS/MS analysis. A significant signal with m/z of 291.1 and the protonated fragments at m/z of 73.0, 130.1, 145.0, and 162.1 were observed in the positive ESI-MS/MS spectrum, which is consistent with those of the standard compound. These results confirm the successful synthesis of GSAC from glutamine and S-allyl-L-cysteine by BlGGT. Copyright © 2015 Elsevier Inc. All rights reserved.

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

    International Nuclear Information System (INIS)

    Mushtaq, Z.; Adnan, A.; Mehmood, Z.

    2014-01-01

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

  15. ORF Alignment: NC_006322 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_006322 gi|52786856 >4uagA 8 419 4 423 5e-47 ... ref|YP_092685.1| MurC [Bacillus li...cheniformis ATCC 14580] gb|AAU41992.1| MurC ... [Bacillus licheniformis DSM 13] sp|Q65G22|MURC_BACLD

  16. Enhanced catalysis of L-asparaginase from Bacillus licheniformis by a rational redesign.

    Science.gov (United States)

    Sudhir, Ankit P; Agarwaal, Viplove V; Dave, Bhaumik R; Patel, Darshan H; Subramanian, R B

    2016-05-01

    L-Asparaginase (3.5.1.1) being antineoplastic in nature are used in the treatment of acute lymphoblastic leukemia (ALL). However glutaminase activity is the cause of various side effects when used as a drug against acute lymphoblastic leukemia (ALL). Therefore, there is a need of a novel L-asparaginase (L-ASNase) with low or no glutaminase activity. Such a property has been observed with L-ASNase from B. licheniformis (BliA). The enzyme being glutaminase free in nature paved the way for its improvement to achieve properties similar to or near to the commercially available L-ASNases. Rational enzyme engineering approach resulted in four mutants: G238N, E232A, D103V and Q112H. Among these the mutant enzyme, D103V, had a specific activity of 597.7IU/mg, which is higher than native (rBliA) (407.65IU/mg). Moreover, when the optimum temperature and in vitro half life were studied and compared with native BliA, D103V mutant BliA was better, showing tolerance to higher temperatures and a 3 fold higher half life. Kinetic studies revealed that the mutant D103V L-ASNase has increased substrate affinity, with Km value of 0.42mM and Vmax of 2778.9μmolmin(-1). Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Sporulation boundaries and spore formation kinetics of Bacillus spp. as a function of temperature, pH and a(w).

    Science.gov (United States)

    Baril, Eugénie; Coroller, Louis; Couvert, Olivier; El Jabri, Mohammed; Leguerinel, Ivan; Postollec, Florence; Boulais, Christophe; Carlin, Frédéric; Mafart, Pierre

    2012-10-01

    Sporulation niches in the food chain are considered as a source of hazard and are not clearly identified. Determining the sporulation environmental boundaries could contribute to identify potential sporulation niches. Spore formation was determined in a Sporulation Mineral Buffer. The effect of incubation temperature, pH and water activity on time to one spore per mL, maximum sporulation rate and final spore concentration was investigated for a Bacillus weihenstephanensis and a Bacillus licheniformis strain. Sporulation boundaries of B. weihenstephanensis and of B. licheniformis were similar to, or included within, the range of temperatures, pH and water activities supporting growth. For instance, sporulation boundaries of B. weihenstephanensis were evaluated at 5°C, 35°C, pH 5.2 and a(w) 0.960 while growth boundaries were observed at 5°C, 37°C, pH 4.9 and a(w) 0.950. Optimum spore formation was determined at 30°C pH 7.2 for B. weihenstephanensis and at 45°C pH 7.2 for B. licheniformis. Lower temperatures and pH delayed the sporulation process. For instance, the time to one spore per mL was tenfold longer when sporulation occurred at 10°C and 20°C, for each strain respectively, than at optimum sporulation temperature. The relative effect of temperature and pH on sporulation rates and on growth rates is similar. This work suggests that the influence of environmental factors on the quantitative changes in sporulation boundaries and rates was similar to their influence on changes in growth rate. Copyright © 2012 Elsevier Ltd. All rights reserved.

  18. Evaluation of Antimicrobial Activity of Bacillus Strains Isolated from Various Resources

    Directory of Open Access Journals (Sweden)

    Mohsen Golnari Maranni

    2017-02-01

    Full Text Available Abstract Background: Prevalence extension of antibiotic resistant bacteria has raised concerns about control of infections especially nosocomial infections. Many attempts have been done to replace antibiotics or limit their use. The use of antimicrobial agents produced by bacteria as antibiotic replacement has been promising in recent years. The goal of this study was to isolate Bacillus strains and evaluate their antimicrobial activity against some standard pathogens and clinical antibiotic resistant strains. Materials and Methods: In the present study, Bacillus strains were isolated from various resources and identified by 16S rDNA PCR method. Then, the phylogenetic tree of the isolates was constructed and antimicrobial activity of the isolates was investigated against some standard pathogens and clinical antibiotic resistant strains using spotting and well diffusion methods. Results: Eight Bacillus strains were isolated from 15 different samples. Based on the molecular identification, the isolates were identified as B.pumilus, B.coagulans, B.licheniformis, B.endophitycus and B.amiloliquefaciens. The results showed that isolates have antimicrobial activity against meticilin-resistant Staphylococcus aureus, vancomycin resistant enterococci, Klebsiella, Acinetobacter, Salmonella, Shigella, Listeria, Streptococcus and Escherichia coli. Conclusion: In this study, isolated Bacillus strains produced antimicrobial agents against pathogens and antibiotic resistant strains and inhibited their growth.

  19. Ampicillin radioprotector effect

    International Nuclear Information System (INIS)

    Padron, E.; Fernandez-Larrea, O.; Rios, F.

    1991-01-01

    This paper deals with the effect of ampicillin during irradiation and recovery of the Bacillus licheniformis RI 75-1 stump with a savage genotype of recovery exposed to ionizing radiations and treated with gamma quantums. Previous research enabled to prove that in Bacillus licheniformis spores suspensions, irradiated of vegetative cells in ampicillin at D-1--0- dose, causes significative lethal increases. In this paper, The irradiation of vegetative cells in presence of 5 mcg/ml of ampicillin increases the viability at doses above 1 kGy. the survival rates was raised when vegetative cells of Bacillus lincheniformis were irradiated at 1.2 kGy and recovered in a nutrient environment during 2 hours (LHR) in presence of ampicillin. Twenty-two stumps of Bacillus licheniformis obtained through different mutagenic treatments were studied in relation to the resistance of ampicillim and at 3 Kgy gamma quantums and a direct correlation between these two was stablished. Previous treatment with ampicillin of the vegetative cells from the Bacillus licheniformis increased the number of resistants to gamma quantums. There was no information about this phenomenom in literature consulted

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

    Directory of Open Access Journals (Sweden)

    Francesco Celandroni

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

  1. Effect of thymol in heating and recovery media on the isothermal and non-isothermal heat resistance of Bacillus spores.

    Science.gov (United States)

    Esteban, Maria-Dolores; Conesa, Raquel; Huertas, Juan-Pablo; Palop, Alfredo

    2015-06-01

    Members of the genus Bacillus include important food-borne pathogen and spoilage microorganisms for food industry. Essential oils are natural products extracted from herbs and spices, which can be used as natural preservatives in many foods because of their antibacterial, antifungal, antioxidant and anti-carcinogenic properties. The aim of this research was to explore the effect of the addition of different concentrations of thymol to the heating and recovery media on the thermal resistance of spores of Bacillus cereus, Bacillus licheniformis and Bacillus subtilis at different temperatures. While the heat resistance was hardly reduced when thymol was present in the heating medium, the effect in the recovery medium was greater, reducing the D100 °C values down to one third for B. subtilis and B. cereus when 0.5 mM thymol was added. This effect was dose dependent and was also observed at other heating temperatures. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. High-level expression of alkaline protease using recombinant ...

    African Journals Online (AJOL)

    AJL

    2012-02-16

    Feb 16, 2012 ... compared with that of wild-type B. licheniformis CICIM B5102. Key word: Alkaline protease, Bacillus amyloliquefaciens, Bacillus licheniformis. INTRODUCTION. Proteases are one of the most important industrial enzyme groups, accounting for approximately 60% of the total enzyme sales (Beg et al., 2003).

  3. Influence of heterologous MreB proteins on cell morphology of Bacillus subtilis.

    Science.gov (United States)

    Schirner, Kathrin; Errington, Jeff

    2009-11-01

    The prokaryotic cytoskeletal protein MreB is thought to govern cell shape by positioning the cell wall synthetic apparatus at growth sites in the cell. In rod-shaped bacteria it forms helical filaments that run around the periphery of the rod during elongation. Gram-positive bacteria often contain more than one mreB gene. Bacillus subtilis has three mreB-like genes, mreB, mbl and mreBH, the first two of which have been shown to be essential under normal growth conditions. Expression of an mreB homologue from the closely related organism Bacillus licheniformis did not have any effect on cell growth or morphology. In contrast, expression of mreB from the phylogenetically more distant bacterium Clostridium perfringens produced shape defects and ultimately cell death, due to disruption of the endogenous MreB cytoskeleton. However, expression of either mreB(B. licheniformis) (mreB(Bl)) or mreB(C. perfringens) (mreB(Cp)) was sufficient to confer a rod shape to B. subtilis deleted for the three mreB isologues, supporting the idea that the three proteins have largely redundant functions in cell morphogenesis. Expression of mreBCD(Bl) could fully compensate for the loss of mreBCD in B. subtilis and led to the formation of rod-shaped cells. In contrast, expression of mreBCD(Cp) was not sufficient to confer a rod shape to B. subtilis Delta mreBCD, indicating that a complex of these three cell shape determinants is not enough for cell morphogenesis of B. subtilis.

  4. Directed evolution improves the fibrinolytic activity of nattokinase from Bacillus natto.

    Science.gov (United States)

    Yongjun, Cai; Wei, Bao; Shujun, Jiang; Meizhi, Weng; Yan, Jia; Yan, Yin; Zhongliang, Zheng; Goulin, Zou

    2011-12-01

    Nattokinase (subtilisin NAT, NK) is a relatively effective microbial fibrinolytic enzyme that has been identified and characterized from Bacillus natto. In the current report, DNA family shuffling was used to improve the fibrinolytic activity of nattokinase. Three homologous genes from B. natto AS 1.107, Bacillus amyloliquefaciens CICC 20164 and Bacillus licheniformis CICC 10092 were shuffled to generate a mutant library. A plate-based method was used to screen the mutant libraries for improved activity. After three rounds of DNA shuffling, one desirable mutant with 16 amino acid substitutions was obtained. The mutant enzyme was purified and characterized. The kinetic measurements showed that the catalytic efficiency of the mutant NK was approximately 2.3 times higher than that of the wild-type nattokinase. In addition, the molecular modeling analysis suggested that the mutations affect the enzymatic function by changing the surface conformation of the substrate-binding pocket. The current study shows that the evolution of nattokinase with improved fibrinolytic activity by DNA family shuffling is feasible and provides useful references to facilitate the application of nattokinase in thrombolytic therapy. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  5. Maintenance metabolism and carbon fluxes in Bacillus species

    Directory of Open Access Journals (Sweden)

    Decasper Seraina

    2008-06-01

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

  6. Oral administration of a select mixture of Bacillus probiotics generates Tr1 cells in weaned F4ab/acR pigs challenged with an F4+ ETEC/VTEC/EPEC strain

    DEFF Research Database (Denmark)

    Zhou, Dong; Zhang, Wei; Wang, Meng-Ling

    2015-01-01

    Although breeding of F4 receptor − negative (F4R−) pigs may prevent post-weaning diarrhea, the underlying immunity is poorly understood. Here, various doses of a Bacillus licheniformis and Bacillus subtilis mixture (BLS-mix) were orally administered to F4ab/acR− pigs for 1 week before F4 (K88) − ...... of BLS-mix during episodes of intestinal inflammation that is caused by enteric pathogens might prohibit clearance of the pathogen. Select probiotic mixtures may allow for tailoring strategies to prevent infectious diseases....

  7. Effects of Temperature on Sensitivity of Bacilus licheniformis RI 75-1 Vegetative Cells at Gamma Quantum

    International Nuclear Information System (INIS)

    Fernandez-Larrea Vega, O.; Rios Brito, F.; Marquez Alvarez, M.; Padron Soler, E.

    1986-01-01

    It is known that strains of E. Coli with wild genotype for reparation, when are irradiated at temperature between 42 0 C and 45 0 C, shown an increase of radioresistance. At the given temperature the number of double strands breaks of DNA decrease. Some authors report that the radioresistance increased is due to the elevation of the irradiation temperature is related to the cell membrane status. The paper includes reports on the effects of increased temperature on the sensitivity - at gamma quantum - of Bacillus licheniformis RI 75-1 vegetative cells. Temperatures of 42 0 C and 60 0 C during irradiation were employed. An increase in radioresistance was found when the temperature of irradiation was increased to 42 0 C. However, a decrease in viability was observed. Heat treatment prior to irradiation showed an increase in the number of radioresistance colonies when compared. (author)

  8. Aerobic Denitrification as an Innovative Method for In-Situ Biological Remediation of Contaminated Subsurface Sites

    Science.gov (United States)

    1989-01-01

    roseus Bacillus stearothermophilus Micrococcus varians Paracoccus denitrificans Bacillus coagulans Paracoccus halodenitrificans Bacillus flrmus Bacillus ...Geodermatophilus Plesiomonas Arachnia Haemophilus Propionibacterium Arthrobacter Halobacteriua Proteus Bacillus Ifalocuccus Pseudoraonas Bacteroides...Mycobacterium peregrinum Nocardia opaca Chromobacterium violaceum Bacillus subtllis Nocardia atlantica Bacillus licheniformis Flavobacterium

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

    Science.gov (United States)

    2003-11-08

    Bacillus anthracis BA0068 Ames Sterne SPS 97.13.213 Bacillus cereus Bacillus coagulans Bacillus licheniformis Bacillus macerans Bacillus ...megaterium Bacillus polymyxa Bacillus sphaericus Bacillus stearothermophilus Bacillus subtilis subsp. niger Bacillus thuringiensis Bacillus popilliae...varicella- zoster virus, and Bacillus anthracis DNA by LightCycler polymerase chain reaction after autoclaving:

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

    Science.gov (United States)

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

    2015-06-01

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

  11. 1

    NARCIS (Netherlands)

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

    2015-01-01

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

  12. Specialization of Bacillus in the Geochemically Challenged Environment of Death Valley

    Science.gov (United States)

    Kopac, S.

    2014-04-01

    Death Valley is the hottest, driest place in North America, a desert with soils containing toxic elements such as boron and lead. While most organisms are unable to survive under these conditions, a diverse community of bacteria survives here. What has enabled bacteria to adapt and thrive in a plethora of extreme and stressful environments where other organisms are unable to grow? The unique environmental adaptations that distinguish ecologically distinct bacterial groups (ecotypes) remain a mystery, in contrast to many animal species (perhaps most notably Darwin's ecologically distinct finch species). We resolve the ecological factors associated with recently diverged ecotypes of the soil bacteria Bacillus subtilis and Bacillus licheniformis, isolated from the dry, geochemically challenging soils of Death Valley, CA. To investigate speciation associated with challenging environmental parameters, we sampled soil transects along a 400m stretch that parallels a decrease in salinity adjacent to a salt flat; transects also encompass gradients in soil B, Cu, Fe, NO3, and P, all of which were quantified in our soil samples. We demarcated strains using Ecotype Simulation, a sequence-based algorithm. Each ecotype's habitat associations were determined with respect to salinity, B, Cu, Fe, NO3, and P. In addition, our sample strains were tested for tolerance of copper, boron and salinity (all known to inhibit growth at high concentrations) by comparing their growth over a 20 hour period. Ecotypes differed in their habitat associations with salinity, boron, copper, iron, and other ecological factors; these environmental dimensions are likely causing speciation of B. subtilis-licheniformis ecotypes at our sample site. Strains also differed in tolerance of boron and copper, providing evidence that our sequence-based demarcations reflect real differences in metabolism. By better understanding the relationship between bacterial speciation and the environment, we can begin to

  13. Estudos sobre a esporulação de uma amostra de bacillus: IV - Outras evidências sobre a atividade do íon Mn[2+] na esporulação endotrófica Studies on the sporulation of a Bacillus strain: IV - Further evidences of the Mn[2+] ion activity in the endotrophic sporulation

    Directory of Open Access Journals (Sweden)

    Leon Rabinovitch

    1973-01-01

    Full Text Available Foram feitas experimentações om o intuito de se buscar mais evidências sobre a participação do íon Mn[2+] no mecanismo esporogenético de uma amostra de Bacillus licheniformis. Quando as formas vegetativas desta bactéria eram depositadas em um meio mineral, carente de fonte de carbono utilizável, em conjunto com um agente seqüestrante de metais como EDTA, a esporulação endotrófica deixava de ocorrer. Entretanto, a esporulação pôde ser protegida quando as células eram previamente saturadas com um excesso de Mn[2+] exógeno. As formas esporuladas obtidas nas condições estudadas mostraram termorresistência a 85ºC durante 20 minutos.In the present paper the authors bring out more evidences concerning the influence of Mn[2+] ion on the endotrophic sporulation of Bacillus licheniformis-2390. Vegetative cells of this bacteria could not sporulate if they were submited to a sufficient concentration of EDTA. Otherwise, this sporulatio ocurred when the vegetative forms were first protected by an excess of exogenous Mn[2+] of Zn[2+], Fe[2+] and Mg[2+]. Those spores obtained shown thermoresistence to 85ºC during 20 minutes.

  14. Potencial in vitro de Bacillus spp. no controle de Curtobacterium flaccumfaciens pv. flaccumfaciens em feijoeiro-comum

    Directory of Open Access Journals (Sweden)

    Evelynne Urzêdo Leão

    Full Text Available RESUMO O feijoeiro-comum (Phaseolus vulgaris é uma cultura de grande relevância na alimentação da população brasileira. A murcha-de-curtobacterium ou murcha bacteriana, causada por Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff. é uma doença vascular que acomete o feijoeiro causando danos significativos. Neste contexto, o objetivo deste trabalho foi avaliar a ação in vitro de Bacillus spp. na inibição de dois isolados de Cff, colonização do sistema radicular e desenvolvimento de plântulas de feijoeiro-comum. Foram realizados dois ensaios in vitro para verificar a atividade antagônica dos isolados Bacillus licheniformis, B. subtilis e B. subtilis + B. lichenformis a dois isolados de Cff. Todos os isolados de Bacillus spp. apresentaram inibição no crescimento dos isolados de Cff. Não foi observada a colonização das raízes das plântulas de feijoeiro-comum, pelos isolados bacterianos avaliados.

  15. An Apple Fruit Fermentation (AFF) Treatment Improves the Composition of the Rhizosphere Microbial Community and Growth of Strawberry (Fragaria × ananassa Duch ‘Benihoppe’) Seedlings

    Science.gov (United States)

    Bu, Yufen; Shao, Wei; Huang, Weijing; Ji, Qianlong; Yao, Yuncong

    2016-01-01

    Plant growth can be promoted by the application of apple fruit fermentation (AFF), despite unclear of the underlying mechanisms, the effects involved in AFF on rhizosphere microorganisms have been hypothesized. We investigated the consequences of applying AFF alone or in combination with Bacillus licheniformis to strawberry tissue culture seedlings in vitro, the analyses of Denaturing Gradient Gel Electrophoresis (DGGE) and 16S rDNA were performed to determine AFF effects on rhizosphere. Moreover, the growth index and antioxidant enzyme activities were determined 30 days after treatments. We identified five dominant bacteria in AFF: Coprinus atramentarius, Bacillus megaterium, Bacillus licheniformis, Weissella and B. subtilis. The greatest number of bacterial species were observed in the rhizosphere of control matrix (water treated), and the lowest diversity appeared in the rhizosphere soil treated with 108 cfu/mL B. licheniformis alone. Combining AFF plus B. licheniformis in one treatment resulted in the largest leaf area, plant height, root length, plant weight, and the markedly higher activities of antioxidant enzymes. We conclude that a combination of AFF plus B. licheniformis treatment to matrix can increase antioxidant enzymes activities in strawberry seedlings, optimize the status of rhizosphere microbial, and promote plant growth. PMID:27755580

  16. Comparative evaluation of the bacteria isolated from decomposing ...

    African Journals Online (AJOL)

    Six (6) bacterial species Bacillus circulans, Bacillus pumilus, Bacillus subtilis, Micrococcus luteus, Streptococcus faecalis and Streptococcus lactis were isolated from decomposing cow milk, while four (4) bacterial species namely Bacillus brevis, Bacillus licheniformis, Lactobacillus casei and Staphylococcus epidermidis ...

  17. Purification and Characterization of a Novel and Robust L-Asparaginase Having Low-Glutaminase Activity from Bacillus licheniformis: In Vitro Evaluation of Anti-Cancerous Properties

    Science.gov (United States)

    Mahajan, Richi V.; Kumar, Vinod; Rajendran, Vinoth; Saran, Saurabh; Ghosh, Prahlad C.; Saxena, Rajendra Kumar

    2014-01-01

    L-asparaginase having low glutaminase has been a key therapeutic agent in the treatment of acute lymphpoblastic leukemia (A.L.L). In the present study, an extracellular L-asparaginase with low glutaminase activity, produced by Bacillus licheniformis was purified to homogeneity. Protein was found to be a homotetramer of 134.8 KDa with monomeric size of 33.7 KDa and very specific for its natural substrate i.e. L-asparagine. The activity of purified L-asparaginase enhanced in presence of cations including Na+ and K+, whereas it was moderately inhibited in the presence of divalent cations and thiol group blocking reagents. The purified enzyme was maximally active over the range of pH 6.0 to 10.0 and temperature of 40°C and enzyme was stable maximum at pH 9.0 and −20°C. CD spectra of L-asparaginase predicted the enzyme to consist of 63.05% α- helix and 3.29% β-sheets in its native form with T222 of 58°C. Fluorescent spectroscopy showed the protein to be stable even in the presence of more than 3 M GdHCl. Kinetic parameters Km, Vmax and kcat of purified enzyme were found as 1.4×10−5 M, 4.03 IU and 2.68×103 s−1, respectively. The purified L-asparaginase had cytotoxic activity against various cancerous cell lines viz. Jurkat clone E6-1, MCF-7 and K-562 with IC50 of 0.22 IU, 0.78 IU and 0.153 IU respectively. However the enzyme had no toxic effect on human erythrocytes and CHO cell lines hence should be considered potential candidate for further pharmaceutical use as an anticancer drug. PMID:24905227

  18. Characteristics of bacillus strains with antifungal activity against phytopathogens

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Young Keun; Senthilkumar, M. [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of)

    2009-12-15

    Four bacterial isolates that showed antifungal activity against Alternaria alternata and other phytopathogens were isolates from bean rhizosphere. 16S rDNA analysis and phylogenetic relationship indicated that these isolates belong to Genus Bacillus. Isolate A1 clustered with Bacillus licheniformis while other isolates A2, A3 and A4 clustered together with B.pumilus. n-Butanol extract of these isolates strongly inhibited the growth of A. alternata while, chloroform extract of isolate A2 and ethyl acetate extract of A1,A3, and A4 inhibited the test fungus partially. All the isolates except A4 produced chitinase enzyme. None of the isolates solubilized mineral phosphate. Radiation sensitivity of isolates A1, A2, A3 and A4 were assessed and the LD{sub 99} values are determined as 0.50, 6.69, 11,60, 1.53 kGy, respectively. Mutant libraries of each isolate were prepared by exposing them to gamma radiation at their respective LD{sub 99} dose. Crude metabolite caused drastic changes on A. alternata hyphal morphology. Appearance of shrunken and collapsed hyphae could be due to the leak of cell wall or changes in membrane permeability.

  19. Characteristics of bacillus strains with antifungal activity against phytopathogens

    International Nuclear Information System (INIS)

    Lee, Young Keun; Senthilkumar, M.

    2009-01-01

    Four bacterial isolates that showed antifungal activity against Alternaria alternata and other phytopathogens were isolates from bean rhizosphere. 16S rDNA analysis and phylogenetic relationship indicated that these isolates belong to Genus Bacillus. Isolate A1 clustered with Bacillus licheniformis while other isolates A2, A3 and A4 clustered together with B.pumilus. n-Butanol extract of these isolates strongly inhibited the growth of A. alternata while, chloroform extract of isolate A2 and ethyl acetate extract of A1,A3, and A4 inhibited the test fungus partially. All the isolates except A4 produced chitinase enzyme. None of the isolates solubilized mineral phosphate. Radiation sensitivity of isolates A1, A2, A3 and A4 were assessed and the LD 99 values are determined as 0.50, 6.69, 11,60, 1.53 kGy, respectively. Mutant libraries of each isolate were prepared by exposing them to gamma radiation at their respective LD 99 dose. Crude metabolite caused drastic changes on A. alternata hyphal morphology. Appearance of shrunken and collapsed hyphae could be due to the leak of cell wall or changes in membrane permeability

  20. Effect of temperature and pH on polygalacturonase production by pectinolytic bacteria Bacillus licheniformis strain GD2a in submerged medium from Raja Nangka (Musa paradisiaca var. formatypica) banana peel waste

    Science.gov (United States)

    Widowati, E.; Utami, R.; Mahadjoeno, E.; Saputro, G. P.

    2017-04-01

    The aim of this research were to determine the effect of temperature (45°C, 55°C, 65°C) and pH (5.0; 6.0; 7.0) on the increase of total cell count and polygalacturonase enzyme activity produced from raja nangka banana (Musa paradisiaca var. formatypica) peel waste by pectinolytic bacterial Bacillus licheniformis strain GD2a. This research applied two sample repetition and one analysis repetition. The result showed temperature and pH affect total cell count. The total cell count on 45°C and pH 7 recorded the highest number at 9.469 log cell/ml. Temperature and pH also affected pectin concentration at the end of fermentation. The lowest pectin concentration recorded at 45°C and pH 7 was 0.425 %. The highest enzyme activity recorded at 65°C and pH 7 was 0.204 U/ml. The highest enzyme protein concentration was recorded at 65°C and resulted as 0.310 mg/ml on pH 6. The highest specific activity was 19.527 U/mg at 65°C and pH 7. By this result, could be concluded that optimum condition process on polygalacturonase production was at 65°C and pH 7 because it gave highest enzyme activity result (0,204 U/ml).

  1. Selection and evaluation of Malaysian Bacillus spp. strains as potential probiotics in cultured tiger grouper (Epinephelus fuscoguttatus).

    Science.gov (United States)

    Yasin, Ina-salwany Md; Razak, Nabilah Fatin; Natrah, F M I; Harmin, Sharr Azni

    2016-07-01

    A total of 58 Gram-positive bacteria strains were isolated from the marine environment and screened for potential probiotics for disease prevention and improving the productivity of tiger grouper Epinephelus fuscoguttatus larvae and juveniles. The bacteria were identified as Bacillus licheniformis, B. subtilis, B. circulans, B. sphaericus, B. cereus, Brevibacillus brevis, Corynebacterium propinquum, Leifsonia aquatica and Paenibacillus macerans. Only 24 strains showed antagonistic activities against four pathogenic strains; Vibrio alginolyticus, V. harveyi, V. parahaemolyticus and Aeromonas hydrophila, where two of the Bacillus strains, B12 and B45 demonstrated intermediate to highest level of inhibitory activity against these pathogenic strains, respectively. Further assessment by co-culture assay showed that Bacillus strain B12 exhibited a total inhibition of V. alginolyticus, while B45 strain displayed no inhibitory activity. Mixed culture of Bacillus B12 and B45 strains to outcompete V. alginolyticus was observed at a cell density of 10(7) CFU ml(-1). Molecular identification and phylogenetic tree analysis have categorized Bacillus strain B12 to the reference strains GQ340480 and JX290193 of? B. amyloliquafaciens, and Bacillus strain B45 with a reference strain JF496522 of B. subtilis. Safety tests of probionts by intraperitoneal administration of B12 and B45 strains at cell densities of 103, 105 and 10(7) CFU ml(-1) revealed no abnormalities and cent percent survival for healthy Epinephelus fuscoguttatus juveniles within 15 days of experimental period. Overall, the study revealed that Bacillus B12 strain possesses tremendous probiotic potential that could be used as a feed supplement in tiger grouper diets. ?

  2. Isolation and characterization of Bacillus subtilis CH16 strain from chicken gastrointestinal tracts for use as a feed supplement to promote weight gain in broilers.

    Science.gov (United States)

    Nguyen, A T V; Nguyen, D V; Tran, M T; Nguyen, L T; Nguyen, A H; Phan, T-N

    2015-06-01

    Spore-forming bacterial strains were isolated from chicken gastrointestinal tracts to develop a heat-stable feed supplement that promotes weight gain in broilers. Seven Bacillus strains having more than 90% sporulation were screened from the isolates and identified to be closely related with Bacillus subtilis and Bacillus licheniformis. Of the seven strains, B. subtilis CH16 was selected to develop a feed supplement for broilers, because it formed 100% heat-stable spores, grew rapidly at 42°C and quickly formed a biofilm. In large-scale trials in broilers (n ≥ 1150 per group), the group fed CH16 (3 × 10(6) CFU g(-1) pellet) showed higher average daily gain (ADG = 61·16) and lower food conversion ratio (FCR = 1·696) than did the group fed B. licheniformis CH22 (ADG = 57·10 and FCR = 1·792), the group fed B. subtilis HU58 (ADG = 51·90 and FCR = 1·868), BioPlus group (ADG = 59·32 and FCR = 1·807) and the control group (ADG = 56·02 and FCR = 1·880). In conclusion, CH16 spores significantly increased ADG by 9·17% and reduced FCR by 9·79% in broilers. The result supports the use of B. subtilis CH16 of chicken intestinal origin as a feed supplement that promote weight gain in broilers. Significance and impact of the study: This study reports screening of Bacillus strains isolated from chicken gastrointestinal tracts for development of a feed supplement that promote weight gain in broilers. Of the seven Bacillus isolates with high sporulation efficiency (≥90%), Bacillus subtilis CH16 strain showed the best growth and biofilm formation at body temperature of broilers (42°C). In large-scale trials in broilers (n ≥ 1150 per group), CH16 spores induced a 9·17% increase in daily weight gain (ADG) and a 9·79% reduction in FCR while the commercial BioPlus(®) YC induced only a 5·89% increase in ADG and a 3·88% reduction in FCR. © 2015 The Society for Applied Microbiology.

  3. Addition of Bacillus sp. inoculums in bedding for swine on a pilot scale: effect on microbial population and bedding temperature.

    Science.gov (United States)

    Corrêa, E K; Ulguim, R R; Corrêa, L B; Castilhos, D D; Bianchi, I; Gil-Turnes, C; Lucia, T

    2012-10-01

    Thermal and microbiological characteristics of beddings for swine were compared according to their depth and of addition of inoculums. Bedding was added to boxes at 0.25 (25D) and 0.50 m (50D), with three treatments: control (no inoculums); T1, with 250 g of Bacillus cereus var. toyoii at 8.4 × 10(7) CFU; and T2, with 250 g of a pool of B. subtilis, Bacillus licheniformis and Bacillus polymyxa at 8.4 × 10(7) CFU (250 g for 25D and 500 g for 50D). Mean temperatures were 28.5 ± 3.9 at the surface and 35.2 ± 8.9 inside the beddings. The most probable number (MPN) of thermophilic bacteria was higher for T1 and T2 than for the control (P<0.05). The MPN of thermophilic bacteria and fungi was greater for D50 than for D25 (P<0.05). The use of 25D without inoculums is recommended due to the reduction of thermophilic microbiota. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Effect of probiotic mixture on some haematological parametres in ...

    African Journals Online (AJOL)

    ml in 1ml normal saline together with daily administration of a mixture of probiotics (Lactobacillus acidophilus, Bacillus pumilus, Bacillus subtilis and Bacillus licheniformis) at 4.5 x10 CFU/ml, Group C: lambs were administered only viable STEC ...

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

    Directory of Open Access Journals (Sweden)

    Sandhya Vardharajula

    2014-08-01

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

  6. Effect of ratio variation of crop wastes on the production of poultry ...

    African Journals Online (AJOL)

    Proteus mirabilis, Proteus morganii, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Bacillus licheniformis, Acetobacter orleanensis, Clostridium butyricum, Micrococcus roseus, Lactobacillus delbrueckii, Clostridium spheroides, and Methanobacillus species. Seven fungal ...

  7. 1373-IJBCS-Article-Lamine Said Babamoussa

    African Journals Online (AJOL)

    KODJIO NORBERT

    Les rhizobactéries Pseudomonas fluorescens ont induit le rendement de maïs le plus élevé, dépassant de ... Bacillus licheniformis, Bacillus lentus, Bacillus circulans, Bacillus firmus et Azospirillum ..... asiciadas al cultivo del maiz. ... bacteria and their inoculation effects on growth and nitrogen uptake of crop plants. J.

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

    Directory of Open Access Journals (Sweden)

    Ladiel Luiz Pedrozo Tavares

    2011-01-01

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

  9. Influence of toluene and salinity on biosurfactant production by Bacillus sp.: scale up from flasks to a bench-scale bioreactor

    Directory of Open Access Journals (Sweden)

    Ellen Cristina Souza

    Full Text Available ABSTRACT To select the best biosurfactant producer, Pseudomonas putida, Bacillus megatherium, Bacillus licheniformis and Bacillus subtilis were cultured in flasks on media with different salinity [low salinity (LS, Bushnell-Haas (BH and artificial sea water (SW media] supplemented or not with toluene as a model pollutant. Toluene inhibited the growth of all microorganisms and stimulated the biosurfactant production. B. subtilis exhibited the best performance, being able to lower the surface tension (ST in the LS medium to 65.5 mN/min in the absence of toluene, and to 46.5 mN/min in the BH medium in the presence of toluene, corresponding to ST reductions of 13.0 and 27.5 mN/m, respectively. Scaling up the process to a bench-scale fermentor, the best results were obtained in the LS medium, where B. subtilis was able to reduce the toluene concentration from 26.0 to 4.3 g/L within 12 h and ST by 17.2 mN/m within 18 h. The results of this study point out that B. subtilis is an interesting biosurfactant producer, which could be used in the bioremediation of toluene-contaminated water.

  10. Evaluation of the effects of Bacillus subtilis and Bacillus Licheni formis promix on blood metabolites and elements and weight gain in rearing calves

    Directory of Open Access Journals (Sweden)

    Gh Moghaddam

    2007-05-01

    Full Text Available Probiotics are beneficial microorganisms which are colonized in the digestive system of livestock and influence some of the blood parameters through special mechanisms. For this purpose, an experiment was conducted to clarify the influence of promix containing living spores of the two bacteria Bacillus subtilis and Bacillus licheniformis on body weight gain, blood metabolites and elements of calves. Twenty male Holstein calves were divided into two groups of ten calves and kept at separate bans. The control group were fed with a standard ration and the treatment group were fed with the same ration plus promix (1.6 × 109 cfu/g feed or 500 g/T feed for 45 days. Calves were weighed in the 1st, 15th, 30th and 45th days and blood samples were collected for measurement of some metabolites and blood elements. The obtained means were compared using the T- test. The results indicated that the average daily weight gain using promix had a non-significant improvement of 4.8% (p>0.05. Also the average daily weight gain showed a non significant improvement in each of the 3 experimental periods (p>0.05. Blood phosphorus and calcium of calves increased significantly influenced by promix with the increase being non significant in the first experimental period and significant in the second and third periods (p

  11. Isolation of four hydrocarbon effluent-degrading Bacillaceae species ...

    African Journals Online (AJOL)

    percentage decreases in total hydrocarbon concentration within 18 days: 98% with Bacillus licheniformis STK08, 87% with Geobacillus stearothermophilus STM04, 80% with Lysinibacillus sphaericus STZ75 and 72% with Bacillus firmus STS84.

  12. Psychrotrophic bacteria isolated from -20°C freezer | Ahmad ...

    African Journals Online (AJOL)

    Three psychrotrophic bacteria, morpho-physiologically, identified as Bacillus subtilis MRLBA7, Bacillus licheniformis MRLBA8 and Bacillus megaterium MRLBA9 were isolated from -20°C freezer of the Microbiology Research Laboratory (MRL), Quaid-i-Azam University, Islamabad, Pakistan. These strains were able to grow ...

  13. Bacteria associated with cultures of psathyrella atroumbonata (Pleger)

    African Journals Online (AJOL)

    These bacteria include Bacillus licheniformis, Bacillus subtilis, Leuconostoc mesenteroides, Pseudomonas aeruginosa, Bacillus cereus and Staphylococcus aureus. The average bacteria count was 1.0 x 106 cfu/ml and these bacteria grew within pH range of 5.0 and 9.0. the optimum temperature range of growth lied ...

  14. Effect of Medium Composition on Commercially Important Alkaline Protease Production by Bacillus licheniformis N-2

    Directory of Open Access Journals (Sweden)

    Javed Iqbal Qazi

    2008-01-01

    Full Text Available Protease production by alkalophilic B. licheniformis N-2 was investigated in 50 mL of the growth medium consisting of (in g/L: glucose 10.0, soybean meal 10.0, K2HPO4 3.0, MgSO4·7H2O 0.5, NaCl 0.5 and CaCl2·2H2O 0.5 at pH=10. Different carbon and nitrogen sources in the form of fine powder of organic, inorganic and defatted meals were studied to select the suitable substrate for alkaline protease production. The highest level of alkaline protease (677.64 U/mL was obtained in the medium containing glucose followed by soluble starch and wheat bran. Among various nitrogen sources, defatted soybean meal was found to be the best inducer of alkaline protease, while inorganic nitrogen sources in the form of ammonium salts repressed the enzyme activity up to 96 %. Thermostability studies showed that the enzyme in the presence of 10 mM Ca2+ ions retained its residual activity up to 80 % even after incubation at 40 °C for 12 h. The enzyme was found stable over a broad range of pH (8–11 and lost 52 % of its residual activity at pH=12. After the treatment with Tween 20, Tween 45, Tween 65, Triton X-405, H2O2 and sodium perborate, each at 1.0 % concentration, the enzyme showed residual activity of 105, 82, 116, 109, 135 and 126 %, respectively. The application of alkaline protease for removal of blood stains from cotton fabric also indicates its potential use in detergent formulations.

  15. Biodegradable Polymeric Substances Produced by a Marine Bacterium from a Surplus Stream of the Biodiesel Industry

    Directory of Open Access Journals (Sweden)

    Sourish Bhattacharya

    2016-11-01

    Full Text Available Crude glycerol is generated as a by-product during transesterification process and during hydrolysis of fat in the soap-manufacturing process, and poses a problem for waste management. In the present approach, an efficient process was designed for simultaneous production of 0.2 g/L extracellular ε-polylysine and 64.6% (w/w intracellular polyhydroxyalkanoate (PHA in the same fermentation broth (1 L shake flask utilizing Jatropha biodiesel waste residues as carbon rich source by marine bacterial strain (Bacillus licheniformis PL26, isolated from west coast of India. The synthesized ε-polylysine and polyhydroxyalkanoate PHA by Bacillus licheniformis PL26 was characterized by thermogravimetric analysis (TGA, differential scanning colorimetry (DSC, Fourier transform infrared spectroscopy (FTIR, and 1H Nuclear magnetic resonance spectroscopy (NMR. The PHA produced by Bacillus licheniformis was found to be poly-3-hydroxybutyrate-co-3-hydroxyvalerate (P3HB-co-3HV. The developed process needs to be statistically optimized further for gaining still better yield of both the products in an efficient manner.

  16. Effect of Bacillus subtilis and Bacillus licheniformis supplementation in diets with low- and high-protein content on ileal crude protein and amino acid digestibility and intestinal microbiota composition of growing pigs.

    Science.gov (United States)

    Kaewtapee, Chanwit; Burbach, Katharina; Tomforde, Georgina; Hartinger, Thomas; Camarinha-Silva, Amélia; Heinritz, Sonja; Seifert, Jana; Wiltafsky, Markus; Mosenthin, Rainer; Rosenfelder-Kuon, Pia

    2017-01-01

    Bacillus spp. seem to be an alternative to antimicrobial growth promoters for improving animals' health and performance. However, there is little information on the effect of Bacillus spp. in combination with different dietary crude protein (CP) levels on the ileal digestibility and microbiota composition. Therefore, the objective of this study was to determine the effect of Bacillus spp. supplementation to low- (LP) and high-protein diets (HP) on ileal CP and amino acid (AA) digestibility and intestinal microbiota composition. Eight ileally cannulated pigs with an initial body weight of 28.5 kg were randomly allocated to a row-column design with 8 pigs and 3 periods of 16 d each. The assay diets were based on wheat-barley-soybean meal with two protein levels: LP (14% CP, as-fed) and HP diet (18% CP, as-fed). The LP and HP diets were supplemented with or without Bacillus spp. at a level of 0.04% (as-fed). The apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of CP and AA was determined. Bacterial community composition from ileal digesta was analyzed by Illumina amplicon sequencing and quantitative real-time PCR. Data were analyzed as a 2 × 2 factorial design using the GLIMMIX procedures of SAS. The supplementation with Bacillus spp. did not affect both AID and SID of CP and AA in growing pigs. Moreover, there was no difference in AID of CP and AA between HP and LP diets, but SID of cystine, glutamic acid, glycine, and proline was lower ( P  digestibility, whereas LP diet may reduce the flow of undigested protein to the large intestine of pigs.

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

    Directory of Open Access Journals (Sweden)

    Yasser R. Abdel-Fattah

    2013-01-01

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

  18. A stochastic model for predicting dextrose equivalent and saccharide composition during hydrolysis of starch by alpha-amylase

    NARCIS (Netherlands)

    Besselink, T.; Baks, T.; Janssen, A.E.M.; Boom, R.M.

    2008-01-01

    A stochastic model was developed that was used to describe the formation and breakdown of all saccharides involved during -amylolytic starch hydrolysis in time. This model is based on the subsite maps found in literature for Bacillus amyloliquefaciens -amylase (BAA) and Bacillus licheniformis

  19. Thermo-aerobic bacteria from geothermal springs in Saudi Arabia ...

    African Journals Online (AJOL)

    Fifteen isolates of thermo-aerobic bacteria were found. Bacillus cereus, B. licheniformis, B. thermoamylovorans, Pseudomonas sp., Pseudomonas aeruginosa and Enterobacter sp. were dominant in hot springs. Genetic relatedness indicated that eleven Bacillus spp. grouped together formed several clusters within one main ...

  20. Production of Extra-Cellular Proteases from Marine Bacillus Sp. Cultured in Media Containing Ammonium Sulfate as the Sole Nitrogen Source

    Directory of Open Access Journals (Sweden)

    Seri Intan, M.

    2005-01-01

    Full Text Available Useful bacterial strains can be used to increase mineralize activity of an aquatic system. These bacteria can specifically degrade targeted compound by producing extra-cellular enzymes. Three species of Bacillus i.e. B. subtilis, B. pumilus and B. licheniformis acquired from shrimp ponds were tested for their ability to utilize ammonia and produce extracellular enzymes. These bacteria were grown in artificial seawater (30 ppt salinity and pH 7.6 supplemented with decreasing yeast extract concentration but increasing ammonium sulfate concentration. All three bacteria grew in artificial seawater containing only 0.01% yeast extract and 1% ammonium sulfate. However, only B. pumilus and B. licheniformis were able to grow in the medium containing only 1% ammonium sulfate as a sole energy source. Bacterialgrowth reduced when alkaline proteases activities was maximum from culture filtrates of all three bacterial cultures during 24 hour culturing in artificial seawater containing 0.01% yeast extract and 1% ammonium sulfate at 30 C when assayed at pH 9. Bacterial growth increased when acid proteases activities was maximum from culture filtrates of all three bacterial cultures during 48 hour culturing in artificial seawater containing 0.01% yeast extract and 1% ammoniumsulfate at 30 C when assayed at pH 5.

  1. Comparison of four feed proteases for improvement of nutritive value of poultry feather meal

    DEFF Research Database (Denmark)

    Pedersen, Mads Brøgger; Yu, S; Plumstead, P

    2012-01-01

    in the production of cost-effective feather by-products for use as feed and fertilizers. The current study examined 4 commercial feed proteases from Bacillus subtilis, Bacillus licheniformis PWD-1, Aspergillus niger, and Serratia proteamaculans HY-3 used to hydrolyze chicken feather under different conditions...

  2. ORF Alignment: NC_006270 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available PEADLAIMQEKE 67 ... RGDLFHELETGIRSTCHLLKKVKESDWSYRPADRMRSLKELASHLTAIPEADLAIM...QEKE Sbjct: 1 ... RGDLFHELETGIRSTCHLLKKVKESDWSYRPADRMRSLKELASHLTAIPEADLAIMQEKE 60 ... ...01287 [Bacillus ... licheniformis DSM 13] ... Length = 147 ... Query: 8 ... RGDLFHELETGIRSTCHLLKKVKESDWSYRPADRMRSLKELASHLTAI

  3. ORF Alignment: NC_006322 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available PEADLAIMQEKE 67 ... RGDLFHELETGIRSTCHLLKKVKESDWSYRPADRMRSLKELASHLTAIPEADLAIM...QEKE Sbjct: 1 ... RGDLFHELETGIRSTCHLLKKVKESDWSYRPADRMRSLKELASHLTAIPEADLAIMQEKE 60 ... ...01287 [Bacillus ... licheniformis DSM 13] ... Length = 147 ... Query: 8 ... RGDLFHELETGIRSTCHLLKKVKESDWSYRPADRMRSLKELASHLTAI

  4. Studies on the occurrence and distribution of heavy metals in ...

    African Journals Online (AJOL)

    The bacteria isolated and identified were. Bacillus subtilis, Bacillus licheniformis, Arthrobacter sp and Achromobacter sp. Fungi isolated were Saccharomyces cerevisiae, Candida utilissima and Sporobolomyces sp. The values for heavy metals for all zones are of public health significance and pose a threat to the survival of ...

  5. Structure based protein engineering of Bacillus stearothermophilus {alpha}-amylase: toward a new substrate specificity

    Energy Technology Data Exchange (ETDEWEB)

    Rasera, Ana Claudia [Sao Paulo Univ., SP (Brazil). Inst. de Ciencias Biomedicas; Iulek, Jorge [Universidade Estadual de Ponta Grossa, PR (Brazil). Inst. de Quimica; Delboni, Luis Fernando; Barbosa, Valma Martins Barbosa [Parana Univ., Curitiba, PR (Brazil). Dept. de Bioquimica

    1997-12-31

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

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

    International Nuclear Information System (INIS)

    Rasera, Ana Claudia; Iulek, Jorge; Delboni, Luis Fernando; Barbosa, Valma Martins Barbosa

    1997-01-01

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

  7. Karakterisasi dan uji aktivitas Bacillus spp. sebagai agensia pengendalian hayati penyakit lincat pada tembakau Temanggung

    Directory of Open Access Journals (Sweden)

    Triwidodo Arwiyanto

    2012-02-01

    Full Text Available Lincat disease of tobacco causing severe losses of the product. Control of the disease with any available measure unlikely giving enough control. A number of Bacillus spp. isolates could suppressed the growth of pathogen in vitro and suppressed the development of lincat disease in the field. This article report the charactheristics of six isolates of Bacillus spp. (Ba-4, Ba-22, Ba-24, Ba-30, Ba-33, dan Ba-41. These isolates proven could suppressed lincat disease in the field. Characterization of the isolates include the morphological, physiological characteristics, and pathogenicity against tobacco plant. The results indicated that the bacterial isolates were belong to the genus Bacillus with the following charactheristics. The bacteria were rod shapes, forming endospore, Gram positive, fermentative, positive reaction in katalase, oksidase, and Voges Proskaeur tests. Negative results were obtained for Methyl Red test, hydrolysis of starch, gelatine, and casein. The present isolates could use citrate and several carbohydrates as carbon sources. Reduce nitrate to nitrite. The isolates could grow in the medium with high osmotic pressure, i.e. could grow in the medium with 7% NaCl. The present isolates grew well in the medium with pH of 4.5 €“10 and could grow in the temperature range of 10 €“50 °C. According to pathogenicity test, the present isolates were not belong to the plant pathogenic bacteria. The present isolates could suppressed the growth of Ralstonia solanacearum in vitro, and could reduce the egg number of Meloidogyne incognita. According to the physiological charactheristics tested, it seem that isolates of Ba-4, Ba-24, Ba-30, dan Ba-33, and Ba-41 having similar charactheristics with Bacillus cereus. The Ba-22 isolate, however, having similar characteristics with B. licheniformis.

  8. Bio sorption of some Rare Earth Elements and Yttrium by Gram Positive and Gram Negative Bacteria

    International Nuclear Information System (INIS)

    Ibrahim, H.A.

    2012-01-01

    The separate bio sorption of the REEs La, Sm, Eu and Dy together with yttrium upon the Gram positive bacteria Bacillus subtilis (B.subtilis) and Bacillus Licheniformis (B. Licheniformis),the Gram negative bacterium Escherichia coli (E. coli ) and Saccharomyces cervisiae (Yeast) was studied. The revelant factors of ph 1-6, contact time (30-180 min), the initial rare earth concentration (50-200 mg/l) have been studied. The amount of the accumulated element was strongly affected by its concentration.In addition, bio sorptive fractionation of Y and the studied REEs from a solution containing a mixture of these elements was also studied. From the obtained data, it was found that Langmuir isotherm model for both B.licheniformis and E.coli gives a best fit for the studied elements over the working range of concentration (50-200 mg/I). Transmission electron microscopy exhibited accumulation throughout the bacterial cell with some granular deposits in both the cell periphery and cytoplasm

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

    International Nuclear Information System (INIS)

    El-Batal, A.I.; Abo-State, M.A.

    2006-01-01

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

  10. Preliminary Study on Keratinase from Two Indonesian Isolates

    Directory of Open Access Journals (Sweden)

    S Rahayu

    2010-01-01

    Full Text Available Keratinases (E.C.3.4.99.11 constitute a group of enzymes capable of disrupting the highly stable keratin structure consisting of disulphide, hydrogen, and hydrophobic bonds in the form of α-helices and β-sheets B. licheniformis MB-2 and Bacillus sp. MTS are two feather-degrading bacteria isolated from Tompaso crater at North Sulawesi and sulfuric land around Tangkuban Perahu in West Java. They were both capable of breaking down whole chicken feathers. In addition both isolates were capable of degrading other proteinous substrates rich in beta structure such as coccon, silk, human hair and fish scales. Result of fermentation experiment implied that addition of nitrogen sources (0.02% yeast extract and 0.02% tryptone to the basal medium increased keratinase production. Our experiments showed that keratinase production of Bacillus sp. MTS was higher and faster than that from B. licheniformis MB-2. Maximum extracellular keratinase activity of the enzyme derived from B. licheniformis was obtained during stationary phase at 72 h, while Bacillus sp. MTS was reached at 48 h. Disulfide reductase activity also detected in the extracellular fluid of Bacillus sp. MTS. The maximum condition for extracellular keratinase activity was 55oC and the enzyme showed two maximum pHs : pH 8.0 and pH 10. The zymogram analysis indicated sixth protein bands of 17, 25, 32, 53, 96 and 122 kD which were able to hydrolyze gelatin substrate in-situ. (Animal Production 12(1: 60-68 (2010Key Words : Bacillus, feather, keratinase, disulfide reductase

  11. Measurement and Prediction of Protein Phase Behaviour and Protein-Protein Interactions

    DEFF Research Database (Denmark)

    Faber, Cornelius

    2006-01-01

    fra Bacillus halmapalus (BHA) og Bacillus licheniformis (BLA), som var blevet oprenset til teknisk grad. Udviklingen af optimerede mikrobielle stammer sammen med fremskreden gæringsteknologi fører til et renere industrielt gæringsmedie med højere produkt koncentration, som vil muliggøre en økonomisk...

  12. Cross-species induction of antimicrobial compounds, biosurfactants and quorum-sensing inhibitors in tropical marine epibiotic bacteria by pathogens and biofouling microorganisms.

    Science.gov (United States)

    Dusane, Devendra H; Matkar, Pratiek; Venugopalan, Valayam P; Kumar, Ameeta Ravi; Zinjarde, Smita S

    2011-03-01

    Enhancement or induction of antimicrobial, biosurfactant, and quorum-sensing inhibition property in marine bacteria due to cross-species and cross-genera interactions was investigated. Four marine epibiotic bacteria (Bacillus sp. S3, B. pumilus S8, B. licheniformis D1, and Serratia marcescens V1) displaying antimicrobial activity against pathogenic or biofouling fungi (Candida albicans CA and Yarrowia lipolytica YL), and bacteria (Pseudomonas aeruginosa PA and Bacillus pumilus BP) were chosen for this study. The marine epibiotic bacteria when co-cultivated with the aforementioned fungi or bacteria showed induction or enhancement in antimicrobial activity, biosurfactant production, and quorum-sensing inhibition. Antifungal activity against Y. lipolytica YL was induced by co-cultivation of the pathogens or biofouling strains with the marine Bacillus sp. S3, B. pumilus S8, or B. licheniformis D1. Antibacterial activity against Ps. aeruginosa PA or B. pumilus BP was enhanced in most of the marine isolates after co-cultivation. Biosurfactant activity was significantly increased when cells of B. pumilus BP were co-cultivated with S. marcescens V1, B. pumilus S8, or B. licheniformis D1. Pigment reduction in the quorum-sensing inhibition indicator strain Chromobacterium violaceum 12472 was evident when the marine strain of Bacillus sp. S3 was grown in the presence of the inducer strain Ps. aeruginosa PA, suggesting quorum-sensing inhibition. The study has important ecological and biotechnological implications in terms of microbial competition in natural environments and enhancement of secondary metabolite production.

  13. Enhanced the energy outcomes from microalgal biomass by the novel biopretreatment

    International Nuclear Information System (INIS)

    He, Shuai; Fan, Xiaolei; Luo, Shengjun; Katukuri, Naveen Reddy; Guo, Rongbo

    2017-01-01

    Highlights: • The micro-aerobic pretreatment was used to improve energy yield of Chlorella sp. • The Bacillus licheniformis was confirmed to damage the cell wall of microalgae. • Obtained energy from Chlorella sp. was improved by 12.3%. • Pretreatment time was decreased from 60 h to 24 h. • The VS degradation efficiency was increased from 75.7% to 82.1%. - Abstract: Microalgae have been considered as one of the most promising biomass for the generation of biofuels. The anaerobic digestion (AD) has been proved to be a promising technique to transfer the microalgal biomass into biofuels. Previous study demonstrated that anaerobic pretreatment of microalgae biomass by Bacillus licheniformis could improve methane production. In this study micro-aerobic bio-pretreatment of microalgal biomass by the facultative anaerobic bacteria Bacillus licheniformis was invested with different loads of oxygen supplied. The bio-hydrogen and biomethane productions were tested to calculate total energy outcomes. The transmission electron microscope (TEM) photographs suggested that the novel micro-aerobic bio-pretreatment (MBP) could effectively damage the firm cell wall of algal cells. The processing time of the novel method (24 h) was less than the previous anaerobic pretreatment (60 h). Results showed that the group with 5 mL oxygen/g VS fed had the highest total energy outcomes, which was 17.6% higher than that of the anaerobic pretreatment.

  14. The use of an economical medium for the production of alkaline ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-05-03

    May 3, 2010 ... marine-processing by-products for the production of alkaline proteases by Bacillus licheniformis NH1. ... meat Sardinelle powder; WSP, whole Sardinelle powder. ... are used as cleaning additives in detergents to facilitate.

  15. 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

  16. A novel ion-beam-mutation effect application in identification of gene involved in bacterial antagonism to fungal infection of ornamental crops

    Science.gov (United States)

    Mahadtanapuk, S.; Teraarusiri, W.; Nanakorn, W.; Yu, L. D.; Thongkumkoon, P.; Anuntalabhochai, S.

    2014-05-01

    This work is on a novel application of ion beam effect on biological mutation. Bacillus licheniformis (B. licheniformis) is a common soil bacterium with an antagonistic effect on Curcuma alismatifolia Gagnep. and Chrysanthemum indicum Linn. In an attempt to control fungal diseases of local crops by utilizing B. licheniformis, we carried out gene analysis of the bacterium to understand the bacterial antagonistic mechanism. The bacterial cells were bombarded to induce mutations using nitrogen ion beam. After ion bombardment, DNA analysis revealed that the modified polymorphism fragment present in the wild type was missing in a bacterial mutant which lost the antifungal activity. The fragments conserved in the wild type but lost in the mutant bacteria was identified to code for the thioredoxin reductase (TrxR) gene. The gene analysis showed that the TrxR gene from B. licheniformis had the expression of the antagonism to fungi in a synchronous time evolution with the fungus inhibition when the bacteria were co-cultivated with the fungi. The collective results indicate the TrxR gene responsible for the antagonism of bacteria B. licheniformis to fungal infection.

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

    Science.gov (United States)

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

    2017-08-01

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

  18. Biodegradation of hydrocarbon compounds in Agbabu natural bitumen

    African Journals Online (AJOL)

    Infrared spectral changes and gravimetric analysis from the preliminary biodegradability study carried out on Agbabu Natural Bitumen showed the vulnerability of the bitumen to some bacteria: Pseudomonas putrefaciens, Pseudomonas nigrificans, Bacillus licheniformis, Pseudomonas fragi and Achromobacter aerogenes.

  19. Probiotics and Probiotic Metabolic Product Improved Intestinal Function and Ameliorated LPS-Induced Injury in Rats.

    Science.gov (United States)

    Deng, Bo; Wu, Jie; Li, Xiaohui; Men, Xiaoming; Xu, Ziwei

    2017-11-01

    In the present study, we sought to determine the effects of Bacillus subtilis (BAS) and Bacillus licheniformis (BAL) in rats after lipopolysaccharide (LPS)-induced acute intestinal inflammation. We also determined whether the B. subtilis metabolic product (BASM) is as effective as the live-cell probiotic. 60 male SD rats were randomly assigned to five groups and administered a diet containing 0.05% B. licheniformis (BAL group), 0.05% B. subtilis (BAS group), 0.5% B. subtilis metabolic product (BASM group), or a basic diet (PC group and NC group) for 40 days. On day 40, BAL, BAS, BASM, and NC groups were injected with 4 mg/kg body weight LPS. 4 h later, all rats were anesthetized and sacrificed. The results showed that the administration of B. licheniformis and B. subtilis improved intestinal function as evidenced by histology, increased enzyme activity, and mucosal thickness. They also increased the number of intraepithelial lymphocytes and decreased mucosal myeloperoxidase activity and plasma TNF-α. In addition, the cecal content of B. subtilis-treated rats had significantly increased microbial diversity, decreased numbers of Firmicutes, and increased numbers of Bacteroidetes as compared to rats fed basic diets. Similar to BAS group, the cecal content of B. licheniformis-treated rats decreased the number of Firmicutes. Administration of B. subtilis metabolic product had similar effects on intestinal function, inflammation response, and microbial diversity as B. subtilis but these effects were attenuated. In conclusion, administration of probiotic strains B. licheniformis or B. subtilis improved intestinal function, ameliorated the inflammation response, and modulated microflora after LPS-induced acute inflammation in rats. Non-living cells also exerted probiotic properties but live cells tended to function better.

  20. Screening of microorganisms for microbial enhanced oil recovery processes

    Energy Technology Data Exchange (ETDEWEB)

    Yonebayashi, H. [Japan National Oil Corp., Tokyo (Japan); Yoshida, S. [Japan Food Research Laboratiories, Tokyo (Japan). Div. of Microbiology; Ono, K. [Japan National Oil Corp., Chiba (Japan). Tech. Research Center; Enomoto, H. [Tohoku University, Sendai (Japan). Dept. of Geoscience and Tech.

    2000-01-01

    The objective of this study is to screen effective microorganisms for the Microbial Enhanced Oil Recovery process (or simply as MEOR). Samples of drilling cuttings, formation water, and soil were collected from domestic drilling sites and oil fields. Moreover, samples of activated-sludge and compost were collected from domestic sewage treatment facility and food treatment facility. At first, microorganisms in samples were investigated by incubation with different media; then they were isolated. By two stage-screening based on metabolizing ability, 4 strains (Bacillus licheniformis TRC-18-2-a, Enterobacter cloacae TRC-322, Bacillus subtilis TRC-4118, and Bacillus subtilis TRC-4126) were isolated as effective microorganisms for oil recovery. B. licheniformis TRC-18-2-a is a multifunctional microorganism possessing excellent surfactant productivity, and in addition it has gas, acid and polymer productivities. E. cloacae TRC-332 has gas and acid producing abilities. B. subtilis TRC-4118 and TRC-4126 are effective biosurfactant producers, and they reduce the interfacial tension to 0.04 and 0.12 dyne/cm, respectively. (author)

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

    Science.gov (United States)

    Baxi, Nandita N

    2014-01-01

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

  2. Aislamiento e identificaci?n de diez cepas bacterianas desnitrificantes a partir de un suelo agr?cola contaminado con abonos nitrogenados proveniente de una finca productora de cebolla en la Laguna de Tota, Boyac?, Colombia

    OpenAIRE

    Ostos Ortiz, Olga Lucia; Benavides L?pez de Mesa, Joaqu?n; Quintero, Gladys

    2006-01-01

    Este articulo presenta los resultados de la versatilidad adaptativa y tolerancia de una consorcio bacteriano constituido por 10 cepas bacterianas end?micas de un suelo agr?cola contaminado, proveniente de una finca productora de cebolla ubicada en la laguna de Tota en Aquitania, Boyac?, Colombia, expuesto al uso incontrolado de fertilizantes nitrogenados. La consorcio bacteriano aislado del suelo est? conformado por: Bacillus megaterium, Bacillus licheniformis, Pseudomonas stutzeri, Acinetoba...

  3. Author Details

    African Journals Online (AJOL)

    Amylase and its Assay using Bacillus Licheniformis Isolated from Excavated Land Sites in Ibadan, Nigeria Abstract PDF. ISSN: 2006-6996. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors · FAQ's ...

  4. Synthesis and processing of escherichia-coli tem-beta-lactamase and bacillus-licheniformis alpha-amylase in escherichia-coli : The role of signal peptidase-i

    NARCIS (Netherlands)

    van Dijl, J M; Smith, H; Bron, Sierd; Venema, Gerard

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

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

    NARCIS (Netherlands)

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

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

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

    Science.gov (United States)

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

  7. Protein-peptide interactions in mixtures of whey peptides and whey proteins

    NARCIS (Netherlands)

    Creusot, N.; Gruppen, H.

    2007-01-01

    The effects of several conditions on the amounts and compositions of aggregates formed in mixtures of whey protein hydrolysate, made with Bacillus licheniformis protease, and whey protein isolate were investigated using response surface methodology. Next, the peptides present in the aggregates were

  8. A novel ion-beam-mutation effect application in identification of gene involved in bacterial antagonism to fungal infection of ornamental crops

    Energy Technology Data Exchange (ETDEWEB)

    Mahadtanapuk, S. [Faculty of Agriculture and Natural Resources, University of Phayao, Maeka, Muang, Phayao 56000 (Thailand); Teraarusiri, W. [Central Laboratory, University of Phayao, Maeka, Muang, Phayao 56000 (Thailand); Nanakorn, W. [The Crown Property Bureau, 173 Nakhonratchasrima Road, Dusit, Bangkok 10300 (Thailand); Yu, L.D., E-mail: yuld@thep-center.org [Plasma and Beam Physics Research Facility, Department of Physics and Materials Science, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Thailand Center of Excellence in Physics, Commission on Higher Education, 328 Si Ayutthaya Road, Bangkok 10400 (Thailand); Thongkumkoon, P. [Plasma and Beam Physics Research Facility, Department of Physics and Materials Science, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Anuntalabhochai, S., E-mail: soanu.1@gmail.com [Molecular Biology Laboratory, Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand)

    2014-05-01

    Highlights: • Ion beam bombardment induced mutation in bacterial B. licheniformis. • A mutant lost antifungal activity. • DNA fingerprint of the mutant was analyzed. • The lost gene was indentified to code for TrxR gene. • TrxR gene from B. licheniformis expressed the flower antagonism to fungi. - Abstract: This work is on a novel application of ion beam effect on biological mutation. Bacillus licheniformis (B. licheniformis) is a common soil bacterium with an antagonistic effect on Curcuma alismatifolia Gagnep. and Chrysanthemum indicum Linn. In an attempt to control fungal diseases of local crops by utilizing B. licheniformis, we carried out gene analysis of the bacterium to understand the bacterial antagonistic mechanism. The bacterial cells were bombarded to induce mutations using nitrogen ion beam. After ion bombardment, DNA analysis revealed that the modified polymorphism fragment present in the wild type was missing in a bacterial mutant which lost the antifungal activity. The fragments conserved in the wild type but lost in the mutant bacteria was identified to code for the thioredoxin reductase (TrxR) gene. The gene analysis showed that the TrxR gene from B. licheniformis had the expression of the antagonism to fungi in a synchronous time evolution with the fungus inhibition when the bacteria were co-cultivated with the fungi. The collective results indicate the TrxR gene responsible for the antagonism of bacteria B. licheniformis to fungal infection.

  9. A novel ion-beam-mutation effect application in identification of gene involved in bacterial antagonism to fungal infection of ornamental crops

    International Nuclear Information System (INIS)

    Mahadtanapuk, S.; Teraarusiri, W.; Nanakorn, W.; Yu, L.D.; Thongkumkoon, P.; Anuntalabhochai, S.

    2014-01-01

    Highlights: • Ion beam bombardment induced mutation in bacterial B. licheniformis. • A mutant lost antifungal activity. • DNA fingerprint of the mutant was analyzed. • The lost gene was indentified to code for TrxR gene. • TrxR gene from B. licheniformis expressed the flower antagonism to fungi. - Abstract: This work is on a novel application of ion beam effect on biological mutation. Bacillus licheniformis (B. licheniformis) is a common soil bacterium with an antagonistic effect on Curcuma alismatifolia Gagnep. and Chrysanthemum indicum Linn. In an attempt to control fungal diseases of local crops by utilizing B. licheniformis, we carried out gene analysis of the bacterium to understand the bacterial antagonistic mechanism. The bacterial cells were bombarded to induce mutations using nitrogen ion beam. After ion bombardment, DNA analysis revealed that the modified polymorphism fragment present in the wild type was missing in a bacterial mutant which lost the antifungal activity. The fragments conserved in the wild type but lost in the mutant bacteria was identified to code for the thioredoxin reductase (TrxR) gene. The gene analysis showed that the TrxR gene from B. licheniformis had the expression of the antagonism to fungi in a synchronous time evolution with the fungus inhibition when the bacteria were co-cultivated with the fungi. The collective results indicate the TrxR gene responsible for the antagonism of bacteria B. licheniformis to fungal infection

  10. Biodegradable Polymeric Substances Produced by a Marine Bacterium from a Surplus Stream of the Biodiesel Industry

    DEFF Research Database (Denmark)

    Bhattacharya, Sourish; Dubey, Sonam; Singh, Priyanka

    2016-01-01

    epsilon-polylysine and 64.6% (w/w) intracellular polyhydroxyalkanoate (PHA) in the same fermentation broth (1 L shake flask) utilizing Jatropha biodiesel waste residues as carbon rich source by marine bacterial strain (Bacillus licheniformis PL26), isolated from west coast of India. The synthesized...

  11. The effect of process conditions on the alpha-amylolytic hydrolysis of amylopectin potato starch : an experimental design approach

    NARCIS (Netherlands)

    Marchal, L.M.; Jonkers, J.; Franke, G.T.; Goooijer, de C.D.; Tramper, J.

    1999-01-01

    The hydrolysis of amylopectin potato starch with Bacillus licheniformis α-amylase (Maxamyl) was studied under industrially relevant conditions (i.e. high dry-weight concentrations). The following ranges of process conditions were chosen and investigated by means of an experimental design: pH [5.6-

  12. Production of a Thermostable Α-Amylase and its Assay using ...

    African Journals Online (AJOL)

    Screening for amylolytic properties from obtained isolates was carried out on starch agar plates while production and characterization of α-amylase was carried out using submerged fermentation. The isolated organism was identified as Bacillus licheniformis. Physiological studies on the isolate showed that temperature of ...

  13. Antibacterial and antioxidant properties of macrocyclic Schiff bases ...

    African Journals Online (AJOL)

    The synthesized compounds were screened for their in vitro growth inhibiting activity against different strains of bacteria viz., Staphylococcus aureus, Bacillus licheniformis, Escherichia coli and Micrococcus luteus (ATCC) and were compared with the standard antibiotic oflaxocin. Also in-vitro antioxidant activity of all ...

  14. Partial purification, characterization and hydrolytic activities of ...

    African Journals Online (AJOL)

    α-Amylase and amyloglucosidase produced by amylolytic Bacillus licheniformis and Aspergillus niger isolated from plantain and yam peels media were partially purified and characterized. Following cultivation of the microbial isolates on the agricultural residue media, crude enzyme solutions were obtained by filtration and ...

  15. Towards the understanding of microbial metabolism in relation to microbial enhanced oil recovery

    DEFF Research Database (Denmark)

    Halim, Amalia Yunita; Nielsen, Sidsel Marie; Nielsen, Kristian Fog

    2017-01-01

    In this study, Bacillus licheniformis 421 was used as a model organism to understand the effects of microbial cell growth and metabolite production under anaerobic conditions in relation to microbial enhanced oil recovery. The bacterium was able to grow anaerobically on different carbon compounds...

  16. Phylogenetic and Comparative Sequence Analysis of Thermostable Alpha Amylases of kingdom Archea, Prokaryotes and Eukaryotes.

    Science.gov (United States)

    Huma, Tayyaba; Maryam, Arooma; Rehman, Shahid Ur; Qamar, Muhammad Tahir Ul; Shaheen, Tayyaba; Haque, Asma; Shaheen, Bushra

    2014-01-01

    Alpha amylase family is generally defined as a group of enzymes that can hydrolyse and transglycosylase α-(1, 4) or α-(1, 6) glycosidic bonds along with the preservation of anomeric configuration. For the comparative analysis of alpha amylase family, nucleotide sequences of seven thermo stable organisms of Kingdom Archea i.e. Pyrococcus furiosus (100-105°C), Kingdom Prokaryotes i.e. Bacillus licheniformis (90-95°C), Geobacillus stearothermophilus (75°C), Bacillus amyloliquefaciens (72°C), Bacillus subtilis (70°C) and Bacillus KSM K38 (55°C) and Eukaryotes i.e. Aspergillus oryzae (60°C) were selected from NCBI. Primary structure composition analysis and Conserved sequence analysis were conducted through Bio Edit tools. Results from BioEdit shown only three conserved regions of base pairs and least similarity in MSA of the above mentioned alpha amylases. In Mega 5.1 Phylogeny of thermo stable alpha amylases of Kingdom Archea, Prokaryotes and Eukaryote was handled by Neighbor-Joining (NJ) algorithm. Mega 5.1 phylogenetic results suggested that alpha amylases of thermo stable organisms i.e. Pyrococcus furiosus (100-105°C), Bacillus licheniformis (90-95°C), Geobacillus stearothermophilus (75°C) and Bacillus amyloliquefaciens (72°C) are more distantly related as compared to less thermo stable organisms. By keeping in mind the characteristics of most thermo stable alpha amylases novel and improved features can be introduced in less thermo stable alpha amylases so that they become more thermo tolerant and productive for industry.

  17. Enhancing RGI lyase thermostability by targeted single point mutations

    DEFF Research Database (Denmark)

    Silva, Inês R.; Larsen, Dorte Møller; Jers, Carsten

    2013-01-01

    Rhamnogalacturonan I lyase (RGI lyase) (EC 4.2.2.-) catalyzes the cleavage of rhamnogalacturonan I in pectins by β-elimination. In this study the thermal stability of a RGI lyase (PL 11) originating from Bacillus licheniformis DSM 13/ATCC14580 was increased by a targeted protein engineering...

  18. Identification, expression, and characterization of a novel bacterial RGI Lyase enzyme for the production of bio-functional fibers

    DEFF Research Database (Denmark)

    da Silva, Ines Isabel Cardoso Rodrigues; Larsen, Dorte Møller; Meyer, Anne S.

    2011-01-01

    A gene encoding a putative rhamnogalacturonan I (RGI) Lyase (EC 4.2.2.-) from Bacillus licheniformis (DSM13) was selected after a homology search and phylogenetic analysis and optimized with respect to codon usage. The designed gene was transformed into Pichia pastoris and the enzyme was produced...

  19. Bacillus velezensis is not a later heterotypic synonym of Bacillus amyloliquefaciens; Bacillus methylotrophicus, Bacillus amyloliquefaciens subsp. plantarum and 'Bacillus oryzicola' are later heterotypic synonyms of Bacillus velezensis based on phylogenomics.

    Science.gov (United States)

    Dunlap, Christopher A; Kim, Soo-Jin; Kwon, Soon-Wo; Rooney, Alejandro P

    2016-03-01

    Bacillus velezensis was previously reported to be a later heterotypic synonym of Bacillus amyloliquefaciens , based primarily on DNA-DNA relatedness values. We have sequenced a draft genome of B. velezensis NRRL B-41580 T . Comparative genomics and DNA-DNA relatedness calculations show that it is not a synonym of B. amyloliquefaciens. It was instead synonymous with Bacillus methylotrophicus. ' Bacillus oryzicola ' is a recently described species that was isolated as an endophyte of rice ( Oryza sativa ). The strain was demonstrated to have plant-pathogen antagonist activity in greenhouse assays, and the 16S rRNA gene was reported to have 99.7 % sequence similarity with Bacillus siamensis and B. methylotrophicus , which are both known for their plant pathogen antagonism. To better understand the phylogenetics of these closely related strains, we sequenced the genome of ' B . oryzicola ' KACC 18228. Comparative genomic analysis showed only minor differences between this strain and the genomes of B. velezensis NRRL B-41580 T , B. methylotrophicus KACC 13015 T and Bacillus amyloliquefaciens subsp. plantarum FZB42 T . The pairwise in silico DNA-DNA hybridization values calculated in comparisons between the strains were all greater than 84 %, which is well above the standard species threshold of 70 %. The results of morphological, physiological, chemotaxonomic and phylogenetic analyses indicate that the strains share phenotype and genotype coherence. Therefore, we propose that B. methylotrophicus KACC 13015 T , B. amyloliquefaciens subsp. plantarum FZB42 T , and ' B. oryzicola' KACC 18228 should be reclassified as later heterotypic synonyms of B. velezensis NRRL B-41580 T , since the valid publication date of B. velezensis precedes the other three strains.

  20. Efforts to identify spore forming bacillus

    Energy Technology Data Exchange (ETDEWEB)

    Zuleiha, M.S.; Hilmy, N. (National Atomic Energy Agency, Jakarta (Indonesia). Pasar Djumat Research Centre)

    1982-04-01

    Efforts to identify 47 species of radioresistant spore forming bacillus sp. isolated from locally produced medical devices have been carried out. The identifications was conducted using 19 kinds of biochemical tests and compared to species to bacillus subtilis W. T.; bacillus pumilus E 601 and bacillus sphaericus Csub(I)A. The results showed that bacillus sp. examined could be divided into 6 groups, i.e. bacillus cereus; bacillus subtilis; bacillus stearothermophylus; bacillus coagulans; bacillus sphaericus and bacillus circulans.

  1. Efforts to identify spore forming bacillus

    International Nuclear Information System (INIS)

    Zuleiha, M.S.; Hilmy, Nazly

    1982-01-01

    Efforts to identify 47 species of radioresistant spore forming bacillus sp. isolated from locally produced medical devices have been carried out. The identifications was conducted using 19 kinds of biochemical tests and compared to species to bacillus subtilis W. T.; bacillus pumilus E 601 and bacillus sphaericus Csub(I)A. The results showed that bacillus sp. examined could be divided into 6 groups, i.e. bacillus cereus; bacillus subtilis; bacillus stearothermophylus; bacillus coagulans; bacillus sphaericus and bacillus circulans. (author)

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

    International Nuclear Information System (INIS)

    Asad, W.; Saleem, F.; Ajaz, M.; Rasool, S.A.

    2014-01-01

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

  3. Possibility of biological control of primocane fruiting raspberry disease caused by Fusarium sambucinum.

    Science.gov (United States)

    Shternshis, Margarita V; Belyaev, Anatoly A; Matchenko, Nina S; Shpatova, Tatyana V; Lelyak, Anastasya A

    2015-10-01

    Biological control agents are a promising alternative to chemical pesticides for plant disease suppression. The main advantage of the natural biocontrol agents, such as antagonistic bacteria compared with chemicals, includes environmental pollution prevention and a decrease of chemical residues in fruits. This study is aimed to evaluate the impact of three Bacillus strains on disease of primocane fruiting raspberry canes caused by Fusarium sambucinum under controlled infection load and uncontrolled environmental factors. Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloliquefaciens were used for biocontrol of plant disease in 2013 and 2014 which differed by environmental conditions. The test suspensions were 10(5) CFU/ml for each bacterial strain. To estimate the effect of biological agents on Fusarium disease, canes were cut at the end of vegetation, and the area of outer and internal lesions was measured. In addition to antagonistic effect, the strains revealed the ability to induce plant resistance comparable with chitosan-based formulation. Under variable ways of cane treatment by bacterial strains, the more effective were B. subtilis and B. licheniformis demonstrating dual biocontrol effect. However, environmental factors were shown to impact the strain biocontrol ability; changes in air temperature and humidity led to the enhanced activity of B. amyloliquefaciens. For the first time, the possibility of replacing chemicals with environmentally benign biological agents for ecologically safe control of the raspberry primocane fruiting disease was shown.

  4. Bacillus Coagulans

    Science.gov (United States)

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

  5. Calcium-induced stabilization of -amylase against guanidine ...

    African Journals Online (AJOL)

    Guanidine hydrochloride (GdnHCl) denaturation of native and Ca- depleted Bacillus licheniformis α-amylase (BLA) was investigated both in the absence and presence of 2 mM calcium chloride (CaCl2) using circular dichroism, fluorescence spectroscopy and biological activity. In both states (Cadepleted and native form), ...

  6. Growth pattern and structural nature of amylases produced by some ...

    African Journals Online (AJOL)

    The growth pattern and microbial biomass formed during metabolic activities of the Bacillus species on starchy substrates was determined. The result showed that the strains B. subtilis (WBS), B. licheniformis (WBL) and B. coagulans (MBC) generally had high growth rate. B. circulans (SBC) and B. coagulans (WBC) has ...

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2005-09-14

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

  8. Biosurfactant and enhanced oil recovery

    Science.gov (United States)

    McInerney, Michael J.; Jenneman, Gary E.; Knapp, Roy M.; Menzie, Donald E.

    1985-06-11

    A pure culture of Bacillus licheniformis strain JF-2 (ATCC No. 39307) and a process for using said culture and the surfactant lichenysin produced thereby for the enhancement of oil recovery from subterranean formations. Lichenysin is an effective surfactant over a wide range of temperatures, pH's, salt and calcium concentrations.

  9. Partial purification and characterization of metalloprotease of ...

    African Journals Online (AJOL)

    USER

    2013-07-31

    Jul 31, 2013 ... The supplementation of partially purified enzyme preparation in detergents such as Rin and Wheel significantly improved their cleansing efficiency as blood and fish curry stains on the cloth disappeared within 15 min (Figure 6). Our finding go hand in hand with earlier findings on Bacillus licheniformis ...

  10. Bulletin of Materials Science | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    A dominant freshwater bacterial isolate, Bacillus licheniformis, which interacted with environmentally relevant concentrations of TiO2 NPs (1 g/mL), was analysed and compared under both light and dark conditions. Aggregation of cells upon NP interaction and adsorption of NPs onto the cell membrane was evident from ...

  11. Growth and Cultivation of the Unusual Generalized Transducing Bacillus Bacteriophage SP-15

    Science.gov (United States)

    Taylor, Martha J.; Goldberg, Ivan D.

    1971-01-01

    Additional properties of SP-15, a generalized transducing bacteriophage notable for the ability to transfer an unusually large fragment of deoxyribonucleic acid (DNA) to Bacillus subtilis and B. licheniformis, are presented together with improved methods that enhance its utility. Simple means have been found to provide the rigid control over moisture that is necessary for the assay of plaque-forming units (PFU). Reproducible procedures for propagating transducing phage, which depend upon an appropriate mixing of PFU with uninfected bacteria, have replaced less reliable methods that utilized infected spores. Transduction of B. subtilis W-23 increased linearly when MgSO4 in recipient cell-SP-15 mixtures was increased from 0.005 to 0.03 m. Methods have been developed that protect SP-15 from the damaging effects of CsCl and of osmotic shock subsequent to dilution. Evidence that the PFU and transducing particles of lysates decay at the same slow rate during extended storage suggests that the decay is a result of damage to protein rather than to DNA. One-step growth experiments, in which SP-15 was propagated on B. subtilis W-23-Sr/1 mg, indicated a latent period of 100 min, a rise period of 60 min, and a burst size of 25 to 34 PFU per infected cell. These findings suggest explanations for some of the technical difficulties SP-15 has presented. PMID:4999971

  12. The use of an economical medium for the production of alkaline ...

    African Journals Online (AJOL)

    The present study is concerned with the selection of new economical media based on agricultural and marine-processing by-products for the production of alkaline proteases by Bacillus licheniformis NH1. Powders from different fish species were prepared and then tested as growth media at a concentration of 10 g/l for ...

  13. Improving the two-step remediation process for CCA-treated wood. Part I, Evaluating oxalic acid extraction

    Science.gov (United States)

    Carol Clausen

    2004-01-01

    In this study, three possible improvements to a remediation process for chromated-copper-arsenate (CCA) treated wood were evaluated. The process involves two steps: oxalic acid extraction of wood fiber followed by bacterial culture with Bacillus licheniformis CC01. The three potential improvements to the oxalic acid extraction step were (1) reusing oxalic acid for...

  14. Mechanism of bactericidal activity of Silver Nitrate - a concentration dependent bi-functional molecule

    Directory of Open Access Journals (Sweden)

    Sureshbabu Ram Kumar Pandian

    2010-10-01

    Full Text Available Silver nitrate imparts different functions on bacteria depending upon its concentration. At lower concentration it induced synthesis of nanoparticles, whereas at higher concentrations it induced cell death. Bacillus licheniformis was used as model system. The MIC was 5 mM, and it induced catalase production, apoptotic body formation and DNA fragmentation.

  15. Stabilization of 5A1 urease by covalent attachement to wool | Ahmed ...

    African Journals Online (AJOL)

    The investigation of five bacterial strains for urease production referred that Bacillus licheniformis 5A1 had the highest urease activity (10.3U/ml/min) after 24h. The enzyme was covalently coupled to different carriers via glutaraldehyde, and wool gave the highest immobilization yield (76.4%) and retained 85% of the original ...

  16. 1033-IJBCS-Article-Lamine Said Baba Moussa

    African Journals Online (AJOL)

    DR GATSING

    1Centre de Recherches Agricoles Sud/Institut National des Recherches ... trois espèces de Pseudomonas, trois espèces de Streptomyces, huit espèces de Bacillus et ..... licheniformis ; B. lentus ; B. circulans et B. ..... asiciadas al cultivo del maiz. ... bacteria and their inoculation effects on growth and nitrogen uptake of crop ...

  17. Identification of bacteria used for microbial enhanced oil recovery process by fluorescence in situ hybridization technique

    Energy Technology Data Exchange (ETDEWEB)

    Fujiwara, K.; Tanaka, S.; Otsuka, M. [Kansai Research Institute, Kyoto (Japan). Lifescience Lab.; Yonebayashi, H. [Japan National Oil Corp., Chiba (Japan). Tech. Research Center; Enomoto, H. [Tohoku University, Sendai (Japan). Dept. of Geoscience and Tech.

    2000-01-01

    A fluorescence in situ hybridization (FISH) technique using 16S rRNA-targeted oligonucleotide probes was developed for rapid detection of microorganisms for use in the microbial enhancement of oil recovery (MEOR) process. Two microorganisms, Enterobacter cloacae TRC-322 and Bacillus licheniformis TRC-18-2-a, were selected from a collection of Enterobacter sp. and Bacillus sp. which were screened in previous studies as candidate microorganisms for injection, and were used for this experiment. Oligonucleotide probes, design based on specific sequences in the 16S rRNA gene were labeled with either fluorescein isothiocyanate (FITC), or 6-car-boxy-X-rhodamine (ROX), and were allowed to hybridize with fixed cells of the two microorganisms noted above. The fluorescence signal emitted from each microorganism cells could clearly be detected by an epifluorescence microscope. Moreover, E. cloacae TRC-322 and B, licheniformis TRC-18-2-a, suspended in actual reservoir brine, including inorganic salts, oil and aboriginal cells of the reservoir brine, could be detected directly by this hybridization method, without the need for cultivation and isolation. (author)

  18. Inhibition of in vitro growth of soil-borne pathogens by compost-inhabiting indigenous bacteria and fungi

    International Nuclear Information System (INIS)

    Ramzan, N.; Noreen, N.; Shahzad, S.

    2014-01-01

    During the present studies, compost-inhabiting microorganisms including 44 fungi and 15 bacteria isolated from different compost samples were evaluated for their in vitro efficacy against soil-borne pathogens viz., Fusarium solani, Macrophomina phaseolina, Pythium aphanidermatum, Rhizoctonia solani, and Sclerotium rolfsii. Compost inhabiting microbes like Trichoderma harzianum, T. virens, Bacillus cereus, B. pumilus, B. subtilis, Micrococcus varians and Pseudomonas fluorescens were found to inhibit all the test pathogens. Acrophialophora fusispora and Penicillium citrinum reduced the mycelial growth of all the test pathogens except Sclerotium rolfsii. Bacillus licheniformis and Bacillus megaterium showed biocontrol activity against all the pathogens except Rhizoctonia solani. Trichoderma harzianum parasitized mycelia of all the tested pathogens and produced coiling around the mycelium. (author)

  19. Mixture of residual fish hydrolysate and fish extract hydrolysate to ...

    African Journals Online (AJOL)

    Bacillus licheniformis 6346 activated in nutrient broth-starch was inoculated to a locally formulated solid medium (paddy husk, rice flour, (NH4)2SO4, cuttle fish powder, murate potash, table salt, triple super phosphate, sesame oil, coconut oil and tap water) and the highest -amylase activity was obtained on day 5 (1075 U g ...

  20. Method of obtaining enhanced oil recovery by the use of microorganisms; Fremgangsmaate for aa oppnaa forbedret oljeutvinning under anvendelse av mikroorganismer

    Energy Technology Data Exchange (ETDEWEB)

    Silver, R.S.; Moon, W.G.; Bunting, P.M.; Acheson, W.P.

    1995-03-27

    The invention deals with an method for improving the recovery from petroleum reservoirs by the use of microorganisms. According to the invention, there is to be injected a suspension of a biological clean breed of the Bacillus licheniformis or spores thereof in order to tighten selectively the relative strong penetrating zones of a reservoir. 2 figs.

  1. 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.

  2. Partial Characterisation of Bacteriocins Produced by Bacillus cereus Isolates from Milk and Milk Products

    Directory of Open Access Journals (Sweden)

    Bojana Bogović Matijašić

    2003-01-01

    Full Text Available Thirty one (19.2 % out of 161 Bacillus cereus isolates from raw milk and milk products were found to produce proteinaceous substances which inhibit the growth of other B. cereus isolates. The detection of antibacterial activity depended on medium and method used. Bactericidal activity was detected in 23 (14 % or 19 (12 % of the tested strains on the triptic soya agar and brain-heart infusion with glucose, respectively, while 11 (7 % of the strains produced bactericidal substances on both media. Nineteen percent of isolates from raw milk and 20 % of isolates from milk products were found to produce bacteriocins. Four B. cereus isolates inhibited the growth of individual test strains belonging to B. licheniformis, B. subtilis, Enterococcus faecalis, Escherichia coli, Staphylococcus aureus, Lactobacillus helveticus and L. casei species. The bacteriocins of four B. cereus isolates were studied in more detail. The production and activity of these substances were detected in stationary- phase of bacterial culture. Two of them were stable after heating at 60 °C, while only one was stable after heating at 75 °C for 15 minutes. All of them were active over a range of pH=3–10. The apparent molecular weights of four bacteriocins detected by SDS-PAGE electrophoresis were in the range of 1 to 8 kDa.

  3. STABILITY IN REAL TIME OF SOME CRYOPRESERVED MICROBIAL STRAINS WITH REFERENCE TO GENETICALLY MODIFIED MICROORGANISMS

    Directory of Open Access Journals (Sweden)

    DANIELA VINTILĂ

    2007-05-01

    Full Text Available The aim of this work is to analyze the viability of microorganisms from Collection of Industrial Microorganisms from Faculty of Animal Science and Biotechnology – Timisoara, during freezing and thawing as part of cryopreservation technique. The stability in real time of 19 strains cryopreserved in 16% glycerol was evaluated during a 6-months period. The strains studied were: Escherichia coli, Lactobacillus acidophilus, Rhizobium meliloti, Saccharomyces cerevisiae, Aspergillus oryzae, Aspergillus niger, Trichoderma viride, Bacillus globigii, Bacillus licheniformis, and 9 strains of Bacillus subtilis. The strains cryopreserved at -20oC and -70oC were activated using the fast thawing protocol. A better cell recovery was achieved with the -70oC protocol reaching an average viability for E. coli of 86,3%, comparing with 78,6% in -20oC protocol. The cell recovery percentages for the other strains were: 92,4% for L. acidophilus, 93,9% for A.niger, 89% for A. oryzae, 86,7% for T. viride, 94,2% for R. meliloti, 82,1% for S. cerevisiae, 89,9% for B. licheniformis. Regarding the viability of genetically modified microorganisms, the values shows a good recovering after freezing and thawing, even after 180 days of cryopreservation. With the -20oC protocol lower viability was observed due probably to the formation of eutectic mixtures and recrystalization processes.

  4. Comparative study of physicochemical analysis of prosopis africana seeds fermented with different starter cultures

    Directory of Open Access Journals (Sweden)

    G. P. Oyeyiola

    2017-01-01

    Full Text Available Prosopis africana (African mesquite is one of the lesser known legume seed crops in Nigeria, which in a fermented state, gives a food condiment. Because of its rich protein content (about 34%, efforts are made to utilize some lesser known legumes to improve the nutritional status of the people. This study was carried out by fermenting Prosopis africana seeds with mono and mixed cultures of bacterial isolates to produce a local condiment called Okpehe. Standard AOAC methods were used to determine the pH, total sugar and crude protein content of the fermented seeds. During the production of Okpehe with mono cultures of bacteria, the pH ranged between 6.80 and 8.92; total sugar between 10.2 and 7.5 mg/g, and crude protein between 34.62 and 41.25%. In the mixed culture inoculated samples, the pH increased from 7.00 to 8.92; total sugar decreased from 9.4 to 7.4 mg/g, while the crude protein increased (35.02 - 44.61% significantly (p < 0.05 as the fermentation progressed. The highest crude protein content of 44.61% was obtained with the combination of Bacillus subtilis and Bacillus licheniformis, while the lowest protein content referred to the combination of Bacillus megaterium and Bacillus pumilus. The result of this study showed that Prosopis africana seeds could be utilized for the production of Okpehe using mixed cultures of B. subtilis and B. licheniformis, so as to increase the protein intake of the populace.

  5. Bacillus velezensis is a later heterotypic synonym of Bacillus amyloliquefaciens.

    Science.gov (United States)

    Wang, Li-Ting; Lee, Fwu-Ling; Tai, Chun-Ju; Kuo, Hsiao-Ping

    2008-03-01

    Strain BCRC 14193, isolated from soil, shared more than 99 % 16S rRNA gene sequence similarity with Bacillus amyloliquefaciens BCRC 11601(T) and Bacillus velezensis BCRC 17467(T). This strain was previously identified as B. amyloliquefaciens, based on DNA-DNA hybridization, but its DNA relatedness value with B. velezensis BCRC 17467(T) was 89 %. To investigate the relatedness of strain BCRC 14193, B. amyloliquefaciens and B. velezensis, the partial sequence of the gene encoding the subunit B protein of DNA gyrase (gyrB) was determined. B. velezensis BCRC 17467(T) shared high gyrB gene sequence similarity with B. amyloliquefaciens BCRC 14193 (98.4 %) and all of the B. amyloliquefaciens strains available (95.5-95.6 %). DNA-DNA hybridization experiments revealed high relatedness values between B. velezensis BCRC 17467(T) and B. amyloliquefaciens BCRC 11601(T) (74 %) and the B. amyloliquefaciens reference strains (74-89 %). Based on these data and the lack of phenotypic distinctive characteristics, we propose Bacillus velezensis as a later heterotypic synonym of Bacillus amyloliquefaciens.

  6. Comparative sequence analyses on the 16S rRNA (rDNA) of Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus and proposal for creation of a new genus, Alicyclobacillus gen. nov

    Science.gov (United States)

    Wisotzkey, J. D.; Jurtshuk, P. Jr; Fox, G. E.; Deinhard, G.; Poralla, K.

    1992-01-01

    Comparative 16S rRNA (rDNA) sequence analyses performed on the thermophilic Bacillus species Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus revealed that these organisms are sufficiently different from the traditional Bacillus species to warrant reclassification in a new genus, Alicyclobacillus gen. nov. An analysis of 16S rRNA sequences established that these three thermoacidophiles cluster in a group that differs markedly from both the obligately thermophilic organisms Bacillus stearothermophilus and the facultatively thermophilic organism Bacillus coagulans, as well as many other common mesophilic and thermophilic Bacillus species. The thermoacidophilic Bacillus species B. acidocaldarius, B. acidoterrestris, and B. cycloheptanicus also are unique in that they possess omega-alicylic fatty acid as the major natural membranous lipid component, which is a rare phenotype that has not been found in any other Bacillus species characterized to date. This phenotype, along with the 16S rRNA sequence data, suggests that these thermoacidophiles are biochemically and genetically unique and supports the proposal that they should be reclassified in the new genus Alicyclobacillus.

  7. Characterization of microsatellite loci in the stick insects Bacillus rossius rossius, Bacillus rossius redtenbacheri and Bacillus whitei (Insecta : Phasmatodea)

    DEFF Research Database (Denmark)

    Andersen, DH; Pertoldi, C; Loeschcke, V

    2005-01-01

    Five microsatellite markers were obtained from a dinucleotide enriched genomic library of the stick insect Bacillus rossius rossius. The markers were tested in three species of Bacillus. All loci were polymorphic when tested across species. The number of alleles at each locus was low (maximum four...

  8. Causes of infectious abortion in the Mediterranean buffalo.

    Directory of Open Access Journals (Sweden)

    G. Galiero

    2010-02-01

    Full Text Available Bacteria and viruses can cause abortion in buffaloes. This review describes the abortigenic infectious agents found in Mediterranean buffalo cows and the microbiological methods used for their diagnosis. The abortigenic agents are: Brucella spp., Arcanobacterium pyogenes, Chlamydophila spp., Coxiella burnetii, Bacillus licheniformis, E.coli, Leptospira spp., Bubaline Herpes Virus-1 (BuHV-1, Bovine Viral Diarrhoea Virus.

  9. Obtaining edaphic biostimulants/biofertilizers from sewage sludge using fermentative processes. Short-time effects on soil biochemical properties.

    Science.gov (United States)

    Rodríguez-Morgado, Bruno; Caballero, Pablo; Paneque, Patricia; Gómez, Isidoro; Parrado, Juan; Tejada, Manuel

    2017-10-28

    In this manuscript, we study the manufacture and effect on soils of different edaphic biostimulants/biofertilizers (BS) obtained from sewage sludge using Bacillus licheniformis as biological tool. These BS consist of different combinations of organic matter, bacteria and enzymes that were subjected to several treatments. These BS were applied in soil in order to observe their influence on the biochemical properties (enzymatic activities and ergosterol content). Dehydrogenase, urease, β-glucosidase, phosphatase activities and ergosterol content were measured at different incubation days. Only dehydrogenase activity and ergosterol content were significantly stimulated after the application of BS1 and BS4. Rest of the extracellular activities were not stimulated probably because B. licheniformis practically has digested all organic substrates during fermentation process.

  10. BacillusRegNet

    DEFF Research Database (Denmark)

    Misirli, Goksel; Hallinan, Jennifer; Röttger, Richard

    2014-01-01

    As high-throughput technologies become cheaper and easier to use, raw sequence data and corresponding annotations for many organisms are becoming available. However, sequence data alone is not sufficient to explain the biological behaviour of organisms, which arises largely from complex molecular...... the associated BacillusRegNet website (http://bacillus.ncl.ac.uk)....

  11. Isolation and Phylogenetic Analysis of Thermophile Community Within Tanjung Sakti Hot Spring, South Sumatera, Indonesia

    Directory of Open Access Journals (Sweden)

    Heni Yohandini

    2015-07-01

    Full Text Available A community of thermophiles within Tanjung Sakti Hot Spring (South Sumatera have been cultivated and identified based on 16S ribosomal RNA gene sequence. The hot spring has temperature 80 °C–91 °C and pH 7–8. We used a simple method for culturing the microbes, by enriching the spring water with nutrient broth media. Phylogenetic analysis showed that the method could recover microbes, which clustered within four distinct taxonomic groups: Anoxybacillus, Geobacillus, Brevibacillus, and Bacillus. These microbes closely related to Anoxybacillus rupiensis, Anoxybacillus flavithermus, Geobacillus pallidus, Brevibacillus thermoruber, Bacillus licheniformis, and Bacillus thermoamylovorans. The 16S ribosomal RNA gene sequence of one isolate only had 96% similarity with Brevibacillus sequence in GenBank.

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

    Directory of Open Access Journals (Sweden)

    Stanojević Olja

    2016-01-01

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

  13. Effects of small peptides, probiotics, prebiotics, and synbiotics on growth performance, digestive enzymes, and oxidative stress in orange-spotted grouper, Epinephelus coioides, juveniles reared in artificial seawater

    Science.gov (United States)

    Wang, Tao; Cheng, Yongzhou; Chen, Xiaoyan; Liu, Zhaopu; Long, Xiaohua

    2017-01-01

    Aquaculture production efficiency may increase by using feed additives. This study investigated the effects of different dietary additives [w/w: 2% small peptides, 0.01% probiotics ( Bacillus licheniformis) and 0.2% prebiotics (inulin)] on growth performance, digestive enzyme activities, and oxidative stress in juvenile Epinephelus coioides reared in artificial seawater of two salt concentrations (13.5 vs. 28.5). Weight gain rate was significantly higher in fish fed the diet supplemented with small peptides, B. licheniformis, inulin, or synbiotics than that in fish fed the basal diet; the greatest weight gain rate was found in fish fed the small peptide treatment [56.0% higher than basal diet]. Higher feed efficiency was detected in fish fed the diet supplemented with small peptides than that of fish in the other dietary treatments. Total protease activity in the stomach and intestines was highest in fish fed the small peptide-treated diet, whereas lipase activity was highest in those fed synbiotics (combination of Bacillus licheniformis and inulin) than that in fish fed the other treatments. Antioxidant enzyme (total superoxide dismutase and catalase) activities and hepatic malondialdehyde content were higher in fish receiving the dietary supplements and maintained in artificial seawater containing 13.5 salinity compared with those in the control (28.5). Hepatic catalase activity in grouper fed the diets with small peptides or synbiotics decreased significantly compared with that in control fish. Overall, the three types of additives improved growth rate of juvenile grouper and digestive enzymes activities to varying degrees but did not effectively improve antioxidant capacity under low-salinity stress conditions.

  14. Dietary supplementation of probiotics affects growth, immune response and disease resistance of Cyprinus carpio fry.

    Science.gov (United States)

    Gupta, Akhil; Gupta, Paromita; Dhawan, Asha

    2014-12-01

    The effects of dietary Bacillus coagulans (MTCC 9872), Bacillus licheniformis (MTCC 6824) and Paenibacillus polymyxa (MTCC 122) supplementation on growth performance, non-specific immunity and protection against Aeromonas hydrophila infection were evaluated in common carp, Cyprinus carpio fry. Laboratory maintained B. coagulans, B. licheniformis and P. polymyxa were used to study antagonistic activity against fish pathogenic bacteria by agar well diffusion assay. Healthy fish fry were challenged by this bacterium for determination of its safety. Fish were fed for 80 days with control basal diet (B0) and experimental diets containing B. coagulans (B1), B. licheniformis (B2) and P. polymyxa (B3) at 10(9) CFU/g diet. Fish fry (mean weight 0.329 ± 0.01 g) were fed these diets and growth performance, various non-specific immune parameters and disease resistance study were conducted at 80 days post-feeding. The antagonism study showed inhibition zone against A. hydrophila and Vibrio harveyi. All the probiotic bacterial strains were harmless to fish fry as neither mortality nor morbidities were observed of the challenge. The growth-promoting influences of probiotic supplemented dietary treatments were observed with fish fry and the optimum survival, growth and feed utilization were obtained with P. polymyxa (B3) supplemented diet. Study of different non-specific innate immunological parameters viz. lysozyme activity, respiratory burst assay and myeloperoxidase content showed significant (p growth, feed utilization, non-specific immune responses and disease resistance of fry common carp, C. carpio. Copyright © 2014. Published by Elsevier Ltd.

  15. Determination of Levan from Bacillus licheniformis by Ultraviolet ...

    African Journals Online (AJOL)

    hydrolysis conditions in the spectrophotometric determination. FODUS method was validated by analyzing its linearity, reproducibility, stability and recovery. Results: Factors influencing absorbance for the determination were confirmed and two regression equations were established. UV absorbance at 320 nm of sulfuric ...

  16. Studies on bioemulsifier production by Bacillus licheniformis PTCC ...

    African Journals Online (AJOL)

    Surfactants are amphipathic molecules which reduce surface and interfacial tensions and widely used in pharmaceutical, cosmetic, food and petroleum industries. Biosurfactants are the structurally diverse group of surface-active molecules synthesized by microorganisms. There are several advantages for biosurfactants in ...

  17. 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.

  18. Host organisms: Bacillus subtilis

    NARCIS (Netherlands)

    Hohman, Hans-Peter; van Dijl, Jan; Krishnappa, Laxmi; Pragai, Zoltan

    2016-01-01

    Bacillus subtilis and its close Bacillus relatives are important bacterial platforms for industrial production of enzymes and fine chemicals such as vitamin B2 and nucleotides. B. subtilis is an attractive bacterial organism for industrial use mainly because of its straightforward genetic

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

    Science.gov (United States)

    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. PMID:27258038

  20. 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.

  1. Assessing the resistance and bioremediation ability of selected bacterial and protozoan species to heavy metals in metal-rich industrial wastewater

    Directory of Open Access Journals (Sweden)

    Kamika Ilunga

    2013-02-01

    Full Text Available Abstract Background Heavy-metals exert considerable stress on the environment worldwide. This study assessed the resistance to and bioremediation of heavy-metals by selected protozoan and bacterial species in highly polluted industrial-wastewater. Specific variables (i.e. chemical oxygen demand, pH, dissolved oxygen and the growth/die-off-rates of test organisms were measured using standard methods. Heavy-metal removals were determined in biomass and supernatant by the Inductively Couple Plasma Optical Emission Spectrometer. A parallel experiment was performed with dead microbial cells to assess the biosorption ability of test isolates. Results The results revealed that the industrial-wastewater samples were highly polluted with heavy-metal concentrations exceeding by far the maximum limits (in mg/l of 0.05-Co, 0.2-Ni, 0.1-Mn, 0.1-V, 0.01-Pb, 0.01-Cu, 0.1-Zn and 0.005-Cd, prescribed by the UN-FAO. Industrial-wastewater had no major effects on Pseudomonas putida, Bacillus licheniformis and Peranema sp. (growth rates up to 1.81, 1.45 and 1.43 d-1, respectively compared to other test isolates. This was also revealed with significant COD increases (p Pseudomonas putida demonstrated the highest removal rates of heavy metals (Co-71%, Ni-51%, Mn-45%, V-83%, Pb-96%, Ti-100% and Cu-49% followed by Bacillus licheniformis (Al-23% and Zn-53% and Peranema sp. (Cd-42%. None of the dead cells were able to remove more than 25% of the heavy metals. Bacterial isolates contained the genes copC, chrB, cnrA3 and nccA encoding the resistance to Cu, Cr, Co-Ni and Cd-Ni-Co, respectively. Protozoan isolates contained only the genes encoding Cu and Cr resistance (copC and chrB genes. Peranema sp. was the only protozoan isolate which had an additional resistant gene cnrA3 encoding Co-Ni resistance. Conclusion Significant differences (p Peranema sp. as a potential candidate for the bioremediation of heavy-metals in wastewater treatment, in addition to Pseudomonas

  2. Fluorene biodegradation potentials of Bacillus strains isolated from ...

    African Journals Online (AJOL)

    Fluorene biodegradation potentials of Bacillus strains isolated from tropical ... Bacillus strains, putatively identified as Bacillus subtilis BM1 and Bacillus amyloliquefaciens BR1 were ... African Journal of Biotechnology, Vol 13(14), 1554-1559 ...

  3. Screen, Identification and Analysis on the Growth-Promoting Ability for the Rice Growth-Promoting Rhizobacteria

    Directory of Open Access Journals (Sweden)

    LIU Ze-ping

    2018-02-01

    Full Text Available Plant growth-promoting rhizobacteria(PGPRcan secrete the growth hormone and promote soil nutrient cycling, thus, is an important germplasm resource of bio -fertilizer. In this study, the PGPR was isolated from the rice rhizosphere. According to 16S rDNA sequences, 10 strains were identifed, including 4 organic phosphorus bacteria (Bacillus pumilus LZP02, Bacillus aryabhattai LZP08, Staphylococcus epidermidis LZP10, Bacillus ginsengisoli LZP05, 3 inorganic phosphorus bacteria(Bacillus megaterium LZP03, Bacillus oryzaecorticis LZP04, Bacillus ginsengisoli LZP07and 3 potassium bacteria(Bacillus aryabhattai LZP01, Bacillus subtilis LZP06, Bacillus licheniformis LZP09. The results from nutrient conversion analysis showed that Bacillus aryabhattai LZP01 and Bacillus subtilis LZP06 performed better on the potassium releasing ability. Bacillus pumilus LZP02 and Bacillus huizhouensis LZP05 performed better on the function of organic phosphorus. Bacillus megaterium LZP03 and Bacillus ginsengisoli LZP07 performed better on the function of inorganic phosphorus. Further, the hormone secretion capacity was measured for these 6 strains. The results showed that all 6 strains could produce auxin and gibberellin, and had the ability to synthesize iron carrier. Moreover, the results showed that Bacillus megaterium LZP03, Bacillus huizhouensis LZP05 and Bacillus subtilis LZP06 had stronger ability to promote the nutrient conversion and hormone secretion. Systematically, we believe that these three strains have great potential application on microbial fertilizer.

  4. In silico exploration of Red Sea Bacillus genomes for natural product biosynthetic gene clusters

    KAUST Repository

    Othoum, Ghofran K; Bougouffa, Salim; Razali, Rozaimi; Bokhari, Ameerah; Alamoudi, Soha; Antunes, André ; Gao, Xin; Hoehndorf, Robert; Arold, Stefan T.; Gojobori, Takashi; Hirt, Heribert; Mijakovic, Ivan; Bajic, Vladimir B.; Lafi, Feras Fawzi; Essack, Magbubah

    2018-01-01

    are better potential sources for novel antibiotics. Moreover, the genome of the Red Sea strain B. paralicheniformis Bac48 is more enriched in modular PKS genes compared to B. licheniformis strains and other B. paralicheniformis strains. This may be linked

  5. Enzymatic Biosynthesis of Novel Resveratrol Glucoside and Glycoside Derivatives

    OpenAIRE

    Pandey, Ramesh Prasad; Parajuli, Prakash; Shin, Ju Yong; Lee, Jisun; Lee, Seul; Hong, Young-Soo; Park, Yong Il; Kim, Joong Su; Sohng, Jae Kyung

    2014-01-01

    A UDP glucosyltransferase from Bacillus licheniformis was overexpressed, purified, and incubated with nucleotide diphosphate (NDP) d- and l-sugars to produce glucose, galactose, 2-deoxyglucose, viosamine, rhamnose, and fucose sugar-conjugated resveratrol glycosides. Significantly higher (90%) bioconversion of resveratrol was achieved with α-d-glucose as the sugar donor to produce four different glucosides of resveratrol: resveratrol 3-O-β-d-glucoside, resveratrol 4′-O-β-d-glucoside, resveratr...

  6. Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus siamensis Form an "Operational Group B. amyloliquefaciens" within the B. subtilis Species Complex.

    Science.gov (United States)

    Fan, Ben; Blom, Jochen; Klenk, Hans-Peter; Borriss, Rainer

    2017-01-01

    The plant growth promoting model bacterium FZB42 T was proposed as the type strain of Bacillus amyloliquefaciens subsp. plantarum (Borriss et al., 2011), but has been recently recognized as being synonymous to Bacillus velezensis due to phylogenomic analysis (Dunlap C. et al., 2016). However, until now, majority of publications consider plant-associated close relatives of FZB42 still as " B. amyloliquefaciens ." Here, we reinvestigated the taxonomic status of FZB42 and related strains in its context to the free-living soil bacterium DSM7 T , the type strain of B. amyloliquefaciens . We identified 66 bacterial genomes from the NCBI data bank with high similarity to DSM7 T . Dendrograms based on complete rpoB nucleotide sequences and on core genome sequences, respectively, clustered into a clade consisting of three tightly linked branches: (1) B. amyloliquefaciens , (2) Bacillus siamensis , and (3) a conspecific group containing the type strains of B. velezensis, Bacillus methylotrophicus , and B. amyloliquefaciens subsp. plantarum . The three monophyletic clades shared a common mutation rate of 0.01 substitutions per nucleotide position, but were distantly related to Bacillus subtilis (0.1 substitutions per nucleotide position). The tight relatedness of the three clusters was corroborated by TETRA, dDDH, ANI, and AAI analysis of the core genomes, but dDDH and ANI values were found slightly below species level thresholds when B. amyloliquefaciens DSM7 T genome sequence was used as query sequence. Due to these results, we propose that the B. amyloliquefaciens clade should be considered as a taxonomic unit above of species level, designated here as "operational group B. amyloliquefaciens " consisting of the soil borne B. amyloliquefaciens , and plant associated B. siamensis and B. velezensis , whose members are closely related and allow identifying changes on the genomic level due to developing the plant-associated life-style.

  7. Insight into the bacterial diversity of fermentation woad dye vats as revealed by PCR-DGGE and pyrosequencing.

    Science.gov (United States)

    Milanović, Vesna; Osimani, Andrea; Taccari, Manuela; Garofalo, Cristiana; Butta, Alessandro; Clementi, Francesca; Aquilanti, Lucia

    2017-07-01

    The bacterial diversity in fermenting dye vats with woad (Isatis tinctoria L.) prepared and maintained in a functional state for approximately 12 months was examined using a combination of culture-dependent and -independent PCR-DGGE analyses and next-generation sequencing of 16S rRNA amplicons. An extremely complex ecosystem including taxa potentially contributing to both indigo reduction and formation, as well as indigo degradation was found. PCR-DGGE analyses revealed the presence of Paenibacillus lactis, Sporosarcina koreensis, Bacillus licheniformis, and Bacillus thermoamylovorans, while Bacillus thermolactis, Bacillus pumilus and Bacillus megaterium were also identified but with sequence identities lower than 97%. Dominant operational taxonomic units (OTUs) identified by pyrosequencing included Clostridium ultunense, Tissierella spp., Alcaligenes faecalis, Erysipelothrix spp., Enterococcus spp., Virgibacillus spp. and Virgibacillus panthothenicus, while sub-dominant OTUs included clostridia, alkaliphiles, halophiles, bacilli, moderately thermophilic bacteria, lactic acid bacteria, Enterobacteriaceae, aerobes, and even photosynthetic bacteria. Based on the current knowledge of indigo-reducing bacteria, it is considered that indigo-reducing bacteria constituted only a small fraction in the unique microcosm detected in the natural indigo dye vats.

  8. Bacillus Strains Most Closely Related to Bacillus nealsonii Are Not Effectively Circumscribed within the Taxonomic Species Definition

    Directory of Open Access Journals (Sweden)

    K. Kealy Peak

    2011-01-01

    Full Text Available Bacillus strains with >99.7% 16S rRNA gene sequence similarity were characterized with DNA:DNA hybridization, cellular fatty acid (CFA analysis, and testing of 100 phenotypic traits. When paired with the most closely related type strain, percent DNA:DNA similarities (% S for six Bacillus strains were all far below the recommended 70% threshold value for species circumscription with Bacillus nealsonii. An apparent genomic group of four Bacillus strain pairings with 94%–70% S was contradicted by the failure of the strains to cluster in CFA- and phenotype-based dendrograms as well as by their differentiation with 9–13 species level discriminators such as nitrate reduction, temperature range, and acid production from carbohydrates. The novel Bacillus strains were monophyletic and very closely related based on 16S rRNA gene sequence. Coherent genomic groups were not however supported by similarly organized phenotypic clusters. Therefore, the strains were not effectively circumscribed within the taxonomic species definition.

  9. Role of fatty acids in Bacillus environmental adaptation

    Directory of Open Access Journals (Sweden)

    Sara Esther Diomande

    2015-08-01

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

  10. Mixed-substrate (glycerol tributyrate and fibrin) zymography for simultaneous detection of lipolytic and proteolytic enzymes on a single gel.

    Science.gov (United States)

    Choi, Nack-Shick; Choi, Jong Hyun; Kim, Bo-Hye; Han, Yun-Jon; Kim, Joong Su; Lee, Seung-Goo; Song, Jae Jun

    2009-06-01

    A new zymography method for simultaneous detection of two different enzymatic activities (lipolytic and proteolytic) using a single SDS-containing or native-conformation gel and a mixed-substrate (glycerol tributyrate and fibrin) (MS)(1) gel was developed. After routine electrophoresis, SDS in the gel was removed by treatment with Triton X-100. Gel proteins were electrotransferred to the MS gel. To visualize lipolytic activity, the MS gel was incubated at 37 degrees C (for 6 or 24 h) until clear bands against an opaque background were observed. To detect proteolytic activity, the same MS gel was stained with Coomassie brilliant blue. Using this method, we show that six lipolytic enzymes from Staphylococcus pasteuri NJ-1 and four proteolytic enzymes from two Bacillus strains, B. licheniformis DJ-2 and B. licheniformis NJ-5, isolated from soil, can be simultaneously detected.

  11. Effect of garlic solution to Bacillus sp. removal

    Science.gov (United States)

    Zainol, N.; Rahim, S. R.

    2018-04-01

    Biofilm is a microbial derived sessile community characterized by cells that are irreversibly attached to a substratum or interface to each other, embedded in a matrix of extracellular polymeric substances that they have produced. Bacillus sp. was used as biofilm model in this study. The purpose of this study is to determine the effect of Garlic solution in term of ratio of water and Garlic solution (W/G) and ratio of Garlic solution to Bacillus sp. (GS/B) on Bacillus sp removal. Garlic solution was used to remove Bacillus sp. In this study, Garlic solution was prepared by crushing the garlic and mixed it with water. the Garlic solution was added into Bacillus sp. mixture and mixed well. The mixture then was spread on nutrient agar. The Bacillus sp. weight on agar plate was measured by using dry weight measurement method. In this study, initially Garlic solution volume and Garlic solution concentration were studied using one factor at time (OFAT). Later two-level-factorial analysis was done to determine the most contributing factor in Bacillus sp. removal. Design Expert software (Version 7) was used to construct experimental table where all the factors were randomized. Bacilus sp removal was ranging between 42.13% to 99.6%. The analysis of the results showed that at W/G of 1:1, Bacillus sp. removal increased when more Garlic solution was added to Bacillus sp. Effect of Garlic solution to Bacillus sp. will be understood which in turn may be beneficial for the industrial purpose.

  12. Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus siamensis Form an “Operational Group B. amyloliquefaciens” within the B. subtilis Species Complex

    Science.gov (United States)

    Fan, Ben; Blom, Jochen; Klenk, Hans-Peter; Borriss, Rainer

    2017-01-01

    The plant growth promoting model bacterium FZB42T was proposed as the type strain of Bacillus amyloliquefaciens subsp. plantarum (Borriss et al., 2011), but has been recently recognized as being synonymous to Bacillus velezensis due to phylogenomic analysis (Dunlap C. et al., 2016). However, until now, majority of publications consider plant-associated close relatives of FZB42 still as “B. amyloliquefaciens.” Here, we reinvestigated the taxonomic status of FZB42 and related strains in its context to the free-living soil bacterium DSM7T, the type strain of B. amyloliquefaciens. We identified 66 bacterial genomes from the NCBI data bank with high similarity to DSM7T. Dendrograms based on complete rpoB nucleotide sequences and on core genome sequences, respectively, clustered into a clade consisting of three tightly linked branches: (1) B. amyloliquefaciens, (2) Bacillus siamensis, and (3) a conspecific group containing the type strains of B. velezensis, Bacillus methylotrophicus, and B. amyloliquefaciens subsp. plantarum. The three monophyletic clades shared a common mutation rate of 0.01 substitutions per nucleotide position, but were distantly related to Bacillus subtilis (0.1 substitutions per nucleotide position). The tight relatedness of the three clusters was corroborated by TETRA, dDDH, ANI, and AAI analysis of the core genomes, but dDDH and ANI values were found slightly below species level thresholds when B. amyloliquefaciens DSM7T genome sequence was used as query sequence. Due to these results, we propose that the B. amyloliquefaciens clade should be considered as a taxonomic unit above of species level, designated here as “operational group B. amyloliquefaciens” consisting of the soil borne B. amyloliquefaciens, and plant associated B. siamensis and B. velezensis, whose members are closely related and allow identifying changes on the genomic level due to developing the plant-associated life-style. PMID:28163698

  13. Otimização da extração enzimática da proteína do feijão - doi: 10.4025/actascitechnol.v32i3.6277

    Directory of Open Access Journals (Sweden)

    Carlos de Oliveira Lopes Junior

    2010-11-01

    Full Text Available Considerando-se a importância do feijão na dieta do brasileiro e, ainda, que este alimento não é permitido na alimentação de fenilcetonúricos, é fundamental promover a extração de suas proteínas, visando à posterior remoção de fenilalanina. No intuito de se otimizar a extração proteica do feijão, dois métodos enzimáticos foram, inicialmente, empregados. Para aquele que produziu o maior rendimento de extração proteica (REP, foi realizado um estudo do efeito de diversos parâmetros, tais como velocidade de centrifugação, tempo de reação, pH inicial e tipo de enzima (uma de Bacillus licheniformis, duas de Bacillus subtilis, uma de Aspergillus sojae e uma de Papaya carica. Os resultados indicaram que o melhor método foi o que empregou uma concentração de matéria-prima de 1:10 (p v-1, pH inicial de 10,5, temperatura de 50°C, e ausência de agitação em ultraturrax. Com relação aos parâmetros testados, o maior REP (93,1% foi obtido ao se utilizar a enzima de Bacillus licheniformis, na velocidade de centrifugação de 10.640 x g, tempo de reação de 3h e pH inicial de 10,5.

  14. 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...

  15. Molecular detection of TasA gene in endophytic Bacillus species ...

    African Journals Online (AJOL)

    Molecular detection of TasA gene in endophytic Bacillus species and characterization of the gene in Bacillus amyloliquefaciens. ... African Journal of Biotechnology ... in Bacillus amyloliquefaciens PEBA20 and 7 strains of Bacillus subtilis, ...

  16. Microbiological and chemical characteristics of tarubá, an indigenous beverage produced from solid cassava fermentation.

    Science.gov (United States)

    Ramos, Cíntia L; de Sousa, Edinaira S O; Ribeiro, Jessimara; Almeida, Tayanny M M; Santos, Claudia Cristina A do A; Abegg, Maxwel A; Schwan, Rosane F

    2015-08-01

    The aim of this work was to identify and characterize the microbiota present during fermentation and in the final beverage, tarubá, by culture-dependent and -independent methods. In addition, target chemical compounds (carbohydrates, organic acids, and ethanol) were evaluated. Lactic acid bacteria (LAB) and mesophilic bacteria were the predominant microorganisms. Among them, Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides, and Bacillus subtilis were frequently isolated and detected by DGGE analysis. Torulaspora delbrueckii was the dominant yeast species. Yeast isolates Pichia exigua, Candida rugosa, T. delbrueckii, Candida tropicalis, Pichia kudriavzevii, Wickerhamomyces anomalus, and Candida ethanolica and bacteria isolates Lb. plantarum, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus sp., and Chitinophaga terrae showed amylolytic activity. Only isolates of P. exigua and T. delbrueckii and all species of the genus Bacillus identified in this work exhibited proteolytic activity. All microbial isolates grew at 38 °C, and only the isolates belonging to Hanseniaspora uvarum species did not grow at 42 °C. These characteristics are important for further development of starter cultures; isolates of T. delbrueckii, P. exigua, and Bacillus species identified in this work displayed all of these properties and are potential strains for use as starter culture in cassava fermented food. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Bacillus and biopolymer: Prospects and challenges

    Directory of Open Access Journals (Sweden)

    Swati Mohapatra

    2017-12-01

    Full Text Available The microbially derived polyhydroxyalkanoates biopolymers could impact the global climate scenario by replacing the conventional non-degradable, petrochemical-based polymer. The biogenesis, characterization and properties of PHAs by Bacillus species using renewable substrates have been elaborated by many for their wide applications. On the other hand Bacillus species are advantageous over other bacteria due to their abundance even in extreme ecological conditions, higher growth rates even on cheap substrates, higher PHAs production ability, and the ease of extracting the PHAs. Bacillus species possess hydrolytic enzymes that can be exploited for economical PHAs production. This review summarizes the recent trends in both non-growth and growth associated PHAs production by Bacillus species which may provide direction leading to future research towards this growing quest for biodegradable plastics, one more critical step ahead towards sustainable development.

  18. Isolation of nitrite-degrading strains from Douchi and their application to degrade high nitrite in Jiangshui.

    Science.gov (United States)

    Guo, Xing; Liu, Bianfang; Gao, Lina; Zhou, Yuan; Shan, Yuanyuan; Lü, Xin

    2018-06-01

    Excessive nitrite in food is potentially harmful to human health because of its carcinogenic effects caused by nitroso-dervivatives. Douchi, which widely distributed throughout the country, is a traditional solid fermented soybean food with low nitrite content. In this study, bacterias which can degrade nitrite were isolated from Douchi and identified according to 16S rDNA sequence. Acinetobacter guillouiae, Acinetobacter bereziniae, Bacillus subtilis, Bacillus tequilensis, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus aryabhattai and Bacillus methylotrophicus were selected. It was shown that all strains have nitrite degradation capability, in which 99.41 % nitrite can be degraded by Bacillus subtilis NDS1. The enzyme activities of these strains were determined at 24 h and 48 h, which corresponded to their nitrite degradation rates. The strains were firstly tried to inoculate in Jiangshui, which is a kind of traditional fermented vegetable in northwest China and often has high nitrite content. It was found that Bacillus subtilis NDS1, Bacillus tequilensis NDS3, Acinetobacter bereziniae NDS4, Bacillus subtilis NDS6, Bacillus subtilis NDS12 can degrade nitrite in Jiangshui more quickly, among which Acinetobacter bereziniae NDS4 degraded almost all nitrite in 48 h while it took 180 h for control. These results indicated that the selected strains have potential to become nitrite degradition agent in food. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  19. Assessing the resistance and bioremediation ability of selected bacterial and protozoan species to heavy metals in metal-rich industrial wastewater.

    Science.gov (United States)

    Kamika, Ilunga; Momba, Maggy N B

    2013-02-06

    Heavy-metals exert considerable stress on the environment worldwide. This study assessed the resistance to and bioremediation of heavy-metals by selected protozoan and bacterial species in highly polluted industrial-wastewater. Specific variables (i.e. chemical oxygen demand, pH, dissolved oxygen) and the growth/die-off-rates of test organisms were measured using standard methods. Heavy-metal removals were determined in biomass and supernatant by the Inductively Couple Plasma Optical Emission Spectrometer. A parallel experiment was performed with dead microbial cells to assess the biosorption ability of test isolates. The results revealed that the industrial-wastewater samples were highly polluted with heavy-metal concentrations exceeding by far the maximum limits (in mg/l) of 0.05-Co, 0.2-Ni, 0.1-Mn, 0.1-V, 0.01-Pb, 0.01-Cu, 0.1-Zn and 0.005-Cd, prescribed by the UN-FAO. Industrial-wastewater had no major effects on Pseudomonas putida, Bacillus licheniformis and Peranema sp. (growth rates up to 1.81, 1.45 and 1.43 d-1, respectively) compared to other test isolates. This was also revealed with significant COD increases (p heavy metals (Co-71%, Ni-51%, Mn-45%, V-83%, Pb-96%, Ti-100% and Cu-49%) followed by Bacillus licheniformis (Al-23% and Zn-53%) and Peranema sp. (Cd-42%). None of the dead cells were able to remove more than 25% of the heavy metals. Bacterial isolates contained the genes copC, chrB, cnrA3 and nccA encoding the resistance to Cu, Cr, Co-Ni and Cd-Ni-Co, respectively. Protozoan isolates contained only the genes encoding Cu and Cr resistance (copC and chrB genes). Peranema sp. was the only protozoan isolate which had an additional resistant gene cnrA3 encoding Co-Ni resistance. Significant differences (p metal-removal and the presence of certain metal-resistant genes indicated that the selected microbial isolates used both passive (biosorptive) and active (bioaccumulation) mechanisms to remove heavy metals from industrial wastewater. This study

  20. Real-Time PCR Assay for a Unique Chromosomal Sequence of Bacillus anthracis

    Science.gov (United States)

    2004-12-01

    13061 Neisseria lactamica .............................................................. 23970 Bacillus coagulans ...NEG Bacillus coagulane 7050 NEG NEG Bacillus cereus 13472 NEG NEG Bacillus licheniforms 12759 NEG NEG Bacillus cereus 13824 NEG NEG Bacillus ...Assay for a Unique Chromosomal Sequence of Bacillus anthracis Elizabeth Bode,1 William Hurtle,2† and David Norwood1* United States Army Medical

  1. Bacillus beijingensis sp. nov. and Bacillus ginsengi sp. nov., isolated from ginseng root.

    Science.gov (United States)

    Qiu, Fubin; Zhang, Xiaoxia; Liu, Lin; Sun, Lei; Schumann, Peter; Song, Wei

    2009-04-01

    Four alkaligenous, moderately halotolerant strains, designated ge09, ge10(T), ge14(T) and ge15, were isolated from the internal tissue of ginseng root and their taxonomic positions were investigated by using a polyphasic approach. Cells of the four strains were Gram-positive-staining, non-motile, short rods. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains ge09 and ge10(T) formed one cluster and strains ge14(T) and ge15 formed another separate cluster within the genus Bacillus. 16S rRNA gene sequence similarities with type strains of other Bacillus species were less than 97 %. Levels of DNA-DNA relatedness among the four strains showed that strains ge09 and ge10(T) and strains ge14(T) and ge15 belonged to two separate species; the mean level of DNA-DNA relatedness between ge10(T) and ge14(T) was only 28.7 %. Their phenotypic and physiological properties supported the view that the two strains represent two different novel species of the genus Bacillus. The DNA G+C contents of strains ge10(T) and ge14(T) were 49.9 and 49.6 mol%, respectively. Strains ge10(T) and ge14(T) showed the peptidoglycan type A4alpha l-Lys-d-Glu. The lipids present in strains ge10(T) and ge14(T) were diphosphatidylglycerol, phosphatidylglycerol, a minor amount of phosphatidylcholine and two unknown phospholipids. Their predominant respiratory quinone was MK-7. The fatty acid profiles of the four novel strains contained large quantities of branched and saturated fatty acids. The predominant cellular fatty acids were iso-C(15 : 0) (42.5 %), anteiso-C(15 : 0) (22.2 %), anteiso-C(17 : 0) (7.3 %) and C(16 : 1)omega7c alcohol (5.7 %) in ge10(T) and iso-C(15 : 0) (50.7 %) and anteiso-C(15 : 0) (20.1 %) in ge14(T). On the basis of their phenotypic properties and phylogenetic distinctiveness, two novel species of the genus Bacillus are proposed, Bacillus beijingensis sp. nov. (type strain ge10(T) =DSM 19037(T) =CGMCC 1.6762(T)) and Bacillus ginsengi sp. nov. (type strain ge14

  2. Antibacterial activity of curcuma long varieties against different strains of bacteria

    International Nuclear Information System (INIS)

    Naz, S.; Jabeen, S.; Ilyas, S.; Aslam, F.; Manzoor, F.; Ali, A.

    2010-01-01

    Crude extracts of curcuminoids and essential oil of Curcuma long varieties Kasur, Faisalabad and Bannu were studied for their antibacterial activity against 4 bacterial strains viz., Bacillus subtilis, Bacillus macerans, Bacillus licheniformis and Azotobacter using agar well diffusion method. Solvents used to determine antibacterial activity were ethanol and methanol. Ethanol was used for the extraction of curcuminoids. Essential oil was extracted by hydrodistillation and diluted in methanol by serial dilution method. Both Curcuminoids and oil showed zone of inhibition against all tested strains of bacteria. Among all the three turmeric varieties, Kasur variety had the most inhibitory effect on the growth of all bacterial strains tested as compared to Faisalabad and Bannu varieties. Among all the bacterial strains B. subtilis was the most sensitive to turmeric extracts of curcuminoids and oil. The MIC value for different strains and varieties ranged from 3.0 to 20.6 mm in diameter. (author)

  3. Change of microflora of two starch samples by gamma irradiation

    International Nuclear Information System (INIS)

    Fretton, R.; Fretton, J.; Delattre, J.M.

    1975-01-01

    Starch is the basic component of a larger number of manufactured foods. The disinfection of such a powder by 60 Co is studied here. Gamma irradiation of two starch samples with different degrees of contamination allows the assumption that, in most cases, good radio-pasteurization can be achieved with 300 krad. The radio-pasteurization doses (varying from 300 to 600 krad) are a function of the initial contamination. Irradiation effects are spectacular with moulds. Activation of spores of some Clostridium species leads us to recommend an irradiation level higher than 200 krad. The most resistant organisms to gamma irradiation are the aerobic and anaerobic sporulated bacteria. The thermophilic forms are the most important. Spores of Bacillus, chiefly Bacillus licheniformis and Bacillus brevis, are the most frequent bacteria. Storage of irradiated starch at room temperature has little effect upon the number of revivable survivors. (orig.) [de

  4. Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils

    OpenAIRE

    Aanniz,Tarik; Ouadghiri,Mouna; Melloul,Marouane; Swings,Jean; Elfahime,Elmostafa; Ibijbijen,Jamal; Ismaili,Mohamed; Amar,Mohamed

    2015-01-01

    The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240) thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5%) represented by B. licheniformis (119), B. aerius (44), B. sonorensis (33), B. ...

  5. In vitro degradation of linamarin by microorganisms isolated from cassava wastewater treatment lagoons

    Directory of Open Access Journals (Sweden)

    S. P Vasconcellos

    2009-12-01

    Full Text Available This study aimed at isolating and characterizing of microorganisms able to use linamarin as sole carbon source. Thirty one microbial strains were isolated from manipueira, a liquid effluent of cassava processing factories. Among these strains, Bacillus licheniformis (isolate 2_2 and Rhodotorulla glutinis (isolate L1 were able to degrade 71% and 95% of added linamarin, respectively, within 7 days, showing high biodegradation activity and great potential for detoxification of cassava processing wastewaters.

  6. Investigation of spore forming bacterial flooding for enhanced oil recovery in a North Sea chalk Reservoir

    DEFF Research Database (Denmark)

    Halim, Amalia Yunita; Nielsen, Sidsel Marie; Eliasson Lantz, Anna

    2015-01-01

    Little has been done to study microbial enhanced oil recovery (MEOR) in chalk reservoirs. The present study focuses on core flooding experiments designed to see microbial plugging and its effect on oil recovery. A pressure tapped core holder was used for this purpose. A spore forming bacteria...... Bacillus licheniformis 421 was used as it was shown to be a good candidate in a previous study. Bacterial spore can penetrate deeper into the chalk rock, squeezing through the pore throats. Our results showed that injection of B. licheniformis 421 as a tertiary oil recovery method, in the residual oil...... saturation state, was able to produce additionally 1.0-2.3% original oil in place (OOIP) in homogeneous cores and 6.9-8.8% OOIP in heterogeneous cores. In addition, the pressure gradient was much higher in the heterogeneous cores, which confirms that bacterial selective plugging plays an important role...

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

    Directory of Open Access Journals (Sweden)

    Nan Jia

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

  8. Germination of Bacillus cereus spores : the role of germination receptors

    NARCIS (Netherlands)

    Hornstra, L.M.

    2007-01-01

    The Bacillus cereus sensu lato group forms a highly homogeneous subdivision of the genus Bacillus and comprises several species that are relevant for humans. Notorious is Bacillus anthracis, the cause of the often-lethal disease anthrax, while the insect pathogen Bacillus

  9. Bacterial, fungal and yeast contamination in six brands of irreversible hydrocolloid impression materials Contaminação por bactérias, fungos e leveduras em seis marcas comerciais de materiais de moldagem à base de hidrocolóide irreversível

    Directory of Open Access Journals (Sweden)

    Luciana Assirati Casemiro

    2007-06-01

    Full Text Available This study assessed the level of contamination of six commercially available irreversible hydrocolloids (two containing chlorhexidine and identified the contamination present in the materials. Petri dishes containing selective and enriched culture media were inoculated with alginate powder (0.06 g, in triplicate. After incubation (37°C/7 days, the colony-forming units (CFU were counted and Gram stained. Biochemical identification of the different morphotypes was also performed. The contamination levels for the materials were: Jeltrate - 389 CFU/g; Jeltrate Plus - 516 CFU/g; Jeltrate Chromatic - 135 CFU/g; Hydrogum - 1,455 CFU/g; Kromopan - 840 CFU/g; and Greengel - 59 CFU/g. Gram staining revealed the presence of Gram-positive bacillus and Gram-positive cocci. The bacteria Staphylococcus epidermidis, Bacillus subtilis, Bacillus sp., Bacillus coagulans, Bacillus licheniformis, Bacillus cereus, Micrococcus luteus, and Nocardia sp.; the filamentous fungi Aspergillus niger, Aspergillus flavus, Rhizopus sp., Neurospora sp.; and the yeast Candida sp. were isolated. The contamination detected in the impression materials points out the need for adopting measures to improve the microbiological quality of these materials. The use of contaminated materials in the oral cavity goes against the basic principles for controlling cross-contamination and may represent a risk for debilitated or immunocompromised patients.Este estudo avaliou o nível de contaminação de seis marcas comerciais de alginato (duas contendo clorexidina e identificou a contaminação presente nesses materiais. Alíquotas de alginato (0,06 g foram semeadas em meios de cultura seletivos e enriquecidos, em triplicata. Após incubação (37°C/7 dias, as unidades formadoras de colônia (UFC foram contadas e foram realizadas as identificações morfotintorial (Gram e bioquímica. Os níveis de contaminação dos materiais foram: Jeltrate - 389 UFC/g; Jeltrate Plus - 516 UFC/g; Jeltrate

  10. Diversity and characterization of ramie-degumming strains

    Directory of Open Access Journals (Sweden)

    Shengwen Duan

    2012-04-01

    Full Text Available Ramie (Boehmeria nivea and Boehmeria tenacissima is a widely used fiber crop. Traditional water retting or chemical boiling method performed in order to extract ramie fiber seriously pollute the environment and severely damage the fiber, so biological method is the general trend of the fiber-extracting industry. Some strains (687, involving 26 genera and 43 species, were collected from the three samples, which produce hydrolyzed circles in the selective culture medium in order to detect the degumming effect and to compare the enzyme activity. Among these strains, 13 of them did not produce cellulase and had a ramie decreasing weight rate above 25 %, which were regarded as efficient ramie-degumming strains named from R1 to R13. R1 to R13 belonged to Amycolata autotrobutylicun, Bacillus subtilis, Clostridium acetobutylicum, Bacillus subtilis, Rhizobium leguminosarum, Bacteroides finegoldii, Streptomyces lividans, Bacillus amyloliquefaciens, Clostridium acetobutylicum, Pseudomonas brassicacearum, Bacillus pumilus, Bacillus licheniformis, Pectobacterium wasabiae respectively. Bacteroides sp., Rhizobium sp. and Pseudomonas sp. were firstly reported to be used in ramie-degumming. At the same time, the pectinase was the key enzyme in the ramie-degumming process.

  11. Drought resistant of bacteria producing exopolysaccharide and IAA in rhizosphere of soybean plant (Glycine max) in Wonogiri Regency Central Java Indonesia

    Science.gov (United States)

    Susilowati, A.; Puspita, A. A.; Yunus, A.

    2018-03-01

    Drought is one of the main problem which limitating the agriculture productivity in most arid region such as in district Eromoko, Wuryantro and SelogiriWonogiri Central Java Indonesia. Bacteria are able to survive under stress condition by producte exopolysaccharide. This study aims to determine the presence of exopolysaccharide-producing drought-resistant bacteria on rhizosphere of soybean (Glycine max) and to determine the species of bacteria based on 16S rRNA gene. Isolation of bacteria carried out by the spread plate method. The decreased of osmotic potential for screening drought tolerant bacteria according to the previous equation [12]. Selection of exopolysaccharide-producing bacteria on solid media ATCC 14 followed by staining the capsule. 16S rRNA gene amplification performed by PCR using primers of 63f and 1387r. The identificationof the bacteria is determined by comparing the results of DNA sequence similarity with bacteria databank in NCBI database. The results showed 11 isolates were exopolysaccharide-producing drought tolerant bacteria. The identity of the bacteria which found are Bacillus sp, Bacillus licheniformis, Bacillus megaterium and Bacillus pumilus.

  12. Chemical synthesis of a dual branched malto-decaose: A potential substrate for alpha-amylases

    DEFF Research Database (Denmark)

    Damager, Iben; Jensen, Morten; Olsen, Carl Erik

    2005-01-01

    A convergent block strategy for general use in efficient synthesis of complex alpha-(1 -> 4)- and alpha-(1 -> 6)-malto-oligosaccharides is demonstrated with the first chemical synthesis of a malto-oligosaccharide, the decasoccharide 6,6""-bis(alpha-maltosyl)-maltohexaose, with two branch points....... Using this chemically defined branched oligosaccharide as a substrate, the cleavage pattern of seven different alpha-amylases were investigated. alpha-Amylases from human saliva, porcine pancreas, barley alpha-amylose 2 and recombinant barley alpha-amylase 1 all hydrolysed the decasaccharide selectively....... This resulted in a branched hexasaccharide and a branched tetrasoccharide. alpha-Amylases from Asperagillus oryzae, Bacillus licheniformis and Bacillus sp. cleaved the decasoccharide at two distinct sites, either producing two branched pentasoccharides, or a branched hexasoccharide and a branched...

  13. Antibacterial protection by enterocin AS-48 in sport and energy drinks with less acidic pH values.

    Science.gov (United States)

    Viedma, Pilar Martinez; Abriouel, Hikmate; Ben Omar, Nabil; López, Rosario Lucas; Valdivia, Eva; Gálvez, Antonio

    2009-04-01

    The low pH and acid content found in sports and energy drinks are a matter of concern in dental health. Raising the pH may solve this problem, but at the same time increase the risks of spoilage or presence of pathogenic bacteria. In the present study, commercial energy drinks were adjusted to pH 5.0 and challenged with Listeria monocytogenes (drinks A to F), Staphylococcus aureus, Bacillus cereus, and Bacillus licheniformis (drink A) during storage at 37 degrees C. L. monocytogenes was able to grow in drink A and survived in drinks D and F for at least 2 days. Addition of enterocin AS-48 (1 microg/ml final concentration) rapidly inactivated L. monocytogenes in all drinks tested. S. aureus and B. cereus also survived quite well in drink A, and were completely inactivated by 12.5 microg/ml enterocin AS-48 after 2 days of storage or by 25 microg/ml bacteriocin after 1 day. B. licheniformis was able to multiply in drink A, but it was completely inactivated by 5 microg/ml enterocin AS-48 after 2 days of storage or by 12.5 microg/ml bacteriocin after 1 day. Results from the present study suggest that enterocin AS-48 could be used as a natural preservative against these target bacteria in less acidic sport and energy drinks.

  14. Isolation and identification of bacteria to improve the strength of concrete.

    Science.gov (United States)

    Krishnapriya, S; Venkatesh Babu, D L; G, Prince Arulraj

    2015-05-01

    The objective of this research work is to isolate and identify calcite precipitating bacteria and to check the suitability of these bacteria for use in concrete to improve its strength. Bacteria to be incorporated in concrete should be alkali resistant to endure the high pH of concrete and endospore forming to withstand the mechanical stresses induced in concrete during mixing. They must exhibit high urease activity to precipitate calcium carbonate in the form of calcite. Bacterial strains were isolated from alkaline soil samples of a cement factory and were tested for urease activity, potential to form endospores and precipitation of calcium carbonate. Based on these results, three isolates were selected and identified by 16S rRNA gene sequencing. They were identified as Bacillus megaterium BSKAU, Bacillus licheniformis BSKNAU and Bacillus flexus BSKNAU. The results were compared with B. megaterium MTCC 1684 obtained from Microbial Type Culture Collection and Gene Bank, Chandigarh, India. Experimental work was carried out to assess the influence of bacteria on the compressive strength and tests revealed that bacterial concrete specimens showed enhancement in compressive strength. The efficiency of bacteria toward crack healing was also tested. Substantial increase in strength and complete healing of cracks was observed in concrete specimens cast with B. megaterium BSKAU, B. licheniformis BSKNAU and B. megaterium MTCC 1684. This indicates the suitability of these bacterial strains for use in concrete. The enhancement of strength and healing of cracks can be attributed to the filling of cracks in concrete by calcite which was visualized by scanning electron microscope. Copyright © 2015 Elsevier GmbH. All rights reserved.

  15. Impacts of Bacillus thuringiensis var. israelensis and Bacillus ...

    African Journals Online (AJOL)

    The study assessed the impact of bio-larvicides- Bacillus thuringiensis var. israelensis (Bti) and B. sphaericus (Bs) on anopheline mosquito larval densities in four selected areas of Lusaka urban district. Larval densities were determined using a standard WHO protocol at each study area prior to and after larviciding.

  16. Use of high-gradient magnetic fishing for reducing proteolysis during fermentation

    DEFF Research Database (Denmark)

    Maury, Trine Lütken; Ottow, Kim Ekelund; Brask, Jesper

    2012-01-01

    Proteolysis during fermentation may have a severe impact on the yield and quality of a secreted product. In the current study, we demonstrate the use of high-gradient magnetic fishing (HGMF) as an efficient alternative to the more conventional methods of preventing proteolytic degradation....... Bacitracin-linked magnetic affinity adsorbents were employed directly in a fermenter during Bacillus licheniformis cultivation to remove trace amounts of unwanted proteases. The constructed magnetic adsorbents had excellent, highly specific binding characteristics in the fermentation broth (K(d) = 1...

  17. Scientific Opinion on the safety and efficacy of Ronozyme RumiStar (alpha-amylase) as a feed additive for dairy cows

    OpenAIRE

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

    2012-01-01

    Ronozyme RumiStar is a feed additive in which the declared enzymatic activity is alpha-amylase. It is produced by a genetically modified Bacillus licheniformis strain. The final enzyme preparations contain no cultivable production organisms or recombinant DNA. Based on the results of a tolerance trial provided by the applicant, it was concluded that Ronozyme RumiStar is safe for use in dairy cows at the maximum proposed dose (400 KNU/kg dry matter of total daily ...

  18. Effects of probiotic supplementation in different nutrient density diets on growth performance, nutrient digestibility, blood profiles, fecal microflora and noxious gas emission in weaning pig.

    Science.gov (United States)

    Lan, Ruixia; Tran, Hoainam; Kim, Inho

    2017-03-01

    Probiotics can serve as alternatives to antibiotics to increase the performance of weaning pigs, and the intake of probiotics is affected by dietary nutrient density. The objective of this study was to evaluate the effects of a probiotic complex in different nutrient density diets on growth performance, digestibility, blood profiles, fecal microflora and noxious gas emission in weaning pigs. From day 22 to day 42, both high-nutrient-density and probiotic complex supplementation diets increased (P probiotic complex supplementation diets had higher (P probiotic complex supplementation diets. Interactive effects on average daily feed intake (ADFI) were observed from day 22 to day 42 and overall, where probiotic complex improved ADFI more dramatically in low-nutrient-density diets. The beneficial effects of probiotic complex (Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis and Clostridium butyricum) supplementation on ADFI is more dramatic with low-nutrient-density diets. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  19. Unearthing Bacillus endophytes from desert plants that enhance growth of Arabidopsis thaliana under abiotic stress conditions

    KAUST Repository

    Bokhari, Ameerah

    2018-01-01

    that these bacteria can confer resilience to plants under salt stress conditions. B. circulans (PK3-15 and PK3-109), B. cereus (PK6-15) B. subtilis (PK3-9) and B. licheniformis (PK5-26) displayed the ability to increased the fresh weight of A. thaliana under salt

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

    Science.gov (United States)

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

    2016-01-01

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

  1. Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model.

    Science.gov (United States)

    Haldar, Lopamudra; Gandhi, D N

    2016-07-01

    To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1) was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2) and (T3) groups received spore biomass of Bacillus coagulans B37 and Bacillus pumilus B9, respectively, suspended in sterilized skim milk at 8-9 log colony-forming units/ml plus basal diet for 28 days, while control group (T4) was supplied with clean water along with basal diet. There was a 14-day post-treatment period. A total of 288 fecal samples (8 fecal collections per rat) were collected at every 7-day interval starting from 0 to 49 days and subjected to the enumeration of the counts of coliforms and lactobacilli and Bacillus spores using respective agar media. In vitro acid and bile tolerance tests on both the strains were performed. The rats those (T2 and T3) received either B. coagulans B37 or B. pumilus B9 spore along with non-fermented skim milk showed decrease (pBacillus spore counts as compared to the control group (T4) and the group fed only skim milk (T1). In vitro study indicated that both the strains were found to survive at pH 2.0 and 3.0 even up to 3 h and tolerate bile up to 2.0% concentration even after 12 h of exposure. This study revealed that oral administration of either B. coagulans B37 or B. pumilus B9 strains might be useful in reducing coliform counts accompanied by concurrent increase in lactobacilli counts in the intestinal flora in rats.

  2. Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model

    Directory of Open Access Journals (Sweden)

    Lopamudra Haldar

    2016-07-01

    Full Text Available Aim: To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. Materials and Methods: An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1 was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2 and (T3 groups received spore biomass of Bacillus coagulans B37 and Bacillus pumilus B9, respectively, suspended in sterilized skim milk at 8-9 log colony-forming units/ml plus basal diet for 28 days, while control group (T4 was supplied with clean water along with basal diet. There was a 14-day post-treatment period. A total of 288 fecal samples (8 fecal collections per rat were collected at every 7-day interval starting from 0 to 49 days and subjected to the enumeration of the counts of coliforms and lactobacilli and Bacillus spores using respective agar media. In vitro acid and bile tolerance tests on both the strains were performed. Results: The rats those (T2 and T3 received either B. coagulans B37 or B. pumilus B9 spore along with non-fermented skim milk showed decrease (p<0.01 in fecal coliform counts and increase (p<0.05 in both fecal lactobacilli and Bacillus spore counts as compared to the control group (T4 and the group fed only skim milk (T1. In vitro study indicated that both the strains were found to survive at pH 2.0 and 3.0 even up to 3 h and tolerate bile up to 2.0% concentration even after 12 h of exposure. Conclusions: This study revealed that oral administration of either B. coagulans B37 or B. pumilus B9 strains might be useful in reducing coliform counts accompanied by concurrent increase in lactobacilli counts in the intestinal flora in rats.

  3. The effect of plant growth promoting rhizobacteria on Datura stramonium L., Abutilon theophrasti Med., Onopordon acanthium L. and Verbascum thapsus L. seed germination

    Directory of Open Access Journals (Sweden)

    Dragana Božić

    2014-09-01

    Full Text Available The effects of several bacterial media [Bacillus licheniformis population 1 (MO1; B. licheniformis population 2 (MO2; B. subtilis (MO3; B. megatherium (MO4; humates (MO5] on seed germination of Datura stramonium L., Abutilon theophrasti Med., Onopordon acanthium L. and Verbascum thapsus L. were tested. Seeds were germinated in Petri dishes containing solutions with different bacterial media. The highest germination percentage in all treatments was recorded for V. thapsus seeds (100.0%. Different treatments had diverse effects (stimulative or inhibitory on seed germination of D. stramonium [from 5% (MO1 to 13.3% (MO3, with 10.0 % in H20], A. theophrasti [from 28.3% (MO3 to 65.0% (MO5, with 43.3 % in H20] and O. acanthium [from 10.0% (MO2 to 13.3% (MO1 and MO3, with 6.7% in H20], depending on the type of media and weed species.

  4. Potential of Bacillus spp produces siderophores insuppressing thewilt disease of banana plants

    Science.gov (United States)

    Kesaulya, H.; Hasinu, J. V.; Tuhumury, G. NC

    2018-01-01

    In nature, different types of siderophore such as hydroxymate, catecholets and carboxylate, are produced by different bacteria. Bacillus spp were isolated from potato rhizospheric soil can produce siderophore of both catecholets and salicylate type with different concentrations. Various strains of Bacillus spp were tested for pathogen inhibition capability in a dual culture manner. The test results showed the ability of inhibition of pathogen isolated from banana wilt disease. From the result tested were found Bacillus niabensis Strain PT-32-1, Bacillus subtilis Strain SWI16b, Bacillus subtilis Strain HPC21, Bacillus mojavensis Strain JCEN3, and Bacillus subtilis Strain HPC24 showed different capabilities in suppressing pathogen.

  5. PRODUCTION OF FIBRINOLYTIC ENZYME (NATTOKINASE) FROM BACILLUS SP.

    OpenAIRE

    Padma Singh, Rekha Negi*, Vani Sharma, Alka Rani, Pallavi and Richa Prasad

    2018-01-01

    During present study Nattokinase which is a novel fibrinolytic enzyme was produced by Bacillus sp. To screen and extract nattokinase enzyme from Bacillus sp. were isolated from soil of different agricultural field by serial dilution method. Out of 10 isolate, one strain i.e. B3 produced nattokinase on screening medium. B3 was identified by biochemical characterization. The caseinolytic activity of Nattokinase was 0.526 U/ml and the selected isolate Bacillus sp. could produce active nattokinas...

  6. Genetic transformation of Bacillus strains close to bacillus subtilis and isolated from the soil

    International Nuclear Information System (INIS)

    Van, C.K.; Kuzin, Yu.Yu.; Kozlovskii, Yu.E.; Prozorov, A.A.

    1986-01-01

    Chromosomal and plasmid transformation was found in five out of 118 Bacillus strains, close or identical to Bacillus subtilis, and isolated from soil in Moscow or in the Moscow district. The efficiency of transformation in these strains was lower than that in derivatives of Bac. subtilis strain 168. In these strains the ability to undergo transformation was dependent on the rate of sporulation and the presence of restrictases. As in the case of Bac. subtilis 168 the strains isolated may be used as models in genetic transformation studies on Bac. subtilis

  7. 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...

  8. In vivo biotinylation of recombinant beta-glucosidase enables simultaneous purification and immobilization on streptavidin coated magnetic particles

    DEFF Research Database (Denmark)

    Alftrén, Johan; Ottow, Kim Ekelund; Hobley, Timothy John

    2013-01-01

    Beta-glucosidase from Bacillus licheniformis was in vivo biotinylated in Escherichia coli and subsequently immobilized directly from cell lysate on streptavidin coated magnetic particles. In vivo biotinylation was mediated by fusing the Biotin Acceptor Peptide to the C-terminal of beta......-glucosidase and co-expressing the BirA biotin ligase. The approach enabled simultaneous purification and immobilization of the enzyme from crude cell lysate on magnetic particles because of the high affinity and strong interaction between biotin and streptavidin. After immobilization of the biotinylated beta...

  9. Bacillus As Potential Probiotics: Status, Concerns, and Future Perspectives

    Directory of Open Access Journals (Sweden)

    Fouad M. F. Elshaghabee

    2017-08-01

    Full Text Available Spore-forming bacilli are being explored for the production and preservation of food for many centuries. The inherent ability of production of large number of secretory proteins, enzymes, antimicrobial compounds, vitamins, and carotenoids specifies the importance of bacilli in food chain. Additionally, Bacillus spp. are gaining interest in human health related functional food research coupled with their enhanced tolerance and survivability under hostile environment of gastrointestinal tract. Besides, bacilli are more stable during processing and storage of food and pharmaceutical preparations, making them more suitable candidate for health promoting formulations. Further, Bacillus strains also possess biotherapeutic potential which is connected with their ability to interact with the internal milieu of the host by producing variety of antimicrobial peptides and small extracellular effector molecules. Nonetheless, with proposed scientific evidences, commercial probiotic supplements, and functional foods comprising of Bacillus spp. had not gained much credential in general population, since the debate over probiotic vs pathogen tag of Bacillus in the research and production terrains is confusing consumers. Hence, it’s important to clearly understand the phenotypic and genotypic characteristics of selective beneficial Bacillus spp. and their substantiation with those having GRAS status, to reach a consensus over the same. This review highlights the probiotic candidature of spore forming Bacillus spp. and presents an overview of the proposed health benefits, including application in food and pharmaceutical industry. Moreover, the growing need to evaluate the safety of individual Bacillus strains as well as species on a case by case basis and necessity of more profound analysis for the selection and identification of Bacillus probiotic candidates are also taken into consideration.

  10. Study of the radiation effect of "9"9Mo/"9"9"mTc generator on Bacillus subtilis and Bacillus pumilus species

    International Nuclear Information System (INIS)

    Fukumori, Neuza T.O.; Endo, Erica M.M.; Felgueiras, Carlos F.; Matsuda, Margareth M.N.; Osso Junior, João A.

    2016-01-01

    In this work, molybdenum-99 loaded columns were challenged with Bacillus subtilis vegetative cells and Bacillus pumilus spores inside and outside the alumina column, and microbial recovery and radiation effect were assessed. Alumina was a barrier for the passage of microorganisms regardless the species, whilst spores were more retained than vegetative cells with a lower microbial recovery, without significant differences between 9.25 and 74 GBq generators. Bacillus pumilus biological indicator showed lower recoveries, suggesting a radiation inactivating effect on microorganisms. - Highlights: • Microorganisms in radionuclide generator may impair the quality of the product. • Killing of Bacillus pumilus was not complete even after 20 days of exposition. • Alumina column was a physical barrier for the microbial recovery. • An alternative biological indicator based on B. pumilus spores is proposed.

  11. Characterization of Emetic Bacillus weihenstephanensis, a New Cereulide-Producing Bacterium

    DEFF Research Database (Denmark)

    Thorsen, Line; Munk Hansen, Bjarne; Nielsen, Kristian Fog

    2006-01-01

    Cereulide production has until now been restricted to the species Bacillus cereus. Here we report on two psychrotolerant Bacillus weihenstephanensis strains, MC67 and MC118, that produce cereulide. The strains are atypical with regard to pheno- and genotypic characteristics normally used for iden......Cereulide production has until now been restricted to the species Bacillus cereus. Here we report on two psychrotolerant Bacillus weihenstephanensis strains, MC67 and MC118, that produce cereulide. The strains are atypical with regard to pheno- and genotypic characteristics normally used...

  12. Antifungal activity of indigenous Bacillus spp. isolated from soil

    Directory of Open Access Journals (Sweden)

    Bjelić Dragana Đ.

    2017-01-01

    Full Text Available Biocontrol using plant growth-promoting rhizobacteria (PGPR represents an alternative approach to disease management, since PGPR are known to promote growth and reduce diseases in various crops. Among the different PGPR, members of the genus Bacillus are prefered for most biotechnological uses due to their capability to form extremely resistant spores and produce a wide variety of metabolites with antimicrobial activity. The objective of this research was to identify antagonistic bacteria for management of the plant diseases. Eleven isolates of Bacillus spp. were obtained from the soil samples collected from different localities in the Province of Vojvodina. The antifungal activity of bacterial isolates against five fungal species was examined using a dual plate assay. Bacillus isolates exhibited the highest antifungal activity against Fusarium proliferatum, Fusarium oxysporum f. sp. cepae and Alternaria padwickii, while they had the least antagonistic effect on Fusarium verticillioides and Fusarium graminearum. Molecular identification showed that effective bacterial isolates were identified as Bacillus safensis (B2, Bacillus pumilus (B3, B11, Bacillus subtilis (B5, B7 and Bacillus megaterium (B8, B9. The highest antagonistic activity was exhibited by isolates B5 (from 39% to 62% reduction in fungal growth and B7 (from 40% to 71% reduction in fungal growth. These isolates of B. subtilis could be used as potential biocontrol agents of plant diseases. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31073

  13. 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

  14. Biosurfactant-biopolymer driven microbial enhanced oil recovery (MEOR) and its optimization by an ANN-GA hybrid technique.

    Science.gov (United States)

    Dhanarajan, Gunaseelan; Rangarajan, Vivek; Bandi, Chandrakanth; Dixit, Abhivyakti; Das, Susmita; Ale, Kranthikiran; Sen, Ramkrishna

    2017-08-20

    A lipopeptide biosurfactant produced by marine Bacillus megaterium and a biopolymer produced by thermophilic Bacillus licheniformis were tested for their application potential in the enhanced oil recovery. The crude biosurfactant obtained after acid precipitation effectively reduced the surface tension of deionized water from 70.5 to 28.25mN/m and the interfacial tension between lube oil and water from 18.6 to 1.5mN/m at a concentration of 250mgL -1 . The biosurfactant exhibited a maximum emulsification activity (E 24 ) of 81.66% against lube oil. The lipopeptide micelles were stabilized by addition of Ca 2+ ions to the biosurfactant solution. The oil recovery efficiency of Ca 2+ conditioned lipopeptide solution from a sand-packed column was optimized by using artificial neural network (ANN) modelling coupled with genetic algorithm (GA) optimization. Three important parameters namely lipopeptide concentration, Ca 2+ concentration and solution pH were considered for optimization studies. In order to further improve the recovery efficiency, a water soluble biopolymer produced by Bacillus licheniformis was used as a flooding agent after biosurfactant incubation. Upon ANN-GA optimization, 45% tertiary oil recovery was achieved, when biopolymer at a concentration of 3gL -1 was used as a flooding agent. Oil recovery was only 29% at optimal conditions predicted by ANN-GA, when only water was used as flooding solution. The important characteristics of biopolymers such as its viscosity, pore plugging capabilities and bio-cementing ability have also been tested. Thus, as a result of biosurfactant incubation and biopolymer flooding under the optimal process conditions, a maximum oil recovery of 45% was achieved. Therefore, this study is novel, timely and interesting for it showed the combined influence of biosurfactant and biopolymer on solubilisation and mobilization of oil from the soil. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Biodegradation of naphthalene and phenanthren by Bacillus subtilis 3KP

    Science.gov (United States)

    Ni'matuzahroh, Trikurniadewi, N.; Pramadita, A. R. A.; Pratiwi, I. A.; Salamun, Fatimah, Sumarsih, Sri

    2017-06-01

    The purposes of this research were to know growth response, degradation ability, and uptake mechanism of naphthalene and phenanthrene by Bacillus subtilis 3KP. Bacillus subtilis 3KP was grown on Mineral Synthetic (MS) medium with addition of 1% yeast extract and naphthalene and phenanthrene respectively 200 ppm in different cultures. Bacillus subtilis 3KP growth response was monitored by Total Plate Count (TPC) method, the degradation ability was monitored by UV-Vis spectrophotometer, and the uptake mechanism of hydrocarbon was monitored by emulsification activity, decrease of surface tension, and activity of Bacterial Adherence to Hydrocarbon (BATH). Bacillus subtilis 3KP was able to grow and show biphasic growth pattern on both of substrates. Naphthalene and phenanthrene were used as a carbon source for Bacillus subtilis 3KP growth that indicated by the reduction of substrate concomitant with the growth. At room temperature conditions (± 30°C) and 90 rpm of agitation for 7 days, Bacillus subtilis 3KP could degrade naphthalene in the amount of 70.5% and phenanthrene in the amount of 24.8%. Based on the analysis of UV-Vis spectrophotometer, three metabolites, 1-hydroxy-2-naphthoic acid, salicylic acid, and pyrocatechol were found in both cultures. The metabolite identification became basis of propose degradation pathway of naphthalene and phenanthrene by Bacillus subtilis 3KP. The results of hydrocarbon uptake mechanism test show that Bacillus subtilis 3KP used all of the mechanism to degrade naphthalene and phenanthrene.

  16. Laser-induced speckle scatter patterns in Bacillus colonies

    Directory of Open Access Journals (Sweden)

    Huisung eKim

    2014-10-01

    Full Text Available Label-free bacterial colony phenotyping technology called BARDOT (BActerial Rapid Detection using Optical scattering Technology provided successful classification of several different bacteria at the genus, species, and serovar level. Recent experiments with colonies of Bacillus species provided strikingly different characteristics of elastic light scatter (ELS patterns, which were comprised of random speckles compared to other bacteria, which are dominated by concentric rings and spokes. Since this laser-based optical sensor interrogates the whole volume of the colony, 3-D information of micro- and macro-structures are all encoded in the far-field scatter patterns. Here, we present a theoretical model explaining the underlying mechanism of the speckle formation by the colonies from Bacillus species. Except for Bacillus polymyxa, all Bacillus spp. produced random bright spots on the imaging plane, which presumably dependent on the cellular and molecular organization and content within the colony. Our scatter model-based analysis revealed that colony spread resulting in variable surface roughness can modify the wavefront of the scatter field. As the center diameter of the Bacillus spp. colony grew from 500 μm to 900 μm, average speckles area decreased 2-fold and the number of small speckles increased 7-fold. In conclusion, as Bacillus colony grows, the average speckle size in the scatter pattern decreases and the number of smaller speckle increases due to the swarming growth characteristics of bacteria within the colony.

  17. Unearthing Bacillus endophytes from desert plants that enhance growth of Arabidopsis thaliana under abiotic stress conditions

    KAUST Repository

    Bokhari, Ameerah M

    2018-04-01

    Here, we embarked a bioprospecting project that focuses on the isolation and characterization of plant root endophytes, collected from the Thar Desert. A total of 381 endophytes were isolated and based on their 16S rRNA gene sequences, genus Bacillus (58 strains) was identified as the major taxon and only endophytes from this genus were isolated from all plant types. Of the 58 Bacillus strains, only 16 strains were selected for screening of plant growth promotion traits such as P and Zn solubilization, indole-3-acetic acid and siderophore production, and antimicrobial activity. Based on the presence of specific plant growth promotion traits 10 strains were shortlisted for further in vitro screening with A. thaliana; to confirm that these bacteria can confer resilience to plants under salt stress conditions. B. circulans (PK3-15 and PK3-109), B. cereus (PK6-15) B. subtilis (PK3-9) and B. licheniformis (PK5-26) displayed the ability to increased the fresh weight of A. thaliana under salt stress conditions by more than 50 % compared to the uninoculated control. An interesting observation was that B. circulans (PK3-109) (shown to produce IAA exopolysaccharide) and B. circulans (PK3-138) (shown to produce IAA) in vitro results were substantially different as B. circulans (PK3-138) decreased the total fresh weight of A. thaliana by 47 %, whilst B. circulans (PK3-109) was one of the best performing strains. Thus, the genomes of these two strains were sequences to unravel the molecular versatility of B. circulans strains, specifically with respect to their interaction with plants. Most of the genome of these strains is identical but the most interesting feature was the presence of 1/ the DegS–DegU two-component system that is known to mediate the salt stress response and DegU also represses toxin wapA similar to antitoxin wapI, and 2/ YxiG, a gene in the unique orthogroup of PK3-109 was found to be linked to WapI. Thus, PK3-138 substantially decreasing the total fresh

  18. Bacterial inoculum enhances keratin degradation and biofilm formation in poultry compost.

    Science.gov (United States)

    Ichida, J M; Krizova, L; LeFevre, C A; Keener, H M; Elwell, D L; Burtt, E H

    2001-11-01

    Native microbial populations can degrade poultry waste, but the process can be hastened by using feather-degrading bacteria. Strains of Bacillus licheniformis and a Streptomyces sp. isolated from the plumage of wild birds were grown in a liquid basal medium and used to inoculate feathers in compost bioreaction vessels. Control vessels had only basal medium added to the feathers, litter and straw. Temperature, ammonia, carbon and nitrogen were monitored for 4 weeks. Scanning electron microscopy of the feather samples showed more complete keratin-degradation, more structural damage, and earlier microbial biofilm formation on inoculated feathers than on uninoculated feathers. A diverse community of aerobic bacteria and fungi were cultured early, but declined rapidly. Thermophilic B. licheniformis and Streptomyces spp. were abundant throughout. Enteric gram-negative bacteria, (e.g., Salmonella, E. coli) originally found on waste feathers were not recovered after day 4. Vessel temperatures reached 64-71 degrees C within 36 h and stabilized at 50 degrees C. When tumble-mixed at day 14, renewed activity peaked at 59 degrees C and quickly dropped as available carbon was used. Feathers soaked in an inoculum of B. licheniformis and Streptomyces degraded more quickly and more completely than feathers that were not presoaked. Inoculation of feather waste could improve composting of the large volume of feather waste generated every year by poultry farms and processing plants.

  19. Heat activation and stability of amylases from Bacillus species | Ajayi ...

    African Journals Online (AJOL)

    Leitch and Collier sporulating Bacillus medium was used to isolate some strains of Bacillus species from soil, wastewater and food sources in Ibadan, Oyo State, Nigeria, by heat activation method. Heat treatment at 80oC allowed the growth of sporulating Bacillus species, in the culture sample source without other bacteria ...

  20. Starter Culture Selection for Making Chinese Sesame-Flavored Liquor Based on Microbial Metabolic Activity in Mixed-Culture Fermentation

    Science.gov (United States)

    Wu, Qun; Ling, Jie

    2014-01-01

    Selection of a starter culture with excellent viability and metabolic activity is important for inoculated fermentation of traditional food. To obtain a suitable starter culture for making Chinese sesame-flavored liquor, the yeast and bacterium community structures were investigated during spontaneous and solid-state fermentations of this type of liquor. Five dominant species in spontaneous fermentation were identified: Saccharomyces cerevisiae, Pichia membranaefaciens, Issatchenkia orientalis, Bacillus licheniformis, and Bacillus amyloliquefaciens. The metabolic activity of each species in mixed and inoculated fermentations of liquor was investigated in 14 different cocultures that used different combinations of these species. The relationships between the microbial species and volatile metabolites were analyzed by partial least-squares (PLS) regression analysis. We found that S. cerevisiae was positively correlated to nonanal, and B. licheniformis was positively associated with 2,3-butanediol, isobutyric acid, guaiacol, and 4-vinyl guaiacol, while I. orientalis was positively correlated to butyric acid, isovaleric acid, hexanoic acid, and 2,3-butanediol. These three species are excellent flavor producers for Chinese liquor. Although P. membranaefaciens and B. amyloliquefaciens were not efficient flavor producers, the addition of them alleviated competition among the other three species and altered their growth rates and flavor production. As a result, the coculture of all five dominant species produced the largest amount of flavor compounds. The result indicates that flavor producers and microbial interaction regulators are important for inoculated fermentation of Chinese sesame-flavored liquor. PMID:24814798

  1. Hydrolysis of wheat B-starch and characterisation of acetylated maltodextrin.

    Science.gov (United States)

    Smrčková, Petra; Horský, Jiří; Šárka, Evžen; Koláček, Jaroslav; Netopilík, Miloš; Walterová, Zuzana; Kruliš, Zdeněk; Synytsya, Andrey; Hrušková, Kateřina

    2013-10-15

    Wheat B-starch was hydrolysed by α-amylase "Liquozyme supra" from Bacillus licheniformis at 90 °C and pH 7. After 2 h, the dextrose equivalent was 18; according to size exclusion chromatography, however, the hydrolysate contained not only dominant malto-oligosaccharides with the degree of polymerisation (DP)40. This non-uniformity of acetylated maltodextrin, both with respect to DP and to DS, must be taken into account in the development of acetylated-maltodextrin applications such as use as plasticisers or compatibilisers in biodegradable composites. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. Not so simple, not so subtle: the interspecies competition between Bacillus simplex and Bacillus subtilis and its impact on the evolution of biofilms

    Science.gov (United States)

    Rosenberg, Gili; Steinberg, Nitai; Oppenheimer-Shaanan, Yaara; Olender, Tsvia; Doron, Shany; Ben-Ari, Julius; Sirota-Madi, Alexandra; Bloom-Ackermann, Zohar; Kolodkin-Gal, Ilana

    2016-01-01

    Bacillus subtilis biofilms have a fundamental role in shaping the soil ecosystem. During this process, they unavoidably interact with neighbour bacterial species. We studied the interspecies interactions between biofilms of the soil-residing bacteria B. subtilis and related Bacillus species. We found that proximity between the biofilms triggered recruitment of motile B. subtilis cells, which engulfed the competing Bacillus simplex colony. Upon interaction, B. subtilis secreted surfactin and cannibalism toxins, at concentrations that were inert to B. subtilis itself, which eliminated the B. simplex colony, as well as colonies of Bacillus toyonensis. Surfactin toxicity was correlated with the presence of short carbon-tail length isomers, and synergistic with the cannibalism toxins. Importantly, during biofilm development and interspecies interactions a subpopulation in B. subtilis biofilm lost its native plasmid, leading to increased virulence against the competing Bacillus species. Overall, these findings indicate that genetic programs and traits that have little effect on biofilm development when each species is grown in isolation have a dramatic impact when different bacterial species interact. PMID:28721238

  3. Effect of Bacillus subtilis on Granite Weathering: A Laboratory Experiment

    Science.gov (United States)

    Song, W.; Ogawa, N.; Oguchi, C. T.; Hatta, T.; Matsukura, Y.

    2006-12-01

    We performed a comparative experiment to investigate how the ubiquitous soil bacterium Bacillus subtilis weathers granite and which granite-forming minerals weather more rapidly via biological processes. Batch type experiments (granite specimen in a 500 ml solution including NaCl, glucose, yeast extract and bacteria Bacillus subtilis at 27°E C) were carried out for 30 days. Granite surfaces were observed by SEM before and after the experiment. Bacillus subtilis had a strong influence on granite weathering by forming pits. There were 2.4 times as many pits and micropores were 2.3 times wider in granite exposed to Bacillus subtilis when compared with bacteria-free samples. Bacillus subtilis appear to preferentially select an optimum place to adhere to the mineral and dissolve essential elements from the mineral to live. Plagioclase was more vulnerable to bacterial weathering than biotite among the granite composing minerals.

  4. Evaluation of antifungal activity from Bacillus strains against ...

    African Journals Online (AJOL)

    In this study, 30 bacterial strains isolated from marine biofilms were screened for their antifungal activity against Rhizoctonia solani by dual culture assay. Two bacterial strains, Bacillus subtilis and Bacillus cereus, showed a clear antagonism against R. solani on potato dextrose agar (PDA) medium. The antagonistic activity ...

  5. Effect of vitamins and bivalent metals on lysine yield in Bacillus ...

    African Journals Online (AJOL)

    The effects of vitamins and bivalent metals on lysine accumulation in Bacillus strains were investigated. Biotin enhanced lysine production in all the Bacillus strains, while folic acid and riboflavin stimulated lysine yields in Bacillus megaterium SP 86 only. All bivalent metals stimulated lysine accumulation in B. megaterium ...

  6. Genome Sequence of Antibiotic-Producing Bacillus amyloliquefaciens Strain KCTC 13012.

    Science.gov (United States)

    Jeong, Haeyoung; Park, Seung-Hwan; Choi, Soo-Keun

    2015-10-01

    We report the 4.0-Mb draft genome sequence of Bacillus amyloliquefaciens (syn. Bacillus velezensis) KCTC 13012, which exhibits a broad spectrum of antagonistic activity against bacteria and fungi and promotes plant growth as well. The genome contains an array of biosynthetic gene clusters for secondary metabolites that are comparable to those in Bacillus amyloliquefaciens subsp. plantarum FZB42(T). Copyright © 2015 Jeong et al.

  7. Screening of Bacillus thuringiensis strains effective against mosquitoes Prospecção de estirpes de Bacillus thuringiensis efetivas contra mosquitos

    Directory of Open Access Journals (Sweden)

    Rose Gomes Monnerat

    2005-02-01

    Full Text Available The objective of this work was to evaluate 210 Bacillus thuringiensis strains against Aedes aegypti and Culex quinquefasciatus larvae to select the most effective. These strains were isolated from different regions of Brazil and are stored in a Bacillus spp. collection at Embrapa Recursos Genéticos e Biotecnologia, Brasília, Brazil. The selected strains were characterized by morphological (microscopy, biochemical (SDS-PAGE 10% and molecular (PCR methods. Six B. thuringiensis strains were identified as mosquito-toxic after the selective bioassays. None of the strains produced the expected PCR products for detection of cry4, cry11 and cyt1A genes. These results indicate that the activity of mosquitocidal Brazilian strains are not related with Cry4, Cry11 or Cyt proteins, so they could be used as an alternative bioinsecticide against mosquitoes.Neste trabalho foram realizados testes de patogenicidade com 210 estirpes de Bacillus thuringiensis contra larvas de Aedes aegypti e Culex quinquefasciatus, a fim de se determinar as mais eficazes. Estas estirpes foram isoladas de diversas regiões do Brasil e estão armazenadas na coleção de Bacillus spp. da Embrapa Recursos Genéticos e Biotecnologia. As estirpes selecionadas foram caracterizadas por métodos morfológicos (microscopia, bioquímicos (SDS-PAGE 10% e moleculares (Reação em Cadeia da Polimerase. Foram selecionadas seis estirpes entomopatogênicas de Bacillus thuringiensis. Nenhuma das estirpes de Bacillus thuringiensis apresentou produtos de PCR esperados para a detecção dos genes cry4, cry11 e cyt1A. A patogenicidade das estirpes não está associada à presença das toxinas Cry4, Cry11 ou Cyt, assim, essas estirpes poderão ser utilizadas para a formatação de um bioinseticida alternativo contra mosquitos.

  8. Metal adsorption capabilities of clinoptilolite and selected strains of bacteria from mine water

    Science.gov (United States)

    Mamba, B. B.; Dlamini, N. P.; Nyembe, D. W.; Mulaba-Bafubiandi, A. F.

    Small-scale mining has socio-economic advantages such as the reduction of unemployment and the general improvement of the economy. However, these operations if not properly managed or controlled have a potential to cause environmental damage, particularly with respect to the contamination of groundwater and water supplies that are not distant from where these mining activities take place. This paper focuses on metal removal from water contaminated by heavy metals emanating from small-scale mining operations using clinoptilolite and bacteria. Removal of As, Ni, Mn, Au, Co, Cu and Fe was carried out on mine water samples using original and HCl-activated (in 0.02 M and 0.04 M) natural clinoptilolite and bacterial strains (a mixed consortia of Bacillus strains ( Bacillus subtilis, Bacillus cereus, Bacillus firmus, Bacillus fusiformis, Bacillus macroides and Bacillus licheniformis), Pseudomonas spp., Shewanella spp. and a mixed consortia of Acidithiobcillus caldus, Leptospirillum spp., Ferroplasma spp. and Sulphobacillus spp.). The purpose of the study was to compare the removal efficiencies of the bacterial strains versus natural clinoptilolite adsorbents for metal cations. The Bacillus consortia removed most of the metals up to 98% metal removal efficiency with the exception of nickel where clinoptilolite showed good removal efficiency. The 0.02 M HCl-activated clinoptilolite also demonstrated excellent removal capabilities with Cu, Co and Fe removal efficiency of up to 98%. Both clinoptilolite and bacteria demonstrated capabilities of removing Cu 2+, Co 2+, Fe 2+, Mn 2+, As 3+ and Au from solution which augurs well for metal recovery from mining and mineral processing solutions, as well as in water decontamination.

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

    Directory of Open Access Journals (Sweden)

    M. Angélica Garrido-Pereira

    2014-11-01

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

  10. Dietary strategies in the diagnosis and treatment of patients with food intolerance

    Directory of Open Access Journals (Sweden)

    O.A. Martynchuk

    2017-11-01

    Full Text Available The most effective way of correcting adverse reactions to food is to completely eliminate the trigger product. The diagnostic capabilities available in the arsenal of modern physician’s are still insufficient, since only certain types of food intolerance can be determined with a high degree of certainty. As a gold standard for the diagnosis and treatment of such reactions, we continue to use various types of dietary interventions. The article examines in detail the diagnostic capabilities of the elimination diet and the controlled diet. As an additional, pathogenetically substantiated direction of therapeutic effect in food intolerance, dysbiosis of the intestine is considered, in the treatment of which adults and children successfully use self-eliminating antagonists of pathogenic bacterial flora. One of the representatives of this group of drugs is Biosporin containing spore-forming bacteria Bacillus subtilis and Bacillus licheniformis.

  11. Synthesis, spectral studies and biological evaluation of 2-aminonicotinic acid metal complexes

    Science.gov (United States)

    Nawaz, Muhammad; Abbasi, Muhammad Waseem; Hisaindee, Soleiman; Zaki, Muhammad Javed; Abbas, Hira Fatima; Mengting, Hu; Ahmed, M. Arif

    2016-05-01

    We synthesized 2-aminonicotinic acid (2-ANA) complexes with metals such as Co(II), Fe(III), Ni(II), Mn(II), Zn(II), Ag(I),Cr(III), Cd(II) and Cu(II) in aqueous media. The complexes were characterized and elucidated using FT-IR, UV-Vis, a fluorescence spectrophotometer and thermo gravimetric analysis (TGA). TGA data showed that the stoichiometry of complexes was 1:2 metal/ligand except for Ag(I) and Mn(II) where the ratio was 1:1. The metal complexes showed varied antibacterial, fungicidal and nematicidal activities. The silver and zinc complexes showed highest activity against Bacillus subtilis and Bacillus licheniformis respectively. Fusarium oxysporum was highly susceptible to nickel and copper complexes whereas Macrophomina phaseolina was completely inert to the complexes. The silver and cadmium complexes were effective against the root-knot nematode Meloidogyne javanica.

  12. Effect of Bacillus subtilis natto on growth performance in Muscovy ducks

    Directory of Open Access Journals (Sweden)

    T Sheng-Qiu

    2013-09-01

    Full Text Available The aim of the present study was to determine whether dietary Bacillus subtilis natto could affect growth performance of Muscovy ducks. A total of 120 hundred Muscovy ducks at the age of 1 day were randomly assigned to four groups (30 Muscovy ducks/group, and fed with diets supplemented with 0% (control group, 0.1%, 0.2%, and 0.4% Bacillus subtilis natto, respectively during the 6-week feeding period. Weight gain, feed intake and feed conversion efficiency of Muscovy ducks were significantly improved by the dietary addition of Bacillus subtilis natto, and the results were more significant in 0.4% dietary Bacillus subtilis natto treatment group; Also, Bacillus subtilis natto reduced Escherichia coli and Salmonella colonies, and increased lactobacilli population in the ileum and the cecum. Biochemical parameters, including total protein, GOT (glutamic oxaloacetic transaminase, GPT (glutamic pyruvic transaminase, AKP (alkaline phosphatase, triiodothyronine (T3 and tetraiodothyronine (T4 contents (pBacillus subtilis natto was added to the diets (p0.05. The results of the present study indicate that diets with 0.4% Bacillus subtilis natto improved the growth performance of Muscovy ducks by increasing the absorption of protein, simulating hormone secretion, suppressing harmful microflora, and improving the duodenal structure and immune functions of Muscovy ducks. It is suggested that Bacillus subtilis natto is a potential candidate to be used use as a probiotic to improve the growth performance of Muscovy ducks.

  13. Optimizing Bacillus circulans Xue-113168 for biofertilizer production ...

    African Journals Online (AJOL)

    Tuoyo Aghomotsegin

    2016-12-28

    Dec 28, 2016 ... In this study, Bacillus circulans Xue-113168 biofertilizer was produced through solid state fermentation ... organic matter, NPK content from 8.83 to 16.16 kg hm2, and reduced chemical ... dependent on the nutritional components. ...... shell fish chitin wastes for the production of Bacillus subtilis W-118.

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

    OpenAIRE

    Yeo, In-Cheol; Lee, Nam Keun; Hahm, Young Tae

    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.

  15. Isolation of bacillus thuringiensis from different samples from Mansehra District

    International Nuclear Information System (INIS)

    Younis, F.; Lodhi, A.F.; Raza, G.

    2009-01-01

    The insecticidal activity of Bacillus thuringiensis has made it very interesting for the control of a variety of agricultural pests and human disease vectors. The present study is an attempt to explore the potential and diversity. of Bacillus thuringiensis. from the local environment for the control of cotton spotted bollworm (Earias sp.), a major pest of cotton. Two hundred and ninety eight samples of soil, grain dust, wild animal dung, birds dropping, decaying leaves and dead insects were collected from different ecological environments of Mansehra District yielding 438 Bacillus thuringiensis isolates that produce parasporal crystalline inclusions. In this study the soil samples were found to be the richest source for Bacillus thuringiensis. (author)

  16. Induced resistance to hydrogen peroxide, UV and gamma radiation in bacillus species

    International Nuclear Information System (INIS)

    Bashandy, A.S.

    2005-01-01

    The catalase activity produced in four bacillus spp.(bacillus cereus, B. laterosporus, B. pumilus and B. subtilis (Escherichia coli was used for comparison) was measured and the sensitivity of these bacteria to hydrogen peroxide was tested. Bacillus spp. had higher resistance to hydrogen peroxide than E. coil. cultures of bacillus spp . When pretreated with sublethal level of hydrogen peroxide, became relatively resistant to the lethal effects of hydrogen than untreated control cultures. These pretreated cells were also resistant to lethality mediated by UV light and gamma radiation. The obtained results suggest that bacillus spp. Possess inducible defense mechanism (s) against the deleterious effects of oxidants and /or ionizing radiation

  17. Construction of novel shuttle expression vectors for gene expression in Bacillus subtilis and Bacillus pumilus.

    Science.gov (United States)

    Shao, Huanhuan; Cao, Qinghua; Zhao, Hongyan; Tan, Xuemei; Feng, Hong

    2015-01-01

    A native plasmid (pSU01) was detected by genome sequencing of Bacillus subtilis strain S1-4. Two pSU01-based shuttle expression vectors pSU02-AP and pSU03-AP were constructed enabling stable replication in B. subtilis WB600. These vectors contained the reporter gene aprE, encoding an alkaline protease from Bacillus pumilus BA06. The expression vector pSU03-AP only possessed the minimal replication elements (rep, SSO, DSO) and exhibited more stability on structure, suggesting that the rest of the genes in pSU01 (ORF1, ORF2, mob, hsp) were unessential for the structural stability of plasmid in B. subtilis. In addition, recombinant production of the alkaline protease was achieved more efficiently with pSU03-AP whose copy number was estimated to be more than 100 per chromosome. Furthermore, pSU03-AP could also be used to transform and replicate in B. pumilus BA06 under selective pressure. In conclusion, pSU03-AP is expected to be a useful tool for gene expression in Bacillus subtilis and B. pumilus.

  18. Screening of Bacillus Species with Potentials of Antibiotics Production

    OpenAIRE

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

    2009-01-01

    Sixteen soil samples were collected from different refuse dump sites in Minna, the capital Niger State, and analysed for the presence of Bacillus species. Physical-chemical analysis of the soil samples revealed the followings: PH value 6.89-8.47; moisture content 1.58 – 21.21% and temperature 27-28ºC. Using both pour plate and streak method of inoculation, total bacterial count in the soil samples ranged from 3.8×104 cfu/g 16.0×104 cfu/g. The identified Bacillus species included: Bacillus cer...

  19. Suitable conditions for xylanases activities from Bacillus sp. GA2(1 and Bacillus sp. GA1(6 and their properties for agricultural residues hydrolysis

    Directory of Open Access Journals (Sweden)

    Sudathip Chantorn

    2016-04-01

    Full Text Available Bacillus sp. GA2(1 and Bacillus sp. GA1(6 were isolated from soybean field in Khon Kaen province, Thailand. Crude enzymes from both isolates showed the activities of cellulase, xylanase, and mannanase at 37°C for 24 h. The highest xylanase activities of Bacillus sp. GA2(1 and Bacillus sp. GA1(6 were 1.58±0.25 and 0.82±0.16 U/ml, respectively. The relative xylanase activities from both strains were more than 60% at pH 5.0 to 8.0. The optimum temperature of xylanases was 50°C in both strains. The residual xylanase activities from both strains were more than 70% at 60°C for 60 min. Five agricultural wastes (AWs, namely coffee residue, soybean meal, potato peel, sugarcane bagasse, and corn cobs, were used as substrates for hydrolysis properties. The highest reducing sugar content of 101±1.32 µg/ml was obtained from soybean meal hydrolysate produced by Bacillus sp. GA2(1 xylanase.

  20. Phosphorescence In Bacillus Spores

    National Research Council Canada - National Science Library

    Reinisch, Lou; Swartz, Barry A; Bronk, Burt V

    2003-01-01

    .... Our present work attempts to build on this approach for environmental applications. We have measured a change in the fluorescence spectra of suspensions of Bacillus bacteria between the vegetative bacteria and their spores at room temperature...

  1. Microbial enhanced oil recovery research. Final report, Annex 5

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, M.M.; Gerogiou, G.

    1993-07-01

    The objective of this project was to develop an engineering framework for the exploitation of microorganisms to enhance oil recovery. An order of magnitude analysis indicated that selective plugging and the production of biosurfactants are the two most likely mechanisms for the mobilization of oil in microbial enhanced oil recovery (MEOR). The latter, biosurfactant production, is easier to control within a reservoir environment and was investigated in some detail. An extensive literature survey indicated that the bacterium Bacillus licheniformis JF-2 produces a very effective surface active agent capable of increasing the capillary number to values sufficiently low for oil mobilization. In addition, earlier studies had shown that growth of this bacterium and biosurfactant production occur under conditions that are typically encountered in MEOR, namely temperatures up to 55{degrees}C, lack of oxygen and salinities of up to 10% w/v. The chemical structure of the surfactant, its interfacial properties and its production by fermentation were characterized in some detail. In parallel, a set of experiments as conducted to measure the transport of Bacillus licheniformis JF-2 in sandpacks. It was shown that the determining parameters for cell transport in porous media are: cell size and degree of coagulation, presence of dispersants, injection velocity and cell concentration. The mechanisms of bacteria retention within the pores of the reservoir were analyzed based on heuristic arguments. A mathematical simulator of MEOR was developed using conservation equations in which the mechanisms of bacteria retention and the growth kinetics of the cells were incorporated. The predictions of the model agreed reasonably well with experimental results.

  2. Micro-Etched Platforms for Thermal Inactivation of Bacillus Anthracis and Bacillus Thuringiensis Spores

    Science.gov (United States)

    2008-03-01

    slips was first coated with a detergent wash. Commercially available Ivory soap shavings were diluted with sterile Millipore® water in a...environments. This removed controllable variability between the Bacillus species and increased the confidence in continued use of such surrogacy

  3. Toxin production ability of Bacillus cereus strains from food product of Ukraine

    Directory of Open Access Journals (Sweden)

    I. Pylypenko

    2017-10-01

    Full Text Available Potential pathogens of foodborne toxic infections – bacterial contaminants Bacillus cereus isolated from plant raw materials and food products from the Ukrainian region were investigated. When determining of the proportion of isolated bacilli from the plant samples, it was established that the epidemiologically significant microorganisms of Bacillus cereus as agents of food poisoning are the second largest. The average value of contaminated samples of Ukrainian plant raw materials and processed products with Bacillus cereus is 36,2 %. The ability of Bacillus cereus strains identified by a complex of morphological, tinctorial, cultural and biochemical properties, to produce specific emetic and enterotoxins was studied. Molecular genetic diagnosis and detection of the toxin-producing ability of isolated 42 Bacillus cereus strains showed both the possibility of their rapid identification and the presence of specific toxicity genes. Multiplex polymerase chain reaction (PCR was carried out with specific primers to detect toxicity determined of various bacilli genes: nheA, hblD, cytK, cesВ. The distribution of toxigenic genes is significantly different among the Bacillus cereus isolates from various sources. The nheA, hblD and cytK enterotoxin genes were detected in 100, 83,3 and 61,9 % of the investigated strains of Bacillus cereus, respectively. The cesB gene encoding emetic toxin was detected in 4,8 % of  strains. Molecular-genetic PCR-method confirmed that all the isolated strains belong to the Bacillus cereus group, and the ability to produce toxins can be attributed to five groups. The main toxins that produce the investigated Bacillus cereus strains were nhe and hbl enterotoxins encoded by the corresponding genes of nheA and hblD. The enterotoxic type of Bacillus cereus was predominant in Ukrainian region.  Studies of domestic plant food raw materials and products have confirmed the need to improve microbiological control of product safety

  4. Feather wastes digestion by new isolated strains Bacillus sp. in ...

    African Journals Online (AJOL)

    Feather wastes digestion by new isolated strains Bacillus sp. in Morocco. ... The most efficient isolated strain selected was compared with Bacillus subtilis ATCC 6633. Results showed ... African Journal of Biotechnology Vol.3(1) 2004: 67-70 ...

  5. Production of Alpha Amylase by Bacillus cereus in Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Helen H. Raplong

    2014-09-01

    Full Text Available Microorganisms have the ability to secrete enzymes when they are grown in the presence of certain substrates. Amylases are among the most important industrial enzymes and are of great significance in biotechnological studies. Bacteria belonging to the genus Bacillus were isolated using mannitol egg yolk polymyxin B (MYP agar a highly selective media for Bacillus cereus isolation. The isolates were tested for α-amylase production on nutrient agar supplemented with starch and in submerged fermentation. The bacteria isolated and identified (using the Microgen Bacillus identification kit were all Bacillus cereus and SB2 had the largest zone of hydrolysis of 12mm on nutrient agar supplemented with starch as well as the highest enzyme activity of 1.62U/ml. Amylase activity of 2.56U/ml was obtained after 24 hours incubation in submerged fermentation. When amylase enzyme production parameters where optimized, maximum amylase activity was obtained at a pH of 6.5, temperature of 350C, incubation time of 24 hours and 4% inoculums concentration. Bacillus cereus SB2 is a potential isolate for alpha-amylase production with soluble starch as the sole carbon source in submerged fermentation.

  6. Synthesis, spectral studies and biological evaluation of 2-aminonicotinic acid metal complexes.

    Science.gov (United States)

    Nawaz, Muhammad; Abbasi, Muhammad Waseem; Hisaindee, Soleiman; Zaki, Muhammad Javed; Abbas, Hira Fatima; Mengting, Hu; Ahmed, M Arif

    2016-05-15

    We synthesized 2-aminonicotinic acid (2-ANA) complexes with metals such as Co(II), Fe(III), Ni(II), Mn(II), Zn(II), Ag(I),Cr(III), Cd(II) and Cu(II) in aqueous media. The complexes were characterized and elucidated using FT-IR, UV-Vis, a fluorescence spectrophotometer and thermo gravimetric analysis (TGA). TGA data showed that the stoichiometry of complexes was 1:2 metal/ligand except for Ag(I) and Mn(II) where the ratio was 1:1. The metal complexes showed varied antibacterial, fungicidal and nematicidal activities. The silver and zinc complexes showed highest activity against Bacillus subtilis and Bacillus licheniformis respectively. Fusarium oxysporum was highly susceptible to nickel and copper complexes whereas Macrophomina phaseolina was completely inert to the complexes. The silver and cadmium complexes were effective against the root-knot nematode Meloidogyne javanica. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. The orotate transporter oroP from Lactococcus lactis can be used both as a very efficient, food-grade selection and counter-selection marker for strain construction in many different organisms

    DEFF Research Database (Denmark)

    Defoor, Els Marie Celine; Martinussen, Jan

    frame oroP on pDBORO necessary for the uptake of orotate was identified. A number of industrial important strains like Lactococcus lactis, Bacillus subtilus, and Bacillus licheniformis have been shown to be unable to metabolize orotate. If the oroP gene was introduced into these strains they acquired...... the ability to utilize orotate. If the strains had a pyrimidine requirement, the oroP gene could function as a selectable marker when growing in the presence of orotate as sole pyrimidine source. In an otherwise resistant strain, oroP was shown to sensitize the strain towards the analog 5-Fluoroorotate....... It was shown that strains who have lost the oroP gene could easily be selected in the presence of 5-Fluoroorotate, thus being an efficient counter-selection marker. pyrimidine-requiring strain (pyr B, C or D) orotate negative Counter-selection marker: wild-type strain! Fluoro-orotate resistant Functional...

  8. Utilization of keratin-containing biowaste to produce biohydrogen

    Energy Technology Data Exchange (ETDEWEB)

    Balint, B.; Rakhely, G.; Kovacs, K.L. [Szeged Univ. (Hungary). Dept. of Biotechnology; Hungarian Academy of Sciences, Szeged (Hungary). Inst. of Biophysics; Bagi, Z.; Perei, K. [Szeged Univ. (Hungary). Dept. of Biotechnology; Toth, A. [Hungarian Academy of Sciences, Szeged (Hungary). Inst. of Biophysics

    2005-12-01

    A two-stage fermentation system was constructed to test and demonstrate the feasibility of biohydrogen generation from keratin-rich biowaste. We isolated a novel aerobic Bacillus strain (Bacillus licheniformis KK1) that displays outstanding keratinolytic activity. The isolated strain was employed to convert keratin-containing biowaste into a fermentation product that is rich in amino acids and peptides. The process was optimized for the second fermentation step, in which the product of keratin fermentation-supplemented with essential minerals-was metabolized by Thermococcus litoralis, an anaerobic hyperthermophilic archaeon. T. litoralis grew on the keratin hydrolysate and produced hydrogen gas as a physiological fermentation byproduct. Hyperthermophilic cells utilized the keratin hydrolysate in a similar way as their standard nutrient, i.e., bacto-peptone. The generalization of the findings to protein-rich waste treatment and production of biohydrogen is discussed and possible means of further improvements are listed. (orig.)

  9. Development of More Effective Biosurfactants for Enhanced Oil Recovery

    Energy Technology Data Exchange (ETDEWEB)

    McInerney, J.J.; Han, S.O.; Maudgalya, S.; Mouttaki, H.; Folmsbee, M.; Knapp, R.; Nagle, D.; Jackson, B.E.; Stuadt, M.; Frey, W.

    2003-01-16

    The objectives of this were two fold. First, core displacement studies were done to determine whether microbial processes could recover residual oil at elevated pressures. Second, the importance of biosurfactant production for the recovery of residual oil was studies. In these studies, a biosurfactant-producing, microorganisms called Bacillus licheniformis strain JF-2 was used. This bacterium produces a cyclic peptide biosurfactant that significantly reduces the interfacial tension between oil and brine (7). The use of a mutant deficient in surfactant production and a mathematical MEOR simulator were used to determine the major mechanisms of oil recovery by these two strains.

  10. Statistically designed optimisation of enzyme catalysed starch removal from potato pulp

    DEFF Research Database (Denmark)

    Thomassen, Lise Vestergaard; Meyer, Anne S.

    2010-01-01

    to obtain dietary fibers is usually accomplished via a three step, sequential enzymatic treatment procedure using a heat stable alpha-amylase, protease, and amyloglucosidase. Statistically designed experiments were performed to investigate the influence of enzyme dose, amount of dry matter, incubation time...... and temperature on the amount of starch released from the potato pulp. The data demonstrated that all the starch could be released from potato pulp in one step when 8% (w/w) dry potato pulp was treated with 0.2% (v/w) (enzyme/substrate (E/S)) of a thermostable Bacillus licheniformis alpha-amylase (Termamyl(R) SC...

  11. Development of More Effective Biosurfactants for Enhanced Oil Recovery/Advanced Recovery Concepts Awards; SEMIANNUAL

    International Nuclear Information System (INIS)

    McInerney, M.J.; Marsh, T.L.; Zhang, X.; Knapp, R.M.; Nagle, Jr. D.P.; Sharma, P.K.; Jackson, B.E.

    2002-01-01

    The objectives of this were two fold. First, core displacement studies were done to determine whether microbial processes could recover residual oil at elevated pressures. Second, the importance of biosurfactant production for the recovery of residual oil was studies. In these studies, a biosurfactant-producing, microorganisms called Bacillus licheniformis strain JF-2 was used. This bacterium produces a cyclic peptide biosurfactant that significantly reduces the interfacial tension between oil and brine (7). The use of a mutant deficient in surfactant production and a mathematical MEOR simulator were used to determine the major mechanisms of oil recovery by these two strains

  12. Development of More Effective Biosurfactants for Enhanced Oil Recovery/Advanced Recovery Concepts Awards

    Energy Technology Data Exchange (ETDEWEB)

    McInerney, M.J.; Marsh, T.L.; Zhang, X.; Knapp, R.M.; Nagle, Jr., D.P.; Sharma, P.K.; Jackson, B.E.

    2002-05-28

    The objectives of this were two fold. First, core displacement studies were done to determine whether microbial processes could recover residual oil at elevated pressures. Second, the importance of biosurfactant production for the recovery of residual oil was studies. In these studies, a biosurfactant-producing, microorganisms called Bacillus licheniformis strain JF-2 was used. This bacterium produces a cyclic peptide biosurfactant that significantly reduces the interfacial tension between oil and brine (7). The use of a mutant deficient in surfactant production and a mathematical MEOR simulator were used to determine the major mechanisms of oil recovery by these two strains.

  13. Increasing the alkaline protease activity of Bacillus cereus and ...

    African Journals Online (AJOL)

    User

    2011-05-09

    May 9, 2011 ... cereus and Bacillus polymyxa simultaneously with the start of sporulation phase as a ... microbial forms to inactivation by chemical or physical agents. .... alkaline pH, 9, 10 and 11 and the pH of the culture media was optimized with .... incubation temperature for alkaline protease production by Bacillus ...

  14. Bacillus spp. Isolated from Puba as a Source of Biosurfactants and Antimicrobial Lipopeptides

    Science.gov (United States)

    Perez, Karla J.; Viana, Jaime dos Santos; Lopes, Fernanda C.; Pereira, Jamile Q.; dos Santos, Daniel M.; Oliveira, Jamil S.; Velho, Renata V.; Crispim, Silvia M.; Nicoli, Jacques R.; Brandelli, Adriano; Nardi, Regina M. D.

    2017-01-01

    Several products of industrial interest are produced by Bacillus, including enzymes, antibiotics, amino acids, insecticides, biosurfactants and bacteriocins. This study aimed to investigate the potential of two bacterial isolates (P5 and C3) from puba, a regional fermentation product from cassava, to produce multiple substances with antimicrobial and surface active properties. Phylogenetic analyses showed close relation of isolates P5 and C3 with Bacillus amyloliquefaciens and Bacillus thuringiensis, respectively. Notably, Bacillus sp. P5 showed antimicrobial activity against pathogens such as Listeria monocytogenes and Bacillus cereus, in addition to antifungal activity. The presence of genes encoding pre-subtilosin (sboA), malonyl CoA transacylase (ituD), and the putative transcriptional terminator of surfactin (sfp) were detected in Bacillus sp. P5, suggesting the production of the bacteriocin subtilosin A and the lipopeptides iturin A and surfactin by this strain. For Bacillus sp. C3 the presence of sboA and spas (subtilin) genes was observed by the first time in members of B. cereus cluster. Bacillus sp. P5 showed emulsifying capability on mineral oil, soybean biodiesel and toluene, while Bacillus sp. C3 showed emulsifying capability only on mineral oil. The reduction of the surface tension in culture medium was also observed for strain P5, confirming the production of surface-active compounds by this bacterium. Monoprotonated molecular species and adducts of sodium and potassium ions of surfactin, iturin, and fengycin were detected in the P5 culture medium. Comparative MS/MS spectra of the peak m/z 1030 (C14 surfactin A or C15 surfactin B [M+Na]+) and peak m/z 1079 (C15 iturin [M+Na]+) showed the same fragmentation profile of standards, confirming the molecular identification. In conclusion, Bacillus sp. P5 showed the best potential for the production of antifungal, antibacterial, and biosurfactant substances. PMID:28197131

  15. Oral administration of a select mixture of Bacillus probiotics generates Tr1 cells in weaned F4ab/acR- pigs challenged with an F4+ ETEC/VTEC/EPEC strain.

    Science.gov (United States)

    Zhou, Dong; Zhu, Yao-Hong; Zhang, Wei; Wang, Meng-Ling; Fan, Wen-Yi; Song, Dan; Yang, Gui-Yan; Jensen, Bent Borg; Wang, Jiu-Feng

    2015-09-17

    Although breeding of F4 receptor - negative (F4R(-)) pigs may prevent post-weaning diarrhea, the underlying immunity is poorly understood. Here, various doses of a Bacillus licheniformis and Bacillus subtilis mixture (BLS-mix) were orally administered to F4ab/acR(-) pigs for 1 week before F4 (K88) - positive ETEC/VTEC/EPEC challenge. Administration of BLS-mix increased the percentage of Foxp3(-)IL-10(+) T cells but not of Foxp3(+)IL-10(+) regulatory T (Treg) cells among peripheral blood CD4(+) T cells. A low dose of BLS-mix feeding resulted in increased the expression of IL-6, TNF-α, IL-10, and the transcription factors Foxp3 and T-bet mRNAs in the jejunum. Administration of either a low or high dose BLS-mix also led to an increase in the percentage of CD4(+)Foxp3(+) Treg cells among intraepithelial lymphocytes and CD4(+)IL-10(+) T cells in the small intestinal Peyer's patches and the lamina propria of F4ab/acR(-) pigs following F4(+) ETEC/VTEC/EPEC challenge. The increased number of IL-10-producing CD4(+) T cells was attributed to an increase in the proportion of Foxp3(-)IL-10(+) Treg cells rather than Foxp3(+)IL-10(+) Treg cells. Our data indicate that oral administration of BLS-mix to newly weaned F4ab/acR(-) pigs ameliorates enteritis in an F4(+) ETEC/VTEC/EPEC model; however, induction of IL-10-producing Foxp3(-) Treg cells by BLS-mix administration cannot account for the protection of newly weaned F4ab/acR(-) pigs from F4(+) ETEC/VTEC/EPEC infection, and that excessive generation of CD4(+)IL-10(+) T cells following consumption of BLS-mix during episodes of intestinal inflammation that is caused by enteric pathogens might prohibit clearance of the pathogen. Select probiotic mixtures may allow for tailoring strategies to prevent infectious diseases.

  16. Identification of Bacillus anthracis by Using Monoclonal Antibody to Cell Wall Galactose-N-Acetylglucosamine Polysaccharide

    Science.gov (United States)

    1990-02-01

    Bacillus circulans ATCC 4513 b - - NR NT NT NT NT Bacillus coagulans ATCC 7050 b - - NR NT NT NT NT Bacillus eugilitis B-61 f - - NR NT NT NT NT...American Society for Microbiology W Identification of Bacillus anthracis by-U-sing Monoclonal Antibody CC to Cell Wall Galactose-N-Acetylglucosamine...Received 22 June 1989/Accepted 31 October 1989 ’ Guanidine extracts of crude Bacillus anthracis cell wall were used to vaccinate BALB/c mice and to

  17. Bacillus: A Biological Tool for Crop Improvement through Bio-Molecular Changes in Adverse Environments

    Science.gov (United States)

    Radhakrishnan, Ramalingam; Hashem, Abeer; Abd_Allah, Elsayed F.

    2017-01-01

    Crop productivity is affected by environmental and genetic factors. Microbes that are beneficial to plants are used to enhance the crop yield and are alternatives to chemical fertilizers and pesticides. Pseudomonas and Bacillus species are the predominant plant growth-promoting bacteria. The spore-forming ability of Bacillus is distinguished from that of Pseudomonas. Members of this genus also survive for a long time under unfavorable environmental conditions. Bacillus spp. secrete several metabolites that trigger plant growth and prevent pathogen infection. Limited studies have been conducted to understand the physiological changes that occur in crops in response to Bacillus spp. to provide protection against adverse environmental conditions. This review describes the current understanding of Bacillus-induced physiological changes in plants as an adaptation to abiotic and biotic stresses. During water scarcity, salinity and heavy metal accumulate in soil, Bacillus spp. produce exopolysaccharides and siderophores, which prevent the movement of toxic ions and adjust the ionic balance and water transport in plant tissues while controlling the pathogenic microbial population. In addition, the synthesis of indole-3-acetic acid, gibberellic acid and1-aminocyclopropane-1-carboxylate (ACC) deaminase by Bacillus regulates the intracellular phytohormone metabolism and increases plant stress tolerance. Cell-wall-degrading substances, such as chitosanase, protease, cellulase, glucanase, lipopeptides and hydrogen cyanide from Bacillus spp. damage the pathogenic bacteria, fungi, nematodes, viruses and pests to control their populations in plants and agricultural lands. The normal plant metabolism is affected by unfavorable environmental stimuli, which suppress crop growth and yield. Abiotic and biotic stress factors that have detrimental effects on crops are mitigated by Bacillus-induced physiological changes, including the regulation of water transport, nutrient up-take and

  18. Bacillus: A Biological Tool for Crop Improvement through Bio-Molecular Changes in Adverse Environments

    Directory of Open Access Journals (Sweden)

    Ramalingam Radhakrishnan

    2017-09-01

    Full Text Available Crop productivity is affected by environmental and genetic factors. Microbes that are beneficial to plants are used to enhance the crop yield and are alternatives to chemical fertilizers and pesticides. Pseudomonas and Bacillus species are the predominant plant growth-promoting bacteria. The spore-forming ability of Bacillus is distinguished from that of Pseudomonas. Members of this genus also survive for a long time under unfavorable environmental conditions. Bacillus spp. secrete several metabolites that trigger plant growth and prevent pathogen infection. Limited studies have been conducted to understand the physiological changes that occur in crops in response to Bacillus spp. to provide protection against adverse environmental conditions. This review describes the current understanding of Bacillus-induced physiological changes in plants as an adaptation to abiotic and biotic stresses. During water scarcity, salinity and heavy metal accumulate in soil, Bacillus spp. produce exopolysaccharides and siderophores, which prevent the movement of toxic ions and adjust the ionic balance and water transport in plant tissues while controlling the pathogenic microbial population. In addition, the synthesis of indole-3-acetic acid, gibberellic acid and1-aminocyclopropane-1-carboxylate (ACC deaminase by Bacillus regulates the intracellular phytohormone metabolism and increases plant stress tolerance. Cell-wall-degrading substances, such as chitosanase, protease, cellulase, glucanase, lipopeptides and hydrogen cyanide from Bacillus spp. damage the pathogenic bacteria, fungi, nematodes, viruses and pests to control their populations in plants and agricultural lands. The normal plant metabolism is affected by unfavorable environmental stimuli, which suppress crop growth and yield. Abiotic and biotic stress factors that have detrimental effects on crops are mitigated by Bacillus-induced physiological changes, including the regulation of water transport

  19. Fungicidal effect of bacteriocins harvested from Bacillus spp.

    Directory of Open Access Journals (Sweden)

    Adetunji, V. O.

    2013-01-01

    Full Text Available Aims: This study investigated the ability of bacteriocins isolated from Bacillus spp. (Bacillus species to inhibit fourdifferent yeast isolates obtained from common food products (nono, yoghurt, ogi and cheese commonly consumed byNigerians with minimal heat treatment.Methodology and results: Forty-five Bacillus spp. was isolated and identified from common food products usingcultural, morphological, physiological and biochemical characteristics. These isolates were tested for antimicrobialactivity against Salmonella enteritidis (3, Micrococcus luteus (1 and Staphylococcus aureus (2. Eight bacteriocinproducing strains were identified from an over- night broth culture centrifugated at 3500 revolutions for five minutes.Fungicidal effects of these bacteriocins were tested against four yeast strains using the Agar Well Diffusion method. Thebacteriocins produced wide zones of inhibition ranging from 5.9±0.000 to 24.00±0.000 mm against the 4 yeast strainstested. There was a significant difference (at p<0.05 between the yeast organisms and the bacteriocins from theBacillus spp.Conclusion, significance and impact of study: The study reveals the antifungal property of bacteriocins from Bacillusspp. and serves therefore as a base for further studies in its use in the control of diseases and extension of shelf-life ofproducts prone to fungi contamination.

  20. Reparation and Immunomodulating Properties of Bacillus sp. Metabolites from Permafrost.

    Science.gov (United States)

    Kalenova, L F; Melnikov, V P; Besedin, I M; Bazhin, A S; Gabdulin, M A; Kolyvanova, S S

    2017-09-01

    An ointment containing metabolites of Bacillus sp. microorganisms isolated from permafrost samples was applied onto the skin wound of BALB/c mice. Metabolites isolated during culturing of Bacillus sp. at 37°C produced a potent therapeutic effect and promoted wound epithelialization by 30% in comparison with the control (ointment base) and by 20% in comparison with Solcoseryl. Treatment with Bacillus sp. metabolites stimulated predominantly humoral immunity, reduced the time of wound contraction and the volume of scar tissue, and promoted complete hair recovery. These metabolites can be considered as modulators of the wound process with predominance of regeneration mechanisms.