WorldWideScience

Sample records for bacillus subtilis strains

  1. Draft Genome Sequence of Bacillus subtilis strain KATMIRA1933

    OpenAIRE

    Karlyshev, Andrey V.; Melnikov, Vyacheslav G.; Chikindas, Michael L.

    2014-01-01

    In this report, we present a draft sequence of Bacillus subtilis KATMIRA1933. Previous studies demonstrated probiotic properties of this strain partially attributed to production of an antibacterial compound, subtilosin. Comparative analysis of this strain’s genome with that of a commercial probiotic strain, B. subtilis Natto, is presented.

  2. Genetic transformation of Bacillus strains close to bacillus subtilis and isolated from the soil

    International Nuclear Information System (INIS)

    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

  3. Enhanced hydrocarbon biodegradation by a newly isolated bacillus subtilis strain

    International Nuclear Information System (INIS)

    The relation between hydrocarbon degradation and biosurfactant (rhamnolipid) production by a new bacillus subtilis 22BN strain was investigated. The strain was isolated for its capacity to utilize n-hexadecane and naphthalene and at the same time to produce surface-active compound at high concentrations (1.5 - 2.0 g l-1). Biosurfactant production was detected by surface tension lowering and emulsifying activity. The strain is a good degrader of both hydrocarbons used with degradability of 98.3 ± 1% and 75 ± 2% for n-hexadecane and naphthalene, respectively. Measurement of cell hydrophobicity showed that the combination of slightly soluble substrate and rhamnolipid developed higher hydrophobicity correlated with increased utilization of both hydrocarbon substrates. To our knowledge, this is the first report of bacillus subtilis strain that degrades hydrophobic compounds and at the same time produces rhamnolipid biosurfactant. (orig.)

  4. Use of bacillus subtilis strains to inhibit postharvest pathogenic fungi

    International Nuclear Information System (INIS)

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

  5. Complete Genome Sequence of Bacillus subtilis Strain CU1050, Which Is Sensitive to Phage SPβ

    OpenAIRE

    Johnson, Christopher M.; Grossman, Alan D.

    2016-01-01

    The Gram-positive bacterium Bacillus subtilis is used as a model organism to study cellular and molecular processes. Here, we announce the complete genomic sequence of B. subtilis strain CU1050, derived from B. subtilis strain 168. CU1050 has historically been used to study suppressor mutations and phage biology, especially the lysogenic phage SPβ.

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

    OpenAIRE

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

    2012-01-01

    Although several strains of B. subtilis with antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a native B. subtilis strain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis 21) demonstrated in vitro antagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected with Rhizoctonia solani and Fusarium verticillioides and treated with B. subtilis 2...

  7. Complete Genome Sequences of Bacillus subtilis subsp. subtilis Laboratory Strains JH642 (AG174) and AG1839

    OpenAIRE

    Smith, Janet L.; Goldberg, Jonathan M.; Grossman, Alan D.

    2014-01-01

    The Gram-positive bacterium Bacillus subtilis is widely used for studies of cellular and molecular processes. We announce the complete genomic sequences of strain AG174, our stock of the commonly used strain JH642, and strain AG1839, a derivative that contains a mutation in the replication initiation gene dnaB and a linked Tn917.

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

    DEFF Research Database (Denmark)

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

    2015-01-01

    Supplementing Bacillus (B.) subtilis mutants selected to overproduce a specific amino acid (AA) may be an alternative method to provide essential AA in pig diets. Two experiments on a B. subtilis strain selected to overproduce Trp were conducted using 8-kg pigs fed Trp-deficient diets for 20 d. B....... subtilis were supplied in a low or high dose in Experiments 1 and 2, respectively. The Trp-deficient diet (0.15 SID Trp:Lys) reduced (p < .05) both gain and feed intake of piglets compared to the positive control diet (0.17 SID Trp:Lys). Supplementation of the B. subtilis strain was not able to...... counterbalance the Trp deficiency in any of the two experiments. No effect of B. subtilis supplementation to piglet diets was observed on the plasma AA profile. In conclusion, this mutant strain of B. subtilis was not able to compensate a Trp deficiency in the tested doses....

  9. Genome Sequence of Bacillus subtilis Strain HUK15, Isolated from Hexachlorocyclohexane-Contaminated Soil

    OpenAIRE

    Gasc , Cyrielle; Richard, Jean-Yves; Peyret, Pierre

    2016-01-01

    Bacillus subtilis strain HUK15 has been isolated from hexachlorocyclohexane (HCH)-long-term-contaminated soil. The genome of strain HUK15 was sequenced to investigate its adaptation toward HCH and its potential capability to degrade the pesticide. Here, we report the annotated draft genome sequence (~4.3 Mbp) of this strain.

  10. Draft Genome Sequence of Bacillus subtilis Strain NKYL29, an Antimicrobial-Peptide-Producing Strain from Soil

    OpenAIRE

    Jiang, Yanbin; Xu, Haijin; Ying LI; Liu, Hongbin; Yu, Lei; Qiao, Mingqiang; Liu, Gang

    2014-01-01

    Bacillus subtilis strain NKYL29 is an antimicrobial-peptide-producing strain isolated from the soil of Ranzhuang Tunnel in Hebei Province, China. Here, we present the draft genome of this strain, which provides the genetic basis for application of the antimicrobial peptide.

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

    Directory of Open Access Journals (Sweden)

    Francisco Fábio Cavalcante Barros

    2013-01-01

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

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

    Science.gov (United States)

    Barros, Francisco Fábio Cavalcante; Simiqueli, Ana Paula Resende; de Andrade, Cristiano José; Pastore, Gláucia Maria

    2013-01-01

    Bacteria in the genus Bacillus are the source of several enzymes of current industrial interest. Hydrolases, such as amylases, proteases, and lipases, are the main enzymes consumed worldwide and have applications in a wide range of products and industrial processes. Fermentation processes by Bacillus subtilis using cassava wastewater as a substrate are reported in the technical literature; however, the same combination of microorganisms and this culture medium is limited or nonexistent. In this paper, the amylase, protease, and lipase production of ten Bacillus subtilis strains previously identified as biosurfactant producers in cassava wastewater was evaluated. The LB1a and LB5a strains were selected for analysis using a synthetic medium and cassava wastewater and were identified as good enzyme producers, especially of amylases and proteases. In addition, the enzymatic activity results indicate that cassava wastewater was better than the synthetic medium for the induction of these enzymes. PMID:23533780

  13. Production and antimicrobial activity of 3-hydroxypropionaldehyde from Bacillus subtilis strain CU12.

    Science.gov (United States)

    Wise, C; Novitsky, L; Tsopmo, A; Avis, T J

    2012-12-01

    Bacillus subtilis strains are known to produce a vast array of antimicrobial compounds. However, some compounds remain to be identified. Disk assays performed in vitro with Bacillus subtilis CU12 showed a significant reduction in mycelial growth of Alternaria solani, Botrytis cinerea, Fusarium sambucinum, and Pythium sulcatum. Crude B. subtilis culture filtrates were subsequently extracted with ethyl acetate and butanol. A bioassay guided purification procedure revealed the presence of one major antifungal compound in the butanol extract. Purification of the compound was performed using a reverse-phase C18 solid phase extraction (SPE) cartridge and flash column chromatography. NMR data showed that the main antimicrobial compound was a cyclic dimer of 3-hydroxypropionaldehyde (HPA). This study demonstrated the antimicrobial activity of B. subtilis strain CU12 against phytopathogenic microorganisms is mediated at least in part by the production of HPA. It also suggests that this B. subtilis strain could be effective at controlling pathogens through protection of its ecological niche by antibiosis. PMID:23179100

  14. Microbial enhanced oil recovery by Bacillus subtilis strains under simulated reservoir conditions

    OpenAIRE

    Gudiña, Eduardo J.; L. R. Rodrigues; J.A. Teixeira; Pereira, J. F.; Coutinho, J.A.P.; Soares, L. P.; Ribeiro, M. T.

    2012-01-01

    Microbial Enhanced Oil Recovery (MEOR) is a tertiary oil recovery process in which microorganisms and their metabolites are used to retrieve unrecoverable oil from mature reservoirs. Stimulation of microorganisms that produce biosurfactants and degrade heavy oil fractions in situ reduces the capillary forces that retain the oil into the reservoir and decreases oil viscosity, thus promoting its flow. As a result, oil production can be increased. In previous work, Bacillus subtilis strains that...

  15. Biosynthesis of hydrolytic enzymes of UV-mutants of Bacillus subtilis B-3 strain

    International Nuclear Information System (INIS)

    The studies were carried out on 12 UV-mutants of Bacillus subtilis B-3 which synthesize from 18 to 105% more hydrolytic enzymes (proteinases, amylases, cellulases and lipases) than the wild strain. After 5 years' storage of the mutants, a 26-fold increase in amylase biosynthesis was observed for a G7 UV-mutant. This mutant showed also other phenotypic differences. The increased ability of mutant G7 to synthesize amylases was controlled by the maltose-substrate induction. (author)

  16. Taxonomy Icon Data: Bacillus subtilis [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available Bacillus subtilis Bacillus subtilis Bacillus_subtilis_L.png Bacillus_subtilis_NL.png Bacillus..._subtilis_S.png Bacillus_subtilis_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus...+subtilis&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus+subtilis&t=NL http://biosciencedbc.j...p/taxonomy_icon/icon.cgi?i=Bacillus+subtilis&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus

  17. [An Efficient Method for Genetic Certification of Bacillus subtilis strains, Prospective Producers of Biopreparations].

    Science.gov (United States)

    Terletskiy, V P; Tyshenko, V I; Novikova, I I; Boikova, I V; Tyulebaev, S D; Shakhtamirov, I Ya

    2016-01-01

    Genetic certification of commercial strains of bacteria antagonistic to phytopathogenic microorganisms guarantees their unequivocal identification and confirmation of safety. In Russia, unlike EU countries, genetic certification of Bacillus subtilis strains is not used. Based on the previously proposed double digestion selective label (DDSL) fingerprinting, a method for genetic identification and certification of B. subtilis strains was proposed. The method was tested on several strains differing in their physiological and biochemical properties and in the composition of secondary metabolites responsible for the spectrum of antibiotic activity. High resolving power of this approach was shown. Optimal restriction endonucleases (SgsI and Eco32I) were determined and validated. A detailed protocol for genetic certification of this bacterial species was developed. DDSL is a universal method, which may be adapted for genetic identification and certification of other bacterial species. PMID:27301128

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

    Science.gov (United States)

    Todorova, Sevdalina; Kozhuharova, Lubka

    2010-07-01

    Antagonistic Bacillus strains were isolated from soil and analyzed for the purpose of determining whether they could be used as natural biological agents. Primary in vitro screening for antagonism of the isolates was performed against five phytopathogenic mould fungi. Strains TS 01 and ZR 02 exhibited the most pronounced inhibitory effects. They were identified as Bacillus subtilis on the basis of their morphological, cultural and physiology-biochemical properties as well as their hierarchical cluster analysis conducted by means of computer program SPSS. The antimicrobial activity of the strains from cultural medium and sterile filtrate were determined in vitro against a great number of predominantly phytopathogenic fungi and bacteria. TS 01 and ZR 02 strains exhibited very broad and at the same time degree varying antibiotic spectra of activities against both Gram-positive and Gram-negative microorganisms. Many of them were tested against sensitivity to the antimicrobial action of B. subtilis for the very first time. B. subtilis TS 01 and ZR 02 showed highest antifungal activity (sterile zone in diameter over 37 mm) against Alternaria solani, Botrytis cinerea, Monilia linhartiana 869, Phytophthora cryptogea 759/1 and Rhizoctonia sp. The most sensitive bacterial species were found to be Pseudomonas syringae pv. tomato Ro and Xanthomonas campestris with sterile zones 48.0 and 50.0 mm in diameter, respectively. The latter draws a conclusion that the isolated and identified Bacillus subtilis strains are promising natural biocontrol agents and should be further studied and tested for control of numerous plant diseases. PMID:24026925

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

    Directory of Open Access Journals (Sweden)

    Yueju Zhao

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

  20. Cyclic lipopeptide profile of three Bacillus subtilis strains; antagonists of Fusarium head blight.

    Science.gov (United States)

    Dunlap, Christopher A; Schisler, David A; Price, Neil P; Vaughn, Steven F

    2011-08-01

    The objective of the study was to identify the lipopetides associated with three Bacillus subtilis strains. The strains are antagonists of Gibberella zeae, and have been shown to be effective in reducing Fusarium head blight in wheat. The lipopeptide profile of three B. subtilis strains (AS43.3, AS43.4, and OH131.1) was determined using mass spectroscopy. Strains AS43.3 and AS43.4 produced the anti-fungal lipopeptides from the iturin and fengycin family during the stationary growth phase. All three strains produced the lipopeptide surfactin at different growth times. Strain OH131.1 only produced surfactin under these conditions. The antifungal activity of the culture supernatant and individual lipopeptides was determined by the inhibition of G. zeae. Cell-free supernatant from strains AS43.3 and AS43.4 demonstrated strong antibiosis of G. zeae, while strain OH131.1 had no antibiosis activity. These results suggest a different mechanism of antagonism for strain OH131.1, relative to AS43.3 and AS43.4. PMID:21887643

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

    Directory of Open Access Journals (Sweden)

    ziba Akbari

    2015-12-01

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

  2. Isolation and characterization of atrazine mineralizing Bacillus subtilis strain HB-6.

    Directory of Open Access Journals (Sweden)

    Jinhua Wang

    Full Text Available Atrazine is a widely used herbicide with great environmental concern due to its high potential to contaminate soil and waters. An atrazine-degrading bacterial strain HB-6 was isolated from industrial wastewater and the 16S rRNA gene sequencing identified HB-6 as a Bacillus subtilis. PCR assays indicated that HB-6 contained atrazine-degrading genes trzN, atzB and atzC. The strain HB-6 was capable of utilizing atrazine and cyanuric acid as a sole nitrogen source for growth and even cleaved the s-triazine ring and mineralized atrazine. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by cooperating with other bacteria, suggesting its huge potential for remediation of atrazine-contaminated sites. To our knowledge, there are few Bacillus subtilis strains reported that can mineralize atrazine, therefore, the present work might provide some new insights on atrazine remediation.

  3. Purification and characterization of keratinase from a new Bacillus subtilis strain

    OpenAIRE

    Cai, Cheng-gang; Chen, Ji-shuang; Qi, Jiong-jiong; Yin, Yun; Zheng, Xiao-dong

    2008-01-01

    The aim of this study was to purify and characterize a keratinase produced by a new isolated Bacillus subtilis KD-N2 strain. The keratinase produced by the isolate was purified using ammonium sulphate precipitation, Sephadex G-75 and DEAE (diethylaminoethyl)-Sepharose chromatographic techniques. The purified enzyme was shown to have a molecular mass of 30.5 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The optimum pH at 50 °C was 8.5 and ...

  4. Draft Genome Sequence of Bacillus subtilis GXA-28, a Thermophilic Strain with High Productivity of Poly-γ-Glutamic Acid

    OpenAIRE

    Zeng, Wei; Chen, Guiguang; Tang, Zhen; Wu, Hao; Shu, Lin; Liang, Zhiqun

    2014-01-01

    Bacillus subtilis GXA-28 is a thermophilic strain that can produce high yield and high molecular weight of poly-γ-glutamic acid under high temperature. Here, we report the draft genome sequence of this strain, which may provide the genomic basis for the high productivity of poly-γ-glutamic acid.

  5. Engineering of a Bacillus subtilis Strain with Adjustable Levels of Intracellular Biotin for Secretory Production of Functional Streptavidin

    OpenAIRE

    Wu, Sau-Ching; Wong, Sui-Lam

    2002-01-01

    Streptavidin is a biotin-binding protein which has been widely used in many in vitro and in vivo applications. Because of the ease of protein recovery and availability of protease-deficient strains, the Bacillus subtilis expression-secretion system is an attractive system for streptavidin production. However, attempts to produce streptavidin using B. subtilis face the problem that cells overproducing large amounts of streptavidin suffer poor growth, presumably because of biotin deficiency. Th...

  6. Keratinase production and keratin degradation by a mutant strain of Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    Cheng-gang CAI; Bing-gan LOU; Xiao-dong ZHENG

    2008-01-01

    A new feather-degrading bacterium was isolated from a local feather waste site and identified as Bacillus subtilis based on morphological, physiochemical, and phylogenetic characteristics. Screening for mutants with elevated keratinolytic activity using N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis resulted in a mutant strain KD-N2 producing keratinolytic activity about 2.5 times that of the wild-type strain. The mutant strain produced inducible keratinase in different substrates of feathers, hair, wool and silk under submerged cultivation. Scanning electron microscopy studies showed the degradation of feathers, hair and silk by the keratinase. The optimal conditions for keratinase production include initial pH of 7.5, inoculum size of 2% (v/v), age of inoculum of 16 h, and cultivation at 23 ℃. The maximum keratinolytic activity of KD-N2 was achieved after 30 h. Essential amino acids like threonine, valine, methionine as well as ammonia were produced when feathers were used as substrates. Strain KD-N2,therefore, shows great promise of finding potential applications in keratin hydrolysis and keratinase production.

  7. 77 FR 1633 - Bacillus Subtilis Strain CX-9060; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2012-01-11

    ..., 2009; 74 FR 15865; FRL-8408-7). The previously submitted studies on Bacillus subtilis MBI 600 include... Register of March 10, 2010 (75 FR 11171) (FRL-8810- 8), EPA issued a notice pursuant to section 408(d)(3...(h) (75 FR 76284, FRL-8853-8) eliminates the option for the expression of tolerances or...

  8. Subtilomycin: A New Lantibiotic from Bacillus subtilis Strain MMA7 Isolated from the Marine Sponge Haliclona simulans

    Directory of Open Access Journals (Sweden)

    Teresa M. Barbosa

    2013-06-01

    Full Text Available Bacteriocins are attracting increased attention as an alternative to classic antibiotics in the fight against infectious disease and multidrug resistant pathogens. Bacillus subtilis strain MMA7 isolated from the marine sponge Haliclona simulans displays a broad spectrum antimicrobial activity, which includes Gram-positive and Gram-negative pathogens, as well as several pathogenic Candida species. This activity is in part associated with a newly identified lantibiotic, herein named as subtilomycin. The proposed biosynthetic cluster is composed of six genes, including protein-coding genes for LanB-like dehydratase and LanC-like cyclase modification enzymes, characteristic of the class I lantibiotics. The subtilomycin biosynthetic cluster in B. subtilis strain MMA7 is found in place of the sporulation killing factor (skf operon, reported in many B. subtilis isolates and involved in a bacterial cannibalistic behaviour intended to delay sporulation. The presence of the subtilomycin biosynthetic cluster appears to be widespread amongst B. subtilis strains isolated from different shallow and deep water marine sponges. Subtilomycin possesses several desirable industrial and pharmaceutical physicochemical properties, including activity over a wide pH range, thermal resistance and water solubility. Additionally, the production of the lantibiotic subtilomycin could be a desirable property should B. subtilis strain MMA7 be employed as a probiotic in aquaculture applications.

  9. Biosurfactant-producing Bacillus subtilis strains isolated from crude oil samples enhance oil recovery at lab scale

    OpenAIRE

    Gudiña, Eduardo J.; L. R. Rodrigues; J.A. Teixeira

    2012-01-01

    Biosurfactant-producing Bacillus subtilis strains isolated from crude oil samples enhance oil recovery at lab scale Eduardo J Gudiña, Lígia R. Rodrigues, José A. Teixeira IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal Microbial Enhanced Oil Recovery (MEOR) is potentially useful to increment oil recovery from reservoirs beyond primary and secondary recovery operations using micro...

  10. Draft Genome Sequence of Bacillus subtilis ALBA01, a Strain with Antagonistic Activity against the Soilborne Fungal Pathogen of Onion Setophoma terrestris

    Science.gov (United States)

    Tobares, Romina A.; Ducasse, Daniel A.; Smania, Andrea M.

    2016-01-01

    Bacillus subtilis is a nonpathogenic bacterium that lives in soil and has long been used as biological control agent in agriculture. Here, we report the genome sequence of a B. subtilis strain isolated from rhizosphere of onion that shows strong biological activity against the soilborne fungal pathogen Setophoma terrestris. PMID:27257193

  11. Purification and characterization of alkaline protease produced by a mutant strain of bacillus subtilis

    International Nuclear Information System (INIS)

    The present study describes the production, purification and characterization of alkaline protease from mutant strain of Bacillus subtilis EMS-8. The enzyme was purified using ammonium sulphate precipitation which gave 2.64 fold purification with 81.5% yield at 70% saturation. The molecular weight of the enzyme was determined using SDS-PAGE and it was found to be 25 KDa. The optimum pH of enzyme activity was 8.5; however the enzyme remained stable up to pH 10 after 24 hrs of incubation. Similarly, the optimum temperature for enzyme activity was 40 degree C, whereas it remained stable up to 90 degree C with greatly reduced activity. Alkaline protease showed highest specificity towards casein. Among different inhibitors, Phenylmethylsulphonyl fluoride (PMSF) completely inhibited the enzyme activity indicating the serine nature of protease. Similarly, the protease activity was greatly reduced in the presence of MnCl/sub 2/, whereas MgCl/sub 2/ enhanced its activity. The shelf life of the protease was also determined and it was found that the activity of the enzyme came to an end after second week, when the enzyme was stored at room temperature. (author)

  12. Purification and characterization of keratinase from a new Bacillus subtilis strain

    Institute of Scientific and Technical Information of China (English)

    Cheng-gang CAI; Ji-shuang CHEN; Jiong-jiong QI; Yun YIN; Xiao-dong ZHENG

    2008-01-01

    The aim of this study was to purify and characterize a keratinase produced by a new isolated Bacillus subtilis KD-N2strain. The keratinase produced by the isolate was purified using ammonium sulphate precipitation, Sephadex G-75 and DEAE (diethylaminoethyl)-Sepharose chromatographic techniques. The purified enzyme was shown to have a molecular mass of 30.5kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The optimum pH at 50℃ was 8.5 and the optimum temperature at pH 8.5 was 55℃. The keratinase was partially inactivated by some metal ions, organic solvents and serine protease inhibitor phenylmethanesulfonyl fluoride (PMSF). Sodium dodecyl sulfate (SDS) and ethylene diamine tetraacetic acid (EDTA) had positive effect on the keratinase activity. Reducing agents including dithiothreitol (DTT),mercaptoethanol, L-cysteine, sodium sulphite, as well as chemicals of SDS, ammonium sulfamate and dimethylsulfoxide (DMSO)stimulated the enzyme activity upon a feather meal substrate. Besides feather keratin, the enzyme is active upon the soluble proteins ovalbumin, bovine serum albumin (BSA), casein and insoluble ones as sheep wool and human hair. Calf hair, silk and collagen could not be hydrolyzed by the keratinase.

  13. [Fermentation of Bacillus subtilis ge25 strain and preliminary study on its antagonistic substances].

    Science.gov (United States)

    Hu, Chen-Yun; Li, Yong; Liu, Min; Ding, Wan-Long; Qin, Min-Jian

    2014-07-01

    Panax ginseng is one of the most important traditional Chinese herbal medicine, soil borne diseases influenced the yield and quality severely. In our previous work, endophytic Bacillus subtilis ge25 strain was isolated from ginseng root, and which showed significant antagonistic activity against several most destructive ginseng phytopathogens. In the present work, crude protein and lipopeptid extracts were prepared from LB and Landy supernate by salting out, acid precipitation methods respectively. The antagonistic activity of crude extracts and stability to temperature and protease digestion were examined by ginseng phytopathogen Alternaria panax. Results showed that, the antagonistic activity of crude protein extracts from LB culture was complete and partially lost when treated by high temperature and proteinase K. However, crude lipopeptid from Landy culture showed significant stabile antagonistic activity to them. Acid-hydrolyzation and TLC-bioautography analysis showed, that the crude lipopeptide contained at least one cyclic lipopeptide. In consideration of the stability and perfect antagonistic activity of ge25, further researches will promote the biocontrol of ginseng diseases in the field. PMID:25272485

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

    Institute of Scientific and Technical Information of China (English)

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

    2009-01-01

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

  15. ISOLATION OF THE ANTIMICROBIAL CYCLIC PEPTIDE SUBTILOSIN A FROM A GUT-ASSOCIATED BACILLUS SUBTILIS STRAIN

    Directory of Open Access Journals (Sweden)

    Ghislain Schyns

    2013-01-01

    Full Text Available The endospore-forming Bacillus subtilis has been used as probiotics over the last 50 years. However, little is known on how Bacillus spp act in the gut compared to other well-characterized probiotics such as lactic acid bacteria. It is believed that the competitive exclusion of pathogens results from different mode of action notably the production of antimicrobial compounds such as bacteriocins. Here, we report the characterization of the unexpected ability of a gut-associated B. subtilis BSP1 to synthetize the cyclic bacteriocin subtilosin A at high level. Our findings suggest that the BSP1 phenotype could be related, at least in part, to a subsequent increased expression level of the subtilosin A biosynthetic gene cluster sbo-alb in response to a higher activity of the stationery and sporulation master regulator Spo0A.

  16. Concomitant production of two proteases and alpha-amylase by a novel strain of Bacillus subtilis in a microprocessor controlled bioreactor

    OpenAIRE

    Hamid Mukhtar; Ikram-ul-Haq,

    2012-01-01

    We describe the simultaneous production of Bacillus subtilis based proteases and alpha amylase using a computer controlled laboratory scale 7.5 L batch bioreactor. The present strain is the first to be reported that concomitantly produces these two industrially important enzymes. The growth and sporulation of Bacillus subtilis was monitored and maximum production of alkaline protease and alpha amylase was found to coincide with maximum sporulation. Two types of proteases were detected in the ...

  17. Ehanced oil recovery under simulated reservoir conditions using an indigenous Bacillus subtilis strain

    OpenAIRE

    Gudiña, Eduardo J.; Pereira, Jorge F. B.; Costa, A R; L. R. Rodrigues; Coutinho, J.A.P.; J.A. Teixeira

    2013-01-01

    Microbial Enhanced Oil Recovery (MEOR) is potentially useful to increment oil recovery from reservoirs beyond primary and secondary recovery operations using microorganisms and their metabolites. In situ stimulation of microorganisms that produce biosurfactants and degrade heavy oil fractions reduces the capillary forces that retain the oil inside the reservoir and decreases oil viscosity, thus promoting its flow and increasing oil production. Bacillus subtilis #573, isolated from crude oil s...

  18. Antibacterial activity and genotypic-phenotypic characteristics of bacteriocin-producing Bacillus subtilis KKU213: potential as a probiotic strain.

    Science.gov (United States)

    Khochamit, Nalisa; Siripornadulsil, Surasak; Sukon, Peerapol; Siripornadulsil, Wilailak

    2015-01-01

    The antimicrobial activity and probiotic properties of Bacillus subtilis strain KKU213, isolated from local soil, were investigated. The cell-free supernatant (CFS) of a KKU213 culture containing crude bacteriocins exhibited inhibitory effects on Gram-positive bacteria, including Bacillus cereus, Listeria monocytogenes, Micrococcus luteus, and Staphylococcus aureus. The antibacterial activity of the CFS precipitated with 40% ammonium sulfate (AS) remained even after treatment at 60 and 100 °C, at pH 4 and 10 and with proteolytic enzymes, detergents and heavy metals. When analyzed by SDS-PAGE and overlaid with the indicator strains B. cereus and S. aureus, the 40% AS precipitate exhibited inhibitory activity on proteins smaller than 10 kDa. However, proteins larger than 25 kDa and smaller than 10 kDa were still observed on a native protein gel. Purified subtilosin A was prepared by Amberlite XAD-16 bead extraction and HPLC and analyzed by Nano-LC-QTOF-MS. Its molecular mass was found to be 3.4 kDa, and it retained its antibacterial activity. These results are consistent with the detection of the anti-listerial subtilosin A gene of the sbo/alb cluster in the KKU213 strain, which is 100% identical to that of B. subtilis subsp. subtilis 168. In addition to stable and cyclic subtilosin A, a mixture of many extracellular antibacterial peptides was also detected in the KKU213 culture. The KKU213 strain produced extracellular amylase, cellulase, lipase and protease, is highly acid-resistant (pH 2) when cultured in inulin and promotes health and reduces infection of intestinally colonized broiler chickens. Therefore, we propose that bacteriocin-producing B. subtilis KKU213 could be used as a potential probiotic strain or protective culture. PMID:25440998

  19. Essential Bacillus subtilis genes

    DEFF Research Database (Denmark)

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

    2003-01-01

    To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximate to4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes were...... predicted to be essential. The vast majority of essential genes were categorized in relatively few domains of cell metabolism, with about half involved in information processing, one-fifth involved in the synthesis of cell envelope and the determination of cell shape and division, and one-tenth related to...... cell energetics. Only 4% of essential genes encode unknown functions. Most essential genes are present throughout a wide range of Bacteria, and almost 70% can also be found in Archaea and Eucarya. However, essential genes related to cell envelope, shape, division, and respiration tend to be lost from...

  20. Characterization of Extracellular Penicilin G Acylase Produced by A New Local Strain of Bacillus subtilis BAC4

    Directory of Open Access Journals (Sweden)

    SUPARTONO

    2008-06-01

    Full Text Available Penicillin G acylase (PGA which catalyses penicillin G hydrolysis reaction is a key enzyme for the industrial production of penicilin G derivatives used in therapeutics. A new local strain of Bacillus subtilis BAC4 was found capable of producing extracellular PGA. However, characteristics of this extracellular PGA are not known. The goal of this research was to characterize the extracellular PGA produced by B. subtilis BAC4. Enzyme production was carried out by batch fermentation, followed by enzyme purification and characterization of the PGA. The PGA activity was determined by the Kornfeld method, with optimal activity for hydrolysing penicillin G observed at 43 oC and pH 8.5. The activation energy of penicillin G hydrolysis by the PGA of B. subtilis BAC4 was determined as 4.9 kcal.mol-1 and Vmax and Km values were found to be 0.7 µmole.min-1.mg-1 and 3.5 mM respectively. PGA catalytic activity was competitively inhibited by phenylacetic acid with an inhibition constant, Ki(PAA, of 347.2 mM. It was concluded that the extracellular PGA of B. subtilis BAC4 can hydrolyse penicillin G efficiently.

  1. Biosurfactant-producing and oil-degrading Bacillus subtilis strains enhance oil recovery in laboratory sand-pack columns.

    Science.gov (United States)

    Gudiña, Eduardo J; Pereira, Jorge F B; Costa, Rita; Coutinho, João A P; Teixeira, José A; Rodrigues, Lígia R

    2013-10-15

    Microbial Enhanced Oil Recovery (MEOR) technology uses microorganisms and their metabolites to retrieve unrecoverable oil from mature reservoirs. In situ stimulation of biosurfactant-producing and oil-degrading microorganisms reduces the capillary forces retaining the oil inside the reservoir and decreases its viscosity, thus promoting oil flow and consequently production. In this work, a sand-pack column model was designed to simulate oil recovery operations and evaluate mobilization of residual oil by the selected microorganisms. Four different hydrocarbon mixtures and three Bacillus subtilis strains isolated from crude oil samples were used. Additional oil recoveries ranged from 6 to 24% depending on the hydrocarbon mixture and microorganism used. Biosurfactant production was observed with all the microorganisms and hydrocarbon mixtures studied. The oils recovered after incubation with B. subtilis isolates showed a reduction in the percentage of long-chain n-alkanes and lower viscosity when compared with the original oils. The results obtained suggest that stimulation of the selected B. subtilis strains in situ can contribute to mobilize entrapped oil in mature reservoirs. PMID:23911831

  2. Bacillus subtilis Deoxyribonucleic Acid Gyrase

    OpenAIRE

    Sugino, A; Bott, K F

    1980-01-01

    Bacillus subtilis 168 was shown to contain a deoxyribonucleic acid (DNA) gyrase activity which closely resembled those of the enzymes isolated from Escherichia coli and Micrococcus luteus in its enzymatic requirements, substrate specificity, and sensitivity to several antibiotics. The enzyme was purified from the wild type and nalidixic acid-resistant and novobiocin-resistant mutants of B. subtilis and was functionally characterized in vitro. The genetic loci nalA and novA but not novB were s...

  3. Powder formulations of two strains of Bacillus subtilis for control of rape seed damping-off caused by Rhizoctonia solani.

    Science.gov (United States)

    Sharifi-Tehrani, A; Ahmadzadeh, M; Farzaneh, M; Sarani, S

    2006-01-01

    Talc-based formulations of Bacillus subtilis strains B1 and B2 were tested as seed and soil treatments separately for their ability to control Rhizoctonia solani, the causal agent of rape seed damping-off, in greenhouse and field trials. In general, the formulated bacteria was more effective to suppress the disease than the suspension of bacterial cells in carboxymethylcellulose solution (1%, w/v), in both greenhouse and field trials. The formulations of strain B1 as soil treatment and strain B2 as seed treatment in greenhouse, and the formulations of strain B2 as seed and soil treatments in field trials had the greatest effect on reducing the rape seed damping-off (66.7%, 73.3%, 41.3%, and 42.4%, respectively). The formulations of strain B1 as soil treatment and strain B2 as seed treatment were the most effective treatments to increase the root dry weights in the infected soil in greenhouse. The formulation of strain B2 as soil treatment had the greatest effect on enhancement of the fresh weight of roots and stem fresh and dry weights. The formulations of strains B1 and B2 stored at 4 degrees C exhibited better shelf life and efficacy in vitro than their counterparts stored at 25 degrees C. Long-term stability of the formulation of strain B1 was found to be better. PMID:17390784

  4. Isolation and characterization of Bacillus subtilis EB-28, an endophytic bacterium strain displaying biocontrol activity against Botrytis cinerea Pers

    Institute of Scientific and Technical Information of China (English)

    Shutong WANG; Tongle HU; Yanling JIAO; Jianjian WEI; Keqiang CAO

    2009-01-01

    The fungal pathogen Botrytis cinerea Pers. causes severe rotting on tomato fruits during storage and shelf life. As a biological control agent, endophytic bacterium was regarded as an effective alternative to chemical control. Out of 238 endophytic bacterial isolates, three strains (EB-15, EB-28, and EB-122) isolated from Lycopersicum esculentum Mill., Speranskia tuberculata (Bge.) Baill, and Dictamnus dasycarpus Turcz. respectively were found to be strongly antagonistic to the pathogen in vitro and were selected for further in vivo tests. One endophytic bacterium strain, encoded EB-28, was selected from the three in vivo tested isolates. The inhibitive rate of EB-28 reached 71.1% in vitro and 52.4% in vivo. EB-28 was identified as Bacillus subtilis according to its morphological, physiological, and biochemical characteristics and 16S rDNA sequence analysis.

  5. Improvement of antifungal and antibacterial antibiotic producing strain of Bacillus subtilis AFCI-69 by radiation and chemical mutagens. Part of a coordinated programme on radiation biology

    International Nuclear Information System (INIS)

    Gamma radiation was used to select higher antibiotic yield mutants of Bacillus subtilis AECL 69. The test organisms were Aspergillus niger RAGENI 70 and Staphylococcus aureus 6571 (16) N.C.T.C. Searches for fermentation, purification and characterization of antibiotics of parent strain and its mutants were carried out

  6. Isolation and characterization of a β-propeller gene containing phosphobacterium Bacillus subtilis strain KPS-11 for growth promotion of potato (Solanum tuberosum L.)

    NARCIS (Netherlands)

    Hanif, Muhammad Kashif; Hameed, Sohail; Imran, Asma; Naqqash, Tahir; Shahid, Muhammad; Van Elsas, Jan D

    2015-01-01

    Phosphate-solubilizing and phytate-mineralizing bacteria collectively termed as phosphobacteria provide a sustainable approach for managing P-deficiency in agricultural soils by supplying inexpensive phosphate to plants. A phosphobacterium Bacillus subtilis strain KPS-11 (Genbank accession no. KP006

  7. Characterization of a bacteriocin-like substance produced from a novel isolated strain of Bacillus subtilis SLYY-3

    Science.gov (United States)

    Li, Junfeng; Li, Hongfang; Zhang, Yuanyuan; Duan, Xiaohui; Liu, Jie

    2014-12-01

    In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY-3, which produced a bacteriocin-like substance (BLS), was characterized to be a strain of Bacillus subtillis by biochemical profiling and 16S rDNA sequence analysis. It is the first time to report that Bacillus subtilis from Jiaozhou Bay sediments could produce a BLS. The BLS of B. subtillis SLYY-3 exhibited strong inhibitory activity against gram-positive bacteria (including Staphylococcus aureus and B. subtillis) and some fungi (including Penicillium glaucum, Aspergillus niger and Aspergillus flavus). The antimicrobial activity was detected from culture in the exponential growth phase and reached its maximum when culture entered into stationary growth phase. It was thermo-tolerant even when being kept at 100°C for 60 min without losing any activity and stable over a wide pH range from 1.0 to 12.0 while being inactivated by proteolytic enzyme and trypsin, indicating the proteinaceous nature of the BLS. The BLS was purified by precipitation with hydrochloric acid (HCl) and gel filteration (Sephadex G-100). SDS-PAGE analysis of the extracellular peptides of SLYY-3 revealed a bacteriocin-like protein with a molecular mass of 66 kDa. Altogether, these characteristics indicate the potential of the BLS for food industry as a protection against pathogenic and spoilage microorganisms.

  8. Characterization of two antimicrobial peptides produced by a halotolerant Bacillus subtilis strain SK.DU.4 isolated from a rhizosphere soil sample

    OpenAIRE

    Baindara, Piyush; Mandal, Santi M.; Chawla, Niharika; Singh, Pradip Kumar; Pinnaka, Anil Kumar; Korpole, Suresh

    2013-01-01

    A bacterial strain producing two antimicrobial peptides was isolated from a rhizosphere soil sample and identified as Bacillus subtilis based on both phenotypic and 16S rRNA gene sequence phylogenetic analysis. It grew optimally up to 14% NaCl and produced antimicrobial peptide within 24 h of growth. The peptides were purified using a combination of chemical extraction and chromatographic techniques. The MALDI-TOF analysis of HPLC purified fractions revealed that the strain SK.DU.4 secreted a...

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

    OpenAIRE

    Anderson, Iain; Sorokin, Alexei; Kapatral, Vinayak; Reznik, Gary; Bhattacharya, Anamitra; Mikhailova, Natalia; Burd, Henry; Joukov, Victor; Kaznadzey, Denis; Walunas, Theresa; D'Souza, Mark; Larsen, Niels; Pusch, Gordon; Liolios, Konstantinos; Grechkin, Yuri

    2005-01-01

    Genome features of the Bacillus cereus group genomes (representative strains of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis sub spp israelensis) were analyzed and compared with the Bacillus subtilis genome. A core set of 1,381 protein families among the four Bacillus genomes, with an additional set of 933 families common to the B. cereus group, was identified. Differences in signal transduction pathways, membrane transporters, cell surface structures, cell wall, and S-...

  10. Draft Genome Sequence of Commercial Textile Dye-Decolorizing and -Degrading Bacillus subtilis Strain C3 Isolated in India

    Science.gov (United States)

    Kunadia, Khushbu; Nathani, Neelam M.; Kothari, Vishal; Kotadia, Rohit J.; Kothari, Charmy R.; Joshi, Anjali; Rank, Jalpa K.; Faldu, Priti R.; Shekar, M. Chandra; Viroja, Mitkumar J.; Patel, Priyank A.; Jadeja, Divyarajsinh; Reddy, Bhaskar; Pal Singh, Ravindra; Koringa, Prakash G.; Joshi, Chaitanya G.

    2016-01-01

    Bacillus subtilis C3, a commercial textile dye-decolorizing and -degrading bacterium, was isolated from the common effluent treatment plant (CEPT) of the Jetpur textile dyeing and printing industrial sector situated in the district of Rajkot, Gujarat, India. Here, we present the annotated 4.18-Mb draft genome sequence of B. subtilis C3, providing information about the metabolic pathways involved in decolorization and degradation of several commercial textile azo dyes. Thus, we confirm B. subtilis C3 as a potential candidate for bioremediation of textile effluents. PMID:26966205

  11. Water surface tension modulates the swarming mechanics of Bacillus subtilis

    OpenAIRE

    Ke, Wan-Ju; Hsueh, Yi-Huang; Cheng, Yu-Chieh; Wu, Chih-Ching; Liu, Shih-Tung

    2015-01-01

    Many Bacillus subtilis strains swarm, often forming colonies with tendrils on agar medium. It is known that B. subtilis swarming requires flagella and a biosurfactant, surfactin. In this study, we find that water surface tension plays a role in swarming dynamics. B. subtilis colonies were found to contain water, and when a low amount of surfactin is produced, the water surface tension of the colony restricts expansion, causing bacterial density to rise. The increased density induces a quorum ...

  12. Comparative Evaluation of Agroindustrial Byproducts for the Production of Alkaline Protease by Wild and Mutant Strains of Bacillus subtilis in Submerged and Solid State Fermentation

    OpenAIRE

    Mukhtar, Hamid; Haq, Ikramul

    2013-01-01

    The present study describes the screening of different agroindustrial byproducts for enhanced production of alkaline protease by a wild and EMS induced mutant strain of Bacillus subtilis IH-72EMS8. During submerged fermentation, different agro-industrial byproducts were tested which include defatted seed meals of rape, guar, sunflower, gluten, cotton, soybean, and gram. In addition to these meals, rice bran, wheat bran, and wheat flour were also evaluated for protease production. Of all the b...

  13. Isolation and characterization of a β-propeller gene containing phosphobacterium Bacillus subtilis strain KPS-11 for growth promotion of potato (Solanum tuberosum L.

    Directory of Open Access Journals (Sweden)

    Kashif eHanif

    2015-06-01

    Full Text Available Phosphate-solubilizing and phytate-mineralizing bacteria collectively termed as phosphobacteria provide a sustainable approach for managing P-deficiency in agricultural soils by supplying inexpensive phosphate to plants. A phosphobacterium Bacillus subtilis strain KPS-11 (Genbank accession no. KP006655 was isolated from potato (Solanum tuberosum L. rhizosphere and characterized for potato plant growth promoting potential. The strain utilized both Ca-phosphate and Na-phytate in vitro and produced 6.48 µg mL-1 indole-3-acetic acid in tryptophan supplemented medium. P-solubilization after 240 h was 66.4 µg mL-1 alongwith the production of 19.3 µg mL-1 gluconic acid and 5.3 µg mL-1 malic acid. The extracellular phytase activity was higher (4.3 x 10-10 kat mg-1 protein than the cell-associated phytase activity (1.6 x 10-10 kat mg-1protein. B. subtilis strain KPS-11 utilized 40 carbon sources and showed resistance against 20 chemicals in GENIII micro-plate system demonstrating its metabolic potential. Phytase-encoding gene β-propeller (BPP showed 92% amino acid similarity to BPP from B. subtilis (accession no.WP_014114128.1 and 83% structural similarity to BPP from B. subtilis (accession no 3AMR_A. Potato inoculation with B. subtilis strain KPS-11 increased the root/shoot length and root/shoot weight of potato as compared to non-inoculated control plants. Moreover, rifampicin-resistant derivative of KPS-11 were able to survive in the rhizosphere and on the roots of potato up to sixty days showing its colonization potential. The study indicates that B. subtilis strain KPS-11 can be a potential candidate for development of potato inoculum in P-deficient soils.

  14. A biosurfactant-producing and oil-degrading Bacillus subtilis strain enhances oil recovery under simulated reservoir conditions

    OpenAIRE

    Gudiña, Eduardo J.; Pereira, J. F.; Costa, Rita; L. R. Rodrigues; Coutinho, João A. P.; J.A. Teixeira

    2013-01-01

    Microbial Enhanced Oil Recovery (MEOR) is potentially useful to increment oil recovery from reservoirs beyond primary and secondary recovery operations using microorganisms and their metabolites. In situ stimulation of microorganisms that produce biosurfactants and degrade heavy oil fractions reduces the capillary forces that retain the oil inside the reservoir and decreases oil viscosity, thus promoting its flow and increasing oil production. Bacillus subtilis #573, isolated from crude oil s...

  15. Screening and characterization of extracelluar L-asparaginase producing Bacillus subtilis strain hswx88, isolated from Taptapani hotspring of Odisha, India

    Institute of Scientific and Technical Information of China (English)

    Biswaprakash Pradhan; Sashi K Dash; Sabuj Sahoo

    2013-01-01

    Objective: To screen and isolate an eco-friendly, a thermophilic and potent L-asparaginase producing bacterium, with novel immunological properties that may obviates hypersensitivity reactions. Methods: In the present study bacterial strain isolated for extracellular L-asparaginase production from hotspring, identified by morphological, biochemical and physiological tests followed by 16S rDNA technology and the L-asparaginase production ability was tested by both semi quantitative and quantitative enzymatic assay. Result:The bacterial strain was identified as Bacillus subtilis strain hswx88 (GenBank Accession Number: JQ237656.1). The extracellular enzyme yielding capacity isolate Bacillus subtilis strain hswx88 (23.8 IU/mL) was found to be 1.7 and 14.5 times higher than the reference organism Pectobacterium carotovorum MTCC 1428 (14.2 IU/mL) and Bacillus sp. BCCS 034 (1.64 IU/mL). Conclusion: The isolate is eco-friendly and useful to produce bulk quantity of extracellular, thermophilic L-asparaginase for the treatment of various tumor cases and for preparation of acrylamide free fry food preparation.

  16. Enhancement of Cellulase Production by Cellulomonas Fimi and Bacillus Subtilis

    International Nuclear Information System (INIS)

    Two bacterial strains identified as Cellulomonas fimi and Baciliius subtilus are cosidered as highly active cellulytic bacteria. Trials for maximizing the cellulolytic activites of the two strains were conducted. A maximum cellulase production was achieved at 1 and 1.5%carboxy methyl cellulose as carbon source, sodium nitrate and yeast as nitrogen source for Cellulomonas fimi and Bacillus subtilis, respectively. Incubation temprature at 30 and 45 degree C, ph at 6 and 7 achieved the highest activity of cellulase for Cellulomonas fimi and bacillus subtilis, respectively

  17. Triple fixation of Bacillus subtilis dormant spores.

    OpenAIRE

    Kozuka, S; Tochikubo, K

    1983-01-01

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

  18. Fast Neutron Radiation Effects on Bacillus Subtili

    Institute of Scientific and Technical Information of China (English)

    CHEN Xiaoming; REN Zhenglong; ZHANG Jianguo; ZHENG Chun; TAN Bisheng; YANG Chengde; CHU Shijin

    2009-01-01

    To examine the sterilizing effect and mechanism of neutron radiation, Bacillus sub-tilis vat. niger, strain (ATCC 9372) spores were irradiated with the fast neutron from the Chinese fast burst reactor Ⅱ(CFBR-Ⅱ). The plate-count results indicated that the D10 value was 384.6 Gy with a neutron radiation dose rate of 7.4 Gy/min. The rudimental catalase activity of the spores declined obviously with the increase in the radiation dose. Meanwhile, under the scanning electron microscope, no visible influence of the neutron radiation on the spore configuration was detected even if the dose was increased to 4 kGy. The content and distribution of DNA double-strand breaks induced by neutron radiation at different doses were measured and quantified by pulsed-field gel electrophoresis (PFGE). Further analysis of the DNA release percentage (PR), the DNA breakage level (L), and the average molecular weight, indicated that DNA fragments were obvi-ously distributed around the 5 kb regions at different radiation doses, which suggests that some points in the DNA molecule were sensitive to neutron radiation. Both PR and L varied regularly to some extent with the increase in radiation dose. Thus neutron radiation has a high sterilization power, and can induce falling enzyme activity and DNA breakage in Bacillus subtilis spores

  19. Degradation of proteins during the fermentation of African locust bean (Parkia biglobosa) by strains of Bacillus subtilis and Bacillus pumilus for production of Soumbala

    DEFF Research Database (Denmark)

    Ouoba, L.I.I.; Rechinger, K.B.; Barkholt, Vibeke;

    2003-01-01

    Aims: To examine isolates of Bacillus subtilis and B. pumilus predominant in Soumbala for their ability to degrade African locust bean proteins (ALBP).Methods and Results: Agar diffusion test in casein and ALBP agar was used for screening of isolates. The profiles of water-soluble proteins and free...... amino acids (FAA) during the fermentation of ALBP by the Bacillus isolates were studied by SDS-PAGE and cation exchange chromatography. The profile of soluble proteins changed with the fermentation time and varied depending on the isolate. The quantity of total FAA and essential FAA such as lysine was...... increased sharply between 24 and 48 h of fermentation and differed among the isolates. Simultaneously, a pH increase was observed. Cysteine, methionine, leucine, isoleucine, tyrosine and phenylalaline appeared during fermentation.Conclusion: The Bacillus isolates studied degraded ALBP leading to a profile...

  20. Comparative Analysis Of The Development Of Swarming Communities Of Bacillus Subtilis In Case Of Pta And ComXP Mutant Strains

    Directory of Open Access Journals (Sweden)

    Kassem Hamze

    2015-08-01

    Full Text Available Abstract Swarming Experiments were carried on with Bacillus subtilis strains to identify the activity of certain genes in the swarming ability and surfactin production. We will examine the effect of comXP as well as pta mutations on the capability of swarming. In different experiments we showed that strain OMG 903 that carries mutation in comXP managed to produce surfactin but showed attenuated defective and random swarming pattern strain OMG 928 that carries mutation in pta gene managed to produce surfactin and showed normal swarming pattern meanwhile double mutation in comXP and pta in strain OMG 929 lead to the absence of surfactin production and didnt manage Thesetoswarmdatashowed. that a threshold of surfactin production is necessary for a normal swarming pattern.

  1. Comparative evaluation of agroindustrial byproducts for the production of alkaline protease by wild and mutant strains of Bacillus subtilis in submerged and solid state fermentation.

    Science.gov (United States)

    Mukhtar, Hamid; Haq, Ikramul

    2013-01-01

    The present study describes the screening of different agroindustrial byproducts for enhanced production of alkaline protease by a wild and EMS induced mutant strain of Bacillus subtilis IH-72(EMS8). During submerged fermentation, different agro-industrial byproducts were tested which include defatted seed meals of rape, guar, sunflower, gluten, cotton, soybean, and gram. In addition to these meals, rice bran, wheat bran, and wheat flour were also evaluated for protease production. Of all the byproducts tested, soybean meal at a concentration of 20 g/L gave maximum production of the enzyme, that is, 5.74  ±  0.26 U/mL from wild and 11.28  ±  0.45 U/mL from mutant strain, during submerged fermentation. Different mesh sizes (coarse, medium, and fine) of the soybean meal were also evaluated, and a finely ground soybean meal (fine mesh) was found to be the best. In addition to the defatted seed meals, their alkali extracts were also tested for the production of alkaline protease by Bacillus subtilis, but these were proved nonsignificant for enhanced production of the enzyme. The production of the enzyme was also studied in solid state fermentation, and different agro-industrial byproducts were also evaluated for enzyme production. Wheat bran partially replaced with guar meal was found as the best substrate for maximum enzyme production under solid state fermentation conditions. PMID:24294129

  2. Comparative Evaluation of Agroindustrial Byproducts for the Production of Alkaline Protease by Wild and Mutant Strains of Bacillus subtilis in Submerged and Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Hamid Mukhtar

    2013-01-01

    Full Text Available The present study describes the screening of different agroindustrial byproducts for enhanced production of alkaline protease by a wild and EMS induced mutant strain of Bacillus subtilis IH-72EMS8. During submerged fermentation, different agro-industrial byproducts were tested which include defatted seed meals of rape, guar, sunflower, gluten, cotton, soybean, and gram. In addition to these meals, rice bran, wheat bran, and wheat flour were also evaluated for protease production. Of all the byproducts tested, soybean meal at a concentration of 20 g/L gave maximum production of the enzyme, that is, 5.74  ±  0.26 U/mL from wild and 11.28  ±  0.45 U/mL from mutant strain, during submerged fermentation. Different mesh sizes (coarse, medium, and fine of the soybean meal were also evaluated, and a finely ground soybean meal (fine mesh was found to be the best. In addition to the defatted seed meals, their alkali extracts were also tested for the production of alkaline protease by Bacillus subtilis, but these were proved nonsignificant for enhanced production of the enzyme. The production of the enzyme was also studied in solid state fermentation, and different agro-industrial byproducts were also evaluated for enzyme production. Wheat bran partially replaced with guar meal was found as the best substrate for maximum enzyme production under solid state fermentation conditions.

  3. [Comparative Sensitivity of the Luminescent Photobacterium phosphoreum, Escherichia coli, and Bacillus subtilis Strains to Toxic Effects of Carbon-Based Nanomaterials and Metal Nanoparticles].

    Science.gov (United States)

    Deryabina, D G; Efremova, L V; Karimov, I F; Manukhov, I V; Gnuchikh, E Yu; Miroshnikov, S A

    2016-01-01

    A comparative analysis of the four commercially available and laboratory luminescent sensor strains to the toxic effect of 10 carbon-based nanomatherials (CBNs) and 10 metal nanoparticles (MNPs) was carried out in this study. The bioluminescence inhibition assays with marine Photobacterium phosphoreum and recombinant Escherichia coli strains were varied in minimal toxic concentrations and EC50 values but led to well correlated biotoxicity evaluation for the most active compounds were ranked as Cu > (MgO, CuO) > (fullerenol, graphene oxide). The novel sensor strain Bacillus subtilis EG 168-1 exhibited the highest sensitivity to CBNs and MNPs that increased significantly number of toxic compounds causing the bacterial bioluminescence inhibition effect. PMID:27476206

  4. Regulation of Polyglutamic Acid Synthesis by Glutamate in Bacillus licheniformis and Bacillus subtilis

    OpenAIRE

    Kambourova, Margarita; Tangney, Martin; Priest, Fergus G.

    2001-01-01

    The synthesis of polyglutamic acid (PGA) was repressed by exogenous glutamate in strains of Bacillus licheniformis but not in strains of Bacillus subtilis, indicating a clear difference in the regulation of synthesis of capsular slime in these two species. Although extracellular γ-glutamyltranspeptidase (GGT) activity was always present in PGA-producing cultures of B. licheniformis under various growth conditions, there was no correlation between the quantity of PGA and enzyme activity. Moreo...

  5. Concomitant production of two proteases and alpha-amylase by a novel strain of Bacillus subtilis in a microprocessor controlled bioreactor

    Directory of Open Access Journals (Sweden)

    Hamid Mukhtar

    2012-09-01

    Full Text Available We describe the simultaneous production of Bacillus subtilis based proteases and alpha amylase using a computer controlled laboratory scale 7.5 L batch bioreactor. The present strain is the first to be reported that concomitantly produces these two industrially important enzymes. The growth and sporulation of Bacillus subtilis was monitored and maximum production of alkaline protease and alpha amylase was found to coincide with maximum sporulation. Two types of proteases were detected in the fermentation broth; a neutral and an alkaline protease most active in a pH range of 7.0-8.0 and 8.0-10, respectively. Maximum production of proteases was observed at an incubation temperature of 37ºC while that of alpha amylase was observed at 40ºC. The optimum aeration and agitation levels for protease production were 0.6 L/L/min and 200rpm, respectively, and for alpha amylase were 0.6 L/L/min and 150 rpm. The kinetic parameters Yp/x and qp were also found to be significant at the given fermentation conditions.

  6. Study of the Antifungal Ability of Bacillus subtilis Strain PY-1 in Vitro and Identification of its Antifungal Substance (Iturin A)

    Institute of Scientific and Technical Information of China (English)

    Meng GONG; Jiang-Dong WANG; Jing ZHANG; Hao YANG; Xiao-Feng LU; Yan PEI; Jing-Qiu CHENG

    2006-01-01

    A Bacillus strain, denoted as PY- 1, was isolated from the vascular bundle of cotton. Biochemical,physiological and 16S rDNA sequence analysis proved that it should belong to Bacillus subtilis. The PY-1 strain showed strong ability against many common plant fungal pathogens in vitro. The antibiotics produced by this strain were stable in neutral and basic conditions, and not sensitive to high temperature. From the culture broth of PY- 1 strain, five antifungal compounds were isolated by acidic precipitation, methanol extraction, gel filtration and reverse-phase HPLC. Advanced identification was performed by mass spectrometry and nuclear magnetic resonance spectroscopy. These five antifungal compounds were proved to be the isomers of iturin A: A2, A3, A4, A6 and A7. In fast atom bombardment mass spectrometry/mass spectrometry collision-induced dissociation spectra, fragmentation ions from two prior linear acylium ions were observed, and the prior ion, Tyr-Asn-Gln-Pro-Asn-Ser-βAA-Asn-CO+, was first reported.

  7. Transformation of Bacillus Subtilis with cloned thymidylate synthetases

    Energy Technology Data Exchange (ETDEWEB)

    Rubin, Edward M.

    1980-01-01

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

  8. Simple, fast and high‐efficiency transformation system for directed evolution of cellulase in Bacillus subtilis

    OpenAIRE

    Zhang, Xiao‐Zhou; Zhang, Y.‐H. Percival

    2010-01-01

    Summary Bacillus subtilis can serve as a powerful platform for directed evolution, especially for secretory enzymes. However, cloning and transformation of a DNA mutant library in B. subtilis are not as easy as they are in Escherichia coli. For direct transformation of B. subtilis, here we developed a new protocol based on supercompetent cells prepared from the recombinant B. subtilis strain SCK6 and multimeric plasmids. This new protocol is simple (restriction enzyme‐, phosphatase‐ and ligas...

  9. Mapping of internal monophosphate 5' ends of Bacillus subtilis messenger RNAs and ribosomal RNAs in wild-type and ribonuclease-mutant strains.

    Science.gov (United States)

    DiChiara, Jeanne M; Liu, Bo; Figaro, Sabine; Condon, Ciarán; Bechhofer, David H

    2016-04-20

    The recent findings that the narrow-specificity endoribonuclease RNase III and the 5' exonuclease RNase J1 are not essential in the Gram-positive model organism,Bacillus subtilis, facilitated a global analysis of internal 5' ends that are generated or acted upon by these enzymes. An RNA-Seq protocol known as PARE (Parallel Analysis of RNA Ends) was used to capture 5' monophosphorylated RNA ends in ribonuclease wild-type and mutant strains. Comparison of PARE peaks in strains with RNase III present or absent showed that, in addition to its well-known role in ribosomal (rRNA) processing, many coding sequences and intergenic regions appeared to be direct targets of RNase III. These target sites were, in most cases, not associated with a known antisense RNA. The PARE analysis also revealed an accumulation of 3'-proximal peaks that correlated with the absence of RNase J1, confirming the importance of RNase J1 in degrading RNA fragments that contain the transcription terminator structure. A significant result from the PARE analysis was the discovery of an endonuclease cleavage just 2 nts downstream of the 16S rRNA 3' end. This latter observation begins to answer, at least forB. subtilis, a long-standing question on the exonucleolytic versus endonucleolytic nature of 16S rRNA maturation. PMID:26883633

  10. Crystallization and preliminary X-ray analysis of the rhamnogalacturonan lyase YesW from Bacillus subtilis strain 168, a member of polysaccharide lyase family 11

    International Nuclear Information System (INIS)

    The crystallization and preliminary X-ray characterization of the polysaccharide lyase family 11 rhamnogalacturonan lyase are presented. Rhamnogalacturonan lyases degrade rhamnogalacturonan I, a major component of pectin, through a β-elimination reaction. YesW from Bacillus subtilis strain 168 is a novel rhamnogalacturonan lyase classified into polysaccharide lyase family 11 (PL-11). The enzyme was crystallized at 293 K using the sitting-drop vapour-diffusion method with 2-methyl-2,4-pentanediol (MPD) as a precipitant. Preliminary X-ray analysis revealed that the YesW crystals belong to space group P21 and diffract to 2.40 Å resolution, with unit-cell parameters a = 56.7, b = 105.6, c = 101.4 Å, β = 94.9°. This is the first report on the crystallization and preliminary X-ray analysis of a family PL-11 rhamnogalacturonan lyase

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

    OpenAIRE

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

    2016-01-01

    Bacillus subtilis strain B7-S screened from18 strains is an aerobic, endospore-forming, model organism of Gram-positive bacteria which is capable to form vanillin during ferulic acid bioconversion. The bioconversion of ferulic acid to vanillin by Bacillus subtilis B7-S (B. subtilis B7-S) was investigated. Based on our results, the optimum bioconversion conditions for the production of vanillin by B. subtilis B7-S can be summarized as follows: temperature 35 °C; initial pH 9.0; inoculum volume...

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

    Directory of Open Access Journals (Sweden)

    ELFI SUSANTI

    2003-01-01

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

  13. Protein-Tyrosine Phosphorylation in Bacillus subtilis

    DEFF Research Database (Denmark)

    Mijakovic, Ivan; Petranovic, Dina; Bottini, N.; Deutscher, J.; Jensen, Peter Ruhdal

    2005-01-01

    phosphorylation, indicating that this post-translational modifi cation could regulate physiological processes ranging from stress response and exopolysaccharide synthesis to DNA metabolism. Some interesting work in this fi eld was done in Bacillus subtilis , and we here present the current state of knowledge on...

  14. Complete Genome of Bacillus subtilis Myophage Grass

    OpenAIRE

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

    2013-01-01

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

  15. 有机溶剂胁迫处理对菌株Bacillus subtilis OST23a产胞外多糖的影响%Effects of n-hexane stress on secretion of antioxidant exopolysaccharides from marine strain Bacillus subtilis OST23a

    Institute of Scientific and Technical Information of China (English)

    房耀维; 刘姝; 王淑军; 吕明生; 焦豫良; 陈丽; 曹纯

    2012-01-01

    [Objective] In this paper, the effects of organic solvent stresses on the yield of an-tioxidant exopolysaccharide (EPS) from Bacillus subtilis was evaluated and the best condition was also determined. [Methods] Marine strain Bacillus subtilis OST23a and mutant UM29, both with the ability to produce antioxidant exopolysaccharide (EPS), were used as the original strain. Based on the detection of the tolerance of the strains to the organic solvent, n-hexane was used to stress treatment. Effects of concentrations and treatment time of n-hexane on the exopolysaccharide excretion from Bacillus subtilis OST23a and strain UD292 were studied. [Results] The productivity of the EPS of Bacillus subtilis OST23a and strain UD292 were 52.97 mg/L and 201.81 mg/L respectively after stress treatment with 3% n-hexane for 6 h. There was no significant difference in the antioxidation activities of the extracellular polysac-charides between strains stress with n-hexane and the original strains. Moreover, the continuous passage experiment showed that the strains have high genetic stability. [Conclusion] The organic solvent stress could improve the productivity of the exopolysaccharide from bacteria, which possesses the potential application in microbial breeding.%[目的] 研究有机溶剂胁迫处理对菌株分泌胞外多糖的影响并确定最佳条件.[方法] 利用分泌抗氧化活性胞外多糖海洋细菌Bacillus subtilis OST23a及其突变菌株UD292为出发菌株,在考察菌株有机溶剂耐受性的基础上,测定不同浓度正己烷胁迫处理不同时间后该菌株抗氧化胞外多糖产量.[结果] 结果表明最佳胁迫处理浓度和时间分别为3%和6h,此时Bacillus subtilis OST23a和菌株UD292胞外多糖分泌量分别从9.02 mg/L和43.92 mg/L显著提高到52.97 mg/L和201.81 mg/L,且胞外多糖的抗氧化性能无显著变化.Bacillus subtilis OST23a和菌株UD292连续传代试验结果表明菌株遗传性状较稳定.[结论] 有机溶剂

  16. Protein-Tyrosine Phosphorylation in Bacillus subtilis

    DEFF Research Database (Denmark)

    Mijakovic, Ivan; Petranovic, Dina; Bottini, N.;

    2005-01-01

    phosphorylation, indicating that this post-translational modifi cation could regulate physiological processes ranging from stress response and exopolysaccharide synthesis to DNA metabolism. Some interesting work in this fi eld was done in Bacillus subtilis , and we here present the current state of knowledge on...... protein-tyrosine phosphorylation in this gram-positive model organism. With its two kinases, two kinase modulators, three phosphatases and at least four different tyrosine-phosphorylated substrates, B. subtilis is the bacterium with the highest number of presently known participants in the global network...

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

    NARCIS (Netherlands)

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

    1997-01-01

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

  18. 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

  19. Decolorization of Distillery Effluent by Thermotolerant Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Soni Tiwari

    2012-01-01

    Full Text Available Problem statement: Ethanol production from sugarcane molasses generate large volume of effluent containing high Biological Oxygen Demand (BOD and Chemical Oxygen Demand (COD along with melanoidin, a color compound generally produced by Millard reaction. Melanodin is a recalcitrant compound degraded by specific microorganisms having ability to produce mono and di-oxygenases peroxidases, phenoxidases and laccases, are mainly responsible for degradation of complex aromatic hydrocarbons like color compound. These compounds causes several toxic effects on living system, therefore may be treated before disposal. Approach: The purpose of this study was to isolate a potential thermotolerant melanoidin decolorizing bacterium from natural resources for treatment of distillery effluent at industrial level. Results: Total 10 isolates were screened on solid medium containing molasses pigments. Three potential melanoidin decolorizing thermotolerant bacterial isolates identified as Bacillus subtilis, Bacillus cereus and Pseudomonas sp. were further optimized for decolorization at different physico-chemical and nutritional level. Out of these three, Bacillus subtilis showed maximum decolorization (85% at 45°C using (w/v 0.1%, glucose; 0.1%, peptone; 0.05%, MgSO4; 0.01%, KH2PO4; pH-6.0 within 24h of incubation under static condition. Conclusion/Recommendations: The strain of Bacillus subtilis can tolerate higher temperature and required very less carbon (0.1%, w/v and nitrogen sources (0.1%, w/v in submerged fermentation. It can be utilized for melanoidin decolorization of distillery effluent at industrial scale.

  20. 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.

  1. Characterization of two antimicrobial peptides produced by a halotolerant Bacillus subtilis strain SK.DU.4 isolated from a rhizosphere soil sample.

    Science.gov (United States)

    Baindara, Piyush; Mandal, Santi M; Chawla, Niharika; Singh, Pradip Kumar; Pinnaka, Anil Kumar; Korpole, Suresh

    2013-01-01

    A bacterial strain producing two antimicrobial peptides was isolated from a rhizosphere soil sample and identified as Bacillus subtilis based on both phenotypic and 16S rRNA gene sequence phylogenetic analysis. It grew optimally up to 14% NaCl and produced antimicrobial peptide within 24 h of growth. The peptides were purified using a combination of chemical extraction and chromatographic techniques. The MALDI-TOF analysis of HPLC purified fractions revealed that the strain SK.DU.4 secreted a bacteriocin-like peptide with molecular mass of 5323.9 Da and a surface-active lipopeptide (m/z 1056 Da). The peptide mass fingerprinting of low-molecular-weight bacteriocin exhibited significant similarity with stretches of secreted lipoprotein of Methylomicrobium album BG8 and displayed 70% sequence coverage. MALDI MS/MS analysis elucidated the lipopeptide as a cyclic lipopeptide with a β-hydroxy fatty acid linked to Ser of a peptide with seven α-amino acids (Asp-Tyr-Asn-Gln-Pro-Asn-Ser) and assigned it to iturin-like group of antimicrobial biosurfactants. However, it differed in amino acid composition with other members of the iturin family. Both peptides were active against Gram-positive bacteria, suggesting that they had an additive effect. PMID:23289832

  2. Effect of decoyinine on the regulation of alpha-amylase synthesis in Bacillus subtilis.

    OpenAIRE

    Nicholson, W L; Chambliss, G H

    1987-01-01

    Decoyinine, an inhibitor of GMP synthetase, allows sporulation in Bacillus subtilis to initiate and proceed under otherwise catabolite-repressing conditions. The effect of decoyinine on alpha-amylase synthesis in B. subtilis, an event which exhibits regulatory features resembling sporulation initiation, was examined. Decoyinine did not overcome catabolite repression of alpha-amylase synthesis in a wild-type strain of B. subtilis but did cause premature and enhanced synthesis in a mutant strai...

  3. Production, purification, and characterization of a-amylase by Bacillus subtilis and its mutant derivates

    OpenAIRE

    DEMİRKAN, Elif

    2011-01-01

    The effects of various carbon and nitrogen sources on production of a-amylase by Bacillus subtilis and its mutant derivates were investigated. The maximum production of a-amylase by all strains was obtained in the presence of mesoinositol as the carbon source. There was no more significant increase in enzyme yield in the case of the supplementation of nitrogen sources, whereas malt extract and tryptone were preferred nitrogen sources for amylase production by Bacillus subtilis and mutant U 2-...

  4. The studies on radiation mutation breeding of Bacillus subtilis with high-yield of amylase

    International Nuclear Information System (INIS)

    The mutagenesis effects on the yield of amylase have been investigated with Bacillus subtilis irradiated by γ-rays and fast neutrons in once or twice irradiation at various dose rates and total irradiation doses. Several parameters such as flat transparent circle, colony diameter, transparent circle diameter and the ratio of flat transparent circle to colony diameter (HC) are used to estimate the radiation mutation of Bacillus subtilis. A series of results has been obtained as (1) Irradiation both with neutrons and γ-rays could make Bacillus subtilis mutationed to produce high-yield amylase effectively. (2) The average colony diameter of Bacillus subtilis irradiated by γ-rays or fast neutrons is smaller than that of control group at various total doses and dose rates. And their colony diameter becomes smaller slightly with the increment of γ-rays irradiation dose. (3) After the second neutrons irradiation, the values of average colony diameter, the biggest colony diameter, average transparent circle diameter and the biggest transparent circle diameter of all mutationed Bacillus subtilis exceed that of original strains greatly. (4) Three kinds of mutationed Bacillus subtilis strains with high-yield amylase have been screened out, in which two strains can produce high-yield amylase steadily after 15 times breeding. Their biggest colony diameter, the biggest transparent circle diameter and the biggest HC value are up to 8.32 mm, 22.38 mm and 5.39 respectively. (authors)

  5. Prodigiosin Induces Autolysins in Actively Grown Bacillus subtilis Cells.

    Science.gov (United States)

    Danevčič, Tjaša; Borić Vezjak, Maja; Tabor, Maja; Zorec, Maša; Stopar, David

    2016-01-01

    Prodigiosin produced by marine bacterium Vibrio ruber DSM 14379 exhibits a potent antimicrobial activity against a broad range of Gram positive and Gram negative bacteria. The mechanism of prodigiosin antimicrobial action, however, is not known. In this work, the effect of prodigiosin on Bacillus subtilis growth, cell membrane leakage, and induction of autolysins was studied. Treating B. subtilis with prodigiosin resulted in rapid decline of optical density and increased cell membrane leakage measured by β-galactosidase activity. Cell lysis was initiated immediately after treatment with prodigiosin in the middle exponential phase and was completed within 2 h. Lytic activity of prodigiosin in mutant strains with impaired autolysin genes lytABCD decreased for 80% compared to the wild type strain, while in lytABCDEF mutant strain prodigiosin had no bacteriolytic but only bacteriostatic effect. Fast prodigiosin lytic activity on individual B. subtilis cells was confirmed by a modified comet assay. The results indicate that prodigiosin autolysin induction in B. subtilis is growth phase dependent. PMID:26858704

  6. Environmental Persistence of Bacillus anthracis and Bacillus subtilis Spores

    Science.gov (United States)

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

    2015-01-01

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

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

    Institute of Scientific and Technical Information of China (English)

    CHEN Tao; CHEN Xun; WANG Jingyu; ZHAO Xueming

    2005-01-01

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

  8. Cannibalism stress response in Bacillus subtilis.

    Science.gov (United States)

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

    2016-01-01

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

  9. Comparison of different Bacillus subtilis expression systems.

    Science.gov (United States)

    Vavrová, Ludmila; Muchová, Katarína; Barák, Imrich

    2010-11-01

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

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

    OpenAIRE

    ziba Akbari; Hashem Nayeri; Keivan Beheshtimaal

    2015-01-01

    Introduction: Amylases are among the most important enzymes and have great significance in present-day biotechnology. Amylase with commercial applications is mainly derived from the genus Bacillus. The main purpose of this study is identification and isolatation amylase enzyme producer Bacillus, determining the amylase enzyme activity and affecting a number of culture medium on amylase enzyme production. Materials and methods: Soil, water and wastewater samples were collected from agricul...

  11. Hyaluronic Acid Production in Bacillus subtilis

    OpenAIRE

    Widner, Bill; Behr, Régine; Von Dollen, Steve; Tang, Maria; Heu, Tia; Sloma, Alan; Sternberg, Dave; DeAngelis, Paul L; Paul H. Weigel; Brown, Steve

    2005-01-01

    The hasA gene from Streptococcus equisimilis, which encodes the enzyme hyaluronan synthase, has been expressed in Bacillus subtilis, resulting in the production of hyaluronic acid (HA) in the 1-MDa range. Artificial operons were assembled and tested, all of which contain the hasA gene along with one or more genes encoding enzymes involved in the synthesis of the UDP-precursor sugars that are required for HA synthesis. It was determined that the production of UDP-glucuronic acid is limiting in...

  12. Bacillus subtilis pur operon expression and regulation.

    OpenAIRE

    Ebbole, D J; Zalkin, H

    1989-01-01

    The Bacillus subtilis pur operon is a 12-gene cluster, purEKB-purC(orf)QLF-purMNH(J)-purD, organized in groups of overlapping coding units separated by intercistronic gaps. Translational fusions of Escherichia coli lacZ were constructed to purE, purC, and purM, the first gene of each group. Analyses of gene fusions integrated into the chromosomal pur operon exclude the possibility of internal promoters in intercistronic regions and support the view that transcription is from the single sigma ...

  13. Bacillus subtilis regulatory protein GerE

    OpenAIRE

    Ducros, V M A; Brannigan, J.A.; Lewis, R J; Wilkinson, A.J.

    1998-01-01

    GerE is the latest-acting of a series of factors which regulate gene expression in the mother cell during sporulation in Bacillus. The gene encoding GerE has been cloned from B. subtilis and overexpressed in Escherichia coli. Purified GerE has been crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The small plate-like crystals belong to the monoclinic space group C2 and diffract beyond 2.2 Angstrom resolution with a synchrotron radiation X-ra...

  14. Homolactic fermentation from glucose and cellobiose using Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Martinez Alfredo

    2009-04-01

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

  15. Hyper production of alkaline protease by mutagenized bacillus subtilis

    International Nuclear Information System (INIS)

    The purpose of this work was to augment the alkaline protease production from Bacillus subtilis by using chemical mutagen (MMS) and UV mutagenesis. A number of mutants were isolated which produce high levels of extra cellular proteases. Analysis of culture supernatants of these mutants had shown that the total amounts of proteolysis activity were increased from 1 to 2 fold over the wild strain. Clones showing promote response were further characterized by analyzing different parameters; like of Temperature, pH substrate concentration and incubation period, to study the activity of protease enzyme. (author)

  16. Nonribosomal Peptide Synthase Gene Clusters for Lipopeptide Biosynthesis in Bacillus subtilis 916 and Their Phenotypic Functions

    OpenAIRE

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

    2014-01-01

    Bacillus cyclic lipopeptides (LPs) have been well studied for their phytopathogen-antagonistic activities. Recently, research has shown that these LPs also contribute to the phenotypic features of Bacillus strains, such as hemolytic activity, swarming motility, biofilm formation, and colony morphology. Bacillus subtilis 916 not only coproduces the three families of well-known LPs, i.e., surfactins, bacillomycin Ls (iturin family), and fengycins, but also produces a new family of LP called loc...

  17. Stoichiometric growth model for riboflavin-producing Bacillus subtilis.

    Science.gov (United States)

    Dauner, M; Sauer, U

    2001-09-01

    Rate equations for measured extracellular rates and macromolecular composition data were combined with a stoichiometric model to describe riboflavin production with an industrial Bacillus subtilis strain using errors in variables regression analysis. On the basis of this combined stoichiometric growth model, we explored the topological features of the B. subtilis metabolic reaction network that was assembled from a large amount of literature. More specifically, we simulated maximum theoretical yields of biomass and riboflavin, including the associated flux regimes. Based on the developed model, the importance of experimental data on building block requirements for maximum yield and flux calculations were investigated. These analyses clearly show that verification of macromolecular composition data is important for optimum flux calculations. PMID:11505383

  18. A part toolbox to tune genetic expression in Bacillus subtilis.

    Science.gov (United States)

    Guiziou, Sarah; Sauveplane, Vincent; Chang, Hung-Ju; Clerté, Caroline; Declerck, Nathalie; Jules, Matthieu; Bonnet, Jerome

    2016-09-01

    Libraries of well-characterised components regulating gene expression levels are essential to many synthetic biology applications. While widely available for the Gram-negative model bacterium Escherichia coli, such libraries are lacking for the Gram-positive model Bacillus subtilis, a key organism for basic research and biotechnological applications. Here, we engineered a genetic toolbox comprising libraries of promoters, Ribosome Binding Sites (RBS), and protein degradation tags to precisely tune gene expression in B. subtilis We first designed a modular Expression Operating Unit (EOU) facilitating parts assembly and modifications and providing a standard genetic context for gene circuits implementation. We then selected native, constitutive promoters of B. subtilis and efficient RBS sequences from which we engineered three promoters and three RBS sequence libraries exhibiting ∼14 000-fold dynamic range in gene expression levels. We also designed a collection of SsrA proteolysis tags of variable strength. Finally, by using fluorescence fluctuation methods coupled with two-photon microscopy, we quantified the absolute concentration of GFP in a subset of strains from the library. Our complete promoters and RBS sequences library comprising over 135 constructs enables tuning of GFP concentration over five orders of magnitude, from 0.05 to 700 μM. This toolbox of regulatory components will support many research and engineering applications in B. subtilis. PMID:27402159

  19. Bacillus subtilis strain deficient for the protein-tyrosine kinase PtkA exhibits impaired DNA replication

    DEFF Research Database (Denmark)

    Petranovic, Dina; Michelsen, Ole; Zahradka, K;

    2007-01-01

    study. We were unable to identify any striking phenotypes related to control of UDP-glucose dehydrogenases, natural competence and DNA lesion repair; however, a very strong phenotype of ΔptkA emerged with respect to DNA replication and cell cycle control, as revealed by flow cytometry and fluorescent...... microscopy. B. subtilis cells lacking the kinase PtkA accumulated extra chromosome equivalents, exhibited aberrant initiation mass for DNA replication and an unusually long D period....... previously shown to regulate the phosphorylation state of UDP-glucose dehydrogenases and single-stranded DNA-binding proteins. This promiscuity towards substrates is reminiscent of eukaryal kinases and has prompted us to investigate possible physiological effects of ptkA and ptpZ gene inactivations in this...

  20. Absence of correlation between rates of cell wall turnover and autolysis shown by Bacillus subtilis mutants.

    OpenAIRE

    Vitković, L; Cheung, H. Y.; Freese, E

    1984-01-01

    Bacillus subtilis mutants with reduced rates of cell wall autolysis reached a constant rate of wall turnover after a longer lag than the standard strain but eventually showed the same turnover rate. In reverse, a turnover-deficient mutant autolysed at a slightly higher rate than the standard strain. Consequently, there is no correlation between the rates of cell wall turnover and autolysis.

  1. Bacillus subtilis as potential producer for polyhydroxyalkanoates

    Directory of Open Access Journals (Sweden)

    Patel Sanjay KS

    2009-07-01

    Full Text Available Abstract Polyhydroxyalkanoates (PHAs are biodegradable polymers produced by microbes to overcome environmental stress. Commercial production of PHAs is limited by the high cost of production compared to conventional plastics. Another hindrance is the brittle nature and low strength of polyhydroxybutyrate (PHB, the most widely studied PHA. The needs are to produce PHAs, which have better elastomeric properties suitable for biomedical applications, preferably from inexpensive renewable sources to reduce cost. Certain unique properties of Bacillus subtilis such as lack of the toxic lipo-polysaccharides, expression of self-lysing genes on completion of PHA biosynthetic process – for easy and timely recovery, usage of biowastes as feed enable it to compete as potential candidate for commercial production of PHA.

  2. Bacillus subtilis Spore Inner Membrane Proteome.

    Science.gov (United States)

    Zheng, Linli; Abhyankar, Wishwas; Ouwerling, Natasja; Dekker, Henk L; van Veen, Henk; van der Wel, Nicole N; Roseboom, Winfried; de Koning, Leo J; Brul, Stanley; de Koster, Chris G

    2016-02-01

    The endospore is the dormant form of Bacillus subtilis and many other Firmicutes. By sporulation, these spore formers can survive very harsh physical and chemical conditions. Yet, they need to go through germination to return to their growing form. The spore inner membrane (IM) has been shown to play an essential role in triggering the initiation of germination. In this study, we isolated the IM of bacterial spores, in parallel with the isolation of the membrane of vegetative cells. With the use of GeLC-MS/MS, over 900 proteins were identified from the B. subtilis spore IM preparations. By bioinformatics-based membrane protein predictions, ca. one-third could be predicted to be membrane-localized. A large number of unique proteins as well as proteins common to the two membrane proteomes were identified. In addition to previously known IM proteins, a number of IM proteins were newly identified, at least some of which are likely to provide new insights into IM physiology, unveiling proteins putatively involved in spore germination machinery and hence putative germination inhibition targets. PMID:26731423

  3. Biological control of Colletotrichum panacicola on Panax ginseng by Bacillus subtilis HK-CSM-1

    OpenAIRE

    Ryu, Hojin; Park, Hoon; Suh, Dong-Sang; Jung, Gun Ho; Park, Kyungseok; Lee, Byung Dae

    2014-01-01

    Background Biological control of plant pathogens using benign or beneficial microorganisms as antagonistic agents is currently considered to be an important component of integrated pest management in agricultural crops. In this study, we evaluated the potential of Bacillus subtilis strain HK-CSM-1 as a biological control agent against Colletotrichum panacicola. Methods The potential of B. subtilis HK-CSM-1 as a biological control agent for ginseng anthracnose was assessed. C. panacicola was i...

  4. Levan from Bacillus subtilis Natto: its effects in normal and in streptozotocin-diabetic rats

    OpenAIRE

    Fernando Cesar Bazani Cabral de Melo; Cássia Thaïs Bussamra Viera Zaia; Maria Antonia Pedrine Colabone Celligoi

    2012-01-01

    Levan is an exopolysaccharide of fructose primarily linked by β-(2→6) glycosidic bonds with some β-(2→1) branched chains. Due to its chemical properties, levan has possible applications in both the food and pharmaceutical industries. Bacillus subtilis is a promising industrial levan producer, as it ferments sucrose and has a high levan-formation capacity. A new strain of B. subtilis was recently isolated from Japanese food natto, and it has produced levan in large quanti...

  5. Pseudosecretion of Escherichia coli chloramphenicol acetyltransferase by Bacillus subtilis.

    OpenAIRE

    Le Grice, S F; Gentz, R; Bannwarth, W; Kocher, H. P.

    1987-01-01

    Bacillus subtilis harboring the vector 25RBSII secrets an Escherichia coli-derived chloramphenicol acetyltransferase into culture supernatants. The secreted enzyme lacks 18 amino acids; these are removed externally rather than during secretion.

  6. Two Genes Encoding Uracil Phosphoribosyltransferase Are Present in Bacillus subtilis

    DEFF Research Database (Denmark)

    Martinussen, Jan; Glaser, Philippe; Andersen, Paal S.;

    1995-01-01

    Uracil phosphoribosyltransferase (UPRTase) catalyzes the key reaction in the salvage of uracil in many microorganisms. Surprisingly, two genes encoding UPRTase activity were cloned from Bacillus subtilis by complementation of an Escherichia coli mutant. The genes were sequenced, and the putative...

  7. Selection of Bacillus subtilis mutants impaired in ammonia assimilation.

    OpenAIRE

    Dean, D R; Aronson, A I

    1980-01-01

    The selection of Bacillus subtilis mutants capable of using D-histidine to fulfill a requirement for L-histidine resulted in mutants with either no glutamate synthase activity or increased amounts of an altered glutamine synthetase.

  8. Degradation of proteins during the fermentation of African locust bean (Parkia biglobosa) by strains of Bacillus subtilis and Bacillus pumilus for production of Soumbala

    DEFF Research Database (Denmark)

    Ouoba, L.I.I.; Rechinger, K.B.; Barkholt, Vibeke; Diawara, B.; Traore, A.S.; Jakobsen, M.

    2003-01-01

    amino acids (FAA) during the fermentation of ALBP by the Bacillus isolates were studied by SDS-PAGE and cation exchange chromatography. The profile of soluble proteins changed with the fermentation time and varied depending on the isolate. The quantity of total FAA and essential FAA such as lysine was...

  9. Expression of UGA-Containing Mycoplasma Genes in Bacillus subtilis

    OpenAIRE

    Kannan, T. R.; Baseman, Joel B.

    2000-01-01

    We used Bacillus subtilis to express UGA-containing Mycoplasma genes encoding the P30 adhesin (one UGA) of Mycoplasma pneumoniae and methionine sulfoxide reductase (two UGAs) of Mycoplasma genitalium. Due to natural UGA suppression, these Mycoplasma genes were expressed as full-length protein products, but at relatively low efficiency, in recombinant wild-type Bacillus. The B. subtilis-expressed Mycoplasma proteins appeared as single bands and not as multiple bands compared to expression in r...

  10. Transformation of Bacillus subtilis by single-stranded plasmid DNA.

    OpenAIRE

    Rudolph, C F; Schmidt, B J; Saunders, C W

    1986-01-01

    The single-stranded form of a pE194-based plasmid transformed Bacillus subtilis protoplasts at least as efficiently as did the double-stranded plasmid, but the single-stranded form did not detectably transform B. subtilis competent cells.

  11. Scientific Opinion on the safety and efficacy of Bacillus subtilis PB6 (Bacillus subtilis) as a feed additive for laying hens and minor poultry species for laying

    OpenAIRE

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

    2015-01-01

    Bacillus subtilis PB6 is the trade name for a feed additive based on viable spores of a strain of Bacillus subtilis. This species is considered by EFSA to be suitable for the qualified presumption of safety approach to safety assessment. This approach requires the identity of the active agent to be established and the absence of toxigenic potential and resistance to antibiotics of human or veterinary clinical significance to be demonstrated. No evidence of toxigenic potential or of resistance...

  12. Biocontrol of Soil Fungi in Tomato with Microencapsulates Containing Bacillus subtilis

    OpenAIRE

    Marcela H. Suarez; Francisco D. Hernandez-Castillo; Gabriel Gallegos-Morales; R. H. Lira-Saldivar; Raul Rodriguez-Herrera; Aguilar, Cristobal N.

    2011-01-01

    Problem statement: An option to reduce pollution by synthetic agro-chemical in root plant disease management is the use of antagonist rhizobacteria belonging to Bacillus genus, because their inhibitory properties, stimulation of plant growth and crop yield increase. Approach: This study was carried out in order to evaluate if Bacillus subtilis strains could play an antagonists role of plant pathogens and if they can be microencapsulated inside a biopolymer matrix. It was adapted an equipment ...

  13. 75 FR 862 - Bacillus subtilis; Registration Review Proposed Decision; Notice of Availability

    Science.gov (United States)

    2010-01-06

    ..., cucurbits, fruiting vegetables, herbs, leafy crops, legumes, ornamental plants and cuttings, peanuts, root... grasses, wheat, barley, corn, and canola. It is also used in greenhouses to treat peat moss and soil..., strawberry, tuber/root and corm vegetables, turf, sod, lawns, trees, and shrubs; Bacillus subtilis strain...

  14. Purification and properties of phosphoribosyl-diphosphate synthetase from Bacillus subtilis

    DEFF Research Database (Denmark)

    Arnvig, Kirsten; Hove-Jensen, Bjarne; Switzer, Robert L.

    1990-01-01

    Phosphoribosyl-diphosphate (PPRibP) synthetase from Bacillus subtiliis has been purified to near homogeneity from an Escherichia coli Δprs strain bearing the cloned B. subtilis prs gene, encoding PPRibP synthentase, on a plasmid. The Mr of the subunit (34,000) and its amino-terminal amino acid...

  15. Involvement of deoxyribonucleic acid polymerase III in W-reactivation in Bacillus subtilis.

    OpenAIRE

    Fields, P I; Yasbin, R E

    1980-01-01

    6-(p-Hydroxyphenylazo)-uracil, a purine analog that is known to specifically inhibit deoxyribonucleic acid polymerase III in gram-positive organisms, inhibited W-reactivation in Bacillus subtilis. On the other hand, W-reactivation proceeded normally in the presence of 6-(p-hydroxyphenylazo)-uracil when a strain possessing a resistant deoxyribonucleic acid polymerase III was used as the host.

  16. Postreplication repair of ultraviolet-irradiated transforming deoxyribonucleic acid in Bacillus subtilis

    International Nuclear Information System (INIS)

    Repair of ultraviolet-irradiated transforming deoxyribonucleic acid (DNA) in several strains of Bacillus subtilis was studied in order to determine the effects of excision repair and postreplication repair on transformation. Two mutations that cause a Uvr- phenotype (uvr-1 and uvr-42) were shown to have strikingly different effects on repair of ultraviolet-irradiated transforming DNA

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

    Directory of Open Access Journals (Sweden)

    Gidi Pelchovich

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

  18. Biocontrol of Soil Fungi in Tomato with Microencapsulates Containing Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Marcela H. Suarez

    2011-01-01

    Full Text Available Problem statement: An option to reduce pollution by synthetic agro-chemical in root plant disease management is the use of antagonist rhizobacteria belonging to Bacillus genus, because their inhibitory properties, stimulation of plant growth and crop yield increase. Approach: This study was carried out in order to evaluate if Bacillus subtilis strains could play an antagonists role of plant pathogens and if they can be microencapsulated inside a biopolymer matrix. It was adapted an equipment and evaluated a technique for microcapsules elaboration, in order to incorporate B. subtilis strains and to analyze their potential as biocontrol agents by determining their antagonistic effect against pathogenic soil fungi; in addition, it was analyzed their effect on tomato plant growth promotion under greenhouse conditions. B. subtilis strains identified as B1, J1, M2 and their mixture were used; microcapsules containing bacterial strains were inoculated to tomato seeds cv. Floradade. When seedlings emerged, a second application of microcapsules containing B. subtilis was performed on the pots, which previously were inoculated with the fungi Rhizoctonia solani and Fusarium oxysporum. Response variables were: Incidence and disease severity, plant growth, aerial and root dry weight, leaf area and fruit yield. Results: The outcome showed that the equipment designed and adapted for microcapsules elaboration was useful to obtain microcapsules containing the bacterial strains. B. subtilis strains exerted apparent biocontrol, since incidence and disease severity was reduced and for that reason inhibited the infective activity of the inoculated plant pathogens, also microcapsules containing Bacillus strains stimulated tomato growth and fruit yield. Conclusion: Microcapsules containing B. subtilis strains could be effective biocontrol agents against soil fungi plant pathogens and could have a potential biofertilizer effect, since they stimulated growth and yield

  19. PRODUCTION OF CALCIUM- AND IRON-BINDING PEPTIDES BY PROBIOTIC STRAINS OF Bacillus subtilis, B. clausii AND B. coagulans GBI-30

    Directory of Open Access Journals (Sweden)

    A.I. Reyes-Méndez

    2015-01-01

    Full Text Available Algunas cepas de Bacillus subtilis, B. clausii y B. coagulans se utilizan comercialmente como probióticos y su actividad proteolítica es de sobra conocida. El objetivo de este trabajo fue la evaluación de estos microorganismos debido a su capacidad para producir péptidos fijadores de calcio y hierro cuando son cultivados en medios con diferentes fuentes de nitrógeno. Los medios para la fermentación incluyeron caldos con soya y triptona (ST, peptona de case´ına (PC, peptona de soya (PS y, un extracto crudo de ficobiliproteína (ECF. Se obtuvieron sobrenadantes libres de células (SLC a partir de los caldos de fermentación y se determinó el grado de hidrólisis (GH y la bioactividad fijadora para calcio y hierro. El GH y la actividad fijadora de minerales fue mayor cuando se emplearon inóculos preparados a partir de un medio diseñado para la producción de proteasas alcalinas (AP. El mejor resultado para el GH y la bioactividad fijadora de calcio se obtuvo con B. subtilis y B. coagulans cultivados en caldo con ECF como fuente de nitrógeno. Se observaron resultados similares para la actividad fijadora de hierro, sin embargo, también se obtuvieron buenos resultados con B. clausii cuando se utilizaron para su crecimiento los caldos ST, PC y PS.

  20. Production of biosurfactant lipopeptides Iturin A, fengycin and surfactin A from Bacillus subtilis CMB32 for control of Colletotrichum gloeosporioides.

    Science.gov (United States)

    Kim, Pyoung Il; Ryu, Jaewon; Kim, Young Hwan; Chi, Youn-Tae

    2010-01-01

    A bacterial strain isolated from soil for its potential to control the anthracnose disease caused by Colletotrichum gloeosporioides was identified as a Bacillus subtilis. Bacillus subtilis CMB32 produced antifungal agents on M9 broth at 30degreesC. Biosurfactant lipopeptides produced by Bacillus subtilis CMB32 were precipitated by adjusting to pH 2 and extracting using chloroform/methanol, and then were purified using column chromatography and reverse-phase HPLC. Molecular masses of the lipopeptides were estimated by MALDI-TOF mass spectrometry as (a) 1080, (b) 1486, and (c) 1044 Da, respectively. They had cyclic structures and amino acid compositions of (a) Pro, Asx, Ser, Tyr, Glx, (b) Glx, Tyr, Thr, Ala, Pro, Ile, and (c) Glx, Leu, Val, Asx, respectively. Further analysis revealed that Bacillus subtilis CMB32 produced three antifungal lipopeptides: (a) iturin A, (b) fengycin, and (c) surfactin A. PMID:20134245

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

    Science.gov (United States)

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

    2015-01-01

    Bacillus cyclic lipopeptides (LPs) have been well studied for their phytopathogen-antagonistic activities. Recently, research has shown that these LPs also contribute to the phenotypic features of Bacillus strains, such as hemolytic activity, swarming motility, biofilm formation, and colony morphology. Bacillus subtilis 916 not only coproduces the three families of well-known LPs, i.e., surfactins, bacillomycin Ls (iturin family), and fengycins, but also produces a new family of LP called locillomycins. The genome of B. subtilis 916 contains four nonribosomal peptide synthase (NRPS) gene clusters, srf, bmy, fen, and loc, which are responsible for the biosynthesis of surfactins, bacillomycin Ls, fengycins, and locillomycins, respectively. By studying B. subtilis 916 mutants lacking production of one, two, or three LPs, we attempted to unveil the connections between LPs and phenotypic features. We demonstrated that bacillomycin Ls and fengycins contribute mainly to antifungal activity. Although surfactins have weak antifungal activity in vitro, the strain mutated in srfAA had significantly decreased antifungal activity. This may be due to the impaired productions of fengycins and bacillomycin Ls. We also found that the disruption of any LP gene cluster other than fen resulted in a change in colony morphology. While surfactins and bacillomycin Ls play very important roles in hemolytic activity, swarming motility, and biofilm formation, the fengycins and locillomycins had little influence on these phenotypic features. In conclusion, B. subtilis 916 coproduces four families of LPs which contribute to the phenotypic features of B. subtilis 916 in an intricate way. PMID:25362061

  2. 40 CFR 180.1128 - Bacillus subtilis MBI 600; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus subtilis MBI 600; exemption... FOOD Exemptions From Tolerances § 180.1128 Bacillus subtilis MBI 600; exemption from the requirement of... biofungicide Bacillus subtilis MBI 600 in or on all food commodities, including residues resulting from...

  3. Fitness Trade-Offs in Competence Differentiation of Bacillus subtilis

    Science.gov (United States)

    Yüksel, Melih; Power, Jeffrey J.; Ribbe, Jan; Volkmann, Thorsten; Maier, Berenike

    2016-01-01

    In the stationary phase, Bacillus subtilis differentiates stochastically and transiently into the state of competence for transformation (K-state). The latter is associated with growth arrest, and it is unclear how the ability to develop competence is stably maintained, despite its cost. To quantify the effect differentiation has on the competitive fitness of B. subtilis, we characterized the competition dynamics between strains with different probabilities of entering the K-state. The relative fitness decreased with increasing differentiation probability both during the stationary phase and during outgrowth. When exposed to antibiotics inhibiting cell wall synthesis, transcription, and translation, cells that differentiated into the K-state showed a selective advantage compared to differentiation-deficient bacteria; this benefit did not require transformation. Although beneficial, the K-state was not induced by sub-MIC concentrations of antibiotics. Increasing the differentiation probability beyond the wt level did not significantly affect the competition dynamics with transient antibiotic exposure. We conclude that the competition dynamics are very sensitive to the fraction of competent cells under benign conditions but less sensitive during antibiotic exposure, supporting the picture of stochastic differentiation as a fitness trade-off. PMID:27375604

  4. Fitness Trade-Offs in Competence Differentiation of Bacillus subtilis.

    Science.gov (United States)

    Yüksel, Melih; Power, Jeffrey J; Ribbe, Jan; Volkmann, Thorsten; Maier, Berenike

    2016-01-01

    In the stationary phase, Bacillus subtilis differentiates stochastically and transiently into the state of competence for transformation (K-state). The latter is associated with growth arrest, and it is unclear how the ability to develop competence is stably maintained, despite its cost. To quantify the effect differentiation has on the competitive fitness of B. subtilis, we characterized the competition dynamics between strains with different probabilities of entering the K-state. The relative fitness decreased with increasing differentiation probability both during the stationary phase and during outgrowth. When exposed to antibiotics inhibiting cell wall synthesis, transcription, and translation, cells that differentiated into the K-state showed a selective advantage compared to differentiation-deficient bacteria; this benefit did not require transformation. Although beneficial, the K-state was not induced by sub-MIC concentrations of antibiotics. Increasing the differentiation probability beyond the wt level did not significantly affect the competition dynamics with transient antibiotic exposure. We conclude that the competition dynamics are very sensitive to the fraction of competent cells under benign conditions but less sensitive during antibiotic exposure, supporting the picture of stochastic differentiation as a fitness trade-off. PMID:27375604

  5. Fitness trade-offs in competence differentiation of Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Melih Yüksel

    2016-06-01

    Full Text Available In the stationary phase, Bacillus subtilis differentiates stochastically and transiently into the state of competence for transformation (K-state. The latter is associated with growth arrest, and it is unclear how the ability to develop competence is stably maintained, despite its cost. To quantify the effect differentiation has on the competitive fitness of B. subtilis, we characterized the competition dynamics between strains with different probabilities of entering the K-state. The relative fitness decreased with increasing differentiation probability both during the stationary phase and during outgrowth. When exposed to antibiotics inhibiting cell wall synthesis, transcription, and translation, cells that differentiated into the K-state showed a selective advantage compared to differentiation-deficient bacteria; this benefit did not require transformation. Although beneficial, the K-state was not induced by sub-MIC concentrations of antibiotics. Increasing the differentiation probability beyond the wt level did not significantly affect the competition dynamics with transient antibiotic exposure. We conclude that the competition dynamics are very sensitive to the fraction of competent cells under benign conditions but less sensitive during antibiotic exposure, supporting the picture of stochastic differentiation as a fitness trade-off.

  6. The antagonistic strain Bacillus subtilis UMAF6639 also confers protection to melon plants against cucurbit powdery mildew by activation of jasmonate- and salicylic acid-dependent defence responses

    OpenAIRE

    Garcia-Gutierrez, Laura; Zeriouh, Houda; Romero, Diego; Cubero, Jaime; de Vicente, Antonio; Përez-García, Alejandro

    2013-01-01

    Biological control of plant diseases has gained acceptance in recent years. Bacillus subtilis UMAF6639 is an antagonistic strain specifically selected for the efficient control of the cucurbit powdery mildew fungus Podosphaera fusca, which is a major threat to cucurbits worldwide. The antagonistic activity relies on the production of the antifungal compounds iturin and fengycin. In a previous study, we found that UMAF6639 was able to induce systemic resistance (ISR) in melon and provide addit...

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

    Science.gov (United States)

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

    2016-01-01

    Bacillus subtilis strain B7-S screened from18 strains is an aerobic, endospore-forming, model organism of Gram-positive bacteria which is capable to form vanillin during ferulic acid bioconversion. The bioconversion of ferulic acid to vanillin by Bacillus subtilis B7-S (B. subtilis B7-S) was investigated. Based on our results, the optimum bioconversion conditions for the production of vanillin by B. subtilis B7-S can be summarized as follows: temperature 35 °C; initial pH 9.0; inoculum volume 5%; ferulic acid concentration 0.6 g/L; volume of culture medium 20%; and shaking speed 200 r/min. Under these conditions, several repeated small-scale batch experiments showed that the maximum conversion efficiency was 63.30% after 3 h of bioconversion. The vanillin products were confirmed by spectral data achieved from UV-vis, inductively coupled plasma atomic emission spectroscope (ICP-AES) and Fourier transform infrared spectrometer (FT-IR) spectra. Scanning electron microscopy (SEM) and transmission electron spectroscopy (TEM) results confirmed that the cell surface of B. subtilis plays a role in the induction of ferulic acid tolerance. These results demonstrate that B. subtilis B7-S has the potential for use in vanillin production through bioconversion of ferulic acid. PMID:26841717

  8. Characterization of an L-arabinose isomerase from Bacillus subtilis

    DEFF Research Database (Denmark)

    Kim, Jin-Ha; Prabhu, Ponnandy; Jeya, Marimuthu;

    2010-01-01

    An isolated gene from Bacillus subtilis str. 168 encoding a putative isomerase was proposed as an L-arabinose isomerase (L-AI), cloned into Escherichia coli, and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1,491 bp, capable of encoding a polypep......An isolated gene from Bacillus subtilis str. 168 encoding a putative isomerase was proposed as an L-arabinose isomerase (L-AI), cloned into Escherichia coli, and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1,491 bp, capable of encoding...

  9. Cucumber Rhizosphere Microbial Community Response to Biocontrol Agent Bacillus subtilis B068150

    OpenAIRE

    Lihua Li; Jincai Ma; A. Mark Ibekwe; Qi Wang; Ching-Hong Yang

    2015-01-01

    Gram-positive bacteria Bacillus subtilis B068150 has been used as a biocontrol agent against the pathogen Fusarium oxysporum cucumerinum. Cucumber was grown in three soils with strain B068150 inoculated in a greenhouse for 90 days, and the colonization ability of strain B068150 in cucumber rhizosphere and non-rhizosphere soils was determined. Changes in total bacteria and fungi community composition and structures using denaturing gradient gel electrophoresis (DGGE) and sequencing were determ...

  10. 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.

  11. Deciphering the conserved genetic loci implicated in plant disease control through comparative genomics of Bacillus amyloliquefaciens subsp. plantarum strains

    OpenAIRE

    Hossain, Mohammad J.; Chao eRan; Ke eLiu; Choong-Min eRyu; Rasmussen-Ivey, Cody R.; Williams, Malachi A.; Mohammad K. Hassan; Soo-Keun eChoi; Haeyoung eJeong; Molli eNewman; Kloepper, Joseph W.; Mark R Liles

    2015-01-01

    To understand the growth-promoting and disease-inhibiting activities of plant growth-promoting rhizobacteria (PGPR) strains, the genomes of 12 Bacillus subtilis group strains with PGPR activity were sequenced and analyzed. These B. subtilis strains exhibited high genomic diversity, whereas the genomes of B. amyloliquefaciens strains (a member of the B. subtilis group) are highly conserved. A pairwise BLASTp matrix revealed that gene family similarity among Bacillus genomes ranges from 32- 90%...

  12. Isolation and Characterization of Phages Infecting Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Anna Krasowska

    2015-01-01

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

  13. Co-producing iturin A and poly-γ-glutamic acid from rapeseed meal under solid state fermentation by the newly isolated Bacillus subtilis strain 3-10.

    Science.gov (United States)

    Yao, Dehui; Ji, Zhixia; Wang, Changjun; Qi, Gaofu; Zhang, Lili; Ma, Xin; Chen, Shouwen

    2012-03-01

    The strain 3-10 was isolated from soil and identified as B. subtilis according to morphological and physiological characteristics and nucleotide sequence of 16S rRNA. It co-produced anti-fungal iturin A and fertilizer synergist of poly-γ-glutamic acid (γ-PGA) under solid state fermentation (SSF) with rapeseed meal. The co-production of iturin A and γ-PGA reached 5.3 and 51.3 g/kg-dry weight culture, respectively, and the number of viable cells reached 1.9 × 10(10) CFU/g-dry weight culture. In pot tests, the shoot length and dry weight of watermelon seedlings treated by the SSF culture improved by 48.0 and 30.8%, respectively compared to the control; and its biocontrol effect on watermelon fusarium wilt achieved 89.6%. These results highlight a novel strategy to exploit the low-cost and widely available rapeseed meal as dual-functional bio-organic fertilizer under SSF by B. subtilis. PMID:22805819

  14. Partial purification, characterization, and kinetic studies of a low-molecular-weight, alkali-tolerant chitinase enzyme from Bacillus subtilis JN032305, A potential biocontrol strain.

    Science.gov (United States)

    Shivakumar, Srividya; Karmali, Anika Nayak; Ruhimbana, Charles

    2014-01-01

    A new alkalophilic low-molecular-mass chitinase of 14 kD from the potent biocontrol agent Bacillus subtilis JN032305 was partially purified and enzymology of the chitinase was studied. The enzyme showed optimal pH of 9.0 and temperature of 50°C. The enzyme was found stable during the 60-min incubation at 50 °C. The chitinase was inhibited by group specific agents like IAA, DAN, TLCK, and SDS and metal ions Mg(2+), Ca(2+), Fe(2+), Mn(2+), Ba(2+), and Hg(2+), whereas Zn(2+) did not show significant inhibitory effect against the chitinase. PMSF partially inhibited the enzyme. Substrates specificity tests indicated that the enzyme showed 75% of relative activity on glycol chitin, 58% on carboxymethylcellulose (CMC), 33% on chitin flakes, and 166% laminarin compared to that on colloidal chitin. The enzyme also hydrolyzed 4-methylumbelliferyl-N-acetyl-D-glucosaminide, indicating its chitobiase activity. The chitinase of this study has broad specificity, which could hydrolyze not only the glycosidic bond in GlcNAc-GlcNAc but also that of related carbohydrates with glycosidic linkages. The partially purified chitinase not only showed antifungal activity against Rhizoctonia solani and Colletotrichum gloeosporioides, two potent phytopathogens of chilli, but also increased the germination of chilli seeds when infected with the two potent phytopathogenic fungi. PMID:24499366

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

    Directory of Open Access Journals (Sweden)

    Danielle eDuanis-Assaf

    2016-01-01

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

  17. Sigma A recognition sites in the Bacillus subtilis genome

    DEFF Research Database (Denmark)

    Jarmer, Hanne Østergaard; Larsen, Thomas Schou; Krogh, Anders Stærmose;

    2001-01-01

    A hidden Markov model of sigma (A) RNA polymerase cofactor recognition sites in Bacillus subtilis, containing either the common or the extended -10 motifs, has been constructed based on experimentally verified sigma (A) recognition sites. This work suggests that more information exists at the ini...

  18. Molecular physiology of weak organic acid stress in Bacillus subtilis

    OpenAIRE

    Brul, S.; Beilen, van, J.W.A.

    2013-01-01

    The mechanism by which weak organic acid (WOA) preservatives inhibit growth of microorganisms may differ between different WOAs and these differences are not well understood. The aim of this thesis has been to obtain a better understanding of the mode of action of these preservatives by which they inhibit the growth of spore-forming bacteria (more specifically Bacillus subtilis).

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

    Institute of Scientific and Technical Information of China (English)

    Guo Hong LI; Yue Mao SHEN; Ke Qin ZHANG

    2005-01-01

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

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

    DEFF Research Database (Denmark)

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

    2009-01-01

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

  1. Sigma A recognition sites in the Bacillus subtilis genome

    DEFF Research Database (Denmark)

    Jarmer, Hanne Østergaard; Larsen, Thomas Schou; Krogh, Anders Stærmose; Saxild, Hans Henrik; Brunak, Søren; Knudsen, Steen

    2001-01-01

    A hidden Markov model of sigma (A) RNA polymerase cofactor recognition sites in Bacillus subtilis, containing either the common or the extended -10 motifs, has been constructed based on experimentally verified sigma (A) recognition sites. This work suggests that more information exists at the...

  2. Inhibition of Bacillus cereus growth by bacteriocin producing Bacillus subtilis isolated from fermented baobab seeds (maari) is substrate dependent.

    Science.gov (United States)

    Kaboré, Donatien; Nielsen, Dennis Sandris; Sawadogo-Lingani, Hagrétou; Diawara, Bréhima; Dicko, Mamoudou Hama; Jakobsen, Mogens; Thorsen, Line

    2013-03-01

    Maari is a spontaneously alkaline fermented food condiment made from baobab tree seeds. Due to the spontaneous nature of maari fermentations growth of the opportunistic human pathogen Bacillus cereus is occasionally observed. Bacillus subtilis strains are important for alkaline seed fermentations because of their enzymatic activities contributing to desirable texture, flavor and pH development. Some B. subtilis strains have antimicrobial properties against B. cereus. In the present work, three bacteriocin producing B. subtilis strains (B3, B122 and B222) isolated from maari were tested. The production of antimicrobial activity by the three strains was found to be greatly influenced by the substrate. All three B. subtilis strains produced antimicrobial activity against B. cereus NVH391-98 in BHI broth as determined by the agar well diffusion assay, whereas no antimicrobial activity was detected in whole cooked baobab seeds and in 10% (w/v) grinded baobab seeds. Incorporation of BHI with up to 5% (w/w) grinded baobab seeds enhanced the antimicrobial activity of B. subtilis compared with pure BHI in a strain dependent manner. Incorporation of BHI with 50% (w/w) baobab grinded seeds decreased the antimicrobial activity. Addition of the inorganic salts FeCl₃, MgSO₄ and MnSO₄ has previously been reported to increase bacteriocin production of B. subtilis, but the addition of these salts to 10% (w/v) grinded baobab seed broth did not cause antimicrobial activity. Survival of B. cereus NVH391-98 in co-culture with B. subtilis was tested in BHI broth, 10% (w/v) grinded baobab seed based broth and during baobab seed fermentation to produce maari. B. cereus NVH391-98 grew well in all three substrates in mono-culture. All the 3 B. subtilis strains were able to decrease B. cereus NVH391-98 to levels below the detection limit (starter culture candidates originating from maari which are able to prevent pathogen outgrowth remain to be identified. PMID:23376785

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

    OpenAIRE

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

    2014-01-01

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

  4. Identification of Bacillus subtilis genes expressed early during sporulation.

    Science.gov (United States)

    Mathiopoulos, C; Sonenshein, A L

    1989-08-01

    Labelled cDNA transcribed in vitro from early-sporulation RNA was enriched for sporulation-specific sequences by subtractive hybridization to an excess of vegetative RNA and used to probe libraries of Bacillus subtilis chromosomal DNA. From the initial collection of clones that coded for RNAs transcribed preferentially during sporulation, several were subcloned and studied in more detail. It was found that two clones contained sequences (dciA and dciB) that had an undetectable level of transcription during vegetative growth but had transcripts that started to appear no later than eight minutes after induction of sporulation. A third DNA segment (dciC) was expressed at a low level in vegetative cells and increased within four minutes after induction of sporulation. The effects of spoO mutations, i.e. mutations that prevent cells from reaching stage I of the sporulation process, were tested. Induction of the dciA and dciB transcripts was significantly reduced in strains carrying mutations in the spoOA and spoOH genes but not in a spoOB mutant strain. In addition, a product of the abrB locus, a locus in which mutations are known to partially overcome the pleiotropic effect of spoOA and spoOB mutations, seemed to be required for dciA and dciB expression. PMID:2481799

  5. Phylogeny and Molecular Taxonomy of the Bacillus subtilis species Complex and the Description of Bacillus subtilis subsp. inaquosorum subsp. nov

    Science.gov (United States)

    The Bacillus subtilis species complex is a tight assemblage of closely related species. For many years, it has been recognized that these species cannot be differentiated on the basis of phenotypic characteristics. Recently, it has been shown that phylogenetic analysis of the 16S ribosomal RNA gen...

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

    DEFF Research Database (Denmark)

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

    2002-01-01

    DNA microarrays were used to analyze the changes in gene expression in Bacillus subtilis strain 168 when nitrogen limiting (glutamate) and nitrogen excess (ammonium plus glutamate) growth conditions were compared. Among more than 100 genes that were significantly induced during nitrogen starvation...... we detected the comG, comF, comE, nin-nucA and comK transcription units together with recA. DNA was added to B. subtilis grown in minimal medium with glutamate as the sole nitrogen source and it was demonstrated that the cells were competent. Based on these observations we propose a simplification of...... previously designed one-step transformation procedures for B. subtilis strain 168....

  7. Genome sequence of Bacillus subtilis subsp. spizizenii gtP20b, isolated from the Indian ocean.

    Science.gov (United States)

    Fan, Longjiang; Bo, Shiping; Chen, Huan; Ye, Wanzhi; Kleinschmidt, Katrin; Baumann, Heike I; Imhoff, Johannes F; Kleine, Michael; Cai, Daguang

    2011-03-01

    Bacillus subtilis is an aerobic spore-forming Gram-positive bacterium that is a model organism and of great industrial significance as the source of diverse novel functional molecules. Here we present, to our knowledge, the first genome sequence of Bacillus subtilis strain gtP20b isolated from the marine environment. A subset of candidate genes and gene clusters were identified, which are potentially involved in production of diverse functional molecules, like novel ribosomal and nonribosomal antimicrobial peptides. The genome sequence described in this paper is due to its high strain specificity of great importance for basic as well as applied researches on marine organisms. PMID:21183663

  8. Isolation and characterization of a novel extracellular metalloprotease from Bacillus subtilis.

    OpenAIRE

    Rufo, G A; Sullivan, B J; Sloma, A; Pero, J

    1990-01-01

    We have isolated and characterized two minor extracellular proteases from culture supernatants of a strain of Bacillus subtilis containing deletion mutations of the genes for the extracellular proteases subtilisin (apr) and neutral protease (npr) and a minor extracellular protease (epr) as well as intracellular serine protease-I (isp-1). Characterization studies have revealed that one of these enzymes is the previously described protease bacillopeptidase F. The second enzyme, the subject of t...

  9. Rapid isolation and sequencing of purified plasmid DNA from Bacillus subtilis.

    OpenAIRE

    Voskuil, M. I.; Chambliss, G H

    1993-01-01

    We report two methods for isolation of plasmid DNA from the gram-positive bacterium Bacillus subtilis. The protoplast alkaline lysis procedure was developed for general use, and the protoplast alkaline lysis magic procedure was developed for isolation of DNA for sequencing. Both procedures yielded large amounts of high-quality DNA in less than 1 h, while current protocols require 4 to 7 h to perform and give lower yields and quality. Plasmid DNA was obtained from strains containing either hig...

  10. Cloning of the Bacillus subtilis recE+ gene and functional expression of recE+ in B. subtilis

    International Nuclear Information System (INIS)

    By use of the Bacillus subtilis bacteriophage cloning vehicle Phi 105J23, B. subtilis chromosomal MboI fragments have been cloned that alleviate the pleiotropic effects of the recE4 mutation. The recombinant bacteriophages Phi 105Rec Phi1 (3.85-kilobase insert) and Phi 105Rec Phi4 (3.3-kilobase insert) both conferred on the recE4 strain YB1015 resistance to ethylmethane sulfonate, methylmethane sulfonate, mitomycin C, and UV irradiation comparable with the resistance observed in recE+ strains. While strain YB1015 (recE4) and its derivatives lysogenized with bacteriophage Phi105J23 were not transformed to prototrophy by B. subtilis chromosomal DNA, strain YB1015 lysogenized with either Phi 105Rec Phi 1 or Phi 105RecPhi 4 was susceptible to transformation with homologous B. subtilis chromosomal DNA. The heteroimmune prophages Phi 105 and SPO2 were essentially uninducible in strain YB1015. Significantly, both recombinant prophages Phi 105RecPhi 1 and Phi 105Rec Phi 4 were fully inducible and allowed the spontaneous and mitomycin C-dependent induction of a coresident SPO2 prophage in a recE4 host. The presence of the recombinant prophages also restored the ability of din genes to be induced in strains carrying the recE4 mutation. Finally, both recombinant bacteriophages elaborated a mitomycin C-inducible, 45-kilodalton protein that was immunoreactive with Escherichia coli recA+ gene product antibodies. Collectively, these data demonstrate that the recE+ gene has been cloned and that this gene elaborates the 45-kilodalton protein that is involved in SOB induction and homologous recombination

  11. Horizontal Transfer of Iturin A Operon, itu, to Bacillus subtilis 168 and Conversion into an Iturin A Producer

    OpenAIRE

    Tsuge, Kenji; Inoue, Satoka; Ano, Takashi; Itaya, Mitsuhiro; Shoda, Makoto

    2005-01-01

    Iturin A and its derivatives are lipopeptide antibiotics produced by Bacillus subtilis and several closely related bacteria. Three iturin group operons (i.e., iturin A, mycosubtilin, and bacillomycin D) of those antibiotic-producing strains have been cloned and sequenced thus far, strongly implying the horizontal transfer of these operons. To examine the nature of such horizontal transfer in terms of antibiotic production, a 42-kb region of the B. subtilis RB14 genome, which contains a comple...

  12. Vacuum-induced Mutations In Bacillus Subtilis Spores

    Science.gov (United States)

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

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

  13. Tip-enhanced Raman scattering of bacillus subtilis spores

    Science.gov (United States)

    Rusciano, G.; Zito, G.; Pesce, G.; Sasso, A.; Isticato, R.; Ricca, E.

    2015-07-01

    Understanding of the complex interactions of molecules at biological interfaces is a fundamental issue in biochemistry, biotechnology as well as biomedicine. A plethora of biological processes are ruled by the molecular texture of cellular membrane: cellular communications, drug transportations and cellular recognition are just a few examples of such chemically-mediated processes. Tip-Enhanced Raman Scattering (TERS) is a novel, Raman-based technique which is ideally suited for this purpose. TERS relies on the combination of scanning probe microscopy and Raman spectroscopy. The basic idea is the use of a metalled tip as a sort of optical nano-antenna, which gives place to SERS effect close to the tip end. Herein, we present the application of TERS to analyze the surface of Bacillus subtilis spores. The choice of this biological systems is related to the fact that a number of reasons support the use of spores as a mucosal delivery system. The remarkable and well-documented resistance of spores to various environmental and toxic effects make them clear potentials as a novel, surface-display system. Our experimental outcomes demonstrate that TERS is able to provide a nano-scale chemical imaging of spore surface. Moreover, we demonstrate that TERS allows differentiation between wilde-type spore and genetically modified strains. These results hold promise for the characterization and optimization of spore surface for drug-delivery applications.

  14. Probing phenotypic growth in expanding Bacillus subtilis biofilms.

    Science.gov (United States)

    Wang, Xiaoling; Koehler, Stephan A; Wilking, James N; Sinha, Naveen N; Cabeen, Matthew T; Srinivasan, Siddarth; Seminara, Agnese; Rubinstein, Shmuel; Sun, Qingping; Brenner, Michael P; Weitz, David A

    2016-05-01

    We develop an optical imaging technique for spatially and temporally tracking biofilm growth and the distribution of the main phenotypes of a Bacillus subtilis strain with a triple-fluorescent reporter for motility, matrix production, and sporulation. We develop a calibration procedure for determining the biofilm thickness from the transmission images, which is based on Beer-Lambert's law and involves cross-sectioning of biofilms. To obtain the phenotype distribution, we assume a linear relationship between the number of cells and their fluorescence and determine the best combination of calibration coefficients that matches the total number of cells for all three phenotypes and with the total number of cells from the transmission images. Based on this analysis, we resolve the composition of the biofilm in terms of motile, matrix-producing, sporulating cells and low-fluorescent materials which includes matrix and cells that are dead or have low fluorescent gene expression. We take advantage of the circular growth to make kymograph plots of all three phenotypes and the dominant phenotype in terms of radial distance and time. To visualize the nonlocal character of biofilm growth, we also make kymographs using the local colonization time. Our technique is suitable for real-time, noninvasive, quantitative studies of the growth and phenotype distribution of biofilms which are either exposed to different conditions such as biocides, nutrient depletion, dehydration, or waste accumulation. PMID:27003268

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

    Science.gov (United States)

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

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

  16. Screening of Bacillus subtilis transposon mutants with altered riboflavin production.

    Science.gov (United States)

    Tännler, Simon; Zamboni, Nicola; Kiraly, Csilla; Aymerich, Stéphane; Sauer, Uwe

    2008-09-01

    To identify novel targets for metabolic engineering of riboflavin production, we generated about 10,000 random, transposon-tagged mutants of an industrial, riboflavin-producing strain of Bacillus subtilis. Process-relevant screening conditions were established by developing a 96-deep-well plate method with raffinose as the carbon source, which mimics, to some extent, carbon limitation in fed batch cultures. Screening in raffinose and complex LB medium identified more efficiently riboflavin overproducing and underproducing mutants, respectively. As expected for a "loss of function" analysis, most identified mutants were underproducers. Insertion mutants in two genes with yet unknown function, however, were found to attain significantly improved riboflavin titers and yields. These genes and possibly further ones that are related to them are promising candidates for metabolic engineering. While causal links to riboflavin production were not obvious for most underproducers, we demonstrated for the gluconeogenic glyceraldehyde-3-phosphate dehydrogenase GapB how a novel, non-obvious metabolic engineering strategy can be derived from such underproduction mutations. Specifically, we improved riboflavin production on various substrates significantly by deregulating expression of the gluconeogenic genes gapB and pckA through knockout of their genetic repressor CcpN. This improvement was also verified under the more process-relevant conditions of a glucose-limited fed-batch culture. PMID:18582593

  17. Administration of Bacillus subtilis strains in the rearing water enhances the water quality, growth performance, immune response, and resistance against Vibrio harveyi infection in juvenile white shrimp, Litopenaeus vannamei.

    Science.gov (United States)

    Zokaeifar, Hadi; Babaei, Nahid; Saad, Che Roos; Kamarudin, Mohd Salleh; Sijam, Kamaruzaman; Balcazar, Jose Luis

    2014-01-01

    In this study, vegetative cell suspensions of two Bacillus subtilis strains, L10 and G1 in equal proportions, was administered at two different doses 10(5) (BM5) and 10(8) (BM8) CFU ml(-1) in the rearing water of shrimp (Litopenaeus vannamei) for eight weeks. Both probiotic groups showed a significant reduction of ammonia, nitrite and nitrate ions under in vitro and in vivo conditions. In comparison to untreated control group, final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR) and digestive enzymatic activity were significantly greater in the BM5 and BM8 groups. Significant differences for survival were recorded in the BM8 group as compared to the control. Eight weeks after the start of experiment, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 80%, whereas cumulative mortality of the shrimp that had been given probiotics was 36.7% with MB8 and 50% with MB5. Subsequently, real-time RT-PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and β-1,3-glucan- binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was only significantly up-regulated in the BM5 group compared to the BM8 and control groups. These results suggest that administration of B. subtilis strains in the rearing water confers beneficial effects for shrimp aquaculture, considering water quality, growth performance, digestive enzymatic activity, immune response and disease resistance. PMID:24161773

  18. High-level expression and characterization of the Bacillus subtilis subsp. subtilis str. BSP1 YwaD aminopeptidase in Pichia pastoris.

    Science.gov (United States)

    Tang, Wei; Li, Zhezhe; Li, Chunhua; Yu, Xianhong; Wang, Fei; Wan, Xin; Wang, Yaping; Ma, Lixin

    2016-06-01

    Aminopeptidases are widely used for creating protein hydrolysates and peptide sequencing. The ywaD gene from a new Bacillus isolate, named Bacillus subtilis subsp. subtilis str. BSP1, was cloned into the yeast expression vector pHBM905A and expressed and secreted by Pichia pastoris strain GS115. The deduced amino acid sequence of the aminopeptidase encoded by the ywaD gene shared up to 98% identity with aminopeptidases from B. subtilis strains 168 and zj016. The yield (3.81 g/l) and specific activity (788 U/mg) of recombinant YwaD in high-density fermentation were extremely high. And 829.83 mg of the purified enzyme (4089.72 U/mg) were harvested. YwaD was glycosylated, and its activity decreased after deglycosylation, which was similar to that of the aminopeptidase from B. subtilis strain zj016. YwaD was most active toward l-arginine-4-nitroanilide. Moreover, it exhibited high resistance to carbamide, which was not true for aminopeptidases from B. subtilis strains 168 and zj016, which could simplify the purification of YwaD. Moreover, the expression and parts of characterization of the aminopeptidase from B. subtilis strain 168 in Pichia pastoris were added as supplementary material. The sequence and other characteristics of YwaD were compared with those of aminopeptidases from B. subtilis strains 168 and zj016, and they will provide a solid foundation for further research on the influence of amino acid mutations on the function of aminopeptidases. PMID:26898926

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

    Indian Academy of Sciences (India)

    CHANDRAKANT B JAGTAP; PRADEEP KUMAR; K KRISHNAMURTHY RAO

    2016-09-01

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

  20. Study on mutagenic breeding of bacillus subtilis and properties of its antifungal substances

    International Nuclear Information System (INIS)

    Bacillus subtilis JA isolated by our laboratory produced a large amount of antifungal substances, which had strong inhibitory activity against various plant pathogenic fungi, such as Rhizoctonia solani, Fusarium graminearum and so on. Ion beam implantation as a new mutagenic methods was applied in our study. After B. subtilis JA was implanted by N+ ions, a strain designated as B. Subtilis JA-026 was screened and obtained, which had a higher ability to produce those antifungal substances. A series of experiments indicated that the antifungal substances were thermostable and partially sensitive to proteinases K and tryproteinase. When the fermentating broth was fractionated with ammonium sulphate of a final saturation of 70%, the precipitate enhanced inhibitory activity while the supernatant lost this activity. It appeared that the antifungal substances were likely to be protein. (authors)

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

    NARCIS (Netherlands)

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

    1995-01-01

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

  2. Structure-function correlation in glycine oxidase from Bacillus subtilis

    OpenAIRE

    Mörtl, Mario; Diederichs, Kay; Welte, Wolfram; Molla, Gianluca; Motteran, Laura; Andriolo, Gabriella; Pilone, Mirella S.; Pollegioni, Loredano

    2004-01-01

    Structure-function relationships of the flavoprotein glycine oxidase (GO), which was recently proposed as the first enzyme in the biosynthesis of thiamine in Bacillus subtilis, has been investigated by a combination of structural and functional studies. The structure of the GO-glycolate complex was determined at 1.8 Å, a resolution at which a sketch of the residues involved in FAD binding and in substrate interaction can be depicted. GO can be considered a member of the amine oxidase class ...

  3. Maltose- und Maltodextrin-Verwertung in Bacillus subtilis

    OpenAIRE

    Schönert, Stefan

    2004-01-01

    In seinem natürlichen Habitat findet das Gram - positive Bodenbakterium Bacillus subtilis hauptsächlich polymere Zuckerformen als Kohlenstoffquelle vor, die aus den von anderen Organismen synthetisierten Speicherstoffen, z.B. Stärke und Glykogen, stammen. Jedoch müssen diese Polysaccharide zuerst extrazellulär zu Maltose und Maltodextrinen hydrolysiert werden, bevor sie aufgenommen werden können.Im Gegensatz zu der sehr gut untersuchten Maltose - und Maltodextrin - Aufnahme in Escherichia col...

  4. Extracellular and membrane-bound proteases from Bacillus subtilis.

    OpenAIRE

    Mäntsälä, P; Zalkin, H

    1980-01-01

    Bacillus subtilis YY88 synthesizes increased amounts of extracellular and membrane-bound proteases. More than 99% of the extracellular protease activity is accounted for by an alkaline serine protease and a neutral metalloprotease. An esterase having low protease activity accounts for less than 1% of the secreted protease. These enzymes were purified to homogeneity. Molecular weights of approximately 28,500 and 39,500 were determined for the alkaline and neutral proteases, respectively. The e...

  5. Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Applied to Quantitative Proteomics of Bacillus subtilis

    DEFF Research Database (Denmark)

    Soufi, Boumediene; Kumar, C.; Gnad, F.;

    2010-01-01

    We applied stable isotope labeling by amino acids in cell culture (SILAC) to large-scale quantitative proteomics analyses of the model bacterium Bacillus subtilis in two physiological conditions: growth on succinate and growth under phosphate starvation. Using a B. subtilis strain auxotrophic for...... most comprehensive quantitative proteomics studies in bacteria, covering more than 75% of the B. subtilis genes expressed in the log phase of growth. Furthermore, we detect and quantify dynamics of 35 Ser/Thr/Tyr phosphorylation sites under growth on succinate, and 10 phosphorylation sites under...

  6. Inhibitory effect of novobiocin on ribonucleic acid synthesis during germination of Bacillus subtilis spores.

    OpenAIRE

    Matsuda, M; Kameyama, T

    1980-01-01

    Novobiocin inhibited ribonculeic acid synthesis during germination of Bacillus subtilis spores. Transcription of certain kinds of genes probably required a preceding conformational change in deoxyribonucleic acid.

  7. Characterization of Bacillus subtilis Colony Biofilms via Mass Spectrometry and Fluorescence Imaging.

    Science.gov (United States)

    Si, Tong; Li, Bin; Zhang, Ke; Xu, Yiran; Zhao, Huimin; Sweedler, Jonathan V

    2016-06-01

    Colony biofilms of Bacillus subtilis are a widely used model for studying cellular differentiation. Here, we applied matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) to examine cellular and molecular heterogeneity in B. subtilis colony biofilms. From B. subtilis cells cultivated on a biofilm-promoting medium, we detected two cannibalistic factors not found in previous MALDI MSI studies of the same strain under different culturing conditions. Given the importance of cannibalism in matrix formation of B. subtilis biofilms, we employed a transcriptional reporter to monitor matrix-producing cell subpopulations using fluorescence imaging. These two complementary imaging approaches were used to characterize three B. subtilis strains, the wild type isolate NCIB3610, and two mutants, Δspo0A and ΔabrB, with defective and enhanced biofilm phenotypes, respectively. Upon deletion of key transcriptional factors, correlated changes were observed in biofilm morphology, signaling, cannibalistic factor distribution, and matrix-related gene expression, providing new insights on cannibalism in biofilm development. This work underscores the advantages of using multimodal imaging to compare spatial patterns of selected molecules with the associated protein expression patterns, obtaining information on cellular heterogeneity and function not obtainable when using a single method to characterize biofilm formation. PMID:27136705

  8. Effects of nitrogen ion irradiation on endoglucanase activity and gene mutation of Bacillus subtilis Bac01

    International Nuclear Information System (INIS)

    Bacillus subtilis Bac01 was mutated by 15 keV N+ ions of 1.5xl016 cm-2. The mutant strain Bac11 with high yield of endoglucanase was isolated using carboxymethylcellulose sodium and congo red indicative plates. It exhibited higher endoglucanase activity (381.89IU) than the original strain Bac01 (93.33IU). Two 1,500 bp endoglucanase gene fragments were obtained with PCR amplification from B. subtilis Bac01 and mutant strain Bac11. BLAST comparison result indicated that 10 nucleotides mutated. Bioinformatics methods were used to analyze the two predicted amino acid sequences, and it was found that 5 amino acid residues changed, being all in the cellulose-binding domain of endoglucanase. (authors)

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

    Directory of Open Access Journals (Sweden)

    Nalisha, I.

    2006-01-01

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

  10. Protective Role of Spore Structural Components in Determining Bacillus subtilis Spore Resistance to Simulated Mars Surface Conditions

    OpenAIRE

    Moeller, Ralf; Schuerger, Andrew C.; Reitz, Günther; Nicholson, Wayne L.

    2012-01-01

    Spores of wild-type and mutant Bacillus subtilis strains lacking various structural components were exposed to simulated Martian atmospheric and UV irradiation conditions. Spore survival and mutagenesis were strongly dependent on the functionality of all of the structural components, with small acid-soluble spore proteins, coat layers, and dipicolinic acid as key protectants.

  11. Heterologous Gene Expression in Lactococcus lactis subsp. lactis : Synthesis, Secretion, and Processing of the Bacillus subtilis Neutral Protease

    NARCIS (Netherlands)

    Guchte, Maarten van de; Kodde, Jan; Vossen, Jos M.B.M. van der; Kok, Jan; Venema, Gerard

    1990-01-01

    The Bacillus subtilis nprE gene lacking its own promoter sequence was inserted in the lactococcal expression vector pMG36e. Upon introduction of the recombinant plasmid into Lactococcus lactis subsp. lactis strain MG1363, neutral protease activity could be visualized by the appearance of large clear

  12. Effect of subinhibitory concentrations of four commonly used biocides on the conjugative transfer of Tn916 in Bacillus subtilis

    DEFF Research Database (Denmark)

    Seier-Petersen, Maria Amalie; Jasni, A.; Aarestrup, Frank Møller;

    2014-01-01

    sodium hypochlorite) on the conjugative transposition of the mobile genetic element Tn916.Methods Conjugation assays were carried out between Bacillus subtilis strains. The donor containing Tn916 was pre-exposed to subinhibitory concentrations of each biocide for a defined length of time, which was...

  13. MOLECULAR-CLONING AND SEQUENCE OF COMK, A GENE REQUIRED FOR GENETIC COMPETENCE IN BACILLUS-SUBTILIS

    NARCIS (Netherlands)

    VANSINDEREN, D; TENBERGE, A; HAYEMA, BJ; HAMOEN, L; VENEMA, G

    1994-01-01

    The transformation-deficient strain E26, isolated as a pHV60 insertion mutant, was used to isolate comK, a novel transcription unit required for genetic competence in Bacillus subtilis. Mutational analysis and sequence determination showed that comK contained one open reading frame (ORF), which coul

  14. Decolourization of 4-chloro-2-nitrophenol by a soil bacterium, Bacillus subtilis RKJ 700.

    Directory of Open Access Journals (Sweden)

    Pankaj Kumar Arora

    Full Text Available A 4-Chloro-2-nitrophenol (4C2NP decolourizing strain RKJ 700 was isolated from soil collected from a pesticide contaminated site of India and identified as Bacillus subtilis on the basis of the 16S rRNA gene sequence analysis. Bacillus subtilis RKJ 700 decolourized 4C2NP up to concentration of 1.5 mM in the presence of additional carbon source. The degradation pathway of 4C2NP was studied and 4-chloro-2-aminophenol, 4-chloro-2-acetaminophenol and 5-chloro-2-methylbenzoxazole (5C2MBZ were identified as metabolites by high performance liquid chromatography and gas chromatography-mass spectrometry. Resting cell studies showed that Bacillus subtilis RKJ 700 depleted 4C2NP completely with stoichiometric formation of 5C2MBZ. This is the first report of (i the degradation of 4C2NP at high concentration (1.5 mM and, (ii the formation of 5C2MBZ by a soil bacterium.

  15. The action of ionizing radiation on Bacillus subtilis spores in a dry and wet system

    International Nuclear Information System (INIS)

    The action of water in combination with ionizing radiation was examined using different strains of Bacillus subtilis spores. The parameter of the experiments was a modification of water content; maximal degree of desiccation was achieved by high vacuum. The Fricke-method for X-ray dosimetry was compared to the ionizing-chamber method. In the dry state spores of both wild and mutant strain appeared to be more sensitive than in the wet state. This contradicts to the opinion of dose enhancement by the indirect action of water. (orig.)

  16. Cell wall mechanical properties as measured with bacterial thread made from Bacillus subtilis.

    OpenAIRE

    Mendelson, N H; Thwaites, J J

    1989-01-01

    Engineering approaches used in the study of textile fibers have been applied to the measurement of mechanical properties of bacterial cell walls by using the Bacillus subtilis bacterial thread system. Improved methods have been developed for the production of thread and for measuring its mechanical properties. The best specimens of thread produced from cultures of strain FJ7 grown in TB medium at 20 degrees C varied in diameter by a factor of 1.09 over a 30-mm thread length. The stress-strain...

  17. 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.

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

    Science.gov (United States)

    Zhang, Kang; Duan, Xuguo; Wu, Jing

    2016-01-01

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

  19. Bacteriocin from Bacillus subtilis as a novel drug against diabetic foot ulcer bacterial pathogens

    Institute of Scientific and Technical Information of China (English)

    Baby Joseph; Berlina Dhas; Vimalin Hena; Justin Raj

    2013-01-01

    Objective:To isolate and identify Bacillus subtilis (B. subtilis) from soil and to characterize and partially purify the bacteriocin. To evaluate the antimicrobial activity against four diabetic foot ulcer bacterial pathogens. Methods:Genotypic identification was done based on Bergey’s manual of systemic bacteriology. Antimicrobial susceptibility test was done by Kirby-Bauer disc diffusion method. Colonies were identified by colony morphology and biochemical characterization and also compared with MTCC 121 strain. Further identification was done by 16S rRNA sequencing. Inhibitory activities of partially purified bacteriocin on all the DFU isolates were done by agar well diffusion method. The strain was identified to produce bacteriocin by stab overlay assay. Bacteriocin was extracted by organic solvent extraction using chloroform, further purified by HPLC and physical, and chemical characterization was performed. Results: The four isolates showed high level of resistance to amoxyclav and sensitivity to ciprofloxacin. HPLC purification revealed that the extracts are bacteriocin. The phylogenetic tree analysis results showed that the isolate was 99%related to B. subtilis BSF01. The results reveled activity to all the four isolates and high level of activity was seen in case of Klebsiella sp. Conclusions:Partially purified bacteriocin was found to have antimicrobial activity against the four diabetic foot ulcer bacterial pathogens, which can thus be applied as a better drug molecule on further studies. The strain B. subtilis are found to be safe for use and these antimicrobial peptides can be used as an antimicrobial in humans to treat DFU bacterial pathogens.

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

    Science.gov (United States)

    Zhang, Kang; Duan, Xuguo; Wu, Jing

    2016-01-01

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

  1. Efficacy of Bacillus subtilis V26 as a biological control agent against Rhizoctonia solani on potato.

    Science.gov (United States)

    Ben Khedher, Saoussen; Kilani-Feki, Olfa; Dammak, Mouna; Jabnoun-Khiareddine, Hayfa; Daami-Remadi, Mejda; Tounsi, Slim

    2015-12-01

    The aim of this study is to evaluate the efficacy of the strain Bacillus subtilis V26, a local isolate from the Tunisian soil, to control potato black scurf caused by Rhizoctonia solani. The in vitro antifungal activity of V26 significantly inhibited R. solani growth compared to the untreated control. Microscopic observations revealed that V26 caused considerable morphological deformations of the fungal hyphae such as vacuolation, protoplast leakage and mycelia crack. The most effective control was achieved when strain V26 was applied 24h prior to inoculation (protective activity) in potato slices. The antagonistic bacterium V26 induced significant suppression of root canker and black scurf tuber colonization compared to untreated controls with a decrease in incidence disease of 63% and 81%, respectively, and promoted plant growth under greenhouse conditions on potato plants. Therefore, B. subtilis V26 has a great potential to be commercialized as a biocontrol agent against R. solani on potato crops. PMID:26563555

  2. Bacillus subtilis spores as vaccine adjuvants: further insights into the mechanisms of action.

    Directory of Open Access Journals (Sweden)

    Renata Damásio de Souza

    Full Text Available Bacillus subtilis spores have received growing attention regarding potential biotechnological applications, including the use as probiotics and in vaccine formulations. B. subtilis spores have also been shown to behave as particulate vaccine adjuvants, promoting the increase of antibody responses after co-administration with antigens either admixed or adsorbed on the spore surface. In this study, we further evaluated the immune modulatory properties of B. subtilis spores using a recombinant HIV gag p24 protein as a model antigen. The adjuvant effects of B. subtilis spores were not affected by the genetic background of the mouse lineage and did not induce significant inflammatory or deleterious effects after parenteral administration. Our results demonstrated that co-administration, but not adsorption to the spore surface, enhanced the immunogenicity of that target antigen after subcutaneous administration to BALB/c and C57BL/6 mice. Spores promoted activation of antigen presenting cells as demonstrated by the upregulation of MHC and CD40 molecules and enhanced secretion of pro-inflammatory cytokines by murine dendritic cells. In addition, in vivo studies indicated a direct role of the innate immunity on the immunomodulatory properties of B. subtilis spores, as demonstrated by the lack of adjuvant effects on MyD88 and TLR2 knockout mouse strains.

  3. Production of the polyketide 6-deoxyerythronolide B in the heterologous host Bacillus subtilis.

    Science.gov (United States)

    Kumpfmüller, Jana; Methling, Karen; Fang, Lei; Pfeifer, Blaine A; Lalk, Michael; Schweder, Thomas

    2016-02-01

    Polyketides, such as erythromycin, are complex natural products with diverse therapeutic applications. They are synthesized by multi-modular megaenzymes, so-called polyketide synthases (PKSs). The macrolide core of erythromycin, 6-deoxyerythronolide B (6dEB), is produced by the deoxyerythronolide B synthase (DEBS) that consists of three proteins each with a size of 330-370 kDa. We cloned and investigated the expression of the corresponding gene cluster from Saccharopolyspora erythraea, which comprises more than 30 kb, in Bacillus subtilis. It is shown that the DEBS genes are functionally expressed in B. subtilis when the native eryAI-III operon was separated into three individual expression cassettes with optimized ribosomal binding sites. A synthesis of 6dEB could be detected by using the acetoin-inducible acoA promoter and a fed-batch simulating EnBase-cultivation strategy. B. subtilis was capable of the secretion of 6dEB into the medium. In order to improve the 6dEB production, several genomic modifications of this production strain were tested. This included the knockout of the native secondary metabolite clusters of B. subtilis for the synthesis of surfactin (26 kb), bacillaene (76 kb), and plipastatin (38 kb). It is revealed that the deletion of the prpBD operon, responsible for propionyl-CoA utilization, resulted in a significant increase of the 6dEB product yield when exogenous propionate is provided. Although the presented B. subtilis 6dEB production strain is not competitive with established Escherichia coli 6dEB production strains, the results of this study indicate that B. subtilis is a suitable heterologous host for the secretory production of a complex polyketide. PMID:26432460

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

    Science.gov (United States)

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

    2014-01-01

    Dolomite is a double salt composed of calcium carbonate (CaCO3) and magnesium carbonate (MgCO3). The heat treatment of CaCO3 and MgCO3 respectively generates calcium oxide (CaO) and magnesium oxide (MgO), which have antimicrobial activity. In this study, heated dolomite powder (HDP) slurry was investigated for its sporicidal activity against Bacillus subtilis ATCC 6633 spores. The B. subtilis spores used in this study were not affected by acidic (pH 1) or alkaline (pH 13) conditions, indicating that they were highly resistant. However, dolomite powder heated to 1000℃ for 1 h could kill B. subtilis spores, even at pH 12.7. Sporicidal activity was only apparent when the dolomite powder was heated to 800℃ or higher, and sporicidal activity increased with increases in the heating temperature. This temperature corresponded to that of the generation of CaO. We determined that MgO did not contribute to the sporicidal activity of HDP. To elucidate the sporicidal mechanism of the HDP against B. subtilis spores, the generation of active oxygen from HDP slurry was examined by chemiluminescence analysis. The generation of active oxygen increased when the HDP slurry concentration rose. The results suggested that, in addition to its alkalinity, the active oxygen species generated from HDP were associated with sporicidal activity. PMID:25252642

  5. Complexity in regulation of tryptophan biosynthesis in Bacillus subtilis.

    Science.gov (United States)

    Gollnick, Paul; Babitzke, Paul; Antson, Alfred; Yanofsky, Charles

    2005-01-01

    Bacillus subtilis uses novel regulatory mechanisms in controlling expression of its genes of tryptophan synthesis and transport. These mechanisms respond to changes in the intracellular concentrations of free tryptophan and uncharged tRNA(Trp). The major B. subtilis protein that regulates tryptophan biosynthesis is the tryptophan-activated RNA-binding attenuation protein, TRAP. TRAP is a ring-shaped molecule composed of 11 identical subunits. Active TRAP binds to unique RNA segments containing multiple trinucleotide (NAG) repeats. Binding regulates both transcription termination and translation in the trp operon, and translation of other coding regions relevant to tryptophan metabolism. When there is a deficiency of charged tRNA(Trp), B. subtilis forms an anti-TRAP protein, AT. AT antagonizes TRAP function, thereby increasing expression of all the genes regulated by TRAP. Thus B. subtilis and Escherichia coli respond to identical regulatory signals, tryptophan and uncharged tRNA(Trp), yet they employ different mechanisms in regulating trp gene expression. PMID:16285852

  6. A Bacillus subtilis dipeptide transport system expressed early during sporulation.

    Science.gov (United States)

    Mathiopoulos, C; Mueller, J P; Slack, F J; Murphy, C G; Patankar, S; Bukusoglu, G; Sonenshein, A L

    1991-08-01

    Two previously identified Bacillus subtilis DNA segments, dciA and dciB, whose transcripts accumulate very rapidly after induction of sporulation, were found in the same 6.2 kb transcription unit, now known as the dciA operon. Analysis of the sequence of the dciA operon showed that its putative products are homologous to bacterial peptide transport systems. The product of the fifth gene, DciAE, is similar to peptide-binding proteins from Escherichia coli and Salmonella typhimurium (DppA and OppA) and B. subtilis (OppA). A null mutation in dciAE abolished the ability of a proline auxotroph to grow in a medium containing the dipeptide Pro-Gly as sole proline source, suggesting that the dciA operon encodes a dipeptide transport system. PMID:1766370

  7. Regulation of the anaerobic metabolism in Bacillus subtilis.

    Science.gov (United States)

    Härtig, Elisabeth; Jahn, Dieter

    2012-01-01

    The Gram-positive soil bacterium Bacillus subtilis encounters changing environmental conditions in its habitat. The access to oxygen determines the mode of energy generation. A complex regulatory network is employed to switch from oxygen respiration to nitrate respiration and various fermentative processes. During adaptation, oxygen depletion is sensed by the [4Fe-4S](2+) cluster containing Fnr and the two-component regulatory system ResDE consisting of the membrane-bound histidine kinase ResE and the cytoplasmic ResD regulator. Nitric oxide is the signal recognized by NsrR. Acetate formation and decreasing pH are measured via AlsR. Finally, Rex is responding to changes in the cellular NAD(+)/NADH ration. The fine-tuned interplay of these regulators at approximately 400 target gene promoters ensures efficient adaptation of the B. subtilis physiology. PMID:23046954

  8. Heat Resistance and Population Stability of Lyophilized Bacillus subtilis Spores

    OpenAIRE

    Odlaug, Theron E.; Caputo, Ross A.; Graham, Gary S.

    1981-01-01

    Bacillus subtilis 5230 spores were lyophilized in 0.067 M phosphate buffer and stored at 2 to 8°C for 9 to 27 months. The lyophilized spores were reconstituted with buffer or 0.9% saline, and the heat resistance was determined in a thermoresistometer. Lyophilization had no effect on the heat resistance of the spores but did result in a slight decrease in population (≤0.3-logarithm reduction). The lyophilized spores maintained heat resistance and population levels over the test periods. The D-...

  9. Unhairing animal hides using probiotic Bacteria bacillus subtilis

    OpenAIRE

    Данилкович, Анатолій Григорович; Гвоздяк, Петро Ілліч; Романюк, Оксана Олександрівна; Ковтуненко, Ольга Василівна

    2013-01-01

    The most efficient technology of processing natural raw materials into skin and fur is the use of enzyme products for soaking and liming processes. Therefore, the use of bacterial products, which produce enzymes of various functional effects, is considered to be very promising for the above mentioned processes.Soaking and liming of flint-dried rabbit hides were carried out using probiotic bacreria Bacillus subtilis on 4 samples in a laboratory centrifuge at soaking temperature 36-38°С and wor...

  10. Cosegregation of cell wall and DNA in Bacillus subtilis.

    OpenAIRE

    Schlaeppi, J M; Karamata, D

    1982-01-01

    Cosegregation of cell wall and DNA of a lysis-negative mutant of Bacillus subtilis was examined by continuously labeling (i) cell wall, (ii) DNA, and (iii) both cell wall and DNA. After four to five generations of chase in liquid media it was found by light microscope autoradiography that the numbers of wall segregation units per cell are 29 and 9 in rich and minimal medium, respectively. Under the same conditions the numbers of segregation units of DNA were almost 50% lower: 15 and 5, respec...

  11. Proteins that interact with GTP during sporulation of Bacillus subtilis

    International Nuclear Information System (INIS)

    During sporulation of Bacillus subtilis, several proteins were shown to interact with GTP in specific ways. UV light was used to cross-link [α-32P]GTP to proteins in cell extracts at different stages of growth. After electrophoresis, 11 bands of radioactivity were found in vegetative cells, 4 more appeared during sporulation, and only 9 remained in mature spores. Based on the labeling pattern with or without UV light to cross-link either [α-32P]GTP or [γ-32P]GTP, 11 bands of radioactivity were apparent guanine nucleotide-binding proteins, and 5 bands appeared to be phosphorylated and/or guanylated. Similar results were found with Bacillus megaterium. Assuming the GTP might be a type of signal for sporulation, it could interact with and regulate proteins by at least three mechanisms

  12. Bacillus subtilis subsp. subtilis CBMDC3f with antimicrobial activity against Gram-positive foodborne pathogenic bacteria: UV-MALDI-TOF MS analysis of its bioactive compounds.

    Science.gov (United States)

    Torres, M J; Petroselli, G; Daz, M; Erra-Balsells, R; Audisio, M C

    2015-06-01

    In this work a new Bacillus sp. strain, isolated from honey, was characterized phylogenetically. Its antibacterial activity against three relevant foodborne pathogenic bacteria was studied; the main bioactive metabolites were analyzed using ultraviolet matrix assisted laser desorption-ionization mass spectrometry (UV-MALDI MS). Bacillus CBMDC3f was phylogenetically characterized as Bacillus subtilis subsp. subtilis after rRNA analysis of the 16S subunit and the gyrA gene (access codes Genbank JX120508 and JX120516, respectively). Its antibacterial potential was evaluated against Listeria monocytogenes (9 strains), B. cereus (3 strains) and Staphylococcus aureus ATCC29213. Its cell suspension and cell-free supernatant (CFS) exerted significant anti-Listeria and anti-S. aureus activities, while the lipopeptides fraction (LF) also showed anti-B. cereus effect. The UV-MALDI-MS analysis revealed surfactin, iturin and fengycin in the CFS, whereas surfactin predominated in the LF. The CFS from CBMDC3f contained surfactin, iturin and fengycin with four, two and four homologues per family, respectively, whereas four surfactin, one iturin and one fengycin homologues were identified in the LF. For some surfactin homologues, their UV-MALDI-TOF/TOF (MS/MS; Laser Induced Decomposition method, LID) spectra were also obtained. Mass spectrometry analysis contributed with relevant information about the type of lipopeptides that Bacillus strains can synthesize. From our results, surfactin would be the main metabolite responsible for the antibacterial effect. PMID:25820813

  13. Molecular Cloning and Production of Recombinant Phytase from Bacillus subtilis ASUIA243 in Pichia pastoris

    OpenAIRE

    Nor Soleha Mohd Dali; Tamrin Nuge; Mohd Hafidz Mahamad Maifiah; Faridah Yusof; Anis Shobirin Meor Hussin; Abd-Elaziem Farouk; and Hamzah Mohd. Salleh

    2011-01-01

    Phytase gene obtained from Bacillus subtilis ASUIA243 was cloned into a medium vector and transformed into E. coli. Restriction enzyme digestion was conducted to get blunt-ended phytase gene and ligated into the Pichia expression vector, pPICZαA. The recombinant vector, pPICZαA-243HPp was then linearized with PmeI and transformed into P. pastoris strain X33. Screening for multi copy gene number of transformants was done by re-plating the selected colonies on increasing concentrati...

  14. Role of GerD in Germination of Bacillus subtilis Spores▿

    OpenAIRE

    Pelczar, Patricia L.; Igarashi, Takao; Setlow, Barbara; Setlow, Peter

    2006-01-01

    Spores of a Bacillus subtilis strain with a gerD deletion mutation (ΔgerD) responded much slower than wild-type spores to nutrient germinants, although they did ultimately germinate, outgrow, and form colonies. Spores lacking GerD and nutrient germinant receptors also germinated slowly with nutrients, as did ΔgerD spores in which nutrient receptors were overexpressed. The germination defect of ΔgerD spores was not suppressed by many changes in the sporulation or germination conditions. Germin...

  15. Identification and Characterization of Mutations Conferring Resistance to d-Amino Acids in Bacillus subtilis

    OpenAIRE

    Leiman, Sara A.; Richardson, Charles; Foulston, Lucy; Elsholz, Alexander K.W.; First, Eric A.; Losick, Richard

    2015-01-01

    Bacteria produce d-amino acids for incorporation into the peptidoglycan and certain nonribosomally produced peptides. However, d-amino acids are toxic if mischarged on tRNAs or misincorporated into protein. Common strains of the Gram-positive bacterium Bacillus subtilis are particularly sensitive to the growth-inhibitory effects of d-tyrosine due to the absence of d-aminoacyl-tRNA deacylase, an enzyme that prevents misincorporation of d-tyrosine and other d-amino acids into nascent proteins. ...

  16. recE4-Independent Recombination Between Homologous Deoxyribonucleic Acid Segments of Bacillus subtilis Plasmids

    OpenAIRE

    Tanaka, T

    1980-01-01

    A plasmid (pLS104) carrying a tandem repetition of the leu region of the Bacillus subtilis chromosome arose spontaneously from pLS103, which carried a single copy of the leu region. Plasmid preparations from strains harboring pLS104 also contained the original plasmid, pLS103, and, in some preparations, plasmids carrying three or four repetitions of the leu region. These plasmids were shown to be generated by recombination between homologous deoxyribonucleic acid (DNA) segments in the tandeml...

  17. Characterization of the decolorizing activity of azo dyes by Bacillus subtilis azoreductase AzoR1

    OpenAIRE

    Montira Leelakriangsak; Sukallaya Borisut

    2012-01-01

    The product of the Bacillus subtilis gene azoR1 is annotated as a putative azoreductase, production of which isinduced in response to thiol-reactive compounds. Here we report on the decolorization of four azo dyes by azoreductaseactivity. The ability of overexpressed AzoR1 strain ORB7106 to catalyze decolorization of azo dyes was investigated on agarplates and in liquid cultures. The decolorization efficacy of a mutant, ORB7106, which has lost negative control of azoR1expression, was signific...

  18. Purification and Characterization of an Extracellular Cholesterol Oxidase of Bacillus subtilis Isolated from Tiger Excreta.

    Science.gov (United States)

    Kumari, Lata; Kanwar, Shamsher S

    2016-01-01

    A mesophilic Bacillus sp. initially isolated from tiger excreta and later identified as a Bacillus subtilis strain was used to produce an extracellular cholesterol oxidase (COX) in cholesterol-enriched broth. This bacterial isolate was studied for the production of COX by manipulation of various physicochemical parameters. The extracellular COX was successfully purified from the cell-free culture broth of B. subtilis by successive salting out with ammonium sulfate, dialysis, and riboflavin-affinity chromatography. The purified COX was characterized for its molecular mass/structure and stability. The enzyme possessed some interesting properties such as high native Mr (105 kDa), multimeric (pentamer of ∼21 kDa protein) nature, organic solvent compatibility, and a half-life of ∼2 h at 37 °C. The bacterial COX exhibited ∼22 % higher activity in potassium phosphate buffer (pH 7.5) in the presence of a nonionic detergent Triton X-100 at 0.05 % (v/v). The K m and V max value of COX of B. subtilis COX were found to be 3.25 mM and 2.17 μmol min ml(-1), respectively. The purified COX showed very little cytotoxicity associated with it. PMID:26453032

  19. Initiation of decay of Bacillus subtilis trp leader RNA.

    Science.gov (United States)

    Deikus, Gintaras; Bechhofer, David H

    2007-07-13

    Transcription termination in the leader region of the Bacillus subtilis trp operon is regulated by binding of the 11-mer TRAP complex to nascent trp RNA, which results in formation of a terminator structure. Rapid decay of trp leader RNA, which is required to release the TRAP complex and maintain a sufficient supply of free TRAP, is mediated by polynucleotide phosphorylase (PNPase). Using purified B. subtilis PNPase, we showed that, when TRAP was present, PNPase binding to the 3' end of trp leader RNA and PNPase digestion of trp leader RNA from the 3' end were inefficient. These results suggested that initiation of trp leader RNA may begin with an endonuclease cleavage upstream of the transcription terminator structure. Such cleavage was observed in vivo. Mutagenesis of nucleotides at the cleavage site abolished processing and resulted in a 4-fold increase in trp leader RNA half-life. This is the first mapping of a decay-initiating endonuclease cleavage site on a native B. subtilis RNA. PMID:17507374

  20. Application of gaseous ozone for inactivation of Bacillus subtilis spores.

    Science.gov (United States)

    Aydogan, Ahmet; Gurol, Mirat D

    2006-02-01

    The effectiveness of gaseous ozone (O3) as a disinfectant was tested on Bacillus subtilis spores, which share the same physiological characteristics as Bacillus anthracis spores that cause the anthrax disease. Spores dried on surfaces of different carrier material were exposed to O3 gas in the range of 500-5000 ppm and at relative humidity (RH) of 70-95%. Gaseous O3 was found to be very effective against the B. subtilis spores, and at O3 concentrations as low as 3 mg/L (1500 ppm), approximately 3-log inactivation was obtained within 4 hr of exposure. The inactivation curves consisted of a short lag phase followed by an exponential decrease in the number of surviving spores. Prehydration of the bacterial spores has eliminated the initial lag phase. The inactivation rate increased with increasing O3 concentration but not >3 mg/L. The inactivation rate also increased with increase in RH. Different survival curves were obtained for various surfaces used to carry spores. Inactivation rates of spores on glass, a vinyl floor tile, and office paper were nearly the same. Whereas cut pile carpet and hardwood flooring surfaces resulted in much lower inactivation rates, another type of carpet (loop pile) showed significant enhancement in the inactivation of the spores. PMID:16568801

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

    DEFF Research Database (Denmark)

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

    2013-01-01

    bikalga were examined for their antimicrobial activity against a panel of 36 indicator organisms including Gram-positive and Gram-negative bacteria and yeasts. The Bacillus spp. isolates showed variable inhibitory abilities depending on the method used. Both Gram-positive and Gram-negative bacteria were...... and Bacillus cereus, while CFS of 2 B. licheniformis (E3 and F9) strains only inhibited M. luteus. The antimicrobial substance(s) produced by B. subtilis subsp. subtilis H4 was further characterized. The antimicrobial substance(s) produced by H4 was detected from mid-exponential growth phase. The...... activity was sensitive to protease and trypsin, but resistant to the proteolytic action of proteinase K and papain. Treatment with α-amylase and lipase II resulted in a complete loss of antimicrobial effect, indicating that a sugar moiety and lipid moiety are necessary for the activity. Treatment with...

  2. Effect of Mono and Di-rhamnolipids on Biofilms Pre-formed by Bacillus subtilis BBK006.

    Science.gov (United States)

    De Rienzo, Mayri A Díaz; Martin, Peter J

    2016-08-01

    Different microbial inhibition strategies based on the planktonic bacterial physiology have been known to have limited efficacy on the growth of biofilms communities. This problem can be exacerbated by the emergence of increasingly resistant clinical strains. Biosurfactants have merited renewed interest in both clinical and hygienic sectors due to their potential to disperse microbial biofilms. In this work, we explore the aspects of Bacillus subtilis BBK006 biofilms and examine the contribution of biologically derived surface-active agents (rhamnolipids) to the disruption or inhibition of microbial biofilms produced by Bacillus subtilis BBK006. The ability of mono-rhamnolipids (Rha-C10-C10) produced by Pseudomonas aeruginosa ATCC 9027 and the di-rhamnolipids (Rha-Rha-C14-C14) produced by Burkholderia thailandensis E264, and phosphate-buffered saline to disrupt biofilm of Bacillus subtilis BBK006 was evaluated. The biofilm produced by Bacillus subtilis BBK006 was more sensitive to the di-rhamnolipids (0.4 g/L) produced by Burkholderia thailandensis than the mono-rhamnolipids (0.4 g/L) produced by Pseudomonas aeruginosa ATCC 9027. Rhamnolipids are biologically produced compounds safe for human use. This makes them ideal candidates for use in new generations of bacterial dispersal agents and useful for use as adjuvants for existing microbial suppression or eradication strategies. PMID:27113589

  3. Identification and characterization of the vanillin dehydrogenase YfmT in Bacillus subtilis 3NA.

    Science.gov (United States)

    Graf, Nadja; Wenzel, Marian; Altenbuchner, Josef

    2016-04-01

    With vanillin as one of the most important flavoring agents, many efforts have been made to optimize its biotechnological production from natural abundant substrates. However, its toxicity against the hosts results in rather low yields and product concentrations. Bacillus subtilis as a soil-dwelling bacterium is a possible lignin-derived compound-degrading microorganism. Therefore, its vanillin and ferulic acid metabolism was investigated. With a rather high tolerance for vanillin up to 20 mM, it is a promising candidate to produce natural vanillin. In this study, the well-studied phenolic acid decarboxylases PadC and BsdBCD could be ascribed to function as the only enzymes in B. subtilis 3NA converting ferulic acid to 4-vinylguaiacol and vanillic acid to guaiacol, respectively. As vanillin also becomes converted to guaiacol, a previous conversion to vanillic acid was assumed. Usage of bioinformatic tools revealed YfmT, which could be shown to function as the only vanillin dehydrogenase in B. subtilis 3NA. Thus, YfmT was further characterized regarding its temperature and pH optima as well as its substrate range. Vanillin and ferulic acid metabolic routes in the tested B. subtilis strain were revealed, a direct conversion of ferulic acid to vanillin, however, could not be found. PMID:26658822

  4. D-amino acids indirectly inhibit biofilm formation in Bacillus subtilis by interfering with protein synthesis.

    Science.gov (United States)

    Leiman, Sara A; May, Janine M; Lebar, Matthew D; Kahne, Daniel; Kolter, Roberto; Losick, Richard

    2013-12-01

    The soil bacterium Bacillus subtilis forms biofilms on surfaces and at air-liquid interfaces. It was previously reported that these biofilms disassemble late in their life cycle and that conditioned medium from late-stage biofilms inhibits biofilm formation. Such medium contained a mixture of D-leucine, D-methionine, D-tryptophan, and D-tyrosine and was reported to inhibit biofilm formation via the incorporation of these D-amino acids into the cell wall. Here, we show that L-amino acids were able to specifically reverse the inhibitory effects of their cognate D-amino acids. We also show that D-amino acids inhibited growth and the expression of biofilm matrix genes at concentrations that inhibit biofilm formation. Finally, we report that the strain routinely used to study biofilm formation has a mutation in the gene (dtd) encoding D-tyrosyl-tRNA deacylase, an enzyme that prevents the misincorporation of D-amino acids into protein in B. subtilis. When we repaired the dtd gene, B. subtilis became resistant to the biofilm-inhibitory effects of D-amino acids without losing the ability to incorporate at least one noncanonical D-amino acid, D-tryptophan, into the peptidoglycan peptide side chain. We conclude that the susceptibility of B. subtilis to the biofilm-inhibitory effects of D-amino acids is largely, if not entirely, due to their toxic effects on protein synthesis. PMID:24097941

  5. selective production and characterization of levan by Bacillus subtilis (Natto) Takahashi.

    Science.gov (United States)

    Shih, Ing-Lung; Yu, Yun-Ti; Shieh, Chwen-Jen; Hsieh, Chien-Yan

    2005-10-19

    To meet the industrial need of an efficient microbial method for increased levan production, Bacillus subtilis (natto) Takahashi, a commercial natto starter for preparing fermented soybeans (natto), was used to produce levan. After cultivation for 21 h, 40-50 mg of levan mL(-1) was produced in medium containing 20% (w/w) sucrose, which was approximately 50% yield on available fructose. The product consisted of two fractions with different molecular masses (1794 and 11 kDa), which were easily separated by fractionation using an ethanol gradient. The products were well characterized by GPC, 13C NMR, and 1H NMR. The various sugars and concentrations, initial pH, fermentation temperature, and agitation speed affected the levan production by B. subtilis (natto) Takahashi. Takahashi strain is the most efficient levan-producing strain among all of the B. subtilis strains tested and, as previously reported, it produced the highest yield of levan in the least time (21 h) under the common cultivation condition. PMID:16218666

  6. Molecular Cloning and Production of Recombinant Phytase from Bacillus subtilis ASUIA243 in Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Nor Soleha Mohd Dali

    2011-12-01

    Full Text Available Phytase gene obtained from Bacillus subtilis ASUIA243 was cloned into a medium vector and transformed into E. coli. Restriction enzyme digestion was conducted to get blunt-ended phytase gene and ligated into the Pichia expression vector, pPICZαA. The recombinant vector, pPICZαA-243HPp was then linearized with PmeI and transformed into P. pastoris strain X33. Screening for multi copy gene number of transformants was done by re-plating the selected colonies on increasing concentration of zeocin. One positive clone, X243HPp#2 was then grown in BMGY media as the starting culture, followed by induction in BMMY media for protein expression study. The supernatant was then analysed by SDS-PAGE and Western blot method to check the protein expression.ABSTRAK: Gen fitase yang didapati daripada Bacillus subtilis ASUIA243 diklonkan sebagai vektor perantara dan berubah menjadi E. coli. Sekatan pencernaan enzim dijalankan untuk mendapatkan gen fitase berhujung tumpul dan diligatkan dengan vektor ekspresi Pichia, pPICZαA. Vektor rekombinan, pPICZαA-243HPp kemudian dilinearkan dengan PmeI dan berubah menjadi P. pastoris strain X33. Penyaringan untuk nombor gen berbilang salinan yang menjalani transformasi genetik dijalankan dengan menyalur semula koloni terpilih dengan penambahan kepekatan zeocin. Satu klon positif, X243HPp#2 kemudian dibiarkan hidup dalam perantara BMGY sebagai kultur permulaan, diikuti dengan aruhan dalam perantara BMMY untuk kajian penglahiran protein. Supernatan kemudian dikaji dengan SDS-PAGE dan kaedah sap Western untuk menyemak penglahiran protein.KEYWORDS:  phytase, Bacillus subtilis, Pichia pastoris, gene cloning.

  7. Modification of the rib operon derived from Bacillus subtilis and its expression in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    Zhang Huitu; Meng Kun; Wang Yaru; Luo Huiying; Yuan Tiezheng; Yang Peilong; Bai Yingguo; Yao Bin; Fan Yunliu

    2007-01-01

    A riboflavin operon(rib operon)derived from Bacillus subtilis 368 was modified on structure and the resulting operons were expressed in various strains of Escherichia coli. The results showed that the optimization of the rib operon and the host strain used for expression are two main factors affecting the riboflavin production. Replacing the promoter l and rfn box of the rib operon with a strong constructive promoter spo l drastically increased the expression of the rib genes. When E. Coli JMl09 was used as the host strain, the highest riboflavin production reached 95.3μg/mL(about eight times higher than that 0f the unmodified rib operon). In addition, when tetracycline(20 μg/mL)was used as the selective pressure, compared with the ampicillin resistant transformants, a higher riboflavin yield Was obtained in tetracycline resistant host strain.

  8. Suppression of Magnaporthe oryzae and interaction between Bacillus subtilis and rice plants in the control of rice blast.

    Science.gov (United States)

    Sha, Yuexia; Wang, Qi; Li, Yan

    2016-01-01

    Magnaporthe oryzae, the causative pathogen of rice blast, has caused extensive losses to rice cultivation worldwide. Strains of the bacterium Bacillus subtilis have been used as biocontrol agents against rice blast. However, little has been reported about the interaction between B. subtilis and the rice plant and its mechanism of action. Here, the colonization process and induced disease resistance by B. subtilis SYX04 and SYX20 in rice plants was examined. Strains of B. subtilis labeled with green fluorescent protein reached population of more than 5 × 10(6) CFU/g after 20 days on mature rice leaves and were detected after 3 days on newly grown leaves. Results showed that SYX04 and SYX20 not only inhibited spore germination, germ tube length, and appressorial formation but also caused a series of alterations in the structures of hyphae and conidia. The cell walls and membrane structures of the fungus showed ultrastructural abnormalities, which became severely degraded as observed through scanning electron microscopy and transmission electron microscopy. The mixture of both B. subtilis and M. oryzae resulted in enhanced activity of peroxidase, and polyphenol oxidase while there was significantly more superoxide dismutase activity in plants that had been sprayed with B. subtilis alone. The present study suggests that colonized SYX04 and SYX20 strains protected rice plants and exhibited antifungal activity and induced systemic resistance, thus indicating their potential biological control agents. PMID:27536521

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

    Institute of Scientific and Technical Information of China (English)

    别小妹; 陆兆新; 吕凤霞; 赵海珍; 杨胜远; 孙力军

    2006-01-01

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

  10. Unusual Biosynthesis and Structure of Locillomycins from Bacillus subtilis 916

    Science.gov (United States)

    Luo, Chuping; Liu, Xuehui; Zhou, Xian; Guo, Junyao; Truong, John; Wang, Xiaoyu; Zhou, Huafei

    2015-01-01

    Three families of Bacillus cyclic lipopeptides—surfactins, iturins, and fengycins—have well-recognized potential uses in biotechnology and biopharmaceutical applications. This study outlines the isolation and characterization of locillomycins, a novel family of cyclic lipopeptides produced by Bacillus subtilis 916. Elucidation of the locillomycin structure revealed several molecular features not observed in other Bacillus lipopeptides, including a unique nonapeptide sequence and macrocyclization. Locillomycins are active against bacteria and viruses. Biochemical analysis and gene deletion studies have supported the assignment of a 38-kb gene cluster as the locillomycin biosynthetic gene cluster. Interestingly, this gene cluster encodes 4 proteins (LocA, LocB, LocC, and LocD) that form a hexamodular nonribosomal peptide synthetase to biosynthesize cyclic nonapeptides. Genome analysis and the chemical structures of the end products indicated that the biosynthetic pathway exhibits two distinct features: (i) a nonlinear hexamodular assembly line, with three modules in the middle utilized twice and the first and last two modules used only once and (ii) several domains that are skipped or optionally selected. PMID:26162886

  11. Crude glycerol from biodiesel industry as substrate for biosurfactant production by Bacillus subtilis ATCC 6633

    Directory of Open Access Journals (Sweden)

    Marylane de Sousa

    2014-04-01

    Full Text Available Glycerol, a co-product of the biodiesel industry, may be a suitable raw material for the production of high added-value compounds by the microorganisms. This study aimed to use the glycerol obtained from the biodiesel production process as the main carbon source for biosurfactant production by Bacillus subtilis ATCC 6633. Results indicated that the strain lowered the surface tension of the cell-free fermented broth to 31.5 ± 1.6 mN/m, indicating the production of biosurfactant. The critical micelle concentration (CMC = 33.6 mN/m obtained was similar to the previously reported for biossurfactants isolated from other Bacillus. The produced biosurfactant was able to emulsify n-hexadecane and soybean oil.

  12. Antimicrobial, antiadhesive and antibiofilm potential of lipopeptides synthesised by Bacillus subtilis, on uropathogenic bacteria.

    Science.gov (United States)

    Moryl, Magdalena; Spętana, Magdalena; Dziubek, Klaudia; Paraszkiewicz, Katarzyna; Różalska, Sylwia; Płaza, Grażyna A; Różalski, Antoni

    2015-01-01

    The aim of this study was to investigate the antimicrobial effect of lipopeptide biosurfactants from surfactin, iturin and fengycin families, synthesised by the Bacillus subtilis I'1a strain, on uropathogenic bacteria, including the effects on planktonic growth, processes of biofilm formation and dislodging. Antimicrobial activity was tested against 32 uropathogenic strains belonging to 12 different species of Gram-negative and Gram-positive bacteria. The sensitivity of 25 tested bacterial strains to the B. subtilis I'1a filtrate was confirmed by an agar diffusion assay. None of the strains seemed to be sensitive to pure surfactin at concentrations ranging from 0.1 mg × ml(-1) to 0.4 mg ml(-1). After the treatment of uropathogens with B. subtilis lipopeptides, the metabolic activity of planktonic cells was inhibited by 88.05±3.96% in the case of 21 studied uropathogens, the process of biofilm formation was reduced by 88.15±4.77% in the case of 24 uropathogens and mature biofilms of 18 strains were dislodged by about 81.20±4.72%. Ten strains of uropathogenic bacteria were selected to study the antimicrobial activity of surfactin (concentrations 0.1, 0.2 and 0.4 mg × ml(-1)). Surfactin had no influence on the metabolic activity of planktonic forms of uropathogens, however, biofilms of 5 tested strains were reduced by 64.77±9.05% in the presence of this biosurfactant at the concentration 0.1 mg × ml(-1). The negative effect of the compound on the biofilm formation process was observed at all concentrations used. The above-described results were fully confirmed by CLSM. It could suggest that synergistic application of biosurfactants could be efficient in uropathogen eradication. PMID:26505130

  13. Detection of Anthrax Simulants with Microcalorimetric Spectroscopy: Bacillus subtilis and Bacillus cereus Spores

    Science.gov (United States)

    Arakawa, Edward T.; Lavrik, Nickolay V.; Datskos, Panos G.

    2003-04-01

    Recent advances in the development of ultrasensitive micromechanical thermal detectors have led to the advent of novel subfemtojoule microcalorimetric spectroscopy (CalSpec). On the basis of principles of photothermal IR spectroscopy combined with efficient thermomechanical transduction, CalSpec provides acquisition of vibrational spectra of microscopic samples and absorbates. We use CalSpec as a method of identifying nanogram quantities of biological micro-organisms. Our studies focus on Bacillus subtilis and Bacillus cereus spores as simulants for Bacillus anthracis spores. Using CalSpec, we measured IR spectra of B. subtilis and B. cereus spores present on surfaces in nanogram quantities (approximately 100 -1000 spores). The spectra acquired in the wavelength range of 690 -4000 cm-1 (2.5 -14.5 μm) contain information-rich vibrational signatures that reflect the different ratios of biochemical makeup of the micro-organisms. The distinctive features in the spectra obtained for the two types of micro-organism can be used to distinguish between the spores of the Bacillus family. As compared with conventional IR and Fourier-transform IR microscopic spectroscopy techniques, the advantages of the present technique include significantly improved sensitivity (at least a full order of magnitude), absence of expensive IR detectors, and excellent potential for miniaturization.

  14. Optimization of medium composition for the production of compounds effective against Xanthomonas campestris by bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Rončević Zorana Z.

    2014-01-01

    Full Text Available The biocontrol agents are a very promising alternative to synthetic pesticides that are presently used to control plant diseases caused by phytopathogenic microorganisms. Members of the Bacillus genera are soil bacteria that produce significant quantities of agriculturally important bioactive compounds. Production of these compounds can be improved by changing the nutritional and environmental conditions. The aim of this study was the optimization of medium composition, using response surface methodology, for the production of compounds effective against Xanthomonas campestris ATCC 13951 by Bacillus subtilis ATCC 6633. To study the production of antimicrobial compounds by selected Bacillus strain, the producing microorganisms were cultivated on nutrient broth. The inhibition zone diameter of 18.0 mm obtained by the diffusion-disc method indicated that the used Bacillus subtilis strain produces compounds with antimicrobial activity against Xanthomonas campestris ATCC 13951. To optimize the composition of the cultivation medium in terms of glycerol, sodium nitrite and phosphates content, experiments were carried out in accordance with Box-Behnken design, and optimization of multiple responses was performed using the concept of desirability function. The developed model predicted that the maximum inhibition zone diameter (26.23 mm against tested phytopathogen is achieved when the initial content of glycerol, sodium nitrite and phosphate were 50.00 g/L, 2.85 g/L and 11.00 g/L, respectively. To minimize the consumption of medium components and costs of effluents processing, additional optimization set was made. The techno-economic analysis of the obtained results has to be done to select optimal medium composition for industrial production of antimicrobial compounds.

  15. Second Messenger Signaling in Bacillus subtilis: Accumulation of Cyclic di-AMP Inhibits Biofilm Formation.

    Science.gov (United States)

    Gundlach, Jan; Rath, Hermann; Herzberg, Christina; Mäder, Ulrike; Stülke, Jörg

    2016-01-01

    The Gram-positive model organism Bacillus subtilis produces the essential second messenger signaling nucleotide cyclic di-AMP. In B. subtilis and other bacteria, c-di-AMP has been implicated in diverse functions such as control of metabolism, cell division and cell wall synthesis, and potassium transport. To enhance our understanding of the multiple functions of this second messenger, we have studied the consequences of c-di-AMP accumulation at a global level by a transcriptome analysis. C-di-AMP accumulation affected the expression of about 700 genes, among them the two major operons required for biofilm formation. The expression of both operons was severely reduced both in the laboratory and a non-domesticated strain upon accumulation of c-di-AMP. In excellent agreement, the corresponding strain was unable to form complex colonies. In B. subtilis, the transcription factor SinR controls the expression of biofilm genes by binding to their promoter regions resulting in transcription repression. Inactivation of the sinR gene restored biofilm formation even at high intracellular c-di-AMP concentrations suggesting that the second messenger acts upstream of SinR in the signal transduction pathway. As c-di-AMP accumulation did not affect the intracellular levels of SinR, we conclude that the nucleotide affects the activity of SinR. PMID:27252699

  16. Second Messenger Signaling in Bacillus subtilis: Accumulation of Cyclic di-AMP Inhibits Biofilm Formation

    Science.gov (United States)

    Gundlach, Jan; Rath, Hermann; Herzberg, Christina; Mäder, Ulrike; Stülke, Jörg

    2016-01-01

    The Gram-positive model organism Bacillus subtilis produces the essential second messenger signaling nucleotide cyclic di-AMP. In B. subtilis and other bacteria, c-di-AMP has been implicated in diverse functions such as control of metabolism, cell division and cell wall synthesis, and potassium transport. To enhance our understanding of the multiple functions of this second messenger, we have studied the consequences of c-di-AMP accumulation at a global level by a transcriptome analysis. C-di-AMP accumulation affected the expression of about 700 genes, among them the two major operons required for biofilm formation. The expression of both operons was severely reduced both in the laboratory and a non-domesticated strain upon accumulation of c-di-AMP. In excellent agreement, the corresponding strain was unable to form complex colonies. In B. subtilis, the transcription factor SinR controls the expression of biofilm genes by binding to their promoter regions resulting in transcription repression. Inactivation of the sinR gene restored biofilm formation even at high intracellular c-di-AMP concentrations suggesting that the second messenger acts upstream of SinR in the signal transduction pathway. As c-di-AMP accumulation did not affect the intracellular levels of SinR, we conclude that the nucleotide affects the activity of SinR. PMID:27252699

  17. Novel broad host range shuttle vectors for expression in Escherichia coli, Bacillus subtilis and Pseudomonas putida.

    Science.gov (United States)

    Troeschel, Sonja Christina; Thies, Stephan; Link, Olga; Real, Catherine Isabell; Knops, Katja; Wilhelm, Susanne; Rosenau, Frank; Jaeger, Karl-Erich

    2012-10-15

    Novel shuttle vectors named pEBP were constructed to allow the gene expression in different bacterial hosts including Escherichia coli, Bacillus subtilis and Pseudomonas putida. These vectors share the inducible promoters P(T7) and P(Xyl) and a cos site to enable packaging of plasmid DNA into phage, and carry different multiple cloning sites and antibiotic resistance genes. Vector pEBP41 generally replicates episomally while pEBP18 replicates episomally in Gram-negative bacteria only, but integrates into the chromosome of B. subtilis. Plasmid copy numbers determined for E. coli and P. putida were in the range of 5-50 per cell. The functionality of pEBP18 and pEBP41 was confirmed by expression of two lipolytic enzymes, namely lipase A from B. subtilis and cutinase from the eukaryotic fungus Fusarium solani pisi in three different host strains. Additionally, we report here the construction of a T7 RNA polymerase-based expression strain of P. putida. PMID:22440389

  18. Incorporation of glycine and serine into sporulating cells of Bacillus subtilis

    International Nuclear Information System (INIS)

    The changes during growth and sporulation in activities of cells of Bacillus subtilis to incorporate various amino acids were investigated with wild-type strain and its asporogenous mutant. In the case of wild type strain the uptake of valine, phenylalanine, and proline was largest during the logarithmic growth period. The uptake of these amino acids decreased rapidly during the early stationary phase. The uptake of valine and cysteine increased again to some extent just prior to the forespore stage. The uptake of glycine and serine, however, was largest at the forespore stage at which the formation of spore coat took place. From these observed phenomena it was assumed that the remarkable incorporation of glycine and serine into the wild type strain during sporulation was closely related to the formation of spore coat. (auth.)

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

    DEFF Research Database (Denmark)

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

    2012-01-01

    known transcription regulation network. Interactions across multiple levels of regulation were involved in adaptive changes that could also be achieved by controlling single genes. Our analysis suggests that global trade-offs and evolutionary constraints provide incentives to favor complex control......Adaptation of cells to environmental changes requires dynamic interactions between metabolic and regulatory networks, but studies typically address only one or a few layers of regulation. For nutritional shifts between two preferred carbon sources of Bacillus subtilis, we combined statistical and...... model-based data analyses of dynamic transcript, protein, and metabolite abundances and promoter activities. Adaptation to malate was rapid and primarily controlled posttranscriptionally compared with the slow, mainly transcriptionally controlled adaptation to glucose that entailed nearly half of the...

  20. The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca

    OpenAIRE

    Romero, Diego; de Vicente, Antonio; Rakotoaly, Rivo H.; Dufour, Samuel E.; Veening, Jan-Willem; Arrebola, Eva; Cazorla, Francisco M.; Kuipers, Oscar P; Paquot, Michel; Perez-Garcia, Alejandro; Stacey, Gary

    2007-01-01

    Podosphaera fusca is the main causal agent of cucurbit powdery mildew in Spain. Four Bacillus subtilis strains, UMAF6614, UMAF6619, UMAF6639, and UMAF8561, with proven ability to suppress the disease on melon in detached leaf and seedling assays, were subjected to further analyses to elucidate the mode of action involved in their biocontrol performance. Cell-free supernatants showed antifungal activities very close to those previously reported for vegetative cells. Identification of three lip...

  1. Separation and identification of endoxylanases from Bacillus subtilis and their actions on wheat bran insoluble dietary fibre

    OpenAIRE

    Xiaoping, Yuan; Jing, Wang; Huiyuan, Yao; Nihorimbere, Venant

    2005-01-01

    A novel and convenient method based on native polyacrylamide gel electrophoresis (PAGE) and homogenization extraction was used for the purification of xylanase from crude enzymes. Two xylanases were purified by this method from the crude enzyme preparation from the selected strain of Bacillus subtilis. Subsequent analysis with thin layer chromatography and high pressure liquid chromatography (HPLC) indicated that these two xylanases were endo-acting enzymes, designated xyl I and xyl II. Both ...

  2. Effect of subinhibitory concentrations of four commonly used biocides on the conjugative transfer of Tn916 in Bacillus subtilis

    OpenAIRE

    Seier-Petersen, Maria Amalie; A. Jasni; Aarestrup, Frank Møller; Vigre, Håkan; Mullany, P.; Roberts, A. P.; Agersø, Yvonne

    2013-01-01

    Objectives Large amounts of biocides are used to reduce and control bacterial growth in the healthcare sector, food production and agriculture. This work explores the effect of subinhibitory concentrations of four commonly used biocides (ethanol, hydrogen peroxide, chlorhexidine digluconate and sodium hypochlorite) on the conjugative transposition of the mobile genetic element Tn916. Methods Conjugation assays were carried out between Bacillus subtilis strains. The donor containing Tn916 was ...

  3. Inactivation of Bacillus Subtilis by Atomic Oxygen Radical Anion

    Institute of Scientific and Technical Information of China (English)

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

    2007-01-01

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

  4. Isolation and characterization of the lacA gene encoding beta-galactosidase in Bacillus subtilis and a regulator gene, lacR.

    OpenAIRE

    Daniel, R.A.; Haiech, J.; Denizot, F; Errington, J

    1997-01-01

    We have isolated transposon insertions in the lacA gene encoding an endogenous beta-galactosidase of Bacillus subtilis. Upstream of the putative operon containing lacA is a negative regulator, lacR, which encodes a product related to a family of regulators that includes the lactose repressor, lacI, of Escherichia coli. New strains with insertions in the lacA gene should be of use in studies using lacZ fusions in B. subtilis.

  5. Transcriptional Regulation and Adaptation to a High-Fiber Environment in Bacillus subtilis HH2 Isolated from Feces of the Giant Panda

    OpenAIRE

    Zhou, Ziyao; Zhou, Xiaoxiao; Li, Jin; Zhong, Zhijun; Li, Wei; Liu, Xuehan; Liu, Furui; Su, Huaiyi; Luo, Yongjiu; Gu, Wuyang; Wang, Chengdong; Zhang, Hemin; Li, Desheng; He, Tingmei; Fu, Hualin

    2015-01-01

    In the giant panda, adaptation to a high-fiber environment is a first step for the adequate functioning of intestinal bacteria, as the high cellulose content of the gut due to the panda's vegetarian appetite results in a harsh environment. As an excellent producer of several enzymes and vitamins, Bacillus subtilis imparts various advantages to animals. In our previous study, we determined that several strains of B. subtilis isolated from pandas exhibited good cellulose decomposition ability, ...

  6. The mycosubtilin synthetase of Bacillus subtilis ATCC6633: A multifunctional hybrid between a peptide synthetase, an amino transferase, and a fatty acid synthase

    OpenAIRE

    Duitman, Erwin H.; Hamoen, Leendert W.; Rembold, Martina; Venema, Gerard; Seitz, Harald; Saenger, Wolfram; Bernhard, Frank; Reinhardt, Richard; Schmidt, Manuel; Ullrich, Christian; Stein, Torsten; Leenders, Frank; Vater, Joachim

    1999-01-01

    Bacillus subtilis strain ATCC6633 has been identified as a producer of mycosubtilin, a potent antifungal peptide antibiotic. Mycosubtilin, which belongs to the iturin family of lipopeptide antibiotics, is characterized by a β-amino fatty acid moiety linked to the circular heptapeptide Asn-Tyr-Asn-Gln-Pro-Ser-Asn, with the second, third, and sixth position present in the D-configuration. The gene cluster from B. subtilis ATCC6633 specifying the biosynthesis of mycosubtilin was identified. The ...

  7. Surfactin production enhances the level of cardiolipin in the cytoplasmic membrane of Bacillus subtilis

    Czech Academy of Sciences Publication Activity Database

    Seydlová, G.; Fišer, R.; Čabala, R.; Kozlík, P.; Svobodová, J.; Pátek, Miroslav

    2013-01-01

    Roč. 1828, č. 11 (2013), s. 2370-2378. ISSN 0005-2736 Institutional support: RVO:61388971 Keywords : Surfactin * Bacillus subtilis * Membrane Subject RIV: EE - Microbiology, Virology Impact factor: 3.431, year: 2013

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

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  9. Impact of a Bacterial Volatile 2,3-Butanediol on Bacillus subtilis Rhizosphere Robustness

    Science.gov (United States)

    Yi, Hwe-Su; Ahn, Yeo-Rim; Song, Geun C.; Ghim, Sa-Youl; Lee, Soohyun; Lee, Gahyung; Ryu, Choong-Min

    2016-01-01

    Volatile compounds, such as short chain alcohols, acetoin, and 2,3-butanediol, produced by certain strains of root-associated bacteria (rhizobacteria) elicit induced systemic resistance in plants. The effects of bacterial volatile compounds (BVCs) on plant and fungal growth have been extensively studied; however, the impact of bacterial BVCs on bacterial growth remains poorly understood. In this study the effects of a well-characterized bacterial volatile, 2,3-butanediol, produced by the rhizobacterium Bacillus subtilis, were examined in the rhizosphere. The nature of 2,3-butanediol on bacterial cells was assessed, and the effect of the molecule on root colonization was also determined. Pepper roots were inoculated with three B. subtilis strains: the wild type, a 2,3-butanediol overexpressor, and a 2,3-butanediol null mutant. The B. subtilis null strain was the first to be eliminated in the rhizosphere, followed by the wild-type strain. The overexpressor mutant was maintained at roots for the duration of the experiment. Rhizosphere colonization by a saprophytic fungus declined from 14 days post-inoculation in roots treated with the B. subtilis overexpressor strain. Next, exudates from roots exposed to 2,3-butanediol were assessed for their impact on fungal and bacterial growth in vitro. Exudates from plant roots pre-treated with the 2,3-butanediol overexpressor were used to challenge various microorganisms. Growth was inhibited in a saprophytic fungus (Trichoderma sp.), the 2,3-butanediol null B. subtilis strain, and a soil-borne pathogen, Ralstonia solanacearum. Direct application of 2,3-butanediol to pepper roots, followed by exposure to R. solanacearum, induced expression of Pathogenesis-Related (PR) genes such as CaPR2, CaSAR8.2, and CaPAL. These results indicate that 2,3-butanediol triggers the secretion of root exudates that modulate soil fungi and rhizosphere bacteria. These data broaden our knowledge regarding bacterial volatiles in the rhizosphere and

  10. Genotypic Diversity among Bacillus cereus and Bacillus thuringiensis Strains

    OpenAIRE

    Carlson, Cathrine Rein; Caugant, Dominique A; Kolstø, Anne-Brit

    1994-01-01

    Twenty-four strains of Bacillus cereus were analyzed by pulsed-field gel electrophoresis (PFGE) and compared with 12 Bacillus thuringiensis strains. In addition, the 36 strains were examined for variation in 15 chromosomal genes encoding enzymes (by multilocus enzyme electrophoresis [MEE]). The genome of each strain had a distinct NotI restriction enzyme digestion profile by PFGE, and the 36 strains could be assigned to 27 multilocus genotypes by MEE. However, neither PFGE nor MEE analysis co...

  11. [Cloning the alpha-amylase gene of Streptococcus bovis and its expression in Bacillus subtilis cells].

    Science.gov (United States)

    Iakorski, P; Kuntsova, M M; Loseva, E F; Khasanov, F K

    1991-06-01

    The gene coding for alpha-amylase from the ruminant bacterium Streptococcus bovis was cloned on the plasmid pMX39 in Bacillus subtilis cells. An alpha-amylase positive colony was isolated in the initial screening of 3900 colonies on the medium containing insoluble starch. The size of the insert was approximately 2.8 kb. The recombinant plasmid was stably maintained in Bacillus subtilis cells under the nonselective conditions. PMID:1944323

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

    NARCIS (Netherlands)

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

    1991-01-01

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

  13. Identification of a sporulation locus in cloned Bacillus subtilis deoxyribonucleic acid.

    OpenAIRE

    Moran, C P; Losick, R; Sonenshein, A L

    1980-01-01

    A cloned deoxyribonucleic acid from the purA-cysA region of the Bacillus subtilis chromosome was shown to contain the spoVC locus, a gene whose product is required for sporulation. This is the first demonstration of a spo locus in cloned B. subtilis deoxyribonucleic acid.

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

    NARCIS (Netherlands)

    Detert Oude Weme, Ruud Gerardus Johannes

    2015-01-01

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

  15. Transfection of Bacillus subtilis protoplasts by bacteriophage phi do7 DNA.

    OpenAIRE

    Perkins, J B; Dean, D H

    1983-01-01

    DNA from the Bacillus subtilis temperate bacteriophage phi do7 was found to efficiently transfect B. subtilis protoplasts; protoplast transfection was more efficient than competent cell transfection by a magnitude of 10(3). Unlike competent cell transfection, protoplast transfection did not require primary recombination, suggesting that phi do7 DNA enters the protoplast as double-stranded molecules.

  16. Identification and characterization of Bacillus subtilis from grass carp (Ctenopharynodon idellus) for use as probiotic additives in aquatic feed.

    Science.gov (United States)

    Guo, Xia; Chen, Dan-Dan; Peng, Kai-Song; Cui, Zheng-Wei; Zhang, Xu-Jie; Li, Shun; Zhang, Yong-An

    2016-05-01

    Bacillus subtilis is widely used as probiotic species in aquaculture for water quality control, growth promoting, or immunity enhancing. The aim of this study is to find novel B. subtilis strains from fish as potential probiotics for aquaculture. Eleven B. subtilis isolates derived from the intestinal tract of grass carp were identified by gene sequencing and biochemical tests. These isolates were classified into 4 groups, and the representatives (GC-5, GC-6, GC-21 and GC-22) of each group were further investigated for antibiotic susceptibility, sporulation rate, biofilm formation, activity against pathogenic bacteria, resistance to stress conditions of intestinal tract (high percentage of bile and low pH) and high temperature, which are important for probiotics to be used as feed additives. Additionally, the adhesion properties of the 4 characterized strains were assessed using Caco-2 cell and gut mucus models. The results showed that the 4 strains differed in their capacities to adhere to intestinal epithelial cells and mucus. Furthermore, the strains GC-21 and GC-22 up-regulated the expression levels of IL-10 and TGF-β but down-regulated IL-1β, suggesting their potential anti-inflammatory abilities. Based on physiological properties of the 4 characterized B. subtilis strains, one or more strains may have potential to be used as probiotics in aquaculture. PMID:26988285

  17. 1株枯草芽胞杆菌的鉴定及其弹性蛋白酶结构研究%Bacillus subtilis Strain Identification and Its Enzyme Structure of Elastase

    Institute of Scientific and Technical Information of China (English)

    王超; 陈启和; 倪辉; 李利君; 蔡慧农; 苏文金

    2012-01-01

    , capable of starch hydrolysis, V-P reaction positive, and identified strain EL32 as Bacillus subtilis. The molecular weight of elastase purified from fermentation broth was 31 ku by SDS-PACE analysis. The elaslase determined by LTQ-MS peplide fingerprint spectrum indicated it was subtilisin, the gene and protein sequence of elastase shared high homology of 99% with subtilisin. The three dimensional structure of the elastase strain EL32 consisted of six a-helices, seven-stranded parallel ?folds and two anti-parallel p-folds, and His, Asp, and Ser were the key groups of active center. The euuuse-producing strain EL32 was identified as Bacillus sublilis, and confirmed ihe elastase to be subtilisin. All these results provided a foundation for the application of the subtilisin.

  18. Levan from Bacillus subtilis Natto: its effects in normal and in streptozotocin-diabetic rats

    Directory of Open Access Journals (Sweden)

    Fernando Cesar Bazani Cabral de Melo

    2012-12-01

    Full Text Available Levan is an exopolysaccharide of fructose primarily linked by β-(2→6 glycosidic bonds with some β-(2→1 branched chains. Due to its chemical properties, levan has possible applications in both the food and pharmaceutical industries. Bacillus subtilis is a promising industrial levan producer, as it ferments sucrose and has a high levan-formation capacity. A new strain of B. subtilis was recently isolated from Japanese food natto, and it has produced levan in large quantities. For future pharmaceutical applications, this study aimed to investigate the effects of levan produced by B. subtilis Natto, mainly as potential hypoglycemic agent, (previously optimized with a molecular weight equal to 72.37 and 4,146 kDa in Wistar male rats with diabetes induced by streptozotocin and non-diabetic rats and to monitor their plasma cholesterol and triacylglycerol levels. After 15 days of experimentation, the animals were sacrificed, and their blood samples were analyzed. The results, compared using analysis of variance, demonstrated that for this type of levan, a hypoglycemic effect was not observed, as there was no improvement of diabetes symptoms during the experiment. However, levan did not affect any studied parameters in normal rats, indicating that the exopolysaccharide can be used for other purposes.

  19. Levan from Bacillus subtilis Natto: its effects in normal and in streptozotocin-diabetic rats.

    Science.gov (United States)

    de Melo, Fernando Cesar Bazani Cabral; Zaia, Cássia Thaïs Bussamra Viera; Celligoi, Maria Antonia Pedrine Colabone

    2012-10-01

    Levan is an exopolysaccharide of fructose primarily linked by β-(2→6) glycosidic bonds with some β-(2→1) branched chains. Due to its chemical properties, levan has possible applications in both the food and pharmaceutical industries. Bacillus subtilis is a promising industrial levan producer, as it ferments sucrose and has a high levan-formation capacity. A new strain of B. subtilis was recently isolated from Japanese food natto, and it has produced levan in large quantities. For future pharmaceutical applications, this study aimed to investigate the effects of levan produced by B. subtilis Natto, mainly as potential hypoglycemic agent, (previously optimized with a molecular weight equal to 72.37 and 4,146 kDa) in Wistar male rats with diabetes induced by streptozotocin and non-diabetic rats and to monitor their plasma cholesterol and triacylglycerol levels. After 15 days of experimentation, the animals were sacrificed, and their blood samples were analyzed. The results, compared using analysis of variance, demonstrated that for this type of levan, a hypoglycemic effect was not observed, as there was no improvement of diabetes symptoms during the experiment. However, levan did not affect any studied parameters in normal rats, indicating that the exopolysaccharide can be used for other purposes. PMID:24031993

  20. [Sporulation or competence development? A genetic regulatory network model of cell-fate determination in Bacillus subtilis].

    Science.gov (United States)

    Lu, Zhenghui; Zhou, Yuling; Zhang, Xiaozhou; Zhang, Guimin

    2015-11-01

    Bacillus subtilis is a generally recognized as safe (GRAS) strain that has been widely used in industries including fodder, food, and biological control. In addition, B. subtilis expression system also plays a significant role in the production of industrial enzymes. However, its application is limited by its low sporulation frequency and transformation efficiency. Immense studies have been done on interpreting the molecular mechanisms of sporulation and competence development, whereas only few of them were focused on improving sporulation frequency and transformation efficiency of B. subtilis by genetic modification. The main challenge is that sporulation and competence development, as the two major developmental events in the stationary phase of B. subtilis, are regulated by the complicated intracellular genetic regulatory systems. In addition, mutual regulatory mechanisms also exist in these two developmental events. With the development of genetic and metabolic engineering, constructing genetic regulatory networks is currently one of the most attractive research fields, together with the genetic information of cell growth, metabolism, and development, to guide the industrial application. In this review, the mechanisms of sporulation and competence development of B. subtilis, their interactions, and the genetic regulation of cell growth were interpreted. In addition, the roles of these regulatory networks in guiding basic and applied research of B. subtilis and its related species were discussed. PMID:26939438

  1. Caracterización de cristales de calcita bioprecipitada por un aislamiento nativo de Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Carolina Montoya

    2007-02-01

    . subtilis isolate collected from a gold mine in Segovia (Antioquia, Colombia. Its calcification capability was assessed by determining the production of CaCO3 crystals using the specific B4 media culture. In addition, mineralogical analyses were conducted, using techniques such as a binocular stereoscopy, plane polarized light optical microscopy (PPLOM, scanning electronic microscopy with energy dispersive X-ray detector (ESEM/EDX and Fourier Transform Infrared spectroscopy (FTIR. These analyses showed that the native isolated strain of B. subtilis produces calcium carbonate (CaCO3 in its low temperature polymorphic form, (calcite.Key words: Bacillus subtilis, calcite, bioprecipitation, applied mineralogy, biomineralogy.

  2. Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The gene coding for a thermophilic neutral protease from Bacillus stearothermophilus was expressed in Bacillus subtilis DB104, under the control of the sacB gene promoter. This was followed by either the native signal peptide sequence of this protease or the signal peptide sequence of the sacB gene. The protease was purified 3.8-fold, with a specific activity of 16530 U mg-1. As analyzed by SDS-PAGE, the molecular mass of the expressed protease was about 35 kDa, and the optimal temperature and pH of the protease were 65℃ and 7.5, respectively. Moreover, it still had about 80% activity after 1 h reaction at 65 ℃ .

  3. Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The gene coding for a thermophilic neutral protease from Bacillus stearothermophilus was expressed in Bacillus subtilis DB104, under the control of the sacB gene promoter. This was followed by either the native signal peptide sequence of this protease or the signal peptide sequence of the sacB gene. The protease was purified 3.8-fold, with a specific activity of 16530 U mg-1. As analyzed by SDS-PAGE, the molecular mass of the expressed protease was about 35 kDa, and the optimal temperature and pH of the protease were 65℃ and 7.5, respectively. Moreover, it still had about 80% activity after 1 h reaction at 65℃.

  4. HIGH-LEVEL PRODUCTION OF A COLD-ACTIVE ƒ-MANNANASE FROM BACILLUS SUBTILIS BS5 AND ITS MOLECULAR CLONING AND EXPRESSION

    OpenAIRE

    Jun, Li; Ling, Xiao; Han, Yan; Shu, Gang; Gui, Xue

    2012-01-01

    Abstract. Mannanases can be useful in the food, feed, pulp and paper industries. In this research a Bacillus subtilis strain (named Bs5) which produced high-level β-mannanase was isolated. Maximum level of β-mannanase (1231.41 U/ml) was reached when Bacillus subtilis Bs5 was grown on konjac powder as the carbon source for nine hours at 32 °C. The β-mannanase was a typical cold-active enzyme and its optimal temperature of 35 °C was the lowest among those of the known mannanases from bacteria. ...

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

    NARCIS (Netherlands)

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

    1996-01-01

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

  6. Bacillus subtilis FZB24® Affects Flower Quantity and Quality of Saffron (Crocus sativus)

    OpenAIRE

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

    2008-01-01

    The effect of Bacillus subtilis FZB24® on saffron (Crocus sativus L.) was studied using saffron corms from Spain and the powdered form of B. subtilis FZB24®. Corms were soaked in water or in B. subtilis FZB24 spore solution for 15min before sowing. Some corms were further soil drenched with the spore solution 6, 10 or 14 weeks after sowing. Growth and saffron stigma chemical composition were measured. Compared to untreated controls, application of B. subtilis FZB24 significantly increased lea...

  7. Heterologous expression, biochemical characterization, and overproduction of alkaline α-amylase from Bacillus alcalophilus in Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Li Jianghua

    2011-10-01

    Full Text Available Abstract Background Alkaline α-amylases have potential applications for hydrolyzing starch under high pH conditions in the starch and textile industries and as ingredients in detergents for automatic dishwashers and laundries. While the alkaline α-amylase gains increased industrial interest, the yield of alkaline α-amylases from wild-type microbes is low, and the combination of genetic engineering and process optimization is necessary to achieve the overproduction of alkaline α-amylase. Results The alkaline α-amylase gene from Bacillus alcalophilus JN21 (CCTCC NO. M 2011229 was cloned and expressed in Bacillus subtilis strain WB600 with vector pMA5. The recombinant alkaline α-amylase was stable at pH from 7.0 to 11.0 and temperature below 40°C. The optimum pH and temperature of alkaline α-amylase was 9.0 and 50°C, respectively. Using soluble starch as the substrate, the Km and Vmax of alkaline α-amylase were 9.64 g/L and 0.80 g/(L·min, respectively. The effects of medium compositions (starch, peptone, and soybean meal and temperature on the recombinant production of alkaline α-amylase in B. subtilis were investigated. Under the optimal conditions (starch concentration 0.6% (w/v, peptone concentration 1.45% (w/v, soybean meal concentration 1.3% (w/v, and temperature 37°C, the highest yield of alkaline α-amylase reached 415 U/mL. The yield of alkaline α-amylase in a 3-L fermentor reached 441 U/mL, which was 79 times that of native alkaline α-amylase from B. alcalophilus JN21. Conclusions This is the first report concerning the heterologous expression of alkaline α-amylase in B. subtilis, and the obtained results make it feasible to achieve the industrial production of alkaline α-amylase with the recombinant B. subtilis.

  8. Molecular cloning and characterisation of the ribC gene from Bacillus subtilis : A point mutation in ribC results in riboflavin overproduction

    NARCIS (Netherlands)

    Coquard, D; Huecas, M; Ott, M; vanDijl, JM; VanLoon, APGM; Hohmann, HP

    1997-01-01

    A mutation leading to roseoflavin resistance and deregulated riboflavin biosynthesis was mapped in the genome of the riboflavin-overproducing Bacillus subtilis strains RB52 and RB50 at map position 147 degrees. The chromosomal location indicates that the deregulating mutation in RB52 and RB50 is an

  9. Complete Genome Sequence of Bacillus subtilis BSn5, an Endophytic Bacterium of Amorphophallus konjac with Antimicrobial Activity for the Plant Pathogen Erwinia carotovora subsp. carotovora ▿

    OpenAIRE

    Deng, Yun; Zhu, Yiguang; Wang, Pengxia; Zhu, Lei; Zheng, Jinshui; Li, Rong; Ruan, Lifang; Peng, Donghai; Sun, Ming

    2011-01-01

    Here, we present the complete genome sequence of Bacillus subtilis strain BSn5, isolated from Amorphophallus konjac calli tissue and showing strong inhibitory activity to Erwinia carotovora subsp. carotovora, which causes Amorphophallus soft rot disease and affects the industry development of this organism.

  10. The mycosubtilin synthetase of Bacillus subtilis ATCC6633 : A multifunctional hybrid between a peptide synthetase, an amino transferase, and a fatty acid synthase

    NARCIS (Netherlands)

    Duitman, EH; Hamoen, LW; Rembold, M; Venema, G; Seitz, H; Saenger, W; Bernhard, F; Reinhardt, R; Schmidt, M; Ullrich, C; Stein, T; Leenders, F; Vater, J

    1999-01-01

    Bacillus subtilis strain ATCC6633 has been identified as a producer of mycosubtilin, a potent antifungal peptide antibiotic. Mycosubtilin, which belongs to the iturin family of lipopeptide antibiotics, is characterized by a p-amino fatty acid moiety linked to the circular heptapeptide Asn-Tyr-Asn-Cl

  11. [Bacillus subtilis and streptomycin resistant mutant growth in the medium with saponite].

    Science.gov (United States)

    Chebotarev, A Iu; Gordienko, A S; Kurdish, I K

    2013-01-01

    The influence of dispersed saponite on growth activity of Bacillus subtilis IMV B-7023 and its streptomycin resistant mutant has been shown. The effectiveness of this process depends on the content of dispersed material and phosphate in the medium. It has been found that when B. subtilis is cultured in the medium containing 0.6 g/l PO4(3-) stimulation of bacteria growth is observed, but at a lower concentration (0.1 g/l PO4(3-)) there is a decline in the culture growth activity. At the same time streptomycin resistant mutant is shown to increase growth activity in the growth medium which contains up to 1.0 g/l saponite, regardless of the concentration of phosphate. It is shown that this effect is a consequence of uniformity of surface properties of streptomycin resistant strain of bacteria and similar parent strain at a concentration of 0.6 g/l PO4(3-). PMID:24479315

  12. Regulation of the tryptophan biosynthetic genes in Bacillus halodurans: common elements but different strategies than those used by Bacillus subtilis.

    Science.gov (United States)

    Szigeti, Reka; Milescu, Mirela; Gollnick, Paul

    2004-02-01

    In Bacillus subtilis, an RNA binding protein called TRAP regulates both transcription and translation of the tryptophan biosynthetic genes. Bacillus halodurans is an alkaliphilic Bacillus species that grows at high pHs. Previous studies of this bacterium have focused on mechanisms of adaptation for growth in alkaline environments. We have characterized the regulation of the tryptophan biosynthetic genes in B. halodurans and compared it to that in B. subtilis. B. halodurans encodes a TRAP protein with 71% sequence identity to the B. subtilis protein. Expression of anthranilate synthetase, the first enzyme in the pathway to tryptophan, is regulated significantly less in B. halodurans than in B. subtilis. Examination of the control of the B. halodurans trpEDCFBA operon both in vivo and in vitro shows that only transcription is regulated, whereas in B. subtilis both transcription of the operon and translation of trpE are controlled. The attenuation mechanism that controls transcription in B. halodurans is similar to that in B. subtilis, but there are some differences in the predicted RNA secondary structures in the B. halodurans trp leader region, including the presence of a potential anti-antiterminator structure. Translation of trpG, which is within the folate operon in both bacilli, is regulated similarly in the two species. PMID:14729709

  13. Activity of essential oils against Bacillus subtilis spores.

    Science.gov (United States)

    Lawrence, Hayley A; Palombo, Enzo A

    2009-12-01

    Alternative methods for controlling bacterial endospore contamination are desired in a range of industries and applications. Attention has recently turned to natural products, such as essential oils, which have sporicidal activity. In this study, a selection of essential oils was investigated to identify those with activity against Bacillus subtilis spores. Spores were exposed to thirteen essential oils, and surviving spores were enumerated. Cardamom, tea tree, and juniper leaf oils were the most effective, reducing the number of viable spores by 3 logs at concentrations above 1%. Sporicidal activity was enhanced at high temperatures (60 degrees C) or longer exposure times (up to one week). Gas chromatography-mass spectrometry analysis identified the components of the active essential oils. However, none of the major oil components exhibited equivalent activity to the whole oils. The fact that oil components, either alone or in combination, did not show the same level of sporicidal activity as the complete oils suggested that minor components may be involved, or that these act synergistically with major components. Scanning electron microscopy was used to examine spores after exposure to essential oils and suggested that leakage of spore contents was the likely mode of sporicidal action. Our data have shown that essential oils exert sporicidal activity and may be useful in applications where bacterial spore reduction is desired. PMID:20075624

  14. Dynamics of Aerial Tower Formation in Bacillus subtilis Biofilms

    Science.gov (United States)

    Sinha, Naveen; Seminara, Agnese; Wilking, James; Brenner, Michael; Weitz, Dave

    2012-02-01

    Biofilms are highly-organized colonies of bacteria that form on surfaces. These colonies form sophisticated structures which make them robust and difficult to remove from environments such as catheters, where they pose serious infection problems. Previous work has shown that sub-mm sized aerial towers form on the surface of Bacillus subtilis colony biofilms. Spore-formation is located preferentially at the tops of these towers, known as fruiting bodies, which aid in the dispersal and propagation of the colony to new sites. The formation of towers is strongly affected by the quorum-sensing molecule surfactin and the cannibalism pathway of the bacteria. In the present work, we use confocal fluorescence microscopy to study the development of individual fruiting bodies, allowing us to visualize the time-dependent spatial distribution of matrix-forming and sporulating bacteria within the towers. With this information, we investigate the physical mechanisms, such as surface tension and polymer concentration gradients, that drive the formation of these structures.

  15. Biocalcifying Bacillus subtilis cells effectively consolidate deteriorated Globigerina limestone.

    Science.gov (United States)

    Micallef, Roderick; Vella, Daniel; Sinagra, Emmanuel; Zammit, Gabrielle

    2016-07-01

    Microbially induced calcite precipitation occurs naturally on ancient limestone surfaces in Maltese hypogea. We exploited this phenomenon and treated deteriorated limestone with biocalcifying bacteria. The limestone was subjected to various mechanical and physical tests to present a statistically robust data set to prove that treatment was indeed effective. Bacillus subtilis conferred uniform bioconsolidation to a depth of 30 mm. Drilling resistance values were similar to those obtained for freshly quarried limestone (9 N) and increased up to 15 N. Treatment resulted in a high resistance to salt deterioration and a slow rate of water absorption. The overall percentage porosity of treated limestone varied by ±6 %, thus the pore network was preserved. We report an eco-friendly treatment that closely resembles the mineral composition of limestone and that penetrates into the porous structure without affecting the limestones' natural properties. The treatment is of industrial relevance since it compares well with stone consolidants available commercially. PMID:27072564

  16. Selectivity in protein degradation during sporulation of Bacillus subtilis

    International Nuclear Information System (INIS)

    The breakdown of cellular protein was investigated in Bacillus subtilis ATCC 6051 labeled with glycine-2-3H or L-phenylalanine-U-14C at the different stages of vegetative growth and sporulation. The growth of the culture was determined by measuring optical density at 660 nm. The heat-resistant spores were scored by plating after heating at 80 deg C for 10 minutes. A question whether the turnover of glycine-labeled protein is similar to that of phenylalanine-labeled protein was experimentally studied. The patterns obtained with the glycine-labeled protein were different from those of phenylalanine-labeled protein. This was not multiple turnover. The cellular protein which was labeled with glycine at an early stage of sporulation showed rapid degradation, but the degradation of the protein labeled with glycine at later stages did not occur at all. Another question whether the labeled glycine incorporated into cells at the different stages of growth and sporulation was present in the spore coat fraction of matured spores was studied. Experiment demonstrated that the glycine incorporated into cells at the late sporulation stage was mainly utilized for the biosynthesis of the spore coat protein. These data suggest that the spore coat protein which contains relatively large amount of glycine is rarely subject to further degradation. (Iwakiri, K.)

  17. Nutrient depletion in Bacillus subtilis biofilms triggers matrix production

    International Nuclear Information System (INIS)

    Many types of bacteria form colonies that grow into physically robust and strongly adhesive aggregates known as biofilms. A distinguishing characteristic of bacterial biofilms is an extracellular polymeric substance (EPS) matrix that encases the cells and provides physical integrity to the colony. The EPS matrix consists of a large amount of polysaccharide, as well as protein filaments, DNA and degraded cellular materials. The genetic pathways that control the transformation of a colony into a biofilm have been widely studied, and yield a spatiotemporal heterogeneity in EPS production. Spatial gradients in metabolites parallel this heterogeneity in EPS, but nutrient concentration as an underlying physiological initiator of EPS production has not been explored. Here, we study the role of nutrient depletion in EPS production in Bacillus subtilis biofilms. By monitoring simultaneously biofilm size and matrix production, we find that EPS production increases at a critical colony thickness that depends on the initial amount of carbon sources in the medium. Through studies of individual cells in liquid culture we find that EPS production can be triggered at the single-cell level by reducing nutrient concentration. To connect the single-cell assays with conditions in the biofilm, we calculate carbon concentration with a model for the reaction and diffusion of nutrients in the biofilm. This model predicts the relationship between the initial concentration of carbon and the thickness of the colony at the point of internal nutrient deprivation. (paper)

  18. Transcriptional regulation of a Bacillus subtilis dipeptide transport operon.

    Science.gov (United States)

    Slack, F J; Mueller, J P; Strauch, M A; Mathiopoulos, C; Sonenshein, A L

    1991-08-01

    The Bacillus subtilis dciA operon, which encodes a dipeptide transport system, was induced rapidly by several conditions that caused the cells to enter stationary phase and initiate sporulation. The in vivo start point of transcription was mapped precisely and shown to correspond to a site of transcription initiation in vitro by the major vegetative form of RNA polymerase. Post-exponential expression was prevented by a mutation in the spo0A gene (whose product is a known regulator of early sporulation genes) but was restored in a spo0A abrB double mutant. This implicated AbrB, another known regulator, as a repressor of dciA. In fact, purified AbrB protein bound to a portion of the dciA promoter region, protecting it against DNase I digestion. Expression of dciA in growing cells was also repressed independently by glucose and by a mixture of amino acids; neither of these effects was mediated by AbrB. PMID:1766371

  19. Novel methyl transfer during chemotaxis in Bacillus subtilis

    International Nuclear Information System (INIS)

    If Bacillus subtilis is incubated in radioactive methionine in the absence of protein synthesis, the methyl-accepting chemotaxis proteins (MCPs) become radioactively methylated. If the bacteria are further incubated in excess nonradioactive methionine (cold-chased) and then given the attractant aspartate, the MCPs lose about half of their radioactivity due to turnover, in which lower specific activity methyl groups from S-adenosylmethionine (AdoMet) replace higher specific activity ones. Due to the cold-chase, the specific activity of the AdoMet pool is reduced at least 2-fold. If, later, the attractant is removed, higher specific activity methyl groups return to the MCPs. Thus, there must exist an unidentified methyl carrier than can reversibly receive methyl groups from the MCPs. In a similar experiment, labeled cells were transferred to a flow cell and exposed to addition and removal of attractant and of repellent. All four kinds of stimuli were found to cause methanol production. Bacterial with maximally labeled MCPs were exposed to many cycles of addition and removal of attractant; the maximum amount of radioactive methanol was evolved on the third, not the first, cycle. This result suggests that there is a precursor-product relationship between methyl groups on the MCPs and on the unidentified carrier, which might be the direct source of methanol. However, since no methanol was produced when a methyltransferase mutant, whose MCPs were unmethylated, was exposed to addition and removal of attractant or repellent, the methanol must ultimately derive from methylated MCPs

  20. A Low Dimensional Approximation For Competence In Bacillus Subtilis.

    Science.gov (United States)

    Nguyen, An; Prugel-Bennett, Adam; Dasmahapatra, Srinandan

    2016-01-01

    The behaviour of a high dimensional stochastic system described by a chemical master equation (CME) depends on many parameters, rendering explicit simulation an inefficient method for exploring the properties of such models. Capturing their behaviour by low-dimensional models makes analysis of system behaviour tractable. In this paper, we present low dimensional models for the noise-induced excitable dynamics in Bacillus subtilis, whereby a key protein ComK, which drives a complex chain of reactions leading to bacterial competence, gets expressed rapidly in large quantities (competent state) before subsiding to low levels of expression (vegetative state). These rapid reactions suggest the application of an adiabatic approximation of the dynamics of the regulatory model that, however, lead to competence durations that are incorrect by a factor of 2. We apply a modified version of an iterative functional procedure that faithfully approximates the time-course of the trajectories in terms of a two-dimensional model involving proteins ComK and ComS. Furthermore, in order to describe the bimodal bivariate marginal probability distribution obtained from the Gillespie simulations of the CME, we introduce a tunable multiplicative noise term in a two-dimensional Langevin model whose stationary state is described by the time-independent solution of the corresponding Fokker-Planck equation. PMID:27045827

  1. Loop grafting of Bacillus subtilis lipase A: inversion of enantioselectivity.

    Science.gov (United States)

    Boersma, Ykelien L; Pijning, Tjaard; Bosma, Margriet S; van der Sloot, Almer M; Godinho, Luís F; Dröge, Melloney J; Winter, Remko T; van Pouderoyen, Gertie; Dijkstra, Bauke W; Quax, Wim J

    2008-08-25

    Lipases are successfully applied in enantioselective biocatalysis. Most lipases contain a lid domain controlling access to the active site, but Bacillus subtilis Lipase A (LipA) is a notable exception: its active site is solvent exposed. To improve the enantioselectivity of LipA in the kinetic resolution of 1,2-O-isopropylidene-sn-glycerol (IPG) esters, we replaced a loop near the active-site entrance by longer loops originating from Fusarium solani cutinase and Penicillium purpurogenum acetylxylan esterase, thereby aiming to increase the interaction surface for the substrate. The resulting loop hybrids showed enantioselectivities inverted toward the desired enantiomer of IPG. The acetylxylan esterase-derived variant showed an inversion in enantiomeric excess (ee) from -12.9% to +6.0%, whereas the cutinase-derived variant was improved to an ee of +26.5%. The enantioselectivity of the cutinase-derived variant was further improved by directed evolution to an ee of +57.4%. PMID:18721749

  2. A four-stranded DNA from Bacillus subtilis which may be an intermediate in genetic recombination

    International Nuclear Information System (INIS)

    DNA of Bacillus subtilis strain UVSS 19-8 M, of high ultraviolet sensitivity, was isolated after cultivating in medium containing bromouracil. Isopycnic banding in CsCl shows an unusual pattern with four bands, including an extra one halfway between those for hybrid and for DNA fully substituted with bromouracil. DNA of this band, amounting to 15-25% of the total DNA mass in one preparation, was isolated and investigated. The characteristics found for this DNA, namely transforming ability, electron microscopic picture, behavior during heat denaturation and gentle shear are in agreement with a fourstranded DNA unit similar to one of the structures postulated by Holliday as intermediates during genetic recombination. The amount of this DNA when the cells were given 5J/m2 of 254 nm UV. UVSS 19-8 M from which this DNA has been isolated is shown to be defective for transformation and transfection, and can be regarded as rec-. (orig./MG)

  3. Cyclic lipopeptides from Bacillus subtilis activate distinct patterns of defence responses in grapevine.

    Science.gov (United States)

    Farace, Giovanni; Fernandez, Olivier; Jacquens, Lucile; Coutte, François; Krier, François; Jacques, Philippe; Clément, Christophe; Barka, Essaid Ait; Jacquard, Cédric; Dorey, Stéphan

    2015-02-01

    Non-self-recognition of microorganisms partly relies on the perception of microbe-associated molecular patterns (MAMPs) and leads to the activation of an innate immune response. Bacillus subtilis produces three main families of cyclic lipopeptides (LPs), namely surfactins, iturins and fengycins. Although LPs are involved in induced systemic resistance (ISR) activation, little is known about defence responses induced by these molecules and their involvement in local resistance to fungi. Here, we showed that purified surfactin, mycosubtilin (iturin family) and plipastatin (fengycin family) are perceived by grapevine plant cells. Although surfactin and mycosubtilin stimulated grapevine innate immune responses, they differentially activated early signalling pathways and defence gene expression. By contrast, plipastatin perception by grapevine cells only resulted in early signalling activation. Gene expression analysis suggested that mycosubtilin activated salicylic acid (SA) and jasmonic acid (JA) signalling pathways, whereas surfactin mainly induced an SA-regulated response. Although mycosubtilin and plipastatin displayed direct antifungal activity, only surfactin and mycosubtilin treatments resulted in a local long-lasting enhanced tolerance to the necrotrophic fungus Botrytis cinerea in grapevine leaves. Moreover, challenge with specific strains overproducing surfactin and mycosubtilin led to a slightly enhanced stimulation of the defence response compared with the LP-non-producing strain of B. subtilis. Altogether, our results provide the first comprehensive view of the involvement of LPs from B. subtilis in grapevine plant defence and local resistance against the necrotrophic pathogen Bo. cinerea. Moreover, this work is the first to highlight the ability of mycosubtilin to trigger an immune response in plants. PMID:25040001

  4. Synergistic effect of surfactin from Bacillus subtilis C4 and Achyrocline satureioides extracts on the viability of Paenibacillus larvae.

    Science.gov (United States)

    Sabaté, D C; Gonzaléz, M J; Porrini, M P; Eguaras, M J; Audisio, M C; Marioli, J M

    2012-04-01

    The aim of this work was to determine the in vitro effect of the mixture between the lipopeptide surfactin, synthesized by Bacillus subtilis C4 (strain isolated from honey) and the most active vegetal extract from Achyrocline satureioides, a traditional medicinal plant, on local strains of Paenibacillus larvae, the agent of American Foulbrood in honeybees. Five P. larvae strains isolated in Córdoba, Argentina, were phenotypically characterized. These and 12 other P. larvae strains from different regions of Argentina were analysed. The antimicrobial activities of the essential oil, hexane (HE) and benzene extracts from A. satureioides were assessed against P. larvae and the HE showed the highest anti-P. larvae activity. A combination of the biosurfactant surfactin, produced by B. subtilis C4, and the HE of A. satureioides revealed a synergistic action on P. larvae. The effective surfactin concentration in the mixture decreased from 32 to 1 μg ml(-1) and the HE concentration from 32 to 4 μg ml(-1), values similar or equal to minimal inhibitory concentrations observed for oxytetracycline. The fractional inhibitory concentration index confirmed synergism in 4 strains and partial synergism in one strain. The combination of surfactin synthesized by B. subtilis C4 and the HE from A. satureioides could be a natural alternative to help beekeepers to combat the American foulbrood agent P. larvae. PMID:22805922

  5. 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

    OpenAIRE

    Ratu SAFITRI; Bambang PRIADIE; Mia MIRANTI; Arum Widi ASTUTI

    2015-01-01

    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...

  6. Cucumber Rhizosphere Microbial Community Response to Biocontrol Agent Bacillus subtilis B068150

    Directory of Open Access Journals (Sweden)

    Lihua Li

    2015-12-01

    Full Text Available Gram-positive bacteria Bacillus subtilis B068150 has been used as a biocontrol agent against the pathogen Fusarium oxysporum cucumerinum. Cucumber was grown in three soils with strain B068150 inoculated in a greenhouse for 90 days, and the colonization ability of strain B068150 in cucumber rhizosphere and non-rhizosphere soils was determined. Changes in total bacteria and fungi community composition and structures using denaturing gradient gel electrophoresis (DGGE and sequencing were determined. Colony counts showed that B068150 colonization in the rhizosphere was significantly higher (p < 0.001 than in non-rhizosphere soils. Based on our data, the introduction of B. bacillus B068150 did not change the diversity of microbial communities significantly in the rhizosphere of three soils. Our data showed that population density of B068150 in clay soil had a significant negative correlation on bacterial diversity in cucumber rhizosphere in comparison to loam and sandy soils, suggesting that the impact of B068150 might be soil specific.

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

    NARCIS (Netherlands)

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

    1994-01-01

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

  8. Enhancement of extracellular expression of Bacillus naganoensis pullulanase from recombinant Bacillus subtilis: Effects of promoter and host.

    Science.gov (United States)

    Song, Wan; Nie, Yao; Mu, Xiao Qing; Xu, Yan

    2016-08-01

    Pullulanase plays an important role in industrial applications of starch processing. However, extracellular production of pullulanase from recombinant Bacillus subtilis is yet limited due to the issues on regulatory elements of B. subtilis expression system. In this study, the gene encoding B. naganoensis pullulanase (PUL) was expressed in B. subtilis WB800 under the promoter PHpaII in the shuttle vector pMA0911. The extracellular activity of expressed pullulanase was 3.9 U ml(-1) from the recombinant B. subtilis WB800/pMA0911-PHpaII-pul. To further enhance the yield of PUL, the promoter PHpaII in pMA0911 was replaced by a stronger constitutive promoter P43. Then the activity was increased to 8.7 U ml(-1) from the recombinant B. subtilis WB800/pMA0911-P43-pul. Effect of host on pullulanase expression was further investigated by comparison between B. subtilis WB600 and B. subtilis WB800. In addition to the available B. subtilis WB800 recombinants, the constructed plasmids pMA0911-PHpaII-pul and pMA0911-P43-pul were transformed into B. subtilis WB600, respectively. Consequently, the extracellular production of PUL was significantly enhanced by B. subtilis WB600/pMA0911-P43-pul, resulting in the extracellular pullulanase activity of 24.5 U ml(-1). Therefore, promoter and host had an impact on pullulanase expression and their optimization would be useful to improve heterologous protein expression in B. subtilis. PMID:27109467

  9. Isolation and Structural Characterization of an Oligosaccharide Produced by Bacillus subtilis in a Maltose-Containing Medium

    Science.gov (United States)

    Shin, Kwang-Soon

    2016-01-01

    Among 116 bacterial strains isolated from Korean fermented foods, one strain (SS-76) was selected for producing new oligosaccharides in a basal medium containing maltose as the sole source of carbon. Upon morphological characterization using scanning electron microscopy, the cells of strain SS-76 appeared rod-shaped; subsequent 16S rRNA gene sequence analysis revealed that strain SS-76 was phylogenetically close to Bacillus subtilis. The main oligosaccharide fraction B extracted from the culture supernatant of B. subtilis SS-76 was purified by high performance liquid chromatography. Subsequent structural analysis revealed that this oligosaccharide consisted only of glucose, and methylation analysis indicated similar proportions of glucopyranosides in the 6-linkage, 4-linkage, and non-reducing terminal positions. Matrix-assisted laser-induced/ionization time-of-flight/mass spectrometry and electrospray ionization-based liquid chromatography-mass spectrometry/mass spectrometry analyses suggested that this oligosaccharide consisted of a trisaccharide unit with 1,6- and 1,4-glycosidic linkages. The anomeric signals in the 1H-nuclear magnetic resonance spectrum corresponded to α-anomeric configurations, and the trisaccharide was finally identified as panose (α-D-glucopyranosyl-1,6-α-D-glucopyranosyl-1,4-D-glucose). These results suggest that B. subtilis SS-76 converts maltose into panose; strain SS-76 may thus find industrial application in the production of panose. PMID:27390729

  10. Isolation and Structural Characterization of an Oligosaccharide Produced by Bacillus subtilis in a Maltose-Containing Medium.

    Science.gov (United States)

    Shin, Kwang-Soon

    2016-06-01

    Among 116 bacterial strains isolated from Korean fermented foods, one strain (SS-76) was selected for producing new oligosaccharides in a basal medium containing maltose as the sole source of carbon. Upon morphological characterization using scanning electron microscopy, the cells of strain SS-76 appeared rod-shaped; subsequent 16S rRNA gene sequence analysis revealed that strain SS-76 was phylogenetically close to Bacillus subtilis. The main oligosaccharide fraction B extracted from the culture supernatant of B. subtilis SS-76 was purified by high performance liquid chromatography. Subsequent structural analysis revealed that this oligosaccharide consisted only of glucose, and methylation analysis indicated similar proportions of glucopyranosides in the 6-linkage, 4-linkage, and non-reducing terminal positions. Matrix-assisted laser-induced/ionization time-of-flight/mass spectrometry and electrospray ionization-based liquid chromatography-mass spectrometry/mass spectrometry analyses suggested that this oligosaccharide consisted of a trisaccharide unit with 1,6- and 1,4-glycosidic linkages. The anomeric signals in the (1)H-nuclear magnetic resonance spectrum corresponded to α-anomeric configurations, and the trisaccharide was finally identified as panose (α-D-glucopyranosyl-1,6-α-D-glucopyranosyl-1,4-D-glucose). These results suggest that B. subtilis SS-76 converts maltose into panose; strain SS-76 may thus find industrial application in the production of panose. PMID:27390729

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

    International Nuclear Information System (INIS)

    Vegetative cells of Bac. thuringiensis var. galleriae (the wild-type strain 351) are much more sensitive to lethal effects of UV light and 60Co-γ-rays than those of Bac. subtilis (the wild-type strain 168). This difference is less pronounced for spores of these strains. By means of repeated γ-irradiation-regrowth cycles radioresistant mutants Bac. thuringiensis Gamsup(r) 14 and Bac. subtilis Gamsup(r) 9 were selected. The vegetative cells of these mutants are correspondingly 19 times and 3.9 times more resistant to lethal effects of γ-radiation than the cells of the parental strains. The resistance of the Gamsup(r) mutant cells to lethal effects of UV light and H2O2 is also increased. The spores of the Gamsup(r) 14 mutant are 1.5-1.7 times more resistant to γ-radiation and UV light than the wild-type spores. The radioresistant mutants and the parental strains do not vary in their capacity for host-cell reactivation of UV- or γ-irradiated phages Tg13 and 105

  12. Isolation and characterization of antagonistic Bacillus strains capable to degrade ethylenethiourea.

    Science.gov (United States)

    Vágvölgyi, Csaba; Sajben-Nagy, Enikő; Bóka, Bettina; Vörös, Mónika; Berki, Adrienn; Palágyi, Andrea; Krisch, Judit; Skrbić, Biljana; Durišić-Mladenović, N; Manczinger, László

    2013-03-01

    In this study, more than 150 bacteria showing antagonistic properties against bacterial and fungal pathogens of the tomato plant were isolated and characterized. The most efficient agents against these phytopathogenic microorganisms belong to the genus Bacillus: the best biocontrol isolates were representatives of Bacillus subtilis, B. mojavensis and B. amyloliquefaciens species. They intensively produced fengycin or/and surfactin depsipeptide antibiotics and also proved to be excellent protease secretors. It was proved, that the selected strains were able to use ethylenethiourea (ETU) as sole nitrogen source. These antagonistic and ETU-degrading Bacillus strains can be applied as biocontrol and also as bioremediation agents. PMID:23143288

  13. Lipopeptides as main ingredients for inhibition of fungal phytopathogens by Bacillus subtilis/amyloliquefaciens

    Science.gov (United States)

    Cawoy, Hélène; Debois, Delphine; Franzil, Laurent; De Pauw, Edwin; Thonart, Philippe; Ongena, Marc

    2015-01-01

    Some isolates of the Bacillus subtilis/amyloliquefaciens species are known for their plant protective activity against fungal phytopathogens. It is notably due to their genetic potential to form an impressive array of antibiotics including non-ribosomal lipopeptides (LPs). In the work presented here, we wanted to gain further insights into the relative role of these LPs in the global antifungal activity of B. subtilis/amyloliquefaciens. To that end, a comparative study was conducted involving multiple strains that were tested against four different phytopathogens. We combined various approaches to further exemplify that secretion of those LPs is a crucial trait in direct pathogen ward off and this can actually be generalized to all members of these species. Our data illustrate that for each LP family, the fungitoxic activity varies in function of the target species and that the production of iturins and fengycins is modulated by the presence of pathogens. Our data on the relative involvement of these LPs in the biocontrol activity and modulation of their production are discussed in the context of natural conditions in the rhizosphere. PMID:25529983

  14. Directed evolution of adenylosuccinate synthetase from Bacillus subtilis and its application in metabolic engineering.

    Science.gov (United States)

    Wang, Xiaoyue; Wang, Guanglu; Li, Xinli; Fu, Jing; Chen, Tao; Wang, Zhiwen; Zhao, Xueming

    2016-08-10

    Adenylosuccinate synthetase (EC. 6.3.4.4) encoded by purA in Bacillus subtilis, catalyzing the first step of the conversion of IMP to AMP, plays an important role in flux distribution in the purine biosynthetic pathway. In this study, we described the use of site saturation mutagenesis to obtain a desired enzyme activity of adenylosuccinate synthetase and its application in flux regulation. Based on sequence alignment and structural modeling, a library of enzyme variants was created by a semi-rational evolution strategy in position Thr238 and Pro242. Other than purA deletion, the leaky mutation purA(P242N) partially reduced the flux towards AMP derived from IMP and increased the riboflavin synthesis precursor GTP, while also kept the requirement of ATP synthesis for cell growth. PurA(P242N) was introduced into an inosine-producing strain and resulted in an approximately 4.66-fold increase in inosine production, from 0.088±0.009g/L to 0.41±0.051g/L, in minimal medium without hypoxanthine accumulation. These results underline that the directed evolution of adenylosuccinate synthetase could tailor its activities and adjust metabolic flux. This mutation may provide a promising application in purine-based product accumulation, like inosine, guanosine and folate which are directly stemming from purine pathway in B. subtilis. PMID:27234879

  15. Biocontrol of Rhizoctonia solani Damping-Off of Tomato with Bacillus subtilis RB14

    OpenAIRE

    Asaka, O.; Shoda, M

    1996-01-01

    Bacillus subtilis RB14, which showed antibiotic activities against several phytopathogens in vitro by producing the antibiotics iturin A and surfactin, was subjected to a pot test to investigate its ability to suppress damping-off of tomato seedlings caused by Rhizoctonia solani. To facilitate recovery from soil, B. subtilis RB14-C, a spontaneous streptomycin-resistant mutant of RB14, was used. Damping-off was suppressed when the culture broth, cell suspension, or cell-free culture broth of R...

  16. Intrinsic Levanase Activity of Bacillus subtilis 168 Levansucrase (SacB)

    OpenAIRE

    Méndez-Lorenzo, Luz; Jaime R Porras-Domínguez; Raga-Carbajal, Enrique; Olvera, Clarita; Rodríguez-Alegría, Maria Elena; Carrillo-Nava, Ernesto; Costas, Miguel; López Munguía, Agustín

    2015-01-01

    Levansucrase catalyzes the synthesis of fructose polymers through the transfer of fructosyl units from sucrose to a growing fructan chain. Levanase activity of Bacillus subtilis levansucrase has been described since the very first publications dealing with the mechanism of levan synthesis. However, there is a lack of qualitative and quantitative evidence regarding the importance of the intrinsic levan hydrolysis of B. subtilis levansucrase and its role in the levan synthesis process. Particul...

  17. Amino acid efflux in response to chemotactic and osmotic signals in Bacillus subtilis.

    OpenAIRE

    Wong, L S; Johnson, M. S.; Sandberg, L. B.; Taylor, B L

    1995-01-01

    We observed a large efflux of nonvolatile radioactivity from Bacillus subtilis in response to the addition of 31 mM butyrate or the withdrawal of 0.1 M aspartate in a flow assay. The major nonvolatile components effluxed were methionine, proline, histidine, and lysine. In studies of the release of volatile radioactivity in chemotaxis by B. subtilis cells that had been labeled with [3H]methionine, the breakdown of methionine to methanethiol can contribute substantially to the volatile radioact...

  18. Methodological approaches to help unravel the intracellular metabolome of Bacillus subtilis

    OpenAIRE

    Meyer, Hanna; Weidmann, Hendrikje; Lalk, Michael

    2013-01-01

    Background Bacillus subtilis (B. subtilis) has become widely accepted as a model organism for studies on Gram-positive bacteria. A deeper insight into the physiology of this prokaryote requires advanced studies of its metabolism. To provide a reliable basis for metabolome investigations, a validated experimental protocol is needed since the quality of the analytical sample and the final data are strongly affected by the sampling steps. To ensure that the sample analyzed precisely reflects the...

  19. Effects of Bacillus subtilis natto and Different Components in Culture on Rumen Fermentation and Rumen Functional Bacteria In Vitro.

    Science.gov (United States)

    Sun, Peng; Li, Jinan; Bu, Dengpan; Nan, Xuemei; Du, Hong

    2016-05-01

    This study was to investigate the effects of live or autoclaved Bacillus subtilis natto, their fermented products and media on rumen fermentation and rumen functional bacteria in vitro. Rumen fluid from three multiparous lactating Holstein cows was combined and transferred into serum bottles after diluted. Fifteen serum bottles were divided into five treatments, which were designed as following: CTR (the fermentation of 0.5 g TMR and ruminal fluids from dairy cows), LBS (CTR plus a minimum of 10(11) cfu live Bacillus subtilis natto), ABS (CTR plus a minimum of 10(11) cfu autoclaved Bacillus subtilis natto), BSC (CTR plus 1 ml Bacillus subtilis natto fermentation products without bacteria), and BSM (CTR plus 1 ml liquid fermentation medium). When separated from the culture, live Bacillus subtilis natto individually increased the concentrations of ammonia-N (P production (P probiotic in dairy ration. PMID:26821238

  20. Effects of genetic modifications and fermentation conditions on 2,3-butanediol production by alkaliphilic Bacillus subtilis.

    Science.gov (United States)

    Białkowska, Aneta M; Jędrzejczak-Krzepkowska, Marzena; Gromek, Ewa; Krysiak, Joanna; Sikora, Barbara; Kalinowska, Halina; Kubik, Celina; Schütt, Fokko; Turkiewicz, Marianna

    2016-03-01

    Two recombinants of alkaliphilic Bacillus subtilis LOCK 1086, constructed via different strategies such as cloning the gene encoding bacterial hemoglobin from Vitreoscilla stercoraria (vhb) and overexpression of the gene encoding acetoin reductase/2,3-butanediol dehydrogenase (bdhA) from B. subtilis LOCK 1086, did not produce more 2,3-butanediol (2,3-BD) than the parental strain. In batch fermentations, this strain synthesized 9.46 g/L in 24 h and 12.80 g/L 2,3-BD in 46 h from sugar beet molasses and an apple pomace hydrolysate, respectively. 2,3-BD production by B. subtilis LOCK 1086 was significantly enhanced in fed-batch fermentations. The highest 2,3-BD concentration (75.73 g/L in 114 h, productivity of 0.66 g/L × h) was obtained in the sugar beet molasses-based medium with four feedings with glucose. In a medium based on the apple pomace hydrolysate with three feedings with sucrose, B. subtilis LOCK 1086 produced up to 51.53 g/L 2,3-BD (in 120 h, productivity of 0.43 g/L × h). PMID:26590588

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

    Directory of Open Access Journals (Sweden)

    Catarina Baptista

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

  2. Bacillus subtilis class Ib ribonucleotide reductase is a dimanganese(III)-tyrosyl radical enzyme.

    Science.gov (United States)

    Zhang, Yan; Stubbe, Joanne

    2011-06-28

    Bacillus subtilis class Ib ribonucleotide reductase (RNR) catalyzes the conversion of nucleotides to deoxynucleotides, providing the building blocks for DNA replication and repair. It is composed of two proteins: α (NrdE) and β (NrdF). β contains the metallo-cofactor, essential for the initiation of the reduction process. The RNR genes are organized within the nrdI-nrdE-nrdF-ymaB operon. Each protein has been cloned, expressed, and purified from Escherichia coli. As isolated, recombinant NrdF (rNrdF) contained a diferric-tyrosyl radical [Fe(III)(2)-Y(•)] cofactor. Alternatively, this cluster could be self-assembled from apo-rNrdF, Fe(II), and O(2). Apo-rNrdF loaded using 4 Mn(II)/β(2), O(2), and reduced NrdI (a flavodoxin) can form a dimanganese(III)-Y(•) [Mn(III)(2)-Y(•)] cofactor. In the presence of rNrdE, ATP, and CDP, Mn(III)(2)-Y(•) and Fe(III)(2)-Y(•) rNrdF generate dCDP at rates of 132 and 10 nmol min(-1) mg(-1), respectively (both normalized for 1 Y(•)/β(2)). To determine the endogenous cofactor of NrdF in B. subtilis, the entire operon was placed behind a Pspank(hy) promoter and integrated into the B. subtilis genome at the amyE site. All four genes were induced in cells grown in Luria-Bertani medium, with levels of NrdE and NrdF elevated 35-fold relative to that of the wild-type strain. NrdE and NrdF were copurified in a 1:1 ratio from this engineered B. subtilis. The visible, EPR, and atomic absorption spectra of the purified NrdENrdF complex (eNrdF) exhibited characteristics of a Mn(III)(2)-Y(•) center with 2 Mn/β(2) and 0.5 Y(•)/β(2) and an activity of 318-363 nmol min(-1) mg(-1) (normalized for 1 Y(•)/β(2)). These data strongly suggest that the B. subtilis class Ib RNR is a Mn(III)(2)-Y(•) enzyme. PMID:21561096

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

    Science.gov (United States)

    Juhas, Mario; Ajioka, James W

    2016-06-01

    Bacillus subtilis is a well-characterized model bacterium frequently used for a number of biotechnology and synthetic biology applications. Novel strategies combining the advantages of B. subtilis with the DNA assembly and editing tools of Escherichia coli are crucial for B. subtilis engineering efforts. We combined Gibson Assembly and λ red recombineering in E. coli with RecA-mediated homologous recombination in B. subtilis for bacterial artificial chromosome-mediated DNA integration into the well-characterized amyE target locus of the B. subtilis chromosome. The engineered integrative bacterial artificial chromosome iBAC(cav) can accept any DNA fragment for integration into B. subtilis chromosome and allows rapid selection of transformants by B. subtilis-specific antibiotic resistance and the yellow fluorescent protein (mVenus) expression. We used the developed iBAC(cav)-mediated system to integrate 10kb DNA fragment from E. coli K12 MG1655 into B. subtilis chromosome. iBAC(cav)-mediated chromosomal integration approach will facilitate rational design of synthetic biology applications in B. subtilis. PMID:27033694

  4. Comprehensive analysis of temporal alterations in cellular proteome of Bacillus subtilis under curcumin treatment.

    Directory of Open Access Journals (Sweden)

    Panga Jaipal Reddy

    Full Text Available Curcumin is a natural dietary compound with antimicrobial activity against various gram positive and negative bacteria. This study aims to investigate the proteome level alterations in Bacillus subtilis due to curcumin treatment and identification of its molecular/cellular targets to understand the mechanism of action. We have performed a comprehensive proteomic analysis of B. subtilis AH75 strain at different time intervals of curcumin treatment (20, 60 and 120 min after the drug exposure, three replicates to compare the protein expression profiles using two complementary quantitative proteomic techniques, 2D-DIGE and iTRAQ. To the best of our knowledge, this is the first comprehensive longitudinal investigation describing the effect of curcumin treatment on B. subtilis proteome. The proteomics analysis revealed several interesting targets such UDP-N-acetylglucosamine 1-carboxyvinyltransferase 1, putative septation protein SpoVG and ATP-dependent Clp protease proteolytic subunit. Further, in silico pathway analysis using DAVID and KOBAS has revealed modulation of pathways related to the fatty acid metabolism and cell wall synthesis, which are crucial for cell viability. Our findings revealed that curcumin treatment lead to inhibition of the cell wall and fatty acid synthesis in addition to differential expression of many crucial proteins involved in modulation of bacterial metabolism. Findings obtained from proteomics analysis were further validated using 5-cyano-2,3-ditolyl tetrazolium chloride (CTC assay for respiratory activity, resazurin assay for metabolic activity and membrane integrity assay by potassium and inorganic phosphate leakage measurement. The gene expression analysis of selected cell wall biosynthesis enzymes has strengthened the proteomics findings and indicated the major effect of curcumin on cell division.

  5. Thermostable levansucrase from Bacillus subtilis BB04, an isolate of Banana peel

    Directory of Open Access Journals (Sweden)

    Viniti D Vaidya

    2012-04-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} Extensive screening resulted in the isolation of Bacillus sp. from Banana peel that produces considerable amount of thermostable levansucrase of molecular size 52kDa. 16S rRNA sequence analysis suggests that it belongs to Bacillus subtilis and was designated as strain BB04. Levansucrase was sucrose inducible, showed optimum activity at 50°C and pH 6.0. It was stable at pH range 6.0 - 7.0. Ca2+ at 1.0 mmol-1 concentration enhanced levansucrase activity by 24%. However levan production was highest at 40°C and pH 6.0. Cane molasses and juice proved to be good sources of sucrose for levan production. B. subtilis BB04 produced relatively more levan using cane molasses (11.32 gl-1 as sucrose source than in cane juice (4.81 gl-1.

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

    DEFF Research Database (Denmark)

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

    2005-01-01

    Both gram-negative and gram-positive bacteria possess protein tyrosine phosphatases (PTPs) with a catalytic Cys residue. In addition, many gram-positive bacteria have acquired a new family of PTPs, whose first characterized member was CpsB from Streptococcus pneumoniae. Bacillus subtilis contains......, we characterize YwqE, compare the activities of the three B. subtilis PTPs (YwqE, YwlE, and YfkJ), and demonstrate that the two B. subtilis class II PTPs do not dephosphorylate the physiological substrates of YwqE....

  7. Draft Genome Sequences of Four Plant Probiotic Bacillus Strains.

    Science.gov (United States)

    Jeong, Haeyoung; Park, Seung-Hwan; Choi, Soo-Keun

    2016-01-01

    Here, we report the whole-genome sequences of four Bacillus strains that exhibit plant probiotic activities. Three of them are the type strains of Bacillus endophyticus, "Bacillus gaemokensis," and Bacillus trypoxylicola, and the other, Bacillus sp. strain KCTC 13219, should be reclassified into a species belonging to the genus Lysinibacillus. PMID:27174273

  8. Antagonistic and Inhibitory Effect of Bacillus subtilis Against Certain Plant Pathogenic Fungi, I

    Directory of Open Access Journals (Sweden)

    S. M. Matar

    2009-01-01

    Full Text Available The antagonistic and inhibitory activity of fourteen Bacillus subtilis isolates (B1 to B14 obtained from different Egyptian sites, were tested against six fungal isolates belonging to four different genera, Rhizoctonia solani, Helminthosporium spp., Alternaria spp. and Fusarium oxysporum. Cultural, morphological and physiological characteristics of these isolates were found to be identical to Bacillus subtilis. When the fourteen B. subtilis isolates were tested as biological control agents for their antagonistic effect on the in vitro growth of the fungal isolates, four B. subtilis isolates B1, B4, B7, B8 had more antagonistic effect on all fungal isolates. Supernatant of B. subtilis isolate B7 had antagonistic effect on 6 fungal isolates but it was more effective on Helminthosporium spp., Alternaria spp. and F. oxysporum. B. subtilis as well as, isolate B7 showed effectiveness in reducing disease incidence and severity levels of tomato plants when added to the F. oxysporum and R. solani-infested soil. Also, it stimulated the growth of tomato plants compared to the other. HPLC analysis of the HCl precipitate of B. subtilis isolate B7 culture supernatant revealed that an identical pattern of five peaks to that of a purified preparation of iturin A was obtained.

  9. The Regularities of Mutagenic Action of gamma-Radiation on Vegetative Bacillus subtilis Cells with Different Repair Genotype

    CERN Document Server

    Boreyko, A V; Krasavin, E A

    2000-01-01

    The regularities of induction of his^-\\to his^+ mutations in vegetative Bacillus subtilis cells with different repair capacity after gamma-irradiation have been studied. The wild type cells, polA1, recE4, recA, recP, add5, recH were used in experiments. It was shown that radiation-induced mutagenesis is determined by a repair genotype of cells. The blocking of different reparation genes is reflected on mutagenesis ratio by the various ways. A frequency of induction mutations in polA strain is higher than in wild type cells and it is characterized by the linearly-quadratic dose curve. The different rec^- strains that belong to various epistatic groups reveal an unequal mutation induction. The add5 and recP strains are characterized by the high-level induction mutations in contrast with the wild type cells. The mutagenesis in recE and recH strains, on the contrary, sharply reduces. The different influence of rec genes inhering to various epistatic groups on mutagenesis in Bacillus subtilis cells probably reflec...

  10. Characterization of the involvement of two compensatory autolysins in mother cell lysis during sporulation of Bacillus subtilis 168.

    OpenAIRE

    Smith, T J; Foster, S. J.

    1995-01-01

    The 30-kDa sporulation-specific peptidoglycan hydrolase CwlC of Bacillus subtilis 168 was purified and characterized. It is an N-acetylmuramoyl-L-alanine amidase (amidase) that is associated with the mother cell wall of sporulating cells, and although it is secreted, it undergoes no N-terminal processing except removal of the initial methionine. It was found that mother cells of a strain insertionally inactivated in cwlC and lytC (the major vegetative amidase gene) did not lyse at the end of ...

  11. ComA, a phosphorylated response regulator protein of Bacillus subtilis, binds to the promoter region of srfA.

    OpenAIRE

    Roggiani, M; Dubnau, D

    1993-01-01

    ComA is a response regulator protein of Bacillus subtilis which is required for the transcription of several genes which are involved in late-growth expression and in responses to environmental stress. Among these genes are degQ, gsiA, and srfA. The last is an operon needed for the development of genetic competence, surfactin production, and normal sporulation. We show here that partially purified ComA protein, isolated from an overproducing Escherichia coli strain, is phosphorylated in vitro...

  12. One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash

    OpenAIRE

    Sanghi, Ashwani; Garg, Neelam; Gupta, V. K.; Mittal, Ashwani; R.C. Kuhad

    2010-01-01

    The present study describes the one-step purification and characterization of an extracellular cellulase-free xylanase from a newly isolated alkalophilic and moderately thermophilic strain of Bacillus subtilis ASH. Xylanase was purified to homogeneity by 10.5-fold with ~43% recovery using ion-exchange chromatography through CM-Sephadex C-50. The purified enzyme revealed a single band on SDS-PAGE gel with a molecular mass of 23 kDa. It showed an optimum pH at 7.0 and was stable over the pH ran...

  13. Effects of Bacillus subtilis KN-42 on Growth Performance, Diarrhea and Faecal Bacterial Flora of Weaned Piglets

    OpenAIRE

    Hu, Yuanliang; Dun, Yaohao; Li, Shenao; Zhao, Shumiao; Peng, Nan; Liang, Yunxiang

    2014-01-01

    This research focused on the effects of different doses of Bacillus subtilis KN-42 on the growth performance, diarrhea incidence, faecal bacterial flora, and the relative number of Lactobacillus and Escherichia coli in faeces of weaned piglets to determine whether the strain can serve as a candidate antimicrobial growth promoter. A total of 360 piglets (initial body weight 7.14±0.63 kg) weaned at 26±2 days of age were randomly allotted to 5 treatment groups (4 pens per treatment with 18 pigs ...

  14. Propriedades emulsificantes e estabilidade do biossurfactante produzido por Bacillus subtilis em manipueira Studies of emulsifying properties and stability of the biosurfactant produced by Bacillus subtilis in cassava wastewater

    Directory of Open Access Journals (Sweden)

    Francisco Fábio Cavalcante Barros

    2008-12-01

    conditions often associated with of such processes and the maintenance of their properties. The aim of this work was to study the stability of the biosurfactant produced by Bacillus subtilis strain LB5a grown in cassava wastewater in a pilot process. Stability studies were carried out by varying the temperature, pH, and salt (NaCl concentration. Another study was the evaluation of their emulsifying index in mixtures of water with hydrocarbons and vegetable oils as well as the stability of the emulsions formed. The results showed that the biosurfactant was stable under all combination of temperature and time tested: 100 °C for 140 minutes and 121 °C for up to 60 minutes. It was also stable in concentrations of NaCl from 2.5 to 20%, and pH from 6 to 10. The biosurfactant showed higher values of emulsion index at 24 hours (EI24 to various cyclical and aliphatic hydrocarbons when compared with SDS. The vegetable oils emulsions were stable despite the fact that the profiles of fatty acid chain in the oils were different. All results described characterize these compounds as potential emulsifiers for different industrial applications.

  15. The spatial architecture of Bacillus subtilis biofilms deciphered using a surface-associated model and in situ imaging.

    Directory of Open Access Journals (Sweden)

    Arnaud Bridier

    Full Text Available The formation of multicellular communities known as biofilms is the part of bacterial life cycle in which bacteria display cooperative behaviour and differentiated phenotypes leading to specific functions. Bacillus subtilis is a Gram-positive bacterium that has served for a decade as a model to study the molecular pathways that control biofilm formation. Most of the data on B. subtilis biofilms have come from studies on the formation of pellicles at the air-liquid interface, or on the complex macrocolonies that develop on semi-solid nutritive agar. Here, using confocal laser scanning microcopy, we show that B. subtilis strains of different origins are capable of forming biofilms on immersed surfaces with dramatically protruding "beanstalk-like" structures with certain strains. Indeed, these structures can reach a height of more than 300 µm with one undomesticated strain from a medical environment. Using 14 GFP-labeled mutants previously described as affecting pellicle or complex colony formation, we have identified four genes whose inactivation significantly impeded immersed biofilm development, and one mutation triggering hyperbiofilm formation. We also identified mutations causing the three-dimensional architecture of the biofilm to be altered. Taken together, our results reveal that B. subtilis is able to form specific biofilm features on immersed surfaces, and that the development of these multicellular surface-associated communities involves regulation pathways that are common to those governing the formation of pellicle and/or complex colonies, and also some specific mechanisms. Finally, we propose the submerged surface-associated biofilm as another relevant model for the study of B. subtilis multicellular communities.

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

    OpenAIRE

    Salcedo L.; Castellanos O.; Grebeshova R.

    1998-01-01

    The catalytic properties of proteases isolated from the filtrate of submerged fermentation of Bacillus subtilis were investigated. Proteases present in the filtrate were determined to be of the serine protease type based on the use of specific protease inhibitors; ethylenediamintetraacetic acid (EDTA) was used as a metalloprotease inhibitor, and phenylmethylsulfonylfluoride (PMSF) was used as a serine protease inhibitor. Protease activity was highly stable in alkaline solutions and at high te...

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

    Institute of Scientific and Technical Information of China (English)

    柴海云; 崔堂兵

    2012-01-01

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

  18. Construction of Phenol Degradation Genetically Engineered Bacteria Bacillus subtilis dqly-2%苯酚降解工程菌Bacillus subtilis dqly-2的构建

    Institute of Scientific and Technical Information of China (English)

    杨庆丽; 刘宇峰; 姬妍茹; 董艳; 高媛

    2012-01-01

    目的:构建苯酚降解工程菌Bacillus Subtilis dqly-2.方法:选取2株苯酚降解菌,分别为铜绿假单胞菌Pseudomonas aeruginosa zllf4和枯草芽孢杆菌Bacillus Subtilis BHf3-4,体外扩增Pseudomonas aeruginosa zllf4的邻苯二酚2,3双加氧酶基因(SYJ),并将此基因转入Bacillus Subtilis BHf3-4中,构建基因工程菌,并对野生菌和工程菌降解能力进行比较.结果:作用96h后,工程菌苯酚降解率为96.18%,显著高于野生菌的84.78%.结论:成功构建高效苯酚降解基因工程菌.

  19. Quantitative Analysis of the Migration and Accumulation of Bacillus subtilis in Asparagus officinalis.

    Science.gov (United States)

    Hao, Bian-Qing; Ma, Li-Ping; Qiao, Xiong-Wu

    2015-09-01

    Bacillus subtilis B96-II is a broad-spectrum biological control strain. It effectively suppresses soil-borne fungal diseases in vegetables. A green fluorescence protein (GFP) was expressed in B96-II to detect migration of B96-II into the root and stem of asparagus. The GFP-tagged B96-II (B96-II-GFP) strain exhibited bright green fluorescence under a fluorescence microscope. GFP was stable and had no apparent effects on the growth of the strain. Asparagus plants were planted in the soil inoculated with B96-II-GFP. Our results showed that B96-II-GFP was detected in both the root and stem 15, 30, and 45 days after the asparagus seedlings were planted. B96-II-GFP was also detected in leaves but at a lower concentration. The highest concentration was detected in 15 days, and the number of bacteria decreased subsequently irrespective of duration of growth or sampling period. The highest concentration of B96-II-GFP was present in the root base suggesting that the root base served as the hub of bacterial migration from the soil to the stem. PMID:26126832

  20. Short-Chain Chromate Ion Transporter Proteins from Bacillus subtilis Confer Chromate Resistance in Escherichia coli▿

    Science.gov (United States)

    Díaz-Magaña, Amada; Aguilar-Barajas, Esther; Moreno-Sánchez, Rafael; Ramírez-Díaz, Martha I.; Riveros-Rosas, Héctor; Vargas, Eréndira; Cervantes, Carlos

    2009-01-01

    Tandem paired genes encoding putative short-chain monodomain protein members of the chromate ion transporter (CHR) superfamily (ywrB and ywrA) were cloned from genomic DNA of Bacillus subtilis strain 168. The transcription of the paired genes, renamed chr3N and chr3C, respectively, was shown to occur via a bicistronic mRNA generated from a promoter upstream of the chr3N gene. The chr3N and chr3C genes conferred chromate resistance when expressed in Escherichia coli strain W3110. The cloned chr3N gene alone did not confer chromate resistance on E. coli, suggesting that both chr3N and chr3C genes are required for function. E. coli cells expressing paired chr3N and chr3C genes demonstrated diminished uptake of chromate compared to that by a vector-only control strain. These results suggest that short-chain CHR proteins form heterodimer transporters which efflux chromate ions from the cytoplasm. PMID:19581367

  1. High-level production of a cold-active B-mannanase from Bacillus subtilis BS5 and its molecular cloning and expression.

    Science.gov (United States)

    Huang, Jun Li; Bao, Ling Xiao; Zou, Han Yan; Che, Shu Gang; Wang, Gui Xue

    2012-01-01

    Mannanases can be useful in the food, feed, pulp and paper industries. In this research a Bacillus subtilis strain (named Bs5) which produced high-level beta-mannanase was isolated. Maximum level of beta-mannanase (1231.41 U/ml) was reached when Bacillus subtilis Bs5 was grown on konjac powder as the carbon source for nine hours at 32 degrees C. The beta-mannanase was a typical cold-active enzyme and its optimal temperature of 35 degrees C was the lowest among those of the known mannanases from bacteria. In addition, the optimal pH was 5.0 and much wide pH range from 3.0-8.0 was also observed in the beta-mannanase. These properties make the beta-mannanase more attractive for biotechnological applications. The DNA sequence coding the beta-mannanase was cloned and the open reading frame consisted of 1089 bp encoding 362 amino acids. A phylogenetic tree of the beta-mannanase based on the similarity of amino acid sequences revealed that the beta-mannanase formed a cluster with the beta-mannanases of Bacillus subtilis, which was separated from the mannanases of fungi and other bacteria. The beta-mannanase gene could be expressed in Escherichia coli and the recombinant beta-mannanase was characterized by Western blot. This study provided a new source of carbohydrate hydrolysis enzyme with novel characteristics from Bacillus subtilis. PMID:23248847

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

    NARCIS (Netherlands)

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

    2008-01-01

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

  3. A positive selection vector for the analysis of structural plasmid instability in Bacillus subtilis

    NARCIS (Netherlands)

    Meima, R; Venema, G; Bron, S

    1996-01-01

    A system for the positive selection of structural plasmid rearrangements in Bacillus subtilis was developed. Random deletions removing a transcription terminator structure in the assay plasmid, designated pGP100, resulted in expression of the cat-86 gene, under control of a constitutive bacteriophag

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

    Directory of Open Access Journals (Sweden)

    Alka Puri

    2015-06-01

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

  5. Transcriptional Profiling in Cotton Associated with Bacillus Subtilis (UFLA285) Induced Biotic-stress Tolerance

    Science.gov (United States)

    Plant growth promoting rhizobacteria (PGPR) confer disease resistance in many agricultural crops. In the case of Bacillus subtilis (UFLA285) isolated from the cotton producing state of Mato Grosso (Brazil), in addition to inducing foliar and root growth, disease resistance against damping-off cause...

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

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  7. Mutational analysis of the regulatory region of the srfA operon in Bacillus subtilis.

    OpenAIRE

    Nakano, M M; Zuber, P

    1993-01-01

    Transcription of the Bacillus subtilis srfA operon is dependent on the transcriptional activator ComA. Mutational analysis of the srfA regulatory region suggests that two regions of dyad symmetry upstream of the srfA promoter may function in transcriptional activation by facilitating a cooperative interaction between ComA dimers.

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

    NARCIS (Netherlands)

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

    2012-01-01

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

  9. Protective effects of SOD on Bacillus subtilis irradiated by γ-rays

    International Nuclear Information System (INIS)

    In order to investigate the protective effect of SOD, Bacillus subtilis vegetative cells treated with SOD of different concentrations were irradiated to different doses by 60Co γ-rays. Cell survival rate was examined with the standard plate-count method. Intercellular SOD activity was measured by SOD kit with xanthine oxidase method, and DNA double-strand breaks was analyzed by pulsed-field gel electrophoresis (PFGE). The results showed that the cell survival rate increased obviously with the SOD-added Bacillus subtilis, though we did not find a clear relation among the intercellular SOD activity, concentration of added SOD and irradiation dose. The DNA release percentage(PR) value decreased evidently with the SOD-treated Bacillus subtilis. Basically,the trend of cell survival rate with concentration of the added SOD and irradiation dose is consistent with the PR value. It can be concluded that exogenous SOD could protect Bacillus subtilis vegetative cell from damage induced by γ-rays, and the protective efficiency of added SOD is related to the concentration. (authors)

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

    DEFF Research Database (Denmark)

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

    2007-01-01

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

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

    DEFF Research Database (Denmark)

    Nicolas, Pierre; Mäder, Ulrike; Dervyn, Etienne;

    2012-01-01

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

  12. Electron transfer reactions, cyanide and O2 binding of truncated hemoglobin from Bacillus subtilis

    DEFF Research Database (Denmark)

    Fernandez, Esther; Larsson, Jonas T.; McLean, Kirsty J.;

    2013-01-01

    The truncated hemoglobin from Bacillus subtilis (trHb-Bs) possesses a surprisingly high affinity for oxygen and resistance to (auto)oxidation; its physiological role in the bacterium is not understood and may be connected with its very special redox and ligand binding reactions. Electron transfer...

  13. A novel halotolerant xylanase from marine isolate Bacillus subtilis cho40: gene cloning and sequencing

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Verma, P.; Deobagkar, D.

    % Composition of amino acid from amino acid sequence of xylanase enzyme from Bacillus subtilis Cho40 Amino acid composition Alanine (Ala (A) 15 7.1% Arginine (Arg) (R) 6 2.8% Asparagine (Asn) (N) 19 9.0% Aspartic acid (Asp) (D...

  14. Differences in cold adaptation of .i.Bacillus subtilis./i. under anaerobic and aerobic conditions

    Czech Academy of Sciences Publication Activity Database

    Beranová, J.; Mansilla, M.C.; de Mendoza, D.; Elhottová, Dana; Konopásek, I.

    2010-01-01

    Roč. 192, č. 16 (2010), s. 4164-4171. ISSN 0021-9193 R&D Projects: GA MŠk LC06066 Institutional research plan: CEZ:AV0Z60660521 Keywords : cold adaptation * Bacillus subtilis * anaerobiosis Subject RIV: EE - Microbiology, Virology Impact factor: 3.726, year: 2010

  15. YbxF, a protein associated with exponential-phase ribosomes in Bacillus subtilis

    Czech Academy of Sciences Publication Activity Database

    Sojka, Luděk; Fučík, Vladimír; Krásný, Libor; Barvík, I.; Jonák, Jiří

    2007-01-01

    Roč. 189, č. 13 (2007), s. 4809-4814. ISSN 0021-9193 R&D Projects: GA AV ČR IAA5052206 Institutional research plan: CEZ:AV0Z50520514 Keywords : ybxF * ymxC * ribosomes * Bacillus subtilis * GFP * growth phase Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.013, year: 2007

  16. Growth rate regulates membrane fluidity and membrane cold adaptation in .i.Bacillus subtilis./i

    Czech Academy of Sciences Publication Activity Database

    Beranová, J.; Jemiola-Rzeminska, M.; Elhottová, Dana; Strzalka, K.; Konopásek, I.

    2007-01-01

    Roč. 274, Suppl. 1 (2007), s. 362. ISSN 1742-464X. [Congress of the Federation-of-European-Biochemical-Societies /32./. 07.07.2007-12.07.2007, Vienna] Institutional research plan: CEZ:AV0Z60660521 Keywords : membrane fluidity * membrane cold adaptation * Bacillus subtilis Subject RIV: EH - Ecology, Behaviour

  17. ISOLATION AND CHARACTERIZATION OF COML, A TRANSCRIPTION UNIT INVOLVED IN COMPETENCE DEVELOPMENT OF BACILLUS-SUBTILIS

    NARCIS (Netherlands)

    VANSINDEREN, D; WITHOFF, S; BOELS, H; VENEMA, G

    1990-01-01

    Using the transformation-deficient mutant M465, which was previously isolated by means of insertional mutagenesis with plasmid pHV60, a transcription unit comL required for genetic competence of Bacillus subtilis was identified. A chromosomal DNA fragment flanking the inserted pHV60 was isolated and

  18. Natural products from Bacillus subtilis with antimicrobial properties☆

    Institute of Scientific and Technical Information of China (English)

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

    2015-01-01

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

  19. Functional analysis of the secretory precursor processing machinery of Bacillus subtilis: identification of a eubacterial homolog of archaeal and eukaryotic signal peptidases

    OpenAIRE

    Tjalsma, Harold; Bolhuis, Albert; Roosmalen, Maarten L. Van; Wiegert, Thomas; Schumann, Wolfgang; Broekhuizen, Cees P.; Quax, Wim J.; Venema, Gerard; Bron, Sierd; van Dijl, Jan Maarten

    1998-01-01

    Approximately 47% of the genes of the Gram-positive bacterium Bacillus subtilis belong to paralogous gene families. The present studies were aimed at the functional analysis of the sip gene family of B. subtilis, consisting of five chromosomal genes, denoted sipS, sipT, sipU, sipV, and sipW. All five sip genes specify type I signal peptidases (SPases), which are actively involved in the processing of secretory preproteins. Interestingly, strains lacking as many as four of these SPases could b...

  20. Catalytic residues Lys197 and Arg199 of Bacillus subtilis phosphoribosyl diphosphate synthase. Alanine-scanning mutagenesis of the flexible catalytic loop

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne; Bentsen, Ann-Kristin K; Harlow, Kenneth W

    2005-01-01

    Eleven of the codons specifying the amino acids of the flexible catalytic loop [KRRPRPNVAEVM(197-208)] of Bacillus subtilis phosphoribosyl diphosphate synthase have been changed individually to specify alanine. The resulting variant enzyme forms, as well as the wildtype enzyme, were produced in an...... Escherichia coli strain lacking endogenous phosphoribosyl diphosphate synthase activity and purified to near homogeneity. The B. subtilis phosphoribosyl diphosphate synthase mutant variants K197A and R199A were studied in detail. The physical properties of the two enzymes were similar to those of the wildtype...

  1. Production of biosurfactant and antifungal compound by fermented food isolate Bacillus subtilis 20B.

    Science.gov (United States)

    Joshi, Sanket; Bharucha, Chirag; Desai, Anjana J

    2008-07-01

    A biosurfactant producing strain, Bacillus subtilis 20B, was isolated from fermented food in India. The strain also showed inhibition of various fungi in in-vitro experiments on Potato Dextrose Agar medium. It was capable of growth at temperature 55 degrees C and salts up to 7%. It utilized different sugars, alcohols, hydrocarbons and oil as a carbon source, with preference for sugars. In glucose based minimal medium it produced biosurfactant which reduced surface tension to 29.5 mN/m, interfacial tension to 4.5 mN/m and gave stable emulsion with crude oil and n-hexadecane. The biosurfactant activity was stable at high temperature, a wide range of pH and salt concentrations for five days. Oil displacement experiments using biosurfactant containing broth in sand pack columns with crude oil showed 30.22% recovery. The possible application of organism as biocontrol agent and use of biosurfactant in microbial enhanced oil recovery (MEOR) is discussed. PMID:17855083

  2. Biosurfactant production by Bacillus subtilis B30 and its application in enhancing oil recovery.

    Science.gov (United States)

    Al-Wahaibi, Yahya; Joshi, Sanket; Al-Bahry, Saif; Elshafie, Abdulkadir; Al-Bemani, Ali; Shibulal, Biji

    2014-02-01

    The fermentative production of biosurfactants by Bacillus subtilis strain B30 and the evaluation of biosurfactant based enhanced oil recovery using core-flood were investigated. Different carbon sources (glucose, sucrose, starch, date molasses, cane molasses) were tested to determine the optimal biosurfactant production. The isolate B30 produced a biosurfactant that could reduce the surface tension and interfacial tension to 26.63±0.45 mN/m and 3.79±0.27 mN/m, respectively in less than 12h in both glucose or date molasses based media. A crude biosurfactant concentration of 0.3-0.5 g/l and critical micelle dilution (CMD) values of 1:8 were observed. The biosurfactants gave stable emulsions with wide range of hydrocarbons including light and heavy crude oil. The biosurfactants were partially purified and identified as a mixture of lipopeptides similar to surfactin, using high performance thin layer chromatography and Fourier transform infrared spectroscopy. The biosurfactants were stable over wide range of pH, salinity and temperatures. The crude biosurfactant preparation enhanced light oil recovery by 17-26% and heavy oil recovery by 31% in core-flood studies. The results are indicative of the potential of the strain for the development of ex situ microbial enhanced oil recovery processes using glucose or date molasses based minimal media. PMID:24240116

  3. Postreplication repair of deoxyribonucleic acid and daughter strand exchange in Uvr- mutants of Bacillus subtilis

    International Nuclear Information System (INIS)

    The fate of pyrimidine dimers in deoxyribonuclei acid (DNA) newly synthesized by Bacillus subtilis after ultraviolet irradiation was monitored by use of a damage-specific endonuclease that introduces single-strand breaks adjacent to nearly all of the dimer sites. Two Uvr- strains, one defective in the initiation of dimer excision and the other defective in a function required for efficient dimer excision, were found to be similar to their wild-type parent in the kinetics and extent of converting low-molecular-weight DNA newly synthesized after ultraviolet irradiation to high molecular weight. In the Uvr- strains large molecules of newly synthesized DNA remained susceptible to nicking by the damage-specific endonuclease even after extended incubation in growth medium, whereas the enzyme-sensitive sites were rapidly removed from both preexisting and newly synthesized DNA in Uvr+ cells. Our results support the hypothesis that postreplication repair in bacteria includes recombination between dimer-containing parental DNA strands and newly synthesized strands

  4. Effect of Bacillus subtilis Natto on Meat Quality and Skatole Content in TOPIGS Pigs.

    Science.gov (United States)

    Sheng, Q K; Zhou, K F; Hu, H M; Zhao, H B; Zhang, Y; Ying, W

    2016-05-01

    This study investigated the effect of Bacillus subtilis (B. subtilis) natto on meat quality and skatole in TOPIGS pigs. Sixty TOPIGS pigs were randomly assigned to 3 groups (including 5 pens per group, with 4 pigs in each pen) and fed with basic diet (control group), basic diet plus 0.1% B. subtilis natto (B group), and basic diet plus 0.1% B. subtilis natto plus 0.1% B. coagulans (BB group), respectively. All pigs were sacrificed at 100 kg. Growth performance, meat quality, serum parameters and oxidation status in the three groups were assessed and compared. Most parameters regarding growth performance and meat quality were not significantly different among the three groups. However, compared with the control group, meat pH24, fat and feces skatole and the content of Escherichia coli (E. Coli), Clostridium, NH3-N were significantly reduced in the B and BB groups, while serum total cholesterol, high density lipoprotein, the levels of liver P450, CYP2A6, and CYP2E1, total antioxidant capability (T-AOC) and glutathione peroxidase and Lactobacilli in feces were significantly increased in the B and BB groups. Further, the combined supplementation of B. subtilis natto and B. coagulans showed more significant effects on the parameters above compared with B. subtilis, and Clostridium, and NH3-N. Our results indicate that the supplementation of pig feed with B. subtilis natto significantly improves meat quality and flavor, while its combination with B. coagulans enhanced these effects. PMID:26954164

  5. Effect of Bacillus subtilis Natto on Meat Quality and Skatole Content in TOPIGS Pigs

    Science.gov (United States)

    Sheng, Q. K.; Zhou, K. F.; Hu, H. M.; Zhao, H. B.; Zhang, Y.; Ying, W.

    2016-01-01

    This study investigated the effect of Bacillus subtilis (B. subtilis) natto on meat quality and skatole in TOPIGS pigs. Sixty TOPIGS pigs were randomly assigned to 3 groups (including 5 pens per group, with 4 pigs in each pen) and fed with basic diet (control group), basic diet plus 0.1% B. subtilis natto (B group), and basic diet plus 0.1% B. subtilis natto plus 0.1% B. coagulans (BB group), respectively. All pigs were sacrificed at 100 kg. Growth performance, meat quality, serum parameters and oxidation status in the three groups were assessed and compared. Most parameters regarding growth performance and meat quality were not significantly different among the three groups. However, compared with the control group, meat pH24, fat and feces skatole and the content of Escherichia coli (E. Coli), Clostridium, NH3-N were significantly reduced in the B and BB groups, while serum total cholesterol, high density lipoprotein, the levels of liver P450, CYP2A6, and CYP2E1, total antioxidant capability (T-AOC) and glutathione peroxidase and Lactobacilli in feces were significantly increased in the B and BB groups. Further, the combined supplementation of B. subtilis natto and B. coagulans showed more significant effects on the parameters above compared with B. subtilis, and Clostridium, and NH3-N. Our results indicate that the supplementation of pig feed with B. subtilis natto significantly improves meat quality and flavor, while its combination with B. coagulans enhanced these effects. PMID:26954164

  6. Characterization of an N-glycosylated Bacillus subtilis leucine aminopeptidase expressed in Pichia pastoris.

    Science.gov (United States)

    Xi, Hongxing; Tian, Yaping; Zhou, Nandi; Zhou, Zhemin; Shen, Wei

    2015-02-01

    Aminopeptidase is an important flavorsome especially in protein hydrolysate debittering by removing hydrophobic amino acid residue at the N-terminal end. Besides, it is also applied to preparation of active peptides and analysis of protein sequence. In this study, leucine aminopeptidase from Bacillus subtilis was cloned and expressed in Pichia pastoris, a widely used heterologous protein expression host. Then it was purified and characterized. After methanol induction for 96 h, the aminopeptidase activity in culture supernatant reached 28.4 U ml(À1) , which was 7.1 times that of wild strain B. subtilis Zj016. The optimal temperature and pH of the purified recombinant enzyme were 60 °C and 8.5, respectively. The purified aminopeptidase was stable within 30-60 °C and pH 8.0-9.0. It was intensively inhibited by Ni(2β) , Ca(2β) , DL-dithiothreitol (DTT) and ethylene diamine tetraacetic acid (EDTA), but activated by Co(2β) . The Km toward leucine-p-nitroanilines (Leu-pNA) of the enzyme was 0.97 mM. The sequence analysis of aminopeptidase indicated three potential N-glycosylation sites and it was further verified via MALDI-TOF-MS analysis. Consequently, the N-glycosylated aminopeptidase exhibited higher thermostability and catalytic efficiency. The purified enzyme exhibited two bands through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) while a single band can be identified when the enzyme was deglycosylated. Circular dichroism spectroscopy indicated that the secondary structure of recombinant aminopeptidase was similar to the wild-type. PMID:25389014

  7. Accelerated Growth Rate and Increased Drought Stress Resilience of the Model Grass Brachypodium distachyon Colonized by Bacillus subtilis B26.

    Directory of Open Access Journals (Sweden)

    François Gagné-Bourque

    Full Text Available Plant growth-promoting bacteria (PGB induce positive effects in plants, for instance, increased growth and reduced abiotic stresses susceptibility. The mechanisms by which these bacteria impact the host plant are numerous, diverse and often specific. Here, we studied the agronomical, molecular and biochemical effects of the endophytic PGB Bacillus subtilis B26 on the full life cycle of Brachypodium distachyon Bd21, an established model species for functional genomics in cereal crops and temperate grasses. Inoculation of Brachypodium with B. subtilis strain B26 increased root and shoot weights, accelerated growth rate and seed yield as compared to control plants. B. subtilis strain B26 efficiently colonized the plant and was recovered from roots, stems and blades as well as seeds of Brachypodium, indicating that the bacterium is able to migrate, spread systemically inside the plant, establish itself in the aerial plant tissues and organs, and is vertically transmitted to seeds. The presence of B. subtilis strain B26 in the seed led to systemic colonization of the next generation of Brachypodium plants. Inoculated Brachypodium seedlings and mature plants exposed to acute and chronic drought stress minimized the phenotypic effect of drought compared to plants not harbouring the bacterium. Protection from the inhibitory effects of drought by the bacterium was linked to upregulation of the drought-response genes, DREB2B-like, DHN3-like and LEA-14-A-like and modulation of the DNA methylation genes, MET1B-like, CMT3-like and DRM2-like, that regulate the process. Additionally, total soluble sugars and starch contents increased in stressed inoculated plants, a biochemical indication of drought tolerance. In conclusion, we show a single inoculation of Brachypodium with a PGB affected the whole growth cycle of the plant, accelerating its growth rates, shortening its vegetative period, and alleviating drought stress effects. These effects are relevant to

  8. Enhancement of Biocontrol Activities and Cyclic Lipopeptides Production by Chemical Mutagenesis of Bacillus subtilis XF-1, a Biocontrol Agent of Plasmodiophora brassicae and Fusarium solani

    OpenAIRE

    Li, Xing-Yu; Yang, Jing-Jing; Mao, Zi-Chao; Ho, Hon-Hing; Wu, Yi-Xing; He, Yue-qiu

    2014-01-01

    Bacillus subtilis XF-1 has been used as a biocontrol agent of clubroot disease of crucifers infected by Plasmodiophora brassicae, an obligate pathogen. In order to maximize the growth inhibition of the pathogen, random mutagenesis using N-methyl-N′-nitro-N-nitrosoguanidine was applied to strain XF-1. The efficacy of 226 selected mutants was assessed against the growth of an indicator fungal pathogen: Fusarium solani using agar plate assay and the disruptive effects on the resting spores of P....

  9. Insights into the Defense-Related Events Occuring in Plant Cells Following Perception of Surfactin-Type Lipopeptide from Bacillus subtilis

    OpenAIRE

    Jourdan, Emmanuel; Henry, Guillaume; Duby, F.; Dommes, Jacques; Barthelemy, Jean-Paul; Thonart, Philippe; Ongena, Marc

    2009-01-01

    Multiple strains of Bacillus subtilis were demonstrated to stimulate plant defense responses, and cyclic lipopeptides may be involved in the elicitation of this induced systemic resistance phenomenon. Here, we further investigated molecular events underlying the interaction between wuch lipopeptides and plant cells. Addition of surfactin but not fengycin or iturin in the micromolar range to tobacco cell suspensions induced defense-related early events such as extracellular medium alkalinizati...

  10. A temperature-sensitive trpS mutation interferes with trp RNA-binding attenuation protein (TRAP) regulation of trp gene expression in Bacillus subtilis.

    OpenAIRE

    Lee, A I; Sarsero, J P; Yanofsky, C

    1996-01-01

    In Bacillus subtilis, the tryptophan-activated trp RNA-binding attenuation protein (TRAP) regulates expression of the seven tryptophan biosynthetic genes by binding to specific repeat sequences in the transcripts of the trp operon and of the folate operon, the operon containing trpG. Steinberg observed that strains containing a temperature-sensitive mutant form of tryptophanyl-tRNA synthetase, encoded by the trpS1 allele, produced elevated levels of the tryptophan pathway enzymes, when grown ...

  11. [Ability of representatives of Pantoea agglomerans, as well as Bacillus subtilis and some Pseudomonas species to suppress the development of phytopathgenic bacteria and micromycetes in regulating plant growth].

    Science.gov (United States)

    Romanenko, V M; Alimov, D M

    2000-01-01

    The ability of representatives of Pantoea agglomerans (Erwinia herbicola (Lohnis) Dye [21]), Bacillus subtilis and some species of Pseudomonas genus to inhibit the growth of phytopathogenic bacteria and micromycetes and to regulate the growth of plants has been comparatively studied. The ability to inhibit the growth of mycellium of phytopathogenic Fusarium avenaceum, F. gibbosum, F. oxysporum was found out in all of 13 investigated strains of P. agglomerans, while the growth of F. culmorum is inhibited by 2 strains and Bipolaris sorokiniana is inhibited by 7 strains. The strains of P. agglomerans and Bacillus subtilis inhibit the growth of mycellium of these mycromycetes to the greater extent than the representatives of Pseudomonas genus. The mycellium growth of B. sorokiniana is better inhibited by B. subtilis and representatives of Pseudomonas genus. Besides the antifungal action 8 strains of P. agglomerans manifested the antagonistic activity in respect to phytopathogenic Agrobacterium tumefaciens and representatives of genera Clavibacter, Erwinia, Pseudomonas, Xanthomonas and also in respect to the microflora which is present in the cabbage and wheat seeds. The strains have been revealed which, parallel with high antagonistic activity in respect to phytopathogenic micromycetes and bacteria, stimulate the seed germination and increase the weight of the cabbage and wheat sprouts. PMID:11421000

  12. Increase of the a-amylase yield by Some Bacillus Strains

    OpenAIRE

    SARIKAYA, Elif

    2000-01-01

    In order to obtain high amounts of a-amylase enzyme from Bacillus bacteria a new medium was developed. In this medium, three strains of Bacillus bacteria were grown in the presence of different carbon, nitrogen and metal ions. The highest a-amylase yield was obtained by addition of Na-citrate, strach and sucrose for the strains of B. subtilis, B. amyloliquefaciens I and B. amyloliquefaciens II, respectively. As a nitrogen source, yeast extract was found to be effective on enzyme yield for...

  13. Bacillus subtilis deoxyribonuclease activity specific for single-stranded deoxyribonucleic acid: cellular site and variations during germination and sporulation.

    OpenAIRE

    Cobianchi, F; Attolini, C; Falaschi, A; Ciarrocchi, G

    1980-01-01

    The endonuclease of Bacillus subtilis specific for single-stranded deoxyribonucleic acid is absent in spores, appears during germination only after the start of deoxyribonucleic acid synthesis, and is located almost exclusively in the periplasm.

  14. Evolution of Bacillus subtilis to enhanced hypobaric growth: global alterations in gene expression

    Science.gov (United States)

    Nicholson, Wayne; Robles-Martinez, Jose; Rivas-Castillo, Andrea; Schuerger, Andrew

    Much astrobiology research is concerned with defining the environmental limits for life in the universe. Because Mars currently is the primary target for life detection missions, it is important to understand how terrestrial microbes might survive, proliferate, and evolve in martian envi-ronments. This issue is relevant in three distinct but related contexts: (i) testing panspermia hypotheses [1], (ii) mitigating the forward contamination of Mars [2], and (iii) understanding the molecular mechanisms leading to microbial growth in extreme extraterrestrial environments [3]. Prime candidates for Earth-to-Mars transfer include bacteria of the genus Bacillus, spores of which are significant contaminants of Mars-bound spacecraft and which are considered good candidates for lithopanspermia [1-4]. It is thus relevant to assess the potential for such microbes to survive and proliferate in the martian environment. The martian atmosphere poses a significant barrier to growth of terrestrial microbes, due to its low pressure (1-10 mbar; average 7 mbar) and anoxic (˜95% CO2) composition. In an earlier study [5] we showed that low pressures approaching those found on the surface of Mars exhibited an inhibitory effect on the germination and vegetative growth of several Bacillus spp. isolated from spacecraft or their assembly facilities. Even in an Earth-like 80%N2/20%O2 atmosphere, growth of B. subtilis cells was nearly completely inhibited at pressures below 35 mbar, well above the highest pressure on the martian surface [5]. The purpose of the present investigation was to use low pressure as a selective agent to test the hypothesis that a terrestrial microorganism, Bacillus subtilis, could evolve the ability for enhanced growth under hypobaric conditions approaching those of Mars. B. subtilis wild-type strains WN624 (SpcR) and WN628 (CmR) have been described previously [6] and were used as ancestral strains. Strains were propagated in LB liquid medium containing the appropriate

  15. CotC-CotU Heterodimerization during Assembly of the Bacillus subtilis Spore Coat▿

    OpenAIRE

    Isticato, Rachele; Pelosi, Assunta; Zilhão, Rita, 1959-; Baccigalupi, Loredana; Henriques, Adriano O.; De Felice, Maurilio; Ricca, Ezio

    2007-01-01

    We report evidence that CotC and CotU, two previously identified components of the Bacillus subtilis spore coat, are produced concurrently in the mother cell chamber of the sporulating cell under the control of σK and GerE and immediately assembled around the forming spore. In the coat, the two proteins interact to form a coat component of 23 kDa. The CotU-CotC interaction was not detected in two heterologous hosts, suggesting that it occurs only in B. subtilis. Monomeric forms of both CotU a...

  16. Effects of microbial loading and sporulation temperature on atmospheric plasma inactivation of Bacillus subtilis spores

    Science.gov (United States)

    Deng, X. T.; Shi, J. J.; Shama, G.; Kong, M. G.

    2005-10-01

    Current inactivation studies of Bacillus subtilis spores using atmospheric-pressure glow discharges (APGD) do not consider two important factors, namely microbial loading at the surface of a substrate and sporulation temperature. Yet these are known to affect significantly microbial resistance to heat and hydrogen peroxide. This letter investigates effects of microbial loading and sporulation temperature on spore resistance to APGD. It is shown that microbial loading can lead to a stacking structure as a protective shield against APGD treatment and that high sporulation temperature increases spore resistance by altering core water content and cross-linked muramic acid content of B. subtilis spores.

  17. Bacillus subtilis genome vector-based complete manipulation and reconstruction of genomic DNA for mouse transgenesis

    OpenAIRE

    Iwata, Tetsuo; Kaneko, Shinya; Shiwa, Yuh; Enomoto, Takayuki; Yoshikawa, Hirofumi; Hirota, Junji

    2013-01-01

    Background The Bacillus subtilis genome (BGM) vector is a novel cloning system for large DNA fragments, in which the entire 4.2 Mb genome of B. subtilis functions as a vector. The BGM vector system has several attractive properties, such as a large cloning capacity of over 3 Mb, stable propagation of cloned DNA and various modification strategies using RecA-mediated homologous recombination. However, genetic modifications using the BGM vector system have not been fully established, and this s...

  18. Conversion of sub-megasized DNA to desired structures using a novel Bacillus subtilis genome vector

    OpenAIRE

    Kaneko, Shinya; Tsuge, Kenji; Takeuchi, Takashi; Itaya, Mitsuhiro

    2003-01-01

    A novel genome vector using the 4215 kb Bacillus subtilis genome provides for precise target cloning and processing of the cloned DNA to the desired structure. Each process highly dependent on homologous recombination in the host B.subtilis is distinguished from the other cloning systems. A 120 kb mouse jumonji (jmj) genomic gene was processed in the genome vector to give a series of truncated sub-megasized DNA. One of these truncated segments containing the first intron was copied in a plasm...

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

    DEFF Research Database (Denmark)

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

    1995-01-01

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

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

    DEFF Research Database (Denmark)

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

    1995-01-01

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

  1. MinCD Proteins Control the Septation Process during Sporulation of Bacillus subtilis

    OpenAIRE

    Barák, Imrich; Prepiak, Peter; Schmeisser, Falko

    1998-01-01

    Mutation of the divIVB locus in Bacillus subtilis causes misplacement of the septum during cell division and allows the formation of anucleate minicells. The divIVB locus contains five open reading frames (ORFs). The last two ORFs (minCD) are homologous to minC and minD of Escherichia coli but a minE homolog is lacking in B. subtilis. There is some similarity between minicell formation and the asymmetric septation that normally occurs during sporulation in terms of polar septum localization. ...

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

    International Nuclear Information System (INIS)

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

  3. Free and attached cells of Bacillus subtilis as starters for production of a soup flavouring (“ogiri egusi”)

    OpenAIRE

    Peter-Ikechukwu, A. I.; Ahaotu, I.; Owuamanam, C. I.; Ogueke, C. C.

    2013-01-01

    Aims: This Bacillus subtilis has been identified to be the main fermenting bacterium during indigenous production of “ogiri egusi”; a traditional soup flavouring rich in protein. Evaluation of the use of starter and broth cultures of this bacterium in the production of ‘ogiri egusi’ was therefore undertaken with the view to improve the fermentation process and quality of product. Methodology and Results: Cowpea granules in association with Bacillus subtilis cells were developed as starter cul...

  4. Effect of berberine on Escherichia coli, Bacillus subtilis, and their mixtures as determined by isothermal microcalorimetry.

    Science.gov (United States)

    Kong, Wei-Jun; Xing, Xiao-Yan; Xiao, Xiao-He; Zhao, Yan-Ling; Wei, Jian-He; Wang, Jia-Bo; Yang, Rui-Chuang; Yang, Mei-Hua

    2012-10-01

    The strong toxicity of pathogenic bacteria has resulted in high levels of morbidity and mortality in the general population. Developing effective antibacterial agents with high efficacy and long activity is in great demand. In this study, the microcalorimetric technique based on heat output of bacterial metabolism was applied to evaluate the effect of berberine on Escherichia coli, Bacillus subtilis, individually and in a mixture of both using a multi-channel microcalorimeter. The differences in shape of the power-time fingerprints and thermokinetic parameters of microorganism growth were compared. The results revealed that low concentration (20 μg/mL) of berberine began to inhibit the growth of E. coli and mixed microorganisms, while promoting the growth of B. subtilis; high concentration of berberine (over 100 μg/mL) inhibited B. subtilis. The endurance of E. coli to berberine was obviously lower than B. subtilis, and E. coli could decrease the endurance of B. subtilis to berberine. The sequence of half-inhibitory concentration (IC(50)) of berberine was: B. subtilis (952.37 μg/mL) > mixed microorganisms (682.47 μg/mL) > E. coli (581.69 μg/mL). Berberine might be a good selection of antibacterial agent used in the future. The microcalorimetric method should be strongly suggested in screening novel antibacterial agents for fighting against pathogenic bacteria. PMID:22878842

  5. Abnormal morphology of Bacillus subtilis ugtP mutant cells lacking glucolipids.

    Science.gov (United States)

    Matsuoka, Satoshi; Chiba, Minako; Tanimura, Yu; Hashimoto, Michihiro; Hara, Hiroshi; Matsumoto, Kouji

    2011-01-01

    Bacillus subtilis Marburg 168 cells with disrupted ugtP, which encodes UDP-glucosyltransferase involved in glucolipid synthesis, were bent and distended. In the ugtP mutant cells, the extracytoplasmic function sigmas SigM, SigV and SigX, were found to be activated. Introduction of a disrupted allele of sigM into the ugtP strain caused even more abnormal morphology, with cells taking on a balloon-like shape; growth of these cells in LB medium was hampered by addition of 1.5% NaCl. Addition of MgSO4 or MnCl2 suppressed the abnormal morphology. In ugtP mutant cells the transcription of the mreB operon from an upstream promoter in maf (designated Pupstream mreB) and PmreBH was 4.3- and 2.3-fold higher, respectively, and localization of GFP-MreB was not in discrete dots (in an apparently helical pattern), but faint and in irregular clusters. GFP-MreB protein was reduced in the ugtP mutant cells. We suggest that glucolipids are important for MreB isoforms to take on the configuration that appears as discrete dots and plays a role in shaping cells into straight rods. PMID:22362028

  6. Constitutive Stringent Response Restores Viability of Bacillus subtilis Lacking Structural Maintenance of Chromosome Protein

    Science.gov (United States)

    Benoist, Camille; Guérin, Cyprien; Noirot, Philippe; Dervyn, Etienne

    2015-01-01

    Bacillus subtilis mutants lacking the SMC-ScpAB complex are severely impaired for chromosome condensation and partitioning, DNA repair, and cells are not viable under standard laboratory conditions. We isolated suppressor mutations that restored the capacity of a smc deletion mutant (Δsmc) to grow under standard conditions. These suppressor mutations reduced chromosome segregation defects and abrogated hypersensitivity to gyrase inhibitors of Δsmc. Three suppressor mutations were mapped in genes involved in tRNA aminoacylation and maturation pathways. A transcriptomic survey of isolated suppressor mutations pointed to a potential link between suppression of Δsmc and induction of the stringent response. This link was confirmed by (p)ppGpp quantification which indicated a constitutive induction of the stringent response in multiple suppressor strains. Furthermore, sublethal concentrations of arginine hydroxamate (RHX), a potent inducer of stringent response, restored growth of Δsmc under non permissive conditions. We showed that production of (p)ppGpp alone was sufficient to suppress the thermosensitivity exhibited by the Δsmc mutant. Our findings shed new light on the coordination between chromosome dynamics mediated by SMC-ScpAB and other cellular processes during rapid bacterial growth. PMID:26539825

  7. Inactivation, mutation induction and repair in Bacillus subtilis spores irradiated with heavy ions

    Science.gov (United States)

    Horneck, G.; Bücker, H.

    Studies on the response of bacterial spores to accelerated heavy ions (HZE particles) help in understanding problems of space radiobiology and exobiology. Layers of spores of Bacillus subtilis strains, differing in repair capabilities, were irradiated with accelerated boron, carbon and neon ions of linear energy transfer (LET) values up to 14000 MeV cm2/g. Inactivation as measured by loss of colony forming ability and induction of mutations as measured by reversion to histidine prototrophy and resistance to 150 μg/ml sodium azide were tested, as well as the influence of repair processes on these effects. For inactivation, the cross-sectional values σ plotted as a function of LET follow a saturation curve. The plateau, which is reached around a LET of 2000 MeV cm2/g, occurs at 2.5 × 10-9 cm2, a value in good agreement with the dimensions of the spore protoplast. Lethal damage produced at LET values < 2000 MeV cm2/g is reparable. Recombination repair is more effective than excision repair. At higher LET values, lethal damage could not be reconstituted by the repair mechanisms studied. In addition, at these high LET values, the frequency of induced mutations was drastically decreased. The data support the assumption of at least two qualitatively different types of lesion, depending on the LET of the affecting heavy ion.

  8. Antifungal activity of Bacillus subtilis 355 against wood-surface contaminant fungi.

    Science.gov (United States)

    Feio, Sonia Savluchinske; Barbosa, Ana; Cabrita, Manuela; Nunes, Lina; Esteves, Alexandra; Roseiro, José Carlos; Curto, Maria João Marcelo

    2004-06-01

    A strain of Bacillus subtilis was examined for antifungal activity against phytopathogenic and wood-surface contaminant fungi. The bacterium was grown in five culture media with different incubation times in order to study cell development, sporulation, and the production of metabolites with antifungal activity. The anti-sapstain and anti-mould activity of the bacterium grown in yeast extract glucose broth (YGB) medium in wood was also evaluated. In YGB, the bacterium inhibited the growth of several fungi and displayed a broader spectrum of activity than in the other media tested. A relationship between bacterial spore production and the formation of metabolites with antifungal activity was detected. YGB medium displayed effective control in wood block tests. YGB medium was extracted with solvents of increasing polarity and the dry residues were applied to silicagel plates, resolved with the appropriate solvent and sprayed with different solutions, detecting the presence, of amines, and higher alcohols. The bioautographic method revealed the presence of at least two active compounds against the blue-stain fungus Cladosporium cucumerinum. PMID:15197600

  9. Augmenting Iron Accumulation in Cassava by the Beneficial Soil Bacterium Bacillus subtilis (GBO3

    Directory of Open Access Journals (Sweden)

    Monica A Freitas

    2015-08-01

    Full Text Available Cassava (Manihot esculenta, a major staple food in the developing world, provides a basic carbohydrate diet for over half a billion people living in the tropics. Despite the iron abundance in most soils, cassava provides insufficient iron for humans as the edible roots contain 3-12 times less iron than other traditional food crops such as wheat, maize, and rice. With the recent identification that the beneficial soil bacterium Bacillus subtilis (strain GB03 activates iron acquisition machinery to increase metal ion assimilation in Arabidopsis, the question arises as to whether this plant-growth promoting rhizobacterium (PGPR also augments iron assimilation to increase endogenous iron levels in cassava. Biochemical analyses reveal that shoot-propagated cassava with GB03-inoculation exhibit elevated iron accumulation after 140 days of plant growth as determined by X-ray microanalysis and total foliar iron analysis. Growth promotion and increased photosynthetic efficiency were also observed for greenhouse-grown plants with GB03-exposure. These results demonstrate the potential of microbes to increase iron accumulation in an important agricultural crop and is consistent with idea that microbial signaling can regulate plant photosynthesis.

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

    OpenAIRE

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

    1993-01-01

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

  11. Greater enhancement of Bacillus subtilis spore yields in submerged cultures by optimization of medium composition through statistical experimental designs.

    Science.gov (United States)

    Chen, Zhen-Min; Li, Qing; Liu, Hua-Mei; Yu, Na; Xie, Tian-Jian; Yang, Ming-Yuan; Shen, Ping; Chen, Xiang-Dong

    2010-02-01

    Bacillus subtilis spore preparations are promising probiotics and biocontrol agents, which can be used in plants, animals, and humans. The aim of this work was to optimize the nutritional conditions using a statistical approach for the production of B. subtilis (WHK-Z12) spores. Our preliminary experiments show that corn starch, corn flour, and wheat bran were the best carbon sources. Using Plackett-Burman design, corn steep liquor, soybean flour, and yeast extract were found to be the best nitrogen source ingredients for enhancing spore production and were studied for further optimization using central composite design. The key medium components in our optimization medium were 16.18 g/l of corn steep liquor, 17.53 g/l of soybean flour, and 8.14 g/l of yeast extract. The improved medium produced spores as high as 1.52 +/- 0.06 x 10(10) spores/ml under flask cultivation conditions, and 1.56 +/- 0.07 x 10(10) spores/ml could be achieved in a 30-l fermenter after 40 h of cultivation. To the best of our knowledge, these results compared favorably to the documented spore yields produced by B. subtilis strains. PMID:19697022

  12. Transfer of nisin gene cluster from Lactococcus lactis ATCC 11454 into the chromosome of Bacillus subtilis 168.

    Science.gov (United States)

    Yuksel, Sahru; Hansen, J Norman

    2007-03-01

    Nisin is an antimicrobial peptide produced by certain strains of Lactococcus lactis. It is a gene-encoded peptide that contains unusual amino acid residues. These novel residues are introduced by posttranslational modification machinery and confer unique chemical and physical properties that are not attainable by regular amino acid residues. To study the modification mechanisms and to create structural analogs with superior properties, it would be advantageous to insert the nisin genes into a bacterial strain that is amenable to genetic manipulation. In this study, we report the cloning and integration of the complete and intact nisin gene cluster into the Bacillus subtilis 168 chromosome. Furthermore, we demonstrate that the nisin genes are transcriptionally active. These results should greatly facilitate the studies of the genes and proteins involved in nisin expression, as well as provide a standard system for the manipulation and expression of genes involved in other members of the lantibiotic family of antimicrobial peptides. PMID:17143619

  13. Engineering of Bacillus subtilis 168 for increased nisin resistance

    DEFF Research Database (Denmark)

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

    2009-01-01

    Nisin is a natural bacteriocin produced commercially by Lactococcus lactis and widely used in the food industry as a preservative because of its broad host spectrum. Despite the low productivity and troublesome fermentation of L. lactis, no alternative cost-effective host has yet been found...... to overcome the nisin sensitivity of B. subtilis by introducing the nisin resistance genes nisFEG and nisI from L. lactis under the control of a synthetic promoter library....

  14. 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.

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

    Science.gov (United States)

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

    2014-10-01

    The present study was undertaken to compare the effect of salinomycin and Bacillus subtilis on growth performance, serum antibody levels against Clostridium spp. and Eimeria spp., and cytokine mRNA expression levels in broiler chickens raised in the used litter. Broiler chickens fed a diet containing salinomycin showed lower (P chickens fed the B. subtilis-enriched diet compared with those on either the salinomycin-fed or control diet-fed chickens. None of the dietary treatments affected (P > 0.05) serum antibody levels against Clostridium perfringens toxins. Both salinomycin and B.subtilis significantly lowered (P chickens. Further study is warranted to investigate the mode of action of salinomycin on host immune response and growth performance in broiler chickens. PMID:25135491

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

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

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

  17. Effect of sporulation conditions on the resistance of Bacillus subtilis spores to heat and high pressure.

    Science.gov (United States)

    Nguyen Thi Minh, Hue; Durand, Alain; Loison, Pauline; Perrier-Cornet, Jean-Marie; Gervais, Patrick

    2011-05-01

    Bacillus subtilis(B. subtilis) cells were placed in various environmental conditions to study the effects of aeration, water activity of the medium, temperature, pH, and calcium content on spore formation and the resulting properties. Modification of the sporulation conditions lengthened the growth period of B. subtilis and its sporulation. In some cases, it reduced the final spore concentration. The sporulation conditions significantly affected the spore properties, including germination capacity and resistance to heat treatment in water (30 min at 97°C) or to high pressure (60 min at 350 MPa and 40°C). The relationship between the modifications of these spore properties and the change in the spore structure induced by different sporulation conditions is also considered. According to this study, sporulation conditions must be carefully taken into account during settling sterilization processes applied in the food industry. PMID:21380515

  18. Structural and genetic analyses of a par locus that regulates plasmid partition in Bacillus subtilis.

    OpenAIRE

    Chang, S.; Chang, S Y; Gray, O

    1987-01-01

    The Bacillus plasmid pLS11 partitions faithfully during cell division. Using a partition-deficient plasmid vector, we randomly cloned DNA fragments of plasmid pLS11 and identified the locus that regulates plasmid partition (par) by cis complementation in Bacillus subtilis. The cloned par gene conferred upon the vector plasmid a high degree of segregational stability. The par locus was mapped to a 167-base-pair segment on pLS11, and its nucleotide sequence was determined. The cloned par fragme...

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

    Directory of Open Access Journals (Sweden)

    Bhunia B

    2012-04-01

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

  20. A cell factory of Bacillus subtilis engineered for the simple bioconversion of myo-inositol to scyllo-inositol, a potential therapeutic agent for Alzheimer's disease

    Directory of Open Access Journals (Sweden)

    Takenaka Shinji

    2011-09-01

    Full Text Available Abstract Background A stereoisomer of inositol, scyllo-inositol, is known as a promising therapeutic agent for Alzheimer's disease, since it prevents the accumulation of beta-amyloid deposits, a hallmark of the disease. However, this compound is relatively rare in nature, whereas another stereoisomer of inositol, myo-inositol, is abundantly available. Results Bacillus subtilis possesses a unique inositol metabolism involving both stereoisomers. We manipulated the inositol metabolism in B. subtilis to permit the possible bioconversion from myo-inositol to scyllo-inositol. Within 48 h of cultivation, the engineered strain was able to convert almost half of 10 g/L myo-inositol to scyllo-inositol that accumulated in the culture medium. Conclusions The engineered B. subtilis serves as a prototype of cell factory enabling a novel and inexpensive supply of scyllo-inositol.

  1. Phosphoribosylpyrophosphate synthetase of Bacillus subtilis. Cloning, characterization and chromosomal mapping of the prs gene

    DEFF Research Database (Denmark)

    Nilsson, Dan; Hove-Jensen, Bjarne

    1987-01-01

    The gene (prs) encoding phosphoribosylpyrophosphate (PRPP) synthetase has been cloned from a library of Bacillus subtilis DNA by complementation of an Escherichia coli prs mutation. Flanking DNA sequences were pruned away by restriction endonuclease and exonuclease BAL 31 digestions, resulting...... in a DNA fragment of approx. 1.8 kb complementing the E. coli prs mutation. Minicell experiments revealed that this DNA fragment coded for a polypeptide, shown to be the PRPP synthetase subunit, with an Mr of approx. 40,000. B. subtilis strains harbouring the prs gene in a multicopy plasmid contained up...... to nine-fold increased PRPP synthetase activity. The prs gene was cloned in an integration vector and the resulting hybrid plasmid inserted into the B. subtilis chromosome by homologous recombination. The integration site was mapped by transduction and the gene order established as purA-guaA-prs-cysA....

  2. Phosphoribosylpyrophosphate synthetase of Bacillus subtilis. Cloning, characterization and chromosomal mapping of the prs gene

    DEFF Research Database (Denmark)

    Nilsson, Dan; Hove-Jensen, Bjarne

    1987-01-01

    The gene (prs) encoding phosphoribosylpyrophosphate (PRPP) synthetase has been cloned from a library of Bacillus subtilis DNA by complementation of an Escherichia coli prs mutation. Flanking DNA sequences were pruned away by restriction endonuclease and exonuclease BAL 31 digestions, resulting in a...... DNA fragment of approx. 1.8 kb complementing the E. coli prs mutation. Minicell experiments revealed that this DNA fragment coded for a polypeptide, shown to be the PRPP synthetase subunit, with an Mr of approx. 40,000. B. subtilis strains harbouring the prs gene in a multicopy plasmid contained up to...... nine-fold increased PRPP synthetase activity. The prs gene was cloned in an integration vector and the resulting hybrid plasmid inserted into the B. subtilis chromosome by homologous recombination. The integration site was mapped by transduction and the gene order established as purA-guaA-prs-cysA....

  3. Localization of the Cortex Lytic Enzyme CwlJ in Spores of Bacillus subtilis

    OpenAIRE

    Bagyan, Irina; Setlow, Peter

    2002-01-01

    The enzyme CwlJ is involved in the depolymerization of cortex peptidoglycan during germination of spores of Bacillus subtilis. CwlJ with a C-terminal His tag was functional and was extracted from spores by procedures that remove spore coat proteins. However, this CwlJ was not extracted from disrupted spores by dilute buffer, high salt concentrations, Triton X-100, Ca2+-dipicolinic acid, dithiothreitol, or peptidoglycan digestion, disappeared during spore germination, and was not present in co...

  4. Influence of phosphate supply on teichoic acid and teichuronic acid content of Bacillus subtilis cell walls.

    OpenAIRE

    Lang, W K; Glassey, K; Archibald, A R

    1982-01-01

    Bacillus subtilis 168 was grown in chemostat culture in fully defined media containing a constant concentration of magnesium and concentrations of phosphate that varied from those giving phosphate-limited growth to those in which phosphate was present in excess and magnesium was limiting. Phosphate-limited bacteria were deficient in wall teichoic acid and contained less than half as much cellular phosphate as did bacteria grown in excess of phosphate. Approximately 70% of the additional phosp...

  5. Sigma factors, asymmetry, and the determination of cell fate in Bacillus subtilis.

    OpenAIRE

    Lewis, P J; Partridge, S R; Errington, J

    1994-01-01

    Soon after the initiation of sporulation, Bacillus subtilis divides asymmetrically to produce sister cells that have very different developmental fates. Recently, it has been proposed that the differential gene expression which begins soon after this division is due to cell-specific activation of the transcription factors sigma F and sigma E in the prespore and the mother cell, respectively. We describe the use of a method for the localization of gene expression in individual sporulating cell...

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

    OpenAIRE

    Xenopoulos, Panagiotis; Piggot, Patrick J.

    2011-01-01

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

  7. Plasmid marker rescue transformation proceeds by breakage-reunion in Bacillus subtilis.

    OpenAIRE

    Weinrauch, Y; Dubnau, D

    1987-01-01

    Bacillus subtilis carrying a plasmid which replicates with a copy number of about 1 was transformed with linearized homologous plasmid DNA labeled with the heavy isotopes 2H and 15N, in the presence of 32Pi and 6-(p-hydroxyphenylazo)-uracil to inhibit DNA replication. Plasmid DNA was isolated from the transformed culture and fractionated in cesium chloride density gradients. The distribution of total and donor plasmid DNA was examined, using specific hybridization probes. The synthesis of new...

  8. Amplification, Sequencing and Cloning of Iranian Native Bacillus subtilis Alpha-amylase Gene in Saccharomyces cerevisiae

    OpenAIRE

    Fahimeh Afzal-Javan; Mohsen Mobini-Dehkordi

    2013-01-01

    Background: Alpha-amylases are digestive enzymes which hydrolyze starch glycosidic bonds to glucose, maltose, maltotriose and dextrin which have diverse applications in a wide range of industries such as food, textile, paper, detergents representing approximately 30% of the world enzyme production.Objectives: In this study, the gene encoding the alpha-amylase enzyme of native isolated Bacillus subtilis was amplified with specific primers containing of NotI and AscI restriction sites by PCR and...

  9. Detection of spore coat protein of Bacillus subtilis by immunological method

    International Nuclear Information System (INIS)

    The spore coat protein of Bacillus subtilis was separated, and the qualitative assay for the spore coat protein was made by use of the immunological technique. The immunological method was found to be useful for judging the maturation of spore coat in the course of sporulation. The spore coat protein antigen appeared at t2 stage of sporulation. The addition of rifampicin at the earlier stages of sporulation inhibited the increase in content of the spore coat antigen. (auth.)

  10. BslA is a self-assembling bacterial hydrophobin that coats the Bacillus subtilis biofilm

    OpenAIRE

    Hobley, Laura; Ostrowski, Adam; Rao, Francesco V.; Bromley, Keith M.; Porter, Michael; Prescott, Alan R.; MacPhee, Cait E.; van Aalten, Daan M F; Nicola R. Stanley-Wall

    2013-01-01

    Biofilms represent the predominant mode of microbial growth in the natural environment. Bacillus subtilis is a ubiquitous Gram-positive soil bacterium that functions as an effective plant growth-promoting agent. The biofilm matrix is composed of an exopolysaccharide and an amyloid fiber-forming protein, TasA, and assembles with the aid of a small secreted protein, BslA. Here we show that natively synthesized and secreted BslA forms surface layers around the biofilm. Biophysical analysis demon...

  11. Peptidoglycan Synthesis in the Absence of Class A Penicillin-Binding Proteins in Bacillus subtilis

    OpenAIRE

    McPherson, Derrell C.; Popham, David L.

    2003-01-01

    Penicillin-binding proteins (PBPs) catalyze the final, essential reactions of peptidoglycan synthesis. Three classes of PBPs catalyze either trans-, endo-, or carboxypeptidase activities on the peptidoglycan peptide side chains. Only the class A high-molecular-weight PBPs have clearly demonstrated glycosyltransferase activities that polymerize the glycan strands, and in some species these proteins have been shown to be essential. The Bacillus subtilis genome sequence contains four genes encod...

  12. Small Regulatory RNA-Induced Growth Rate Heterogeneity of Bacillus subtilis

    OpenAIRE

    Mars, Ruben A. T.; Nicolas, Pierre; Ciccolini, Mariano; Reilman, Ewoud; Reder, Alexander; Schaffer, Marc; Maeder, Ulrike; Voelker, Uwe; van Dijl, Jan Maarten; Denham, Emma L.

    2015-01-01

    Author Summary Bacterial cells that share the same genetic information can display very different phenotypes, even if they grow under identical conditions. Despite the relevance of this population heterogeneity for processes like drug resistance and development, the molecular players that induce heterogenic phenotypes are often not known. Here we report that in the Gram-positive model bacterium Bacillus subtilis a small regulatory RNA (sRNA) can induce heterogeneity in growth rates by increas...

  13. Multiple response optimization of Bacillus subtilis EA-CB0015 culture and identification of antifungal metabolites

    OpenAIRE

    Ord??z, Sergio; Mosquera, Sandra; Gonz??lez Jaramillo, Lina Mar??a; Villegas Escobar, Valeska

    2014-01-01

    The low yields of biomass and antimicrobial metabolites obtained in fermentation processes are limiting factors for implementing biological control agents in the field -- In this context, optimization of the culture medium for the biological control agent Bacillus subtilis EA-CB0015 was conducted in submerged culture to maximize the biomass production and antifungal activity -- Additionally, the active metabolites against the phytopathogen Mycosphaerella fijiensis produced under optimized con...

  14. Biomechanics of bacterial walls: studies of bacterial thread made from Bacillus subtilis.

    OpenAIRE

    Thwaites, J J; Mendelson, N H

    1985-01-01

    Bacterial threads of up to 1 m in length have been produced from filaments of separation-suppressed mutants of Bacillus subtilis. Individual threads may contain 20,000 cellular filaments in parallel alignment. The tensile properties of bacterial threads have been examined by using conventional textile engineering techniques. The kinetics of elongation at constant load are indicative of a viscoelastic material. Both Young's modulus and breaking stress are highly dependent upon relative humidit...

  15. Comparative Study of Pressure-Induced Germination of Bacillus subtilis Spores at Low and High Pressures

    OpenAIRE

    Wuytack, Elke Y.; Boven, Steven; Michiels, Chris W.

    1998-01-01

    We have studied pressure-induced germination of Bacillus subtilis spores at moderate (100 MPa) and high (500 to 600 MPa) pressures. Although we found comparable germination efficiencies under both conditions by using heat sensitivity as a criterion for germination, the sensitivity of pressure-germinated spores to some other agents was found to depend on the pressure used. Spores germinated at 100 MPa were more sensitive to pressure (>200 MPa), UV light, and hydrogen peroxide than were those g...

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

    OpenAIRE

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

    2015-01-01

    The germination of spore-forming bacteria in high-salinity environments is of applied interest for food microbiology and soil ecology. It has previously been shown that high salt concentrations detrimentally affect Bacillus subtilis spore germination, rendering this process slower and less efficient. The mechanistic details of these salt effects, however, remained obscure. Since initiation of nutrient germination first requires germinant passage through the spores' protective integuments, the...

  17. Comparative Study of Pressure- and Nutrient-Induced Germination of Bacillus subtilis Spores

    OpenAIRE

    Wuytack, Elke Y.; Soons, Johan; Poschet, Filip; Michiels, Chris W.

    2000-01-01

    Germination experiments with specific germination mutants of Bacillus subtilis, including a newly isolated mutant affected in pressure-induced germination, suggest that a pressure of 100 MPa triggers the germination cascades that are induced by the nutrient germinant alanine (Ala) and by a mixture of asparagine, glucose, fructose, and potassium ions (AGFK), by activating the receptors for alanine and asparagine, GerA and GerB, respectively. As opposed to germination at 100 MPa, germination at...

  18. Undecaprenyl Pyrophosphate Involvement in Susceptibility of Bacillus subtilis to Rare Earth Elements

    OpenAIRE

    Inaoka, Takashi; Ochi, Kozo

    2012-01-01

    The rare earth element scandium has weak antibacterial potency. We identified a mutation responsible for a scandium-resistant phenotype in Bacillus subtilis. This mutation was found within the uppS gene, which encodes undecaprenyl pyrophosphate synthase, and designated uppS86 (for the Thr-to-Ile amino acid substitution at residue 86 of undecaprenyl pyrophosphate synthase). The uppS86 mutation also gave rise to increased resistance to bacitracin, which prevents cell wall synthesis by inhibitin...

  19. Identification of host genes that affect acquisition of an integrative and conjugative element in Bacillus subtilis

    OpenAIRE

    Johnson, Christopher M; Grossman, Alan D.

    2014-01-01

    Conjugation, a major type of horizontal gene transfer in bacteria, involves transfer of DNA from a donor to a recipient using donor-encoded conjugation machinery. Using a high throughput screen (Tn-seq), we identified genes in recipients that contribute to acquisition of the integrative and conjugative element ICEBs1 by Bacillus subtilis. We found that null mutations in some genes caused an increase, and others a decrease in conjugation efficiency. Some mutations affected conjugation only whe...

  20. Bacillithiol is a major buffer of the labile zinc pool in Bacillus subtilis

    OpenAIRE

    Ma, Zhen; Chandrangsu, Pete; Helmann, Tyler C.; Romsang, Adisak; Gaballa, Ahmed; Helmann, John D.

    2014-01-01

    Intracellular zinc levels are tightly regulated since zinc is an essential cofactor for numerous enzymes, yet can be toxic when present in excess. The majority of intracellular zinc is tightly associated with proteins and is incorporated during synthesis from a poorly defined pool of kinetically labile zinc. In Bacillus subtilis, this labile pool is sensed by equilibration with the metalloregulator Zur, as an indication of zinc sufficiency, and by CzrA, as an indication of zinc excess. Here, ...

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

    OpenAIRE

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

    1991-01-01

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

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

    KAUST Repository

    Li, Yongxin

    2012-02-03

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

  3. Crystallization of the Effector-Binding Domain of Repressor DeoR from Bacillus subtilis

    Czech Academy of Sciences Publication Activity Database

    Písačková, Jana; Procházková, Kateřina; Fábry, Milan; Řezáčová, Pavlína

    2013-01-01

    Roč. 13, č. 2 (2013), s. 844-848. ISSN 1528-7483 R&D Projects: GA MŠk ME08016; GA MŠk(CZ) LK11205; GA ČR GA203/09/0820 Institutional research plan: CEZ:AV0Z50520514; CEZ:AV0Z40550506 Keywords : X-ray crystallography * deoxyribonucleoside regulator * Bacillus subtilis * thermofluor assay Subject RIV: CE - Biochemistry Impact factor: 4.558, year: 2013

  4. Modeling the inactivation of Bacillus subtilis spores by ethylene oxide processing

    OpenAIRE

    Mendes, G. C.; Brandão, T. R. S.; Silva, C. L. M.

    2011-01-01

    Ethylene oxide is currently a dominant agent in medical device sterilization. This work intends to study the main effects and interactions of temperature, ethylene oxide concentration, and relative humidity on commercial spore strips of Bacillus subtilis, var. niger (ATCC 9372) inactivation, the most common microorganism used in controlling the efficacy of the process. Experiments were carried out using a full factorial experimental design at two levels (23 factorial design). Limit targ...

  5. Modelling the inactivation of Bacillus subtilis spores by ethylene oxide processing

    OpenAIRE

    Mendes, G. C.; Brandão, T. R. S.; Silva, C. L. M.

    2009-01-01

    Ethylene oxide is currently a dominant agent in medical devices sterilization. This work intends to study the main effects and interactions of temperature (T), ethylene oxide (EO) concentration and relative humidity (RH) on commercial spore strips of Bacillus subtilis, var. niger (ATCC 9372) inactivation, the most common microorganism used in controlling the efficacy of the process. Experiments were carried out using a full factorial experimental design at two levels (23 factorial desig...

  6. Biofilm formation by Bacillus subtilis: new insights into regulatory strategies and assembly mechanisms

    OpenAIRE

    Cairns, Lynne S; Hobley, Laura; Stanley-Wall, Nicola R.

    2014-01-01

    Biofilm formation is a social behaviour that generates favourable conditions for sustained survival in the natural environment. For the Gram-positive bacterium Bacillus subtilis the process involves the differentiation of cell fate within an isogenic population and the production of communal goods that form the biofilm matrix. Here we review recent progress in understanding the regulatory pathways that control biofilm formation and highlight developments in understanding the composition, func...

  7. Biofilm formation by Bacillus subtilis: new insights into regulatory strategies and assembly mechanisms.

    Science.gov (United States)

    Cairns, Lynne S; Hobley, Laura; Stanley-Wall, Nicola R

    2014-08-01

    Biofilm formation is a social behaviour that generates favourable conditions for sustained survival in the natural environment. For the Gram-positive bacterium Bacillus subtilis the process involves the differentiation of cell fate within an isogenic population and the production of communal goods that form the biofilm matrix. Here we review recent progress in understanding the regulatory pathways that control biofilm formation and highlight developments in understanding the composition, function and structure of the biofilm matrix. PMID:24988880

  8. On the effect of N-methyl-bis (3-mesyloxypropyl) amine hydroxychloride on Bacillus subtilis cells.

    Science.gov (United States)

    Shimi, I R; Shoukry, S

    1975-06-01

    N-Methyl-bis (3-mesyloxypropyl)amine hydrochloride is now in use as an antitumer drug. In view of its activity against some bacteria the present work was conducted to study its mode of action of Bacillus subtilis. The compound was found to induce irreversible damage to bacterial DNA whereas its effect on RNA was temporary and depending on maintenance of effective concentrations of the compound. PMID:168172

  9. Differential Actions of Chlorhexidine on the Cell Wall of Bacillus subtilis and Escherichia coli

    OpenAIRE

    Cheung, Hon-Yeung; Wong, Matthew Man-Kin; Cheung, Sau-Ha; Liang, Longman Yimin; Lam, Yun-Wah; Chiu, Sung-Kay

    2012-01-01

    Chlorhexidine is a chlorinated phenolic disinfectant used commonly in mouthwash for its action against bacteria. However, a comparative study of the action of chlorhexidine on the cell morphology of Gram-positive and Gram-negative bacteria is lacking. In this study, the actions of chlorhexidine on the cell morphology were identified with the aids of electron microscopy. After exposure to chlorhexidine, numerous spots of indentation on the cell wall were found in both Bacillus subtilis and Esc...

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

    Directory of Open Access Journals (Sweden)

    Andrew J.

    2014-03-01

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

  11. Cell division of cycle of Bacillus subtilis: evidence of variability in period D.

    OpenAIRE

    Holmes, M.; Rickert, M; Pierucci, O

    1980-01-01

    In Bacillus subtilis the deoxyribonucleic acid content and the extent of cell division during inhibition of chromosome replication increased as a function of the average cell mass, independent of the growth rate. At each growth rate, mass, deoxyribonucleic acid, and residual division varied in different cultures. The variation is consistent with a large variability in the D period. At growth rates higher than 1.5 doublings per h at 37 degrees C, the change in D accounts for the growth rate de...

  12. Cell wall synthesis and initiation of deoxyribonucleic acid replication in Bacillus subtilis.

    OpenAIRE

    Sandler, N.; Keynan, A

    1981-01-01

    We have observed a connection between cell wall synthesis and the initiation of chromosome replication in Bacillus subtilis. Initiation of chromosome replication was prevented in synchronous cultures in the presence of the cell wall synthesis inhibitor vancomycin. When vancomycin was added to the cultures after initiation of chromosome replication, one round of replication was completed but no reinitiation occurred. Similar results were obtained when cell wall synthesis was inhibited by risto...

  13. Scale-down and parallel operation of a riboflavin production process with Bacillus subtilis

    OpenAIRE

    Knorr, Bettina

    2007-01-01

    Novel parallel bioreactor systems at a milliliter scale were recently developed for the design and improvement of biological cultivations. The objective of this work was to identify the reaction parameters that were necessary for a representative scale-down of an industrial manufacturing process to be carried out with the new technology. The process for the production of riboflavin with Bacillus subtilis, operated in a controlled fed-batch mode, served as an example for investigations in stir...

  14. Energy and calcium ion dependence of proteolysis during sporulation of Bacillus subtilis cells.

    OpenAIRE

    O'Hara, M B; Hageman, J H

    1990-01-01

    Bacterial cells degrade intracellular proteins at elevated rates during starvation and can selectively degrade proteins by energy-dependent processes. Sporulating bacteria can degrade protein with apparent first-order rate constants of over 0.20 h-1. We have shown, with an optimized [14C]leucine-labeling and chasing procedure, in a chemically defined sporulation medium, that intracellular protein degradation in sporulating cells of Bacillus subtilis 168 (trpC2) is apparently energy dependent....

  15. Production and Characteristics of Raw-Potato-Starch-Digesting α-Amylase from Bacillus subtilis 65

    OpenAIRE

    Hayashida, Shinsaku; Teramoto, Yuji; Inoue, Takehiro

    1988-01-01

    A newly isolated bacterium, identified as Bacillus subtilis 65, was found to produce raw-starch-digesting α-amylase. The electrophoretically homogeneous preparation of enzyme (molecular weight, 68,000) digested and solubilized raw corn starch to glucose and maltose with small amounts of maltooligosaccharides ranging from maltotriose to maltoheptaose. This enzyme was different from other amylases and could digest raw potato starch almost as fast as it could corn starch, but it showed no adsorb...

  16. Cloning, Sequencing, and Disruption of the Bacillus subtilis psd Gene Coding for Phosphatidylserine Decarboxylase

    OpenAIRE

    Matsumoto, Kouji; Okada, Masahiro; Horikoshi, Yuko; Matsuzaki, Hiroshi; Kishi, Tsutomu; Itaya, Mitsuhiro; Shibuya, Isao

    1998-01-01

    The psd gene of Bacillus subtilis Marburg, encoding phosphatidylserine decarboxylase, has been cloned and sequenced. It encodes a polypeptide of 263 amino acid residues (deduced molecular weight of 29,689) and is located just downstream of pss, the structural gene for phosphatidylserine synthase that catalyzes the preceding reaction in phosphatidylethanolamine synthesis (M. Okada, H. Matsuzaki, I. Shibuya, and K. Matsumoto, J. Bacteriol. 176:7456–7461, 1994). Introduction of a plasmid contain...

  17. Episodic Selection and the Maintenance of Competence and Natural Transformation in Bacillus subtilis

    OpenAIRE

    Johnsen, P. J.; Dubnau, D; Levin, B. R.

    2009-01-01

    We present a new hypothesis for the selective pressures responsible for maintaining natural competence and transformation. Our hypothesis is based in part on the observation that in Bacillus subtilis, where transformation is widespread, competence is associated with periods of nongrowth in otherwise growing populations. As postulated for the phenomenon of persistence, the short-term fitness cost associated with the production of transiently nongrowing bacteria can be compensated for and the c...

  18. Metabolic engineering of Bacillus subtilis for the efficient biosynthesis of uniform hyaluronic acid with controlled molecular weights.

    Science.gov (United States)

    Jia, Yuning; Zhu, Jing; Chen, Xiaofei; Tang, Dongyang; Su, Ding; Yao, Wenbing; Gao, Xiangdong

    2013-03-01

    Bacillus subtilis was engineered into an efficient hyaluronic acid (HA) producer by introducing two inducible artificial operons carrying HA synthase gene from Pasteurella multocida and precursor genes encoding enzymes involved in synthesis of the sugar precursors. A two-stage induction strategy was established for metabolic engineering of recombinant B. subtilis to efficiently produce uniform HA with controlled molecular weights. Strain TPG223 produced larger HA molecules (yield=6.8 g/L; molecular weight=4.5 MDa) than strain PG6181 (yield=2.4 g/L; molecular weight=13 KDa), indicating that the enzymes involved in the synthesis of UDP-glucuronic acid are essential for HA biosynthesis. Strain TPG223 was able to synthesize HA molecules ranging in molecular weight from 8 KDa to 5.4 MDa indicating that size control is achievable in vivo through appropriate tools. The work reported here not only advanced mechanisms research of size control in vivo, but also could be an attractive alternative for commercial preparation of uniform size-defined HA. PMID:23433979

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

    KAUST Repository

    Li, Yongxin

    2015-03-24

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

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

    Science.gov (United States)

    Li, Yongxin; Li, Zhongrui; Yamanaka, Kazuya; Xu, Ying; Zhang, Weipeng; Vlamakis, Hera; Kolter, Roberto; Moore, Bradley S.; Qian, Pei-Yuan

    2015-03-01

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

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

    Directory of Open Access Journals (Sweden)

    Salcedo L.

    1998-06-01

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

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

    DEFF Research Database (Denmark)

    Jensen, Kaj Frank

    1978-01-01

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

  3. Multifactorial Resistance of Bacillus subtilis Spores to High-Energy Proton Radiation: Role of Spore Structural Components and the Homologous Recombination and Non-Homologous End Joining DNA Repair Pathways

    OpenAIRE

    Moeller, Ralf; Reitz, Günther; Li, Zuofeng; Klein, Stuart; Nicholson, Wayne L.

    2012-01-01

    The space environment contains high-energy charged particles (e.g., protons, neutrons, electrons, α-particles, heavy ions) emitted by the Sun and galactic sources or trapped in the radiation belts. Protons constitute the majority (87%) of high-energy charged particles. Spores of Bacillus species are one of the model systems used for astro- and radiobiological studies. In this study, spores of different Bacillus subtilis strains were used to study the effects of high energetic proton irradiati...

  4. Translation Control of Swarming Proficiency in Bacillus subtilis by 5-Amino-pentanolylated Elongation Factor P.

    Science.gov (United States)

    Rajkovic, Andrei; Hummels, Katherine R; Witzky, Anne; Erickson, Sarah; Gafken, Philip R; Whitelegge, Julian P; Faull, Kym F; Kearns, Daniel B; Ibba, Michael

    2016-05-20

    Elongation factor P (EF-P) accelerates diprolyl synthesis and requires a posttranslational modification to maintain proteostasis. Two phylogenetically distinct EF-P modification pathways have been described and are encoded in the majority of Gram-negative bacteria, but neither is present in Gram-positive bacteria. Prior work suggested that the EF-P-encoding gene (efp) primarily supports Bacillus subtilis swarming differentiation, whereas EF-P in Gram-negative bacteria has a more global housekeeping role, prompting our investigation to determine whether EF-P is modified and how it impacts gene expression in motile cells. We identified a 5-aminopentanol moiety attached to Lys(32) of B. subtilis EF-P that is required for swarming motility. A fluorescent in vivo B. subtilis reporter system identified peptide motifs whose efficient synthesis was most dependent on 5-aminopentanol EF-P. Examination of the B. subtilis genome sequence showed that these EF-P-dependent peptide motifs were represented in flagellar genes. Taken together, these data show that, in B. subtilis, a previously uncharacterized posttranslational modification of EF-P can modulate the synthesis of specific diprolyl motifs present in proteins required for swarming motility. PMID:27002156

  5. Heat resistance of spore-forming microorganisms (Bacillus sporothermodurans, Bacillus subtilis and Geobacillus stearothermophilus) under isothermal and non-iiothermal conditions

    OpenAIRE

    Gómez Jódar, Isabel

    2015-01-01

    [SPA]El principal género de microorganismos esporulados altamente resistentes al calor involucrados en el deterioro de alimentos es Bacillus. Este género causa problemas de no esterilidad en alimentos enlatados y reduce la vida comercial de muchos alimentos procesados. En este estudio se determinó la termorresistencia de Bacillus sporothermodurans IIC65, Bacillus subtilis IC9 y Geobacillus stearothermophilus T26 mediante un termorresistómetro Mastia (Conesa et al., 2009). Las determinaciones ...

  6. Biosynthesis of Polymyxins B, E, and P Using Genetically Engineered Polymyxin Synthetases in the Surrogate Host Bacillus subtilis.

    Science.gov (United States)

    Kim, Se-Yu; Park, Soo-Young; Choi, Soo-Keun; Park, Seung-Hwan

    2015-07-01

    The development of diverse polymyxin derivatives is needed to solve the toxicity and resistance problems of polymyxins. However, no platform has generated polymyxin derivatives by genetically engineering a polymyxin synthetase, which is a nonribosomal peptide synthetase. In this study, we present a two-step approach for the construction of engineered polymyxin synthetases by substituting the adenylation (A) domains of polymyxin A synthetase, which is encoded by the pmxABCDE gene cluster of Paenibacillus polymyxa E681. First, the seventh L-threonine-specific A-domain region in pmxA was substituted with the Lleucine- specific A-domain region obtained from P. polymyxa ATCC21830 to make polymyxin E synthetase, and then the sixth D-leucine-specific A-domain region (A6-D-Leu-domain) was substituted with the D-phenylalanine-specific A-domain region (A6-D-Phe-domain) obtained from P. polymyxa F4 to make polymyxin B synthetase. This step was performed in Escherichia coli on a pmxA-containing fosmid, using the lambda Red recombination system and the sacB gene as a counter-selectable marker. Next, the modified pmxA gene was fused to pmxBCDE on the chromosome of Bacillus subtilis BSK4dA, and the resulting recombinant strains BSK4-PB and BSK4-PE were confirmed to produce polymyxins B and E, respectively. We also succeeded in constructing the B. subtilis BSK4-PP strain, which produces polymyxin P, by singly substituting the A6-D-Leu-domain with the A6-D-Phe-domain. This is the first report in which polymyxin derivatives were generated by genetically engineering polymyxin synthetases. The two recombinant B. subtilis strains will be useful for improving the commercial production of polymyxins B and E, and they will facilitate the generation of novel polymyxin derivatives. PMID:26059516

  7. Antimicrobial gageomacrolactins characterized from the fermentation of the marine-derived bacterium Bacillus subtilis under optimum growth conditions.

    Science.gov (United States)

    Tareq, Fakir Shahidullah; Kim, Ji Hye; Lee, Min Ah; Lee, Hyi-Seung; Lee, Jong-Seok; Lee, Yeon-Ju; Shin, Hee Jae

    2013-04-10

    Marine bacteria are a potential source of structurally diversified bioactive secondary metabolites that are not found in terrestrial sources. In our continuous effort to search for new antimicrobial agents from marine-derived bacteria, we isolated bacterial strain 109GGC020 from a marine sediment sample collected from Gageocho, Republic of Korea. The strain was identified as Bacillus subtilis based on a 16s rRNA sequence analysis. After a 7-day fermentation of the B. subtilis strain under optimum growth conditions three new and four known secondary metabolites were discovered using chromatographic procedures, and their biological activities were evaluated against both bacteria and crop-devastating fungi. The discovered metabolites were confirmed by extensive 2D NMR and high-resolution ESI-MS data analyses to have the structures of new macrolactin derivatives gageomacrolactins 1-3 and known macrolactins A (4), B (5), F (6), and W (7). The stereoconfigurations of 1-3 were assigned based on coupling constant values, chemical derivatization studies, and a literature review. The coupling constants were very crucial to determine the relative geometries of olefins in 1-3 because of overlap of the ¹H NMR signals. The NMR data of these compounds were recorded in different solvents to overcome this problem and obtain accurate coupling constant values. The new macrolactin derivatives 1-3 displayed good antibiotic properties against both Gram-positive (S. aureus, B. subtilis, and B. cereus) and Gram-negative (E. coli, S. typhi, and P. aeruginosa) bacteria with minimum inhibitory concentration (MIC) values of 0.02-0.05 μM. Additionally, the antifungal activities of 1-7 were evaluated against pathogenic fungi and found to inhibit mycelial growth of A. niger, B. cinerea, C. acutatum, C. albicans, and R. solani with MIC values of 0.04-0.3 μM, demonstrating that these compounds were good fungicides. PMID:23488669

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

    OpenAIRE

    Pandey, R

    2014-01-01

    Spores of Gram-positive bacteria such as Bacillus and Clostridium cause huge economic losses to the food industry. In food products, spores survive under food preservation conditions and subsequent germination and outgrowth eventually causes food spoilage. Therefore efforts are being made to eliminate or inactivate these bacterial spores in foods. In this regard food industry uses different preservation methods such as thermal-treatment, weak acids, antimicrobial compounds etc. Complete therm...

  9. A high-efficiency recombineering system with PCR-based ssDNA in Bacillus subtilis mediated by the native phage recombinase GP35.

    Science.gov (United States)

    Sun, Zhaopeng; Deng, Aihua; Hu, Ting; Wu, Jie; Sun, Qinyun; Bai, Hua; Zhang, Guoqiang; Wen, Tingyi

    2015-06-01

    Bacillus subtilis and its closely related species are important strains for industry, agriculture, and medicine. However, it is difficult to perform genetic manipulations using the endogenous recombination machinery. In many bacteria, phage recombineering systems have been employed to improve recombineering frequencies. To date, an efficient phage recombineering system for B. subtilis has not been reported. Here, we, for the first time, identified that GP35 from the native phage SPP1 exhibited a high recombination activity in B. subtilis. On this basis, we developed a high-efficiency GP35-meditated recombineering system. Taking single-stranded DNA (ssDNA) as a recombineering substrate, ten recombinases from diverse sources were investigated in B. subtilis W168. GP35 showed the highest recombineering frequency (1.71 ± 0.15 × 10(-1)). Besides targeting the purine nucleoside phosphorylase gene (deoD), we also demonstrated the utility of GP35 and Beta from Escherichia coli lambda phage by deleting the alpha-amylase gene (amyE) and uracil phosphoribosyltransferase gene (upp). In all three genetic loci, GP35 exhibited a higher frequency than Beta. Moreover, a phylogenetic tree comparing the kinship of different recombinase hosts with B. subtilis was constructed, and the relationship between the recombineering frequency and the kinship of the host was further analyzed. The results suggested that closer kinship to B. subtilis resulted in higher frequency in B. subtilis. In conclusion, the recombinase from native phage or prophage can significantly promote the genetic recombineering frequency in its host, providing an effective genetic tool for constructing genetically engineered strains and investigating bacterial physiology. PMID:25750031

  10. A non-destructive method for characterizing phenotypes and growth of a Bacillus subtilis biofilm using fluorescence microscopy

    Science.gov (United States)

    Koehler, Stephan; Wang, Xiaoling; Wilking, James; Weitz, Dave

    2015-11-01

    We develop an imaging technique for characterizing growth of biofilms using a triple fluorescent labeled strain for the three main phenotypes of a Bacillus subtilis biofilm on an agar substrate. We find that the biofilm does not flow across the substrate and thus growth is due to colonization at the periphery and thickening of the interior regions. We obtain local height and its composition of the three main phenotypes, which are motile, matrix-producing and sporulating, as well as the non-fluorescent material, which can be spores, dormant or dead cells or extracellular matrix. This technique is suitable for the study of biofilm growth and inhibition for different conditions such as biocides or bioremediation.

  11. Stepwise optimization of a low-temperature Bacillus subtilis expression system for "difficult to express" proteins.

    Science.gov (United States)

    Welsch, Norma; Homuth, Georg; Schweder, Thomas

    2015-08-01

    In order to improve the overproduction of "difficult to express" proteins, a low-temperature expression system for Bacillus subtilis based on the cold-inducible promoter of the desaturase-encoding des gene was constructed. Selected regulatory DNA sequence elements from B. subtilis genes known to be cold-inducible were fused to different model genes. It could be demonstrated that these regulatory elements are able to mediate increased heterologous gene expression, either by improved translation efficiency or by higher messenger RNA (mRNA) stability. In case of a cold-adapted β-galactosidase from Pseudoalteromonas haloplanktis TAE79A serving as the model, significantly higher expression was achieved by fusing its coding sequence to the so-called "downstream box" sequence of cspB encoding the major B. subtilis cold-shock protein. The combination of this fusion with a cspB 5'-UTR stem-loop structure resulted in further enhancement of the β-galactosidase expression. In addition, integration of the transcription terminator of the B. subtilis cold-inducible bkd operon downstream of the target genes caused a higher mRNA stability and enabled thus a further significant increase in expression. Finally, the fully optimized expression system was validated by overproducing a B. subtilis xylanase as well as an α-glucosidase from Saccharomyces cerevisiae, the latter known for tending to form inclusion bodies. These analyses verified the applicability of the engineered expression system for extracellular and intracellular protein synthesis in B. subtilis, thereby confirming the suitability of this host organism for the overproduction of critical, poorly soluble proteins. PMID:25851716

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

    Directory of Open Access Journals (Sweden)

    Marciniak Bogumiła C

    2012-05-01

    Full Text Available Abstract Background Bacillus subtilis is a favorable host for the production of industrially relevant proteins because of its capacity of secreting proteins into the medium to high levels, its GRAS (Generally Recognized As Safe status, its genetic accessibility and its capacity to grow in large fermentations. However, production of heterologous proteins still faces limitations. Results This study aimed at the identification of bottlenecks in secretory protein production by analyzing the response of B. subtilis at the transcriptome level to overproduction of eight secretory proteins of endogenous and heterologous origin and with different subcellular or extracellular destination: secreted proteins (NprE and XynA of B. subtilis, Usp45 of Lactococcus lactis, TEM-1 β-lactamase of Escherichia coli, membrane proteins (LmrA of L. lactis and XylP of Lactobacillus pentosus and lipoproteins (MntA and YcdH of B. subtilis. Responses specific for proteins with a common localization as well as more general stress responses were observed. The latter include upregulation of genes encoding intracellular stress proteins (groES/EL, CtsR regulated genes. Specific responses include upregulation of the liaIHGFSR operon under Usp45 and TEM-1 β-lactamase overproduction; cssRS, htrA and htrB under all secreted proteins overproduction; sigW and SigW-regulated genes mainly under membrane proteins overproduction; and ykrL (encoding an HtpX homologue specifically under membrane proteins overproduction. Conclusions The results give better insights into B. subtilis responses to protein overproduction stress and provide potential targets for genetic engineering in order to further improve B. subtilis as a protein production host.

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

    DEFF Research Database (Denmark)

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

    2016-01-01

    Mutants of Bacillus subtilis can be developed to overproduce Val in vitro. It was hypothesized that addition of Bacillus subtilis mutants to pig diets can be a strategy to supply the animal with Val. The objective was to investigate the effect of Bacillus subtilis mutants on growth performance and...... blood amino acid (AA) concentrations when fed to piglets. Experiment 1 included 18 pigs (15.0±1.1 kg) fed one of three diets containing either 0.63 or 0.69 standardized ileal digestible (SID) Val : Lys, or 0.63 SID Val : Lys supplemented with a Bacillus subtilis mutant (mutant 1). Blood samples were...... obtained 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding and analyzed for AAs. In Experiment 2, 80 piglets (9.1±1.1 kg) were fed one of four diets containing 0.63 or 0.67 SID Val : Lys, or 0.63 SID Val : Lys supplemented with another Bacillus subtilis mutant (mutant 2) or its parent wild...

  14. T-box-mediated control of the anabolic proline biosynthetic genes of Bacillus subtilis.

    Science.gov (United States)

    Brill, Jeanette; Hoffmann, Tamara; Putzer, Harald; Bremer, Erhard

    2011-04-01

    Bacillus subtilis possesses interlinked routes for the synthesis of proline. The ProJ-ProA-ProH route is responsible for the production of proline as an osmoprotectant, and the ProB-ProA-ProI route provides proline for protein synthesis. We show here that the transcription of the anabolic proBA and proI genes is controlled in response to proline limitation via a T-box-mediated termination/antitermination regulatory mechanism, a tRNA-responsive riboswitch. Primer extension analysis revealed mRNA leader transcripts of 270 and 269 nt for the proBA and proI genes, respectively, both of which are synthesized from SigA-type promoters. These leader transcripts are predicted to fold into two mutually exclusive secondary mRNA structures, forming either a terminator or an antiterminator configuration. Northern blot analysis allowed the detection of both the leader and the full-length proBA and proI transcripts. Assessment of the level of the proBA transcripts revealed that the amount of the full-length mRNA species strongly increased in proline-starved cultures. Genetic studies with a proB-treA operon fusion reporter strain demonstrated that proBA transcription is sensitively tied to proline availability and is derepressed as soon as cellular starvation for proline sets in. Both the proBA and the proI leader sequences contain a CCU proline-specific specifier codon prone to interact with the corresponding uncharged proline-specific tRNA. By replacing the CCU proline specifier codon in the proBA T-box leader with UUC, a codon recognized by a Phe-specific tRNA, we were able to synthetically re-engineer the proline-specific control of proBA transcription to a control that was responsive to starvation for phenylalanine. PMID:21233158

  15. Determination and modeling the optimum conditions of beta glucanase Bacillus subtilis B5d activity with potential used as feed additive

    Directory of Open Access Journals (Sweden)

    Sholeh Dahpahlevan

    2016-06-01

    Full Text Available Introduction: Microbial β - 1, 4 glucanase is one of the key enzymes in glucan hydrolysis. Among bacteria, Bacillus species is a main source for industrial glucanse production. According to interaction between environmental factors on the enzymatic catalytic properties, in this study the potential of β-glucanase production by native strain B. subtilis B5d isolated from apple phyllosphere orchards and determination and modeling the optimum conditions of enzyme activity was surveyed. Materials and methods: In this study, the potential of β-glucanase production by B. subtilis B5d was confirmed using qualitative method. The B. subtilis B5d isolate was identified through biochemical and molecular methods. Determination of optimum condition for beta glucanase activity was evaluated by response surface method and Central Composite Design. Investigated variables included temperature (20- 80 °C, pH (3.5- 8 and concentration of substrate (0.1- 1%. Results: According to the molecular and biochemical analysis, the strain B. subtilis B5d belonges to B.subtilis species. Based on the Central Composite Design results, the quadratic model was the most suitable model for predicting β-glucanase activity in experimental temperature, pH and substrate concentration range. Furthermore, the optimization of enzyme activity showed that the β-glucanase produced by strain B5d has highest catalytic activity at pH (5.5- 7 and temperature (50- 65°C and the high concentration of lechinan. However, the enzyme activity is reduced in 80 and 20 °C as well as low concentration of substrate. Validation test of the prediction model of optimization obtained by response surface methodology showed a good agreement between experimental and predicted data. Discussion and conclusion: According to the optimum catalytic characteristics, the β-glucanase produced by B. subtilis B5d has a potential to be used as a feed additive.

  16. Use of a Novel Report Protein to Study the Secretion Signal of Flagellin in Bacillus subtilis.

    Science.gov (United States)

    Wang, Guangqiang; Xia, Yongjun; Xiong, Zhiqiang; Zhang, Hui; Ai, Lianzhong

    2016-08-01

    Flagellin (also called Hag) is the main component of bacterial flagellum and is transported across the cytoplasmic membrane by flagellar secretion apparatus. Because flagella play an essential role in the pathogenesis of numerous pathogens, the flagellins of Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Campylobacter jejuni, and Vibrio cholerae have been intensively studied; however, very few studies have focused on the flagellin of Bacillus subtilis, which is considered to be a model organism with which to study the secretion of bacteria and is used on an industrial scale for the secretion of proteins. The signal of B. subtilis flagellin is still debated. This study was performed to seek the export signals of flagellin from B. subtilis. The naturally nonsecretory, intrinsically disordered domain of nucleoskeletal-like protein (Nsp) was used as the reporter protein. Our results demonstrate that the export signal is contained within the first 50 amino acids of B. subtilis flagellin. Nsp is easily degraded inside the cell and can be exported into culture medium with the aid of the signal of flagellin. This method provides a new potential strategy for the expression of proteins with high proteolytic susceptibility via fusion to export signals. PMID:27154466

  17. Identification of a soluble diacylglycerol kinase required for lipoteichoic acid production in Bacillus subtilis.

    Science.gov (United States)

    Jerga, Agoston; Lu, Ying-Jie; Schujman, Gustavo E; de Mendoza, Diego; Rock, Charles O

    2007-07-27

    Diacylglycerol kinases (DagKs) are key enzymes in lipid metabolism that function to reintroduce diacylglycerol formed from the hydrolysis of phospholipids into the biosynthetic pathway. Bacillus subtilis is a prototypical Gram-positive bacterium with a lipoteichoic acid structure containing repeating units of sn-glycerol-1-P groups derived from phosphatidylglycerol head groups. The B. subtilis homolog of the prokaryotic DagK gene family (dgkA; Pfam01219) was not a DagK but rather was an undecaprenol kinase. The three members of the soluble DagK protein family (Pfam00781) in B. subtilis were tested by complementation of an E. coli dgkA mutant, and only the essential yerQ gene possessed DagK activity. This gene was dubbed dgkB, and the soluble protein product was purified, and its DagK activity was verified in vitro. Conditional inactivation of dgkB led to the accumulation of diacylglycerol and the cessation of lipoteichoic acid formation in B. subtilis. This study identifies a soluble protein encoded by the dgkB (yerQ) gene as an essential kinase in the diacylglycerol cycle that drives lipoteichoic acid production. PMID:17535816

  18. Preliminary X-ray crystallographic studies of Bacillus subtilis SpeA protein

    International Nuclear Information System (INIS)

    In order to further illustrate the catalytic mechanism of arginine decarboxylase by determining the three-dimensional structure of the enzyme the speA gene was amplified from B. subtilis genomic DNA and cloned. The enzyme was expressed in Escherichia coli and purified to homogeneity by nickel-chelation chromatography followed by size-exclusion chromatography. High-quality crystals were obtained using the hanging-drop vapour-diffusion method at 298 K. The speA gene in Bacillus subtilis encodes arginine decarboxylase, which catalyzes the conversion of arginine to agmatine. Arginine decarboxylase is an important enzyme in polyamine metabolism in B. subtilis. In order to further illustrate the catalytic mechanism of arginine decarboxylase by determining the three-dimensional structure of the enzyme, the speA gene was amplified from B. subtilis genomic DNA and cloned into the expression vector pET-28a(+). SpeA was expressed in Escherichia coli and purified to homogeneity by nickel-chelation chromatography followed by size-exclusion chromatography. High-quality crystals were obtained using the hanging-drop vapour-diffusion method at 289 K. The best crystal diffracted to 2.0 Å resolution and belonged to space group P21, with unit-cell parameters a = 86.4, b = 63.3 c = 103.3 Å, β = 113.9°

  19. The Antimicrobial Properties of Silver Nanoparticles in Bacillus subtilis Are Mediated by Released Ag+ Ions.

    Directory of Open Access Journals (Sweden)

    Yi-Huang Hsueh

    Full Text Available The superior antimicrobial properties of silver nanoparticles (Ag NPs are well-documented, but the exact mechanisms underlying Ag-NP microbial toxicity remain the subject of intense debate. Here, we show that Ag-NP concentrations as low as 10 ppm exert significant toxicity against Bacillus subtilis, a beneficial bacterium ubiquitous in the soil. Growth arrest and chromosomal DNA degradation were observed, and flow cytometric quantification of propidium iodide (PI staining also revealed that Ag-NP concentrations of 25 ppm and above increased membrane permeability. RedoxSensor content analysis and Phag-GFP expression analysis further indicated that reductase activity and cytosolic protein expression decreased in B. subtilis cells treated with 10-50 ppm of Ag NPs. We conducted X-ray absorption near-edge structure (XANES and extended X-ray absorption fine structure (EXAFS analyses to directly clarify the valence and fine structure of Ag atoms in B. subtilis cells placed in contact with Ag NPs. The results confirmed the Ag species in Ag NP-treated B. subtilis cells as Ag2O, indicating that Ag-NP toxicity is likely mediated by released Ag+ ions from Ag NPs, which penetrate bacterial cells and are subsequently oxidized intracellularly to Ag2O. These findings provide conclusive evidence for the role of Ag+ ions in Ag-NP microbial toxicity, and suggest that the impact of inappropriately disposed Ag NPs to soil and water ecosystems may warrant further investigation.

  20. Bacillus subtilis ZH168多酚氧化酶分离纯化研究%Purification of polyphenol oxidase from the Bacillus subtilis ZH168

    Institute of Scientific and Technical Information of China (English)

    张丽香

    2015-01-01

    从一株产黑色素Bacillus subtilis ZH168发酵液中提取多酚氧化酶,通过硫酸铵盐析,超滤,阴离子交换层析,活性和变性电泳确定该酶有2条同工酶带,分离纯化到其中分子量较大的同工酶,为101.5 ku,并将纯化同工酶带作基质辅助激光吸附-离子化飞行时间质谱(MALDI-TOF-MS)获得蛋白肽指纹谱,通过一二级质谱检索结果确定该蛋白与枯草茅孢杆菌芽孢衣蛋白有极高相似性.

  1. Duodenal histology and carcass quality of feedlot cattle supplemented with calcium butyrate and Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Thiago Simas de Oliveira Moreira

    2016-01-01

    Full Text Available The experiment was carried out at the Comigo Technology Center, in Rio Verde, State of Goiás, Brazil, with the objective of evaluating the effects of supplementation with calcium butyrate, as a growth promoting agent for the duodenal mucosa and Bacillus subtilis as a probiotic performance enhancer in feedlot cattle. Calcium butyrate (5 and 10 g per animal per day and Bacillus (10 g per animal per day were added to a basal diet. There were used 85 Nelore bulls, with average weight of 315 ± 7 kg. The experiment lasted 118 days, including the adaptation period, until slaughter at 30 months of age. Diets were distributed in a completely randomized design with four treatments, where: T1 = control (basal diet; T2 = basal diet + 5 g calcium butyrate; T3 = basal diet + 10 g calcium butyrate and T4 = basal diet + 10 g calcium butyrate + 10 g probiotic with four replications and five to six animals per replication. It was used a forage: concentrate ratio of 30:70, the roughage used was the corn silage. Height and width measurements of intestinal villi were taken, and carcass and meat quality were evaluated. The supplementation of calcium butyrate and Bacillus subtilis positively influenced (p < 0.05 the carcass marbling level and calcium butyrate increased the villus height in the small intestine.

  2. Plasmid Mediated Antibiotic and Heavy Metal Resistance in Bacillus Strains Isolated From Soils in Rize, Turkey

    Directory of Open Access Journals (Sweden)

    Elif SEVİM

    2015-09-01

    Full Text Available Fifteen Bacillus strains which were isolated from soil samples were examined for resistance to 17 different antibiotics (ampicillin, methicillin, erythromycin, norfloxacin, cephalotine, gentamycin, ciprofloxacin, streptomycin, tobramycin, chloramphenicol, trimethoprim-sulfamethoxazole, tetracycline, vancomycin, oxacilin, neomycin, kanamycin and, novabiocin and to 10 different heavy metals (copper, lead, cobalt, chrome, iron, mercury, zinc, nickel, manganese and, cadmium and for the presence of plasmid DNA. A total of eleven strains (67% were resistant to at least one antibiotic. The most common resistance was observed against methicillin and oxacillin. The most resistance strains were found as Bacillus sp. B3 and Bacillus sp. B11. High heavy metal resistance against copper, chromium, zinc, iron and nickel was detected, but mercury and cobalt resistance was not detected, except for 3 strains (B3, B11, and B12 which showed mercury resistance. It has been determined that seven Bacillus strains have plasmids. The isolated plasmids were transformed into the Bacillus subtilis W168 and it was shown that heavy metal and antibiotic resistance determinants were carried on these plasmids. These results showed that there was a correlation between plasmid content and resistance for both antibiotic and heavy metal resistance

  3. Engineering of Bacillus subtilis for the Production of 2,3-Butanediol from Sugarcane Molasses.

    Science.gov (United States)

    Deshmukh, Apoorva Nandkumar; Nipanikar-Gokhale, Padmaja; Jain, Rishi

    2016-05-01

    2,3-butanediol is known to be a platform chemical with several potential industrial applications. Sustainable industrial scale production can be attained by using a sugarcane molasses based fermentation process using Bacillus subtilis. However, the accumulation of acetoin needs to be reduced to improve process efficiency. In this work, B. subtilis was genetically modified in order to increase the yield of 2,3-butanediol. Metabolic engineering strategies such as cofactor engineering and overexpression of the key enzyme butanediol dehydrogenase were attempted. Both the strategies individually led to a statistically significant increase in the 2,3-butanediol yields for sugarcane molasses based fermentation. Cofactor engineering led to a 26 % increase in 2,3-butanediol yield and overexpression of bdhA led to a 11 % increase. However, the combination of the two strategies did not lead to a synergistic increase in 2,3-butanediol yield. PMID:26825987

  4. Influence of Silica Nanoparticles on Antioxidant Potential of Bacillus subtilis IMV B-7023

    Science.gov (United States)

    Skorochod, Iryna O.; Roy, Alla O.; Kurdish, Ivan K.

    2016-03-01

    It was found that if introduced into a nutrient medium of 0.05-1 g/L nano-SiO2, the oxidant activity (OA) of the culture medium (CM) of bacilli increased by 43.2-60.1 % and the antioxidant activity (AA) decreased by 4.5-11.8 %. SiO2 nanoparticles had different effects on antiradical activity (ARA) of the CM of Bacillus subtilis IMV B-7023. In particular, nano-SiO2 had no significant effect on the ability of the CM of bacilli to inactivate the 2.2-diphenyl-1-picrylhydrazyl (DPPH·) free radical. However, for the content of the nanomaterial of 0.01-1 g/L decreased hydroxyl radical scavenging in the CM of B. subtilis IMV B-7023 on 7.2-17.6 % compared with a control. Low doses of silica nanoparticles stimulated the reducing power of the CM of bacteria and then highly suppressed it.

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

    Directory of Open Access Journals (Sweden)

    Sirec Teja

    2012-08-01

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

  6. Tyrosine 601 of Bacillus subtilis DnaK undergoes phosphorylation and is crucial for chaperone activity and heat shock survival

    Directory of Open Access Journals (Sweden)

    Lei eShi

    2016-04-01

    Full Text Available In order to screen for cellular substrates of the Bacillus subtilis BY-kinase PtkA, and its cognate phosphotyrosine-protein phosphatase PtpZ, we performed a triple SILAC-based quantitative phosphoproteome analysis. Detected tyrosine phosphorylation sites for which the phosphorylation level decreased in the ΔptkA strain and increased in the ΔptpZ strain, compared to the wild type, were considered as potential substrates of PtkA/PtpZ. One of those sites was the residue tyrosine 601 of the molecular chaperone DnaK. We confirmed that DnaK is a substrate of PtkA and PtpZ by in vitro phosphorylation and dephosphorylation assays. In vitro, DnaK Y601F mutant exhibited impaired interaction with its co-chaperones DnaJ and GrpE, along with diminished capacity to hydrolyze ATP and assist the re-folding of denatured proteins. In vivo, loss of DnaK phosphorylation in the mutant strain dnaK Y601F, or in the strain overexpressing the phosphatase PtpZ, led to diminished survival upon heat shock, consistent with the in vitro results. The decreased survival of the mutant dnaK Y601F at an elevated temperature could be rescued by complementing with the wild type dnaK allele expressed ectopically. We concluded that the residue tyrosine 601 of DnaK can be phosphorylated and dephosphorylated by PtkA and PtpZ, respectively. Furthermore, Y601 is important for DnaK chaperone activity and heat shock survival of B. subtilis.

  7. DegQ regulates the production of fengycins and biofilm formation of the biocontrol agent Bacillus subtilis NCD-2.

    Science.gov (United States)

    Wang, Peipei; Guo, Qinggang; Ma, Yinan; Li, Shezeng; Lu, Xiuyun; Zhang, Xiaoyun; Ma, Ping

    2015-09-01

    Bacillus subtilis NCD-2 is an excellent biocontrol agent for tomato gray mold and cotton soil-borne diseases. The fengycin lipopeptides serve as a major role in its biocontrol ability. A previous study revealed that insertion of degQ with the mini-Tn10 transposon decreased the antifungal activity of strain NCD-2 against the growth of Botrytis cinerea. To clarify the regulation of degQ on the production of fengycin, we deleted degQ by in-frame mutagenesis. Compared with the wild-type strain NCD-2, the degQ-null mutant had decreased extracellular protease and cellulase activities as well as antifungal ability against the growth of B. cinerea in vitro. The lipopeptides from the degQ-null mutant also had significantly decreased antifungal activity against B. cinerea in vitro and in vivo. This result was confirmed by the decreased fengycin production in the degQ-null mutant that was detected by fast protein liquid chromatography analysis. Quantitative reverse transcription PCR further demonstrated that degQ positively regulated the expression of the fengycin synthetase gene. In addition, the degQ-null mutant also had a flatter colony phenotype and significantly decreased biofilm formation ability relative to the wild-type strain. All of those characteristics from degQ-null mutant could be restored to the strain NCD-2 wild-type level by complementation of intact degQ in the mutant. Therefore, DegQ may be an important regulator of fengycin production and biofilm formation in B. subtilis NCD-2. PMID:26302846

  8. Direct surfactin-gramicidin S antagonism supports detoxification in mixed producer cultures of Bacillus subtilis and Aneurinibacillus migulanus.

    Science.gov (United States)

    Rautenbach, Marina; Eyéghé-Bickong, Hans André; Vlok, Nicolas Maré; Stander, Marietjie; de Beer, Abré

    2012-12-01

    Antibiotic production as a defence mechanism is a characteristic of a wide variety of organisms. In natural evolutionary adaptation, cellular events such as sporulation, biofilm formation and resistance to antibiotics enable some micro-organisms to survive environmental and antibiotic stress conditions. The two antimicrobial cyclic peptides in this study, gramicidin S (GS) from Aneurinibacillus migulanus and the lipopeptide surfactin (Srf) from Bacillus subtilis, have been shown to affect both membrane and intercellular components of target organisms. Many functions, other than that of antimicrobial activity, have been assigned to Srf. We present evidence that an additional function may exist for Srf, namely that of a detoxifying agent that protects its producer from the lytic activity of GS. We observed that Srf producers were more resistant to GS and could be co-cultured with the GS producer. Furthermore, exogenous Srf antagonized the activity of GS against both Srf-producing and non-producing bacterial strains. A molecular interaction between the anionic Srf and the cationic GS was observed with circular dichroism and electrospray MS. Our results indicate that the formation of an inactive complex between GS and Srf supports resistance towards GS, with the anionic Srf forming a chemical barrier to protect its producer. This direct detoxification combined with the induction of protective stress responses in B. subtilis by Srf confers resistance toward GS from A. migulanus and allows survival in mixed cultures. PMID:23103974

  9. Enhancement of Bacillus subtilis Lipopeptide Biosurfactants Production through Optimization of Medium Composition and Adequate Control of Aeration

    Directory of Open Access Journals (Sweden)

    Dhouha Ghribi

    2011-01-01

    Full Text Available Interest in biosurfactants has increased considerably in recent years, as they are potentially used in many commercial applications in petroleum, pharmaceuticals, biomedical, and food processing industries. Since improvement of their production was of great importance to reduce the final coast, cultural conditions were analyzed to optimize biosurfactants production from Bacillus subtilis SPB1 strain. A high yield of biosurfactants was obtained from a culture of B. subtilis using carbohydrate substrate as a carbon source; among carbohydrates, glucose enhanced the best surfactin production. The optimum glucose concentration was 40 g/L. Higher amount of biosurfactants was obtained using 5 g/L of urea as organic nitrogen source and applying C/N ratio of 7 with ammonium chloride as inorganic nitrogen source. The highest amount of biosurfactants was recorded with the addition of 2% kerosene. Moreover, it was shown, using an automated full-controlled 2.6 L fermenter, that aeration of the medium, which affected strongly the growth regulated biosurfactants synthesis by the producing cell. So that, low or high aerations lead to a decrease of biosurfactants synthesis yields. It was found that when using dissolved oxygen saturation of the medium at 30%, biosurfactants production reached 4.92 g/L.

  10. A community-curated consensual annotation that is continuously updated: the Bacillus subtilis centred wiki SubtiWiki.

    OpenAIRE

    Flórez, Lope A.; Roppel, Sebastian F.; Schmeisky, Arne G.; Lammers, Christoph R.; Stülke, Jörg

    2009-01-01

    Bacillus subtilis is the model organism for Gram-positive bacteria, with a large amount of publications on all aspects of its biology. To facilitate genome annotation and the collection of comprehensive information on B. subtilis, we created SubtiWiki as a community-oriented annotation tool for information retrieval and continuous maintenance. The wiki is focused on the needs and requirements of scientists doing experimental work. This has implications for the design of the interface and for ...

  11. Analysis of Peptidoglycan Structure from Vegetative Cells of Bacillus subtilis 168 and Role of PBP 5 in Peptidoglycan Maturation

    OpenAIRE

    Atrih, Abdelmadjid; Bacher, Gerold; Allmaier, Günter; Williamson, Michael P; Foster, Simon J.

    1999-01-01

    The composition and fine structure of the vegetative cell wall peptidoglycan from Bacillus subtilis were determined by analysis of its constituent muropeptides. The structures of 39 muropeptides, representing 97% of the total peptidoglycan, were elucidated. About 99% analyzed muropeptides in B. subtilis vegetative cell peptidoglycan have the free carboxylic group of diaminopimelic acid amidated. Anhydromuropeptides and products missing a glucosamine at the nonreducing terminus account for 0.4...

  12. Production of polyhydroxyalkanoates (PHAs) by Bacillus subtilis and Escherichia coli grown on cane molasses fortified with ethanol

    OpenAIRE

    Eman Zakaria Gomaa

    2014-01-01

    The aim of this work was to study the production of polyhydroxyalkanoates (PHAs) by Bacillus subtilis and Escherichia coli isolated from the industrial contaminated soil samples using cane molasses as an inexpensive substrate. The amount of PHA accumulated followed a similar pattern to its growth for each of treatment indicating a growth-related production, yielding maximum PHA production of 54.1 and 47.16% for B. subtilis and E. coli, respectively after 96 h cultivation in the medium contain...

  13. An inducible recA expression Bacillus subtilis genome vector for stable manipulation of large DNA fragments

    OpenAIRE

    Ogawa, Takafumi; Iwata, Tetsuo; Kaneko, Shinya; Itaya, Mitsuhiro; Hirota, Junji

    2015-01-01

    Background The Bacillus subtilis genome (BGM) vector is a novel cloning system based on the natural competence that enables B. subtilis to import extracellular DNA fragments into the cell and incorporate the recombinogenic DNA into the genome vector by homologous recombination. The BGM vector system has several attractive properties, such as a megabase cloning capacity, stable propagation of cloned DNA inserts, and various modification strategies using RecA-mediated homologous recombination. ...

  14. AN INTEGRATED APPROACH USING Bacillus subtilis B26 AND ESSENTIAL OILS TO LIMIT FUNGAL DISCOLORATION OF WOOD

    OpenAIRE

    Yu Wang; Jianmin Chang,; Jeffrey J. Morrell,; Camille M. Freitag,; Joseph J. Karchesy

    2012-01-01

    Bacillus subtilis and essential oils have been explored separately for their ability to limit colonization by wood stain and mold fungi, but neither approach has been completely effective. One alternative strategy would be to combine the bacterial biocontrol with one or more natural products extracts. In this report, the ability of combinations of B. subtilis B26 and 20 essential oils to limit fungal stain was explored on Douglas-fir sapwood wafers under controlled laboratory conditions. A nu...

  15. Temporal Expression of the Bacillus subtilis secA Gene, Encoding a Central Component of the Preprotein Translocase

    OpenAIRE

    Herbort, Markus; Klein, Michael; Manting, Erik H.; Driessen, Arnold J. M.; Freudl, Roland

    1999-01-01

    In Bacillus subtilis, the secretion of extracellular proteins strongly increases upon transition from exponential growth to the stationary growth phase. It is not known whether the amounts of some or all components of the protein translocation apparatus are concomitantly increased in relation to the increased export activity. In this study, we analyzed the transcriptional organization and temporal expression of the secA gene, encoding a central component of the B. subtilis preprotein transloc...

  16. Role of Bacillus subtilis BacB in the Synthesis of Bacilysin

    OpenAIRE

    Rajavel, Malligarjunan; Mitra, Ashima; Gopal, Balasubramanian

    2009-01-01

    Bacilysin is a non-ribosomally synthesized dipeptide antibiotic that is active against a wide range of bacteria and some fungi. Synthesis of bacilysin (L-alanine-[2,3-epoxycyclohexano-4]-L-alanine) is achieved by proteins in the bac operon, also referred to as the bacABCDE (ywfBCDEF) gene cluster in B. subtilis. Extensive genetic analysis from several strains of B. subtilis suggests that the bacABC gene cluster encodes all the proteins that synthesize the epoxyhexanone ring of L-anticapsin...

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2009-06-15

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

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

    International Nuclear Information System (INIS)

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

  19. Antagonismo de Trichoderma SPP. E Bacillus subtilis (UFV3918 a Fusarium sambucinum em Pinus elliottii engelm

    Directory of Open Access Journals (Sweden)

    Caciara Gonzatto Maciel

    2014-06-01

    Full Text Available Pinus elliottii é uma espécie de importância no setor florestal e apresenta vulnerabilidade na qualidade sanitária de suas sementes, especialmente pela associação de Fusarium spp., responsável por perdas de plântulas no viveiro. Este trabalho teve como objetivo avaliar a ação antagonista in vitro e in vivo dos agentes Trichoderma spp. e Bacillus subtilis (UFV3918 no controle de Fusarium sambucinum, responsável por danos em plântulas de Pinus elliottii. O controle in vitro foi avaliado através da inibição do crescimento micelial (confronto pareado de culturas, após a incubação a 25±2 ºC e fotoperíodo de 12 h. Para os testes in vivo (desenvolvidos em condições de viveiro, as sementes inicialmente foram inoculadas com o patógeno e, na sequência, microbiolizadas com os agentes antagônicos, para posterior semeadura. Utilizaram-se as técnicas de contato com o biocontrolador em meio BDA por 48 h e peliculização, como formas de microbiolização. Tanto Trichoderma spp. quanto Bacillus subtilis (UFV3918 foram eficientes no controle in vitro de F. sambucinum, e no teste de biocontrole in vivo o produto Bacillus subtilis (UFV3918 destacou-se, reduzindo as perdas de plântulas causadas pelo patógeno, assim como potencializando as variáveis de comprimento de plântula, massa verde e massa seca.

  20. Xylitol production from xylose mother liquor: a novel strategy that combines the use of recombinant Bacillus subtilis and Candida maltosa

    Science.gov (United States)

    2011-01-01

    Background Xylose mother liquor has high concentrations of xylose (35%-40%) as well as other sugars such as L-arabinose (10%-15%), galactose (8%-10%), glucose (8%-10%), and other minor sugars. Due to the complexity of this mother liquor, further isolation of xylose by simple method is not possible. In China, more than 50,000 metric tons of xylose mother liquor was produced in 2009, and the management of sugars like xylose that present in the low-cost liquor is a problem. Results We designed a novel strategy in which Bacillus subtilis and Candida maltosa were combined and used to convert xylose in this mother liquor to xylitol, a product of higher value. First, the xylose mother liquor was detoxified with the yeast C. maltosa to remove furfural and 5-hydromethylfurfural (HMF), which are inhibitors of B. subtilis growth. The glucose present in the mother liquor was also depleted by this yeast, which was an added advantage because glucose causes carbon catabolite repression in B. subtilis. This detoxification treatment resulted in an inhibitor-free mother liquor, and the C. maltosa cells could be reused as biocatalysts at a later stage to reduce xylose to xylitol. In the second step, a recombinant B. subtilis strain with a disrupted xylose isomerase gene was constructed. The detoxified xylose mother liquor was used as the medium for recombinant B. subtilis cultivation, and this led to L-arabinose depletion and xylose enrichment of the medium. In the third step, the xylose was further reduced to xylitol by C. maltosa cells, and crystallized xylitol was obtained from this yeast transformation medium. C. maltosa transformation of the xylose-enriched medium resulted in xylitol with 4.25 g L-1·h-1 volumetric productivity and 0.85 g xylitol/g xylose specific productivity. Conclusion In this study, we developed a biological method for the purification of xylose from xylose mother liquor and subsequent preparation of xylitol by C. maltosa-mediated biohydrogenation of xylose

  1. Xylitol production from xylose mother liquor: a novel strategy that combines the use of recombinant Bacillus subtilis and Candida maltosa

    Directory of Open Access Journals (Sweden)

    Jiang Mingguo

    2011-02-01

    Full Text Available Abstract Background Xylose mother liquor has high concentrations of xylose (35%-40% as well as other sugars such as L-arabinose (10%-15%, galactose (8%-10%, glucose (8%-10%, and other minor sugars. Due to the complexity of this mother liquor, further isolation of xylose by simple method is not possible. In China, more than 50,000 metric tons of xylose mother liquor was produced in 2009, and the management of sugars like xylose that present in the low-cost liquor is a problem. Results We designed a novel strategy in which Bacillus subtilis and Candida maltosa were combined and used to convert xylose in this mother liquor to xylitol, a product of higher value. First, the xylose mother liquor was detoxified with the yeast C. maltosa to remove furfural and 5-hydromethylfurfural (HMF, which are inhibitors of B. subtilis growth. The glucose present in the mother liquor was also depleted by this yeast, which was an added advantage because glucose causes carbon catabolite repression in B. subtilis. This detoxification treatment resulted in an inhibitor-free mother liquor, and the C. maltosa cells could be reused as biocatalysts at a later stage to reduce xylose to xylitol. In the second step, a recombinant B. subtilis strain with a disrupted xylose isomerase gene was constructed. The detoxified xylose mother liquor was used as the medium for recombinant B. subtilis cultivation, and this led to L-arabinose depletion and xylose enrichment of the medium. In the third step, the xylose was further reduced to xylitol by C. maltosa cells, and crystallized xylitol was obtained from this yeast transformation medium. C. maltosa transformation of the xylose-enriched medium resulted in xylitol with 4.25 g L-1·h-1 volumetric productivity and 0.85 g xylitol/g xylose specific productivity. Conclusion In this study, we developed a biological method for the purification of xylose from xylose mother liquor and subsequent preparation of xylitol by C. maltosa

  2. Purine salvage pathways of Bacillus subtilis and effect of guanine on growth of GMP reductase mutants.

    Science.gov (United States)

    Endo, T; Uratani, B; Freese, E

    1983-07-01

    We have isolated numerous mutants containing mutations in the salvage pathways of purine synthesis. The mutations cause deficiencies in adenine phosphoribosyltransferase (adeF), in hypoxanthine-guanine phosphoribosyltransferase (guaF), in adenine deaminase (adeC), in inosine-guanosine phosphorylase, (guaP), and in GMP reductase (guaC). The physiological properties of mutants containing one or more of these mutations and corresponding enzyme measurements have been used to derive a metabolic chart of the purine salvage pathway of Bacillus subtilis. PMID:6408059

  3. Simultaneous and selective production of levan and poly(gamma-glutamic acid) by Bacillus subtilis.

    Science.gov (United States)

    Shih, Ing-Lung; Yu, Yun-Ti

    2005-01-01

    Bacillus subtilis(natto) Takahashi, used to prepare the fermented soybean product natto, was grown in a basal medium containing 5% (w/w) sucrose and 1.5% (w/w) L-glutamate and produced 58% (w/w) poly(gamma-glutamic acid) and 42% (w/w) levan simultaneously. After 21 h, 40-50 mg levan ml-1 had been produced in medium containing 20% (w/w) sucrose but without L-glutamate. In medium containing L-glutamic acid but without sucrose, mainly poly(gamma-glutamic acid) was produced. PMID:15703872

  4. SacY, a Transcriptional Antiterminator from Bacillus subtilis, Is Regulated by Phosphorylation In Vivo†

    OpenAIRE

    Idelson, Maria; Amster-Choder, Orna

    1998-01-01

    SacY antiterminates transcription of the sacB gene in Bacillus subtilis in response to the presence of sucrose in the growth medium. We have found that it can substitute for BglG, a homologous protein, in antiterminating transcription of the bgl operon in Escherichia coli. We therefore sought to determine whether, similarly to BglG, SacY is regulated by reversible phosphorylation in response to the availability of the inducing sugar. We show here that two forms of SacY, phosphorylated and non...

  5. Different agroresidues used in solid substrate fermentation for alpha- amylase production by bacillus subtilis-329

    International Nuclear Information System (INIS)

    The best mass ratio for agroresidue fermentation for a-amylase production by locally isolated Bacillus subtilis-239 was found to be wheat bran to rice bran 2:1 with 70% initial moisture content for 60 h incubation time. Among different inorganic nitrogen sources supplemented, sodium nitrate and ammonium chloride (0.5% w/w) increased the enzyme yield upto 178 U/ml and 176 U/ml, respectively, whereas all the organic nitrogen sources decreased the enzyme production. Addition of glucose (1% w/w) as a carbon source enhanced a-amylase synthesis to 185 U/ml as compared to the control (134 U/ml). (author)

  6. Protease obtention using Bacillus subtilis 3411 and amaranth seed meal medium at different aeration rates

    Directory of Open Access Journals (Sweden)

    Pastor Maria Delia

    2001-01-01

    Full Text Available The influence of the addition of Amaranthus cruenthus seed meal to the medium, as nutrient and growth factor, on protease production by Bacillus subtilis 3411 was studied. Tests were carried out in a rotary shaker and in mechanically stirred fermenters. The influence of aeration was also evaluated. The addition of amaranth in a concentration of 20 g/L resulted in 400% increase in protease production. Aeration up to 750 r.p.m. and 1 L/L.min had a favorable effect.

  7. The Bacillus subtilis Primosomal Protein DnaD Untwists Supercoiled DNA

    OpenAIRE

    Zhang, Wenke; Allen, Stephanie; Roberts, Clive J.; Soultanas, Panos

    2006-01-01

    The essential Bacillus subtilis DnaD and DnaB proteins have been implicated in the initiation of DNA replication. Recently, DNA remodeling activities associated with both proteins were discovered that could provide a link between global or local nucleoid remodeling and initiation of replication. DnaD forms scaffolds and opens up supercoiled plasmids without nicking to form open circular complexes, while DnaB acts as a lateral compaction protein. Here we show that DnaD-mediated opening of supe...

  8. Regulation of the Bacillus subtilis ytmI Operon, Involved in Sulfur Metabolism

    OpenAIRE

    Burguière, Pierre; Fert, Juliette; Guillouard, Isabelle; Auger, Sandrine; Danchin, Antoine; Martin-Verstraete, Isabelle

    2005-01-01

    The YtlI regulator of Bacillus subtilis activates the transcription of the ytmI operon encoding an l-cystine ABC transporter, a riboflavin kinase, and proteins of unknown function. The expression of the ytlI gene and the ytmI operon was high with methionine and reduced with sulfate. Using deletions and site-directed mutagenesis, a cis-acting DNA sequence important for YtlI-dependent regulation was identified upstream from the −35 box of ytmI. Gel mobility shift assays confirmed that YtlI spec...

  9. Localization of the Germination Protein GerD to the Inner Membrane in Bacillus subtilis Spores▿

    OpenAIRE

    Pelczar, Patricia L.; Setlow, Peter

    2008-01-01

    GerD of Bacillus subtilis is a protein essential for normal spore germination with either l-alanine or a mixture of l-asparagine, d-glucose, d-fructose, and potassium ions. GerD's amino acid sequence suggests that it may be a lipoprotein, indicating a likely location in a membrane. Location in the spore's outer membrane seems unlikely, since removal of this membrane does not result in a gerD spore germination phenotype, suggesting that GerD is likely in the spore's inner membrane. In order to...

  10. DNA segregation by the bacterial actin AlfA during Bacillus subtilis growth and development

    OpenAIRE

    Becker, Eric; Herrera, Nick C; Gunderson, Felizza Q.; Derman, Alan I.; Dance, Amber L; Sims, Jennifer; Larsen, Rachel A.; Pogliano, Joe

    2006-01-01

    We here identify a protein (AlfA; actin like filament) that defines a new family of actins that are only distantly related to MreB and ParM. AlfA is required for segregation of Bacillus subtilis plasmid pBET131 (a mini pLS32-derivative) during growth and sporulation. A 3-kb DNA fragment encoding alfA and a downstream gene (alfB) is necessary and sufficient for plasmid stability. AlfA-GFP assembles dynamic cytoskeletal filaments that rapidly turn over (t1/2

  11. Growth and sporulation of Bacillus subtilis under microgravity (7-IML-1)

    Science.gov (United States)

    Mennigmann, Horst-Dieter

    1992-01-01

    The experiment was aimed at measuring the growth and sporulation of Bacillus subtilis under microgravity. The hardware for the experiment consists of a culture chamber (15 ml) made from titanium and closed by a membrane permeable for gases but not for water. Two variants of this basic structure were built which fit into the standard Biorack container types 1 and 2 respectively. Growth of the bacteria will be monitored by continuously measuring the optical density with a built-in miniaturized photometer. Other parameters (viability, sporulation, fine structure, size distribution of cells and spores, growth kinetics, etc.) will be measured on the fixed samples and on those where metabolism was temporarily halted, respectively.

  12. The Bacillus subtilis DnaD and DnaB Proteins Exhibit Different DNA Remodelling Activities

    OpenAIRE

    Zhang, Wenke; Carneiro, Maria J. V. M.; Turner, Ian J.; ALLEN, Stephanie; Roberts, Clive J.; Soultanas, Panos

    2005-01-01

    Primosomal protein cascades load the replicative helicase onto DNA. In Bacillus subtilis a putative primosomal cascade involving the DnaD-DnaB-DnaI proteins has been suggested to participate in both the DnaA and PriA-dependent loading of the replicative helicase DnaC onto the DNA. Recently we discovered that DnaD has a global remodelling DNA activity suggesting a more widespread role in bacterial nucleoid architecture. Here, we show that DnaB forms a “square-like” tetramer with a hole in the ...

  13. Characterization of BshA, bacillithiol glycosyltransferase from Staphylococcus aureus and Bacillus subtilis

    OpenAIRE

    Upton, Heather; Newton, Gerald L.; Gushiken, Melissa; Lo, Kelly; Holden, D; Fahey, Robert C.; Rawat, Mamta

    2012-01-01

    The first step during bacillithiol (BSH) biosynthesis involves the formation of N-acetylglucosaminylmalate from UDP-N-acetylglucosamine and L-malate and is catalyzed by a GT4 class glycosyltransferase enzyme (BshA). Recombinant Staphylococcus aureus and Bacillus subtilis BshA were highly specific and active with L-malate but the former showed low activity with D-glyceric acid and the latter with D-malate. We show that BshA is inhibited by BSH and similarly that MshA (first enzyme of mycothiol...

  14. In vivo recognition of Bacillus subtilis by desorption electrospray ionization mass spectrometry (DESI-MS).

    Science.gov (United States)

    Song, Yishu; Talaty, Nari; Datsenko, Kirill; Wanner, Barry L; Cooks, R Graham

    2009-05-01

    Desorption electrospray ionization mass spectrometry (DESI-MS) of culture of the bacterium Bacillus subtilis as a biofilm growing on agar nutrient gives simple, high quality mass spectra dominated in both the positive and negative ion modes by signals due to the cyclic lipopeptide, Surfactin(C15). This in vivo experiment, performed by direct analysis of untreated microorganism samples under ambient conditions, allows rapid identification of this microorganism and the antibiotics that it produces. The result is suggestive of the capabilities of DESI-MS for in vivo microorganism characterization in general and for monitoring fermentation processes for the production of antibiotics and other biochemicals. PMID:19381372

  15. d-Amino Acids Indirectly Inhibit Biofilm Formation in Bacillus subtilis by Interfering with Protein Synthesis

    OpenAIRE

    Leiman, Sara A.; May, Janine M.; Lebar, Matthew D.; Kahne, Daniel; Kolter, Roberto; Losick, Richard

    2013-01-01

    The soil bacterium Bacillus subtilis forms biofilms on surfaces and at air-liquid interfaces. It was previously reported that these biofilms disassemble late in their life cycle and that conditioned medium from late-stage biofilms inhibits biofilm formation. Such medium contained a mixture of d-leucine, d-methionine, d-tryptophan, and d-tyrosine and was reported to inhibit biofilm formation via the incorporation of these d-amino acids into the cell wall. Here, we show that l-amino acids were ...

  16. Small regulatory RNA-induced growth rate heterogeneity of Bacillus subtilis.

    OpenAIRE

    Mars, Ruben A. T.; Pierre Nicolas; Mariano Ciccolini; Ewoud Reilman; Alexander Reder; Marc Schaffer; Ulrike Mäder; Uwe Völker; Jan Maarten van Dijl; Denham, Emma L.

    2015-01-01

    Isogenic bacterial populations can consist of cells displaying heterogeneous physiological traits. Small regulatory RNAs (sRNAs) could affect this heterogeneity since they act by fine-tuning mRNA or protein levels to coordinate the appropriate cellular behavior. Here we show that the sRNA RnaC/S1022 from the Gram-positive bacterium Bacillus subtilis can suppress exponential growth by modulation of the transcriptional regulator AbrB. Specifically, the post-transcriptional abrB-RnaC/S1022 inter...

  17. Sekvencijsko optimiranje prinosa poli(γ-glutaminske kiseline) iz novoizoliranoga soja Bacillus subtilis

    OpenAIRE

    Bajaj, Ishwar B.; Singhal, Rekha S.

    2009-01-01

    Provedbom taksonomske i 16S rRNA studije identificiran je novi soj bakterije Bacillus subtilis, podrijetlom iz mora, koji proizvodi poli(γ-glutaminsku kiselinu) (engl. PGA). Za poboljšanje proizvodnje PGA primijenjeno je sekvencijsko optimiranje. Jednofaktorskom metodom istražen je utjecaj izvora ugljika i dušika te pH-vrijednosti na proizvodnju PGA. Primjenom Plackett-Burmanovog dizajna odabrana su hranjiva koja najviše utječu na prinos, a metodom odzivnih površina razvijen je matematički mo...

  18. Mutations that relieve nutritional repression of the Bacillus subtilis dipeptide permease operon.

    OpenAIRE

    Slack, F J; Mueller, J P; Sonenshein, A L

    1993-01-01

    The Bacillus subtilis dciA operon encodes a dipeptide transport complex that is induced rapidly as cells enter stationary phase and initiate sporulation. Expression of this operon in growing cells is repressed by glucose, by a mixture of amino acids, and by the AbrB protein. A genetic screen was devised to identify mutations that allow inappropriate expression from the dciA promoter during growth. These mutations resulted in increased dciA transcription during growth in nutrient broth, in min...

  19. Substrate induction of siderophore transport in Bacillus subtilis mediated by a novel one-component regulator

    OpenAIRE

    Gaballa, Ahmed; Helmann, John D.

    2007-01-01

    When iron is scarce, Bacillus subtilis expresses genes involved in the synthesis and uptake of the siderophore bacillibactin (BB) and uptake systems to pirate other microbial siderophores. Here, we demonstrate that transcriptional induction of the feuABCybbA operon, encoding the Fe-BB uptake system, is mediated by Btr (formerly YbbB) which is encoded by the immediately upstream gene. Btr contains an AraC-type DNA binding domain fused to a substrate binding protein (SBP) domain related to FeuA...

  20. Septation, dephosphorylation, and the activation of σF during sporulation in Bacillus subtilis

    OpenAIRE

    King, Nicole; Dreesen, Oliver; Stragier, Patrick; Pogliano, Kit; Losick, Richard

    1999-01-01

    Cell-specific activation of transcription factor σF during sporulation in Bacillus subtilis requires the formation of the polar septum and the activity of a serine phosphatase (SpoIIE) located in the septum. The SpoIIE phosphatase indirectly activates σF by dephosphorylating a protein (SpoIIAA-P) in the pathway that controls the activity of the transcription factor. By use of a SpoIIE–GFP fusion protein in time-course and time-lapse experiments and by direct visualization of septa in living c...

  1. Structure of the Phosphatase Domain of the Cell Fate Determinant SpoIIE from Bacillus subtilis

    OpenAIRE

    Levdikov, Vladimir M; Blagova, Elena V.; Rawlings, Andrea E.; Jameson, Katie; Tunaley, James; Hart, Darren J.; Barak, Imrich; Wilkinson, Anthony J.

    2012-01-01

    Sporulation in Bacillus subtilis begins with an asymmetric cell division producing two genetically identical cells with different fates. SpoIIE is a membrane protein that localizes to the polar cell division sites where it causes FtsZ to relocate from mid-cell to form polar Z-rings. Following polar septation, SpoIIE establishes compartment-specific gene expression in the smaller forespore cell by dephosphorylating the anti-sigma factor antagonist SpoIIAA, leading to the release of the RNA pol...

  2. PRODUCTION OPTIMIZATION OF EXTRACELLULAR L-ASPARAGINASE THROUGH SOLID- STATE FERMENTATION BY ISOLATED BACILLUS SUBTILIS.

    Directory of Open Access Journals (Sweden)

    Susmita Shukla

    2013-02-01

    Full Text Available L-asparaginase has been used as anti-tumor agent for the treatment of acute lymphoblastic leukemia and food processing aid to reduce the formation of cancer causing acrylamide. Extracellular Lasparaginase production was optimized through solid state fermentation using ground nut cake by isolated Bacillus subtilis. which was not reported in literature.Optimum production of L-asparaginase enzyme (18.4U/ml was obtained after 48h of incubation at 370C moisture content of 70% and at pH 7.

  3. A Catalytic Mechanism Revealed by the Crystal Structures of the Imidazolonepropionase from Bacillus subtilis.

    OpenAIRE

    Yu, Y.; Liang, Y.H.; Brostromer, E.; Quan, J. M.; PANJIKAR, S; Dong, Y. H.; Su, X. D.

    2006-01-01

    Imidazolonepropionase (EC 3.5.2.7) catalyzes the third step in the universal histidine degradation pathway, hydrolyzing the carbon-nitrogen bonds in 4-imidazolone-5-propionic acid to yield N-formimino-l-glutamic acid. Here we report the crystal structures of the Bacillus subtilis imidazolonepropionase and its complex at 2.0-A resolution with substrate analog imidazole-4-acetic acid sodium (I4AA). The structure of the native enzyme contains two domains, a TIM (triose-phosphate isomerase) barre...

  4. Cloning, nucleotide sequence, and expression of the Bacillus subtilis lon gene.

    OpenAIRE

    Riethdorf, S.; Völker, U; Gerth, U.; Winkler, A; Engelmann, S; Hecker, M.

    1994-01-01

    The lon gene of Escherichia coli encodes the ATP-dependent serine protease La and belongs to the family of sigma 32-dependent heat shock genes. In this paper, we report the cloning and characterization of the lon gene from the gram-positive bacterium Bacillus subtilis. The nucleotide sequence of the lon locus, which is localized upstream of the hemAXCDBL operon, was determined. The lon gene codes for an 87-kDa protein consisting of 774 amino acid residues. A comparison of the deduced amino ac...

  5. Localization of UvrA and Effect of DNA Damage on the Chromosome of Bacillus subtilis

    OpenAIRE

    Smith, Bradley T.; Grossman, Alan D.; Walker, Graham C.

    2002-01-01

    We found that the nucleotide excision repair protein UvrA, which is involved in DNA damage recognition, localizes to the entire chromosome both before and after damage in living Bacillus subtilis cells. We suggest that the UvrA2B damage recognition complex is constantly scanning the genome, searching for lesions in the DNA. We also found that DNA damage induces a dramatic reconfiguration of the chromosome such that it no longer fills the entire cell as it does during normal growth. This recon...

  6. Isolation and characterization of topological specificity mutants of minD in Bacillus subtilis

    OpenAIRE

    Karoui, M E; Errington, J

    2001-01-01

    In rod-shaped bacteria such as Bacillus subtilis, division site selection is mediated by MinC and MinD, which together function as a division inhibitor. Topological specificity is imposed by DivIVA, which ensures that MinCD specifically inhibits division close to the cell poles, while allowing division at mid-cell. MinD plays a central role in this process, as it positions and activates MinC and is dependent on DivIVA for its own positioning at the poles. To investigate MinD activities furthe...

  7. Single-stranded plasmid DNA in Bacillus subtilis and Staphylococcus aureus.

    OpenAIRE

    te Riele, H; Michel, B.; Ehrlich, S D

    1986-01-01

    Plasmid pC194 was found to exist in a double-stranded and a single-stranded DNA form in Bacillus subtilis and Staphylococcus aureus. This single-stranded DNA was found as a circular molecule of the same size as the parental monomer and corresponded to only one of the two DNA strands. It represented one-third of plasmid copies. Single- and double-stranded DNA copies in similar proportions to the above were detected for five other S. aureus plasmids (pC221, pC223, pE194, pT127, and pT181) and o...

  8. Plasmid transduction by Bacillus subtilis bacteriophage SPP1: effects of DNA homology between plasmid and bacteriophage.

    OpenAIRE

    Deichelbohrer, I; Alonso, J.C.; Lüder, G; Trautner, T A

    1985-01-01

    Any SPP1 DNA restriction fragment cloned into Bacillus subtilis plasmid pC194 or pUB110 increased the transduction frequency of the plasmid by SPP1 100- to 1,000-fold over the transduction level of the plasmid alone. This increment was observed irrespective of whether a fragment contained the SPP1 packaging origin (pac). Furthermore, an SPP1 derivative into whose genome pC194 DNA had been integrated transduced pC194 DNA with a greatly enhanced frequency. Transduction enhancement mediated by D...

  9. Cloning, sequencing, and expression of Bacillus subtilis genes involved in ATP-dependent nuclease synthesis.

    OpenAIRE

    Kooistra, J; Venema, G

    1991-01-01

    The genes encoding the subunits of the Bacillus subtilis ATP-dependent nuclease (add genes) have been cloned. The genes were located on an 8.8-kb SalI-SmaI chromosomal DNA fragment. Transformants of a recBCD deletion mutant of Escherichia coli with plasmid pGV1 carrying this DNA fragment showed ATP-dependent nuclease activity. Three open reading frames were identified on the 8.8-kb SalI-SmaI fragment, which could encode three proteins with molecular masses of 135 (AddB protein), 141 (AddA pro...

  10. Role of Branched-Chain Amino Acid Transport in Bacillus subtilis CodY Activity

    OpenAIRE

    Belitsky, Boris R.

    2015-01-01

    CodY is a branched-chain amino acid-responsive transcriptional regulator that controls the expression of several dozen transcription units in Bacillus subtilis. The presence of isoleucine, valine, and leucine in the growth medium is essential for achieving high activity of CodY and for efficient regulation of the target genes. We identified three permeases—BcaP, BraB, and BrnQ—that are responsible for the bulk of isoleucine and valine uptake and are also involved in leucine uptake. At least o...

  11. Efficient biosynthesis of polysaccharides chondroitin and heparosan by metabolically engineered Bacillus subtilis.

    Science.gov (United States)

    Jin, Peng; Zhang, Linpei; Yuan, Panhong; Kang, Zhen; Du, Guocheng; Chen, Jian

    2016-04-20

    Chondroitin and heparosan, important polysaccharides and key precursors of chondroitin sulfate and heparin/heparan sulfate, have drawn much attention due to their wide applications in many aspects. In this study, we designed two independent synthetic pathways of chondroitin and heparosan in food-grade Bacillus subtilis, integrating critical synthases genes derived from Escherichia coli into B. subtilis genome. By RT-PCR analysis, we confirmed that synthases genes transcripted an integral mRNA chain, suggesting co-expression. In shaken flask, chondroitin and heparosan were produced at a level of 1.83gL(-1) and 1.71gL(-1), respectively. Since B. subtilis endogenous tuaD gene encodes the limiting factor of biosynthesis, overexpressing tuaD resulted in enhanced chondroitin and heparosan titers, namely 2.54gL(-1) and 2.65gL(-1). Moreover, production reached the highest peaks of 5.22gL(-1) and 5.82gL(-1) in 3-L fed-batch fermentation, respectively, allowed to double the production that in shaken flask. The weight-average molecular weight of chondroitin and heparosan from B. subtilis E168C/pP43-D and E168H/pP43-D were 114.07 and 67.70kDa, respectively. This work provided alternative safer synthetic pathways for metabolic engineering of chondroitin and heparosan in B. subtilis and a useful approach for enhancing production, which can be optimized for further improvement. PMID:26876870

  12. Antimicrobial activity of surfactants produced by Bacillus subtilis R14 against multidrug-resistant bacteria Atividade antimicrobiana de surfactantes produzidos por Bacillus subtilis R14 frente a bacterias multidroga-resistentes

    Directory of Open Access Journals (Sweden)

    Paulo André Vicente Fernandes

    2007-12-01

    Full Text Available Lipopeptides represent a class of microbial surfactants with increasing scientific, therapeutic and biotechnological interests. The genus Bacillus is a producer of these active compounds, and among them B. subtilis produces surfactin, the most potent biosurfactant known. These compounds can act as antibiotics, antivirals, antitumorals, immunomodulators and enzyme inhibitors. In this work, the antimicrobial activity of biosurfactants obtained by cultivation of B. subtilis R14 was investigated against multidrug-resistant bacteria. During cultivation in defined medium, the surface tension of the medium was reduced from 54 mN/m in the beginning of the microbial growth to 30 mN/m after 20 hours. A crude surfactant concentration of 2.0 g/L was obtained after 40 hours of cultivation. A preliminary characterization suggested that two surfactants were produced. The evaluation of the antimicrobial activity of these compounds was carried out against 29 bacteria. Enterococcus faecalis (11 strains, Staphylococcus aureus (6 strains and Pseudomonas aeruginosa (7 strains and Escherichia coli CI 18 (1 strain displayed a profile of well defined drug resistance. All strains were sensitive to the surfactants, in particular Enterococcus faecalis. The results demonstrated that lipopeptides have a broad spectrum of action, including antimicrobial activity against microorganisms with multidrug-resistant profiles.Os lipopeptídeos representam uma classe de surfactantes microbiológicos com crescente interesse científico, terapêutico e biotecnológico. O gênero Bacillus é um dos maiores produtores destes compostos ativos. Dentre as espécies produtoras de biossurfactante, B. subtilis produz surfactina um dos mais conhecidos. Estes compostos atuam como antibióticos, antivirais, agente antitumorais, imunomoduladores e inibidores enzimáticos. O objetivo deste trabalho foi determinar a atividade antimicrobiana de biossurfactantes, obtidos pelo cultivo de B. subtilis R

  13. Partial characterization of bacitracin like inhibitory substance from bacillus subtilis BS15, a local soil isolate

    International Nuclear Information System (INIS)

    The aim of this study was to investigate the production of bacteriocin/bacteriocin-like inhibitory substances (BLIS) from Bacillus subtilis BS15, isolated from soil. The inhibitory substance was partially purified and characterized as BLIS with a molecular-weight of 3-5 kDa, as determined by SDS-PAGE. Its production was observed during the late exponential phase or at the beginning of stationary-phase. It retained its activity up to 80 deg. C and over a wide range of pH i.e., 3-9. It was found active against several clinically important bacterial species such as Listeria monocytogenes, Staphylococcus aureus, Bacillus cereus, Salmonella typhi and also against the food-spoilage causing microbes, and may be considered as future food preservative. (author)

  14. A Bacitracin-Resistant Bacillus subtilis Gene Encodes a Homologue of the Membrane-Spanning Subunit of the Bacillus licheniformis ABC Transporter

    OpenAIRE

    Ohki, Reiko; Tateno, Kozue; Okada, Youji; Okajima, Haruo; Asai, Kei; Sadaie, Yoshito; Murata, Makiko; Aiso, Toshiko

    2003-01-01

    Bacitracin is a peptide antibiotic nonribosomally produced by Bacillus licheniformis. The bcrABC genes which confer bacitracin resistance to the bacitracin producer encode ATP binding cassette (ABC) transporter proteins, which are hypothesized to pump out bacitracin from the cells. Bacillus subtilis 168, which has no bacitracin synthesizing operon, has several genes homologous to bcrABC. It was found that the disruption of ywoA, a gene homologous to bcrC, resulted in hypersensitivity to bacit...

  15. The iturin-like lipopeptides are essential components in the biological control arsenal of Bacillus subtilis against bacterial diseases of cucurbits.

    Science.gov (United States)

    Zeriouh, Houda; Romero, Diego; Garcia-Gutierrez, Laura; Cazorla, Francisco M; de Vicente, Antonio; Perez-Garcia, Alejandro

    2011-12-01

    The antibacterial potential of four strains of Bacillus subtilis, UMAF6614, UMAF6619, UMAF6639, and UMAF8561, previously selected on the basis of their antifungal activity and efficacy against cucurbit powdery mildew, was examined. Among these strains, UMAF6614 and UMAF6639 showed the highest antibacterial activity in vitro, especially against Xanthomonas campestris pv. cucurbitae and Pectobacterium carotovorum subsp. carotovorum. These strains produced the three families of lipopeptide antibiotics known in Bacillus spp.: surfactins, iturins, and fengycins. Using thin-layer chromatography analysis and direct bioautography, the antibacterial activity could be associated with iturin lipopeptides. This result was confirmed by mutagenesis analysis using lipopeptide-defective mutants. The antibacterial activity was practically abolished in iturin-deficient mutants, whereas the fengycin mutants retained certain inhibitory capabilities. Analyses by fluorescence and transmission electron microscopy revealed the cytotoxic effect of these compounds at the bacterial plasma membrane level. Finally, biological control assays on detached melon leaves demonstrated the ability of UMAF6614 and UMAF6639 to suppress bacterial leaf spot and soft rot; accordingly, the biocontrol activity was practically abolished in mutants deficient in iturin biosynthesis. Taken together, our results highlight the potential of these B. subtilis strains as biocontrol agents against fungal and bacterial diseases of cucurbits and the versatility of iturins as antifungal and antibacterial compounds. PMID:22066902

  16. Survival of the biocontrol agents Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 on the spikes of barley in the field

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xin; ZHANG Bing-xin; ZHANG Zhen; SHEN Wei-feng; YANG Ching-hong; YU Jing-quan; ZHAO Yu-hua

    2005-01-01

    Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents, Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116, on the spikes of barley and their effect on suppression of FHB. Survival and colonization of the Brevibacillus brevis ZJY- 1 and Bacillus subtilis ZJY- 116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley FHB disease was resulted from the treatment of barley spikes with biocontrol agents before inoculation with F. graminearum.

  17. Purification and identification of Bacillus subtilis SPB1 lipopeptide biosurfactant exhibiting antifungal activity against Rhizoctonia bataticola and Rhizoctonia solani.

    Science.gov (United States)

    Mnif, Inès; Grau-Campistany, Ariadna; Coronel-León, Jonathan; Hammami, Inès; Triki, Mohamed Ali; Manresa, Angeles; Ghribi, Dhouha

    2016-04-01

    This study reports the potential of a soil bacterium, Bacillus subtilis strain SPB1, to produce lipopeptide biosurfactants. Firstly, the crude lipopeptide mixture was tested for its inhibitory activity against phytopathogenic fungi. A minimal inhibitory concentration (MIC), an inhibitory concentration at 50 % (IC50 %), and an inhibitory concentration at 90 % (IC90 %) values were determined to be 0.04, 0.012, and 0.02 mg/ml, respectively, for Rhizoctonia bataticola with a fungistatic mode of action. For Rhizoctonia solani, a MIC, an IC50 %, and IC90 % values were determined to be 4, 0.25, and 3.3 mg/ml, respectively, with a fungicidal mode of action. For both of the fungi, a loss of sclerotial integrity, granulation and fragmentation of hyphal mycelia, followed by hyphal shriveling and cell lysis were observed with the treatment with SPB1 biosurfactant fraction. After extraction, separation, and purification, different lipopeptide compounds were identified in the culture filtrate of strain SPB1. Mass spectroscopic analysis confirmed the presence of different lipopeptide compounds consisting of surfactin isoforms with molecular weights of 1007, 1021, and 1035 Da; iturin isoforms with molecular weights of 1028, 1042, and 1056 Da; and fengycin isoforms with molecular weights of 1432 and 1446 Da. Two new clusters of lipopeptide isoforms with molecular weights of 1410 and 1424 Da and 973 and 987 Da, respectively, were also detected. This study reported the ability of a B. subtilis strain to co-produce lipopeptide isoforms with potential use as antifungal compounds. PMID:26645234

  18. Functional analysis of 14 genes that constitute the purine catabolic pathway in Bacillus subtilis and evidence for a novel regulon controlled by the PucR transcription activator

    DEFF Research Database (Denmark)

    Schultz, Anna Charlotte; Nygaard, P.; Saxild, Hans Henrik

    2001-01-01

    The soil bacterium Bacillus subtilis has developed a highly controlled system for the utilization of a diverse array of low molecular-weight compounds as a nitrogen source when the preferred nitrogen sources, e.g., glutamate plus ammonia, are exhausted. We have identified such a system for the...... utilization of purines as nitrogen source in B. subtilis. Based on growth studies of strains with knockout mutations in genes, complemented with enzyme analysis, we could ascribe functions to 14 genes encoding enzymes or proteins of the purine degradation pathway. A functional xanthine dehydrogenase requires...... by pucF. In a pucR mutant, the level of expression was low for all genes tested, indicating that PucR is a positive regulator of puc gene expression. All 14 genes except pucI are located in a gene cluster at 284 to 285 degrees on the chromosome and are contained in six transcription units, which are...

  19. Ectopic integration vectors for generating fluorescent promoter fusions in Bacillus subtilis with minimal dark noise.

    Directory of Open Access Journals (Sweden)

    Stephanie Trauth

    Full Text Available Fluorescent protein promoter reporters are important tools that are widely used for diverse purposes in microbiology, systems biology and synthetic biology and considerable engineering efforts are still geared at improving the sensitivity of the reporter systems. Here we focus on dark noise, i.e. the signal that is generated by the empty vector control. We quantitatively characterize the dark noise of a few common bacterial reporter systems by single cell microscopy. All benchmarked reporter systems generated significant amounts of dark noise that exceed the cellular autofluorescence to different extents. We then reengineered a multicolor set of fluorescent ectopic integration vectors for Bacillus subtilis by introducing a terminator immediately upstream of the promoter insertion site, resulting in an up to 2.7-fold reduction of noise levels. The sensitivity and dynamic range of the new high-performance pXFP_Star reporter system is only limited by cellular autofluorescence. Moreover, based on studies of the rapE promoter of B. subtilis we show that the new pXFP_Star reporter system reliably reports on the weak activity of the rapE promoter whereas the original reporter system fails because of transcriptional interference. Since the pXFP_Star reporter system properly isolates the promoter from spurious transcripts, it is a particularly suitable tool for quantitative characterization of weak promoters in B. subtilis.

  20. Regulation of proteolysis in Bacillus subtilis: effects of calcium ions and energy poisons

    International Nuclear Information System (INIS)

    Bacillus subtilis cells carry out extensive intracellular proteolysis (k = 0.15-0.23/h) during sporulation. Protein degradation was measured in cells growing in chemically defined sporulation medium, by following the release of [14C]-leucine from the cells during spore formation. Sodium arsenate, carbonyl cyanide 3-chlorophenyl hydrazone, and sodium azide strongly inhibited proteolysis without altering cell viability greatly, which suggested that bulk proteolysis in B. subtilis is energy dependent. The authors have tested the hypothesis that the energy requirement may be for pumping in Ca2+. When [Ca2+] was -6, rates of proteolysis in sporulating cells were reduced 4-8 times that in cells in calcium ion- sufficient medium. Further, omission of Ca2+ from the medium prevented the increase in the activity of the major intracellular serine protease. However, the presence of energy poisons in the media at levels which inhibited proteolysis, had no detectable effect on the uptake of by cells [45Ca]. The authors concluded that B. subtilis cells required both metabolic energy and calcium ions for normal proteolysis

  1. Effect of medium components and culture conditions in Bacillus subtilis EA-CB0575 spore production.

    Science.gov (United States)

    Posada-Uribe, Luisa F; Romero-Tabarez, Magally; Villegas-Escobar, Valeska

    2015-10-01

    Bacillus subtilis spores have important biotechnological applications; however, achieving both, high spore cell densities and sporulation efficiencies in fermentation, is poorly reported. In this study, medium components and culture conditions were optimized with different statistical methods to increase spore production of the plant growth promoting rhizobacteria B. subtilis EA-CB0575. Key medium components were determined with Plackett-Burman (PB) design, and the optimum concentration levels of two components (glucose, MgSO4·7H2O) were optimized with a full factorial and central composite design, achieving 1.37 × 10(9) CFU/mL of spore cell density and 93.5 % of sporulation efficiency in shake flask. The optimized medium was used to determine the effect of culture conditions on spore production at bioreactor level, finding that maintaining pH control did not affect significantly spore production, while the interaction of agitation and aeration rates had a significant effect on spore cell density. The overall optimization generated a 17.2-fold increase in spore cell density (8.78 × 10(9) CFU/mL) and 1.9-fold increase in sporulation efficiency (94.2 %) compared to that of PB design. These results indicate the potential of B. subtilis EA-CB0575 to produce both, high spore cell densities and sporulation efficiencies, with very low nutrient requirements and short incubation period which can represent savings of process production. PMID:26135004

  2. The interaction of Bacillus subtilis sigmaA with RNA polymerase.

    Science.gov (United States)

    Johnston, Elecia B; Lewis, Peter J; Griffith, Renate

    2009-11-01

    RNA polymerase (RNAP) is an essential and highly conserved enzyme in all organisms. The process of transcription initiation is fundamentally different between prokaryotes and eukaryotes. In prokaryotes, initiation is regulated by sigma factors, making the essential interaction between sigma factors and RNAP an attractive target for antimicrobial agents. Our objective was to achieve the first step in the process of developing novel antimicrobial agents, namely to prove experimentally that the interaction between a bacterial RNAP and an essential sigma factor can be disrupted by introducing carefully designed mutations into sigma(A) of Bacillus subtilis. This disruption was demonstrated qualitatively by Far-Western blotting. Design of mutant sigmas was achieved by computer-aided visualization of the RNAP-sigma interface of the B. subtilis holoenzyme (RNAP + sigma) constructed using a homology modeling approach with published crystal structures of bacterial RNAPs. Models of the holoenzyme and the core RNAP were rigorously built, evaluated, and validated. To allow a high-quality RNAP-sigma interface model to be constructed for the design of mutations, a crucial error in the B. subtilis sigma(A) sequence in published databases at amino acid 165 had to be corrected first. The new model was validated through determination of RNAP-sigma interactions using targeted mutations. PMID:19735077

  3. Small regulatory RNA-induced growth rate heterogeneity of Bacillus subtilis.

    Science.gov (United States)

    Mars, Ruben A T; Nicolas, Pierre; Ciccolini, Mariano; Reilman, Ewoud; Reder, Alexander; Schaffer, Marc; Mäder, Ulrike; Völker, Uwe; van Dijl, Jan Maarten; Denham, Emma L

    2015-03-01

    Isogenic bacterial populations can consist of cells displaying heterogeneous physiological traits. Small regulatory RNAs (sRNAs) could affect this heterogeneity since they act by fine-tuning mRNA or protein levels to coordinate the appropriate cellular behavior. Here we show that the sRNA RnaC/S1022 from the Gram-positive bacterium Bacillus subtilis can suppress exponential growth by modulation of the transcriptional regulator AbrB. Specifically, the post-transcriptional abrB-RnaC/S1022 interaction allows B. subtilis to increase the cell-to-cell variation in AbrB protein levels, despite strong negative autoregulation of the abrB promoter. This behavior is consistent with existing mathematical models of sRNA action, thus suggesting that induction of protein expression noise could be a new general aspect of sRNA regulation. Importantly, we show that the sRNA-induced diversity in AbrB levels generates heterogeneity in growth rates during the exponential growth phase. Based on these findings, we hypothesize that the resulting subpopulations of fast- and slow-growing B. subtilis cells reflect a bet-hedging strategy for enhanced survival of unfavorable conditions. PMID:25790031

  4. Small regulatory RNA-induced growth rate heterogeneity of Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Ruben A T Mars

    2015-03-01

    Full Text Available Isogenic bacterial populations can consist of cells displaying heterogeneous physiological traits. Small regulatory RNAs (sRNAs could affect this heterogeneity since they act by fine-tuning mRNA or protein levels to coordinate the appropriate cellular behavior. Here we show that the sRNA RnaC/S1022 from the Gram-positive bacterium Bacillus subtilis can suppress exponential growth by modulation of the transcriptional regulator AbrB. Specifically, the post-transcriptional abrB-RnaC/S1022 interaction allows B. subtilis to increase the cell-to-cell variation in AbrB protein levels, despite strong negative autoregulation of the abrB promoter. This behavior is consistent with existing mathematical models of sRNA action, thus suggesting that induction of protein expression noise could be a new general aspect of sRNA regulation. Importantly, we show that the sRNA-induced diversity in AbrB levels generates heterogeneity in growth rates during the exponential growth phase. Based on these findings, we hypothesize that the resulting subpopulations of fast- and slow-growing B. subtilis cells reflect a bet-hedging strategy for enhanced survival of unfavorable conditions.

  5. Protein-tyrosine phosphorylation in Bacillus subtilis: a 10-year retrospective

    Directory of Open Access Journals (Sweden)

    Josef eDeutscher

    2015-01-01

    Full Text Available The discovery of tyrosine-phosphorylated proteins in Bacillus subtilis in the year 2003 was followed by a decade of intensive research activity. Here we provide an overview of the lessons learned in that period. While the number of characterized kinases and phosphatases involved in reversible protein-tyrosine phosphorylation in B. subtilis has remained essentially unchanged, the number of proteins known to be targeted by this post-translational modification has increased dramatically. This is mainly due to phosphoproteomics and interactomics studies, which were instrumental in identifying new tyrosine-phosphorylated proteins. Despite their structural similarity, the two B. subtilis protein-tyrosine kinases (BY-kinases, PtkA and PtkB (EpsB, seem to accomplish different functions in the cell. The PtkB is encoded by a large operon involved in exopolysaccharide production, and its main role appears to be the control of this process. The PtkA seems to have a more complex role; it phosphorylates and regulates a large number of proteins involved in the DNA, fatty acid and carbon metabolism and engages in physical interaction with other types of kinases (Ser/Thr kinases, leading to mutual phosphorylation. PtkA also seems to respond to several activator proteins, which direct its activity towards different substrates. In that respect PtkA seems to function as a highly connected signal integration device.

  6. Activation of pur Gene Expression by a Homologue of the Bacillus subtilis PurR repressor:

    DEFF Research Database (Denmark)

    Kilstrup, Mogens; Martinussen, Jan

    1998-01-01

    R encoded repressor from Bacillus subtilis. The wildtype purR gene complements the purine auxotrophy of a purR::Iss1mutant, and it was shown that the purR::Iss1 mutation lowers transcription from the purine regulated L. lactis purD promoter. In a parallel study on the regulation of purC and purD expression....... We have identified a PurBox sequence overlapping the -35 region of the L. lactis purR promoter and found, by studies of a purR-lacLM fusion plasmid, that purR is autoregulated. Because of the high similarity of the PurR proteins from B. subtilis and L. lactis, we looked for PurBox sequences in the...... promoter regions of the PurR regulated genes in B. subtilis, and identified a perfectly matching PurBox in the purA promoter region, and slightly degenerate PurBox like sequences in the promoter regions for the pur operon and the purR gene....

  7. Enhanced and Secretory Expression of Human Granulocyte Colony Stimulating Factor by Bacillus subtilis SCK6

    Science.gov (United States)

    Bashir, Shaista; Sadaf, Saima; Ahmad, Sajjad; Akhtar, Muhammad Waheed

    2015-01-01

    This study describes a simplified approach for enhanced expression and secretion of a pharmaceutically important human cytokine, that is, granulocyte colony stimulating factor (GCSF), in the culture supernatant of Bacillus subtilis SCK6 cells. Codon optimized GCSF and pNWPH vector containing SpymwC signal sequence were amplified by prolonged overlap extension PCR to generate multimeric plasmid DNA, which was used directly to transform B. subtilis SCK6 supercompetent cells. Expression of GCSF was monitored in the culture supernatant for 120 hours. The highest expression, which corresponded to 17% of the total secretory protein, was observed at 72 hours of growth. Following ammonium sulphate precipitation, GCSF was purified to near homogeneity by fast protein liquid chromatography on a QFF anion exchange column. Circular dichroism spectroscopic analysis showed that the secondary structure contents of the purified GCSF are similar to the commercially available GCSF. Biological activity, as revealed by the regeneration of neutrophils in mice treated with ifosfamine, was also similar to the commercial preparation of GCSF. This, to our knowledge, is the first study that reports secretory expression of human GCSF in B. subtilis SCK6 with final recovery of up to 96 mg/L of the culture supernatant, without involvement of any chemical inducer. PMID:26881203

  8. Three biotechnical processes using Ashbya gossypii, Candida famata, or Bacillus subtilis compete with chemical riboflavin production.

    Science.gov (United States)

    Stahmann, K P; Revuelta, J L; Seulberger, H

    2000-05-01

    Chemical riboflavin production, successfully used for decades, is in the course of being replaced by microbial processes. These promise to save half the costs, reduce waste and energy requirements, and use renewable resources like sugar or plant oil. Three microorganisms are currently in use for industrial riboflavin production. The hemiascomycetes Ashbya gossypii, a filamentous fungus, and Candida famata, a yeast, are naturally occurring overproducers of this vitamin. To obtain riboflavin production with the gram-positive bacterium Bacillus subtilis requires at least the deregulation of purine synthesis and a mutation in a flavokinase/FAD-synthetase. It is common to all three organisms that riboflavin production is recognizable by the yellow color of the colonies. This is an important tool for the screening of improved mutants. Antimetabolites like itaconate, which inhibits the isocitrate lyase in A. gossypii, tubercidin, which inhibits purine biosynthesis in C. famata, or roseoflavin, a structural analog of riboflavin used for B. subtilis, have been applied successfully for mutant selections. The production of riboflavin by the two fungi seems to be limited by precursor supply, as was concluded from feeding and gene-overexpression experiments. Although flux studies in B. subtilis revealed an increase both in maintenance metabolism and in the oxidative part of the pentose phosphate pathway, the major limitation there seems to be the riboflavin pathway. Multiple copies of the rib genes and promoter replacements are necessary to achieve competitive productivity. PMID:10855708

  9. Effect of ethanol perturbation on viscosity and permeability of an inner membrane in Bacillus subtilis spores.

    Science.gov (United States)

    Loison, Pauline; Gervais, Patrick; Perrier-Cornet, Jean-Marie; Kuimova, Marina K

    2016-09-01

    In this work, we investigated how a combination of ethanol and high temperature (70°C), affect the properties of the inner membrane of Bacillus subtilis spores. We observed membrane permeabilization for ethanol concentrations ≥50%, as indicated by the staining of the spores' DNA by the cell impermeable dye Propidium Iodide. The loss of membrane integrity was also confirmed by a decrease in the peak corresponding to dipicolinic acid using infrared spectroscopy. Finally, the spore refractivity (as measured by phase contrast microscopy) was decreased after the ethanol-heat treatment, suggesting a partial rehydration of the protoplast. Previously we have used fluorescent lifetime imaging microscopy (FLIM) combined with the fluorescent molecular rotor Bodipy-C12 to study the microscopic viscosity in the inner membrane of B. subtilis spores, and showed that at normal conditions it is characterized by a very high viscosity. Here we demonstrate that the ethanol/high temperature treatment led to a decrease of the viscosity of the inner membrane, from 1000cP to 860cP for wild type spores at 50% of ethanol. Altogether, our present work confirms the deleterious effect of ethanol on the structure of B. subtilis spores, as well as demonstrates the ability of FLIM - Bodipy-C12 to measure changes in the microviscosity of the spores upon perturbation. PMID:27267704

  10. Simulation of Bacillus subtilis biofilm growth on agar plate by diffusion–reaction based continuum model

    Science.gov (United States)

    Zhang, Xianlong; Wang, Xiaoling; Nie, Kai; Li, Mingpeng; Sun, Qingping

    2016-08-01

    Various species of bacteria form highly organized spatially-structured aggregates known as biofilms. To understand how microenvironments impact biofilm growth dynamics, we propose a diffusion–reaction continuum model to simulate the formation of Bacillus subtilis biofilm on an agar plate. The extended finite element method combined with level set method are employed to perform the simulation, numerical results show the quantitative relationship between colony morphologies and nutrient depletion over time. Considering that the production of polysaccharide in wild-type cells may enhance biofilm spreading on the agar plate, we inoculate mutant colony incapable of producing polysaccharide to verify our results. Predictions of the glutamate source biofilm’s shape parameters agree with the experimental mutant colony better than that of glycerol source biofilm, suggesting that glutamate is rate limiting nutrient for Bacillus subtilis biofilm growth on agar plate, and the diffusion-limited is a better description to the experiment. In addition, we find that the diffusion time scale is of the same magnitude as growth process, and the common-employed quasi-steady approximation is not applicable here.

  11. Induced sensitivity of Bacillus subtilis colony morphology to mechanical media compression

    Directory of Open Access Journals (Sweden)

    Jessica K. Polka

    2014-09-01

    Full Text Available Bacteria from several taxa, including Kurthia zopfii, Myxococcus xanthus, and Bacillus mycoides, have been reported to align growth of their colonies to small features on the surface of solid media, including anisotropies created by compression. While the function of this phenomenon is unclear, it may help organisms navigate on solid phases, such as soil. The origin of this behavior is also unknown: it may be biological (that is, dependent on components that sense the environment and regulate growth accordingly or merely physical. Here we show that B. subtilis, an organism that typically does not respond to media compression, can be induced to do so with two simple and synergistic perturbations: a mutation that maintains cells in the swarming (chained state, and the addition of EDTA to the growth media, which further increases chain length. EDTA apparently increases chain length by inducing defects in cell separation, as the treatment has only marginal effects on the length of individual cells. These results lead us to three conclusions. First, the wealth of genetic tools available to B. subtilis will provide a new, tractable chassis for engineering compression sensitive organisms. Second, the sensitivity of colony morphology to media compression in Bacillus can be modulated by altering a simple physical property of rod-shaped cells. And third, colony morphology under compression holds promise as a rapid, simple, and low-cost way to screen for changes in the length of rod-shaped cells or chains thereof.

  12. Point-cycle bistability and stochasticity in a regulatory circuit for Bacillus subtilis competence.

    Science.gov (United States)

    Xi, Hongguang; Duan, Lixia; Turcotte, Marc

    2013-08-01

    Bacillus subtilis is a very well-studied organism in biology. Recent results show that an evolutionary plausible alternative competence regulation circuit for this bacterium, despite presenting equivalent functionality, exhibits physiologically important differences. Thus, it is not a priori clear why Nature only selects a specific gene regulation circuit other than a plethora of equivalent others. Here, we use simulations to study this question further. Based on the wild-type Bacillus subtilis circuit, we add a positive autoregulation feedback loop to the intermediate gene comS. We use bifurcation theory to study the dynamical features of the hypothetical gene circuit versus the feedback strength of the added loop, and we rely on stochastic simulations to perform in silico experiments. We discover the existence of a bistable system: a stable limit cycle and a stable fixed point separated by an unstable limit cycle with a varying height of underlying stochastic potential. This structure is absent from the wild type. The coexistence of the unstable limit cycle with stochastic noise endows the circuit with an ability to confine, prevent or switch between its two stable attractors. PMID:23693123

  13. Deciphering the conserved genetic loci implicated in plant disease control through comparative genomics of Bacillus amyloliquefaciens subsp. plantarum strains

    Directory of Open Access Journals (Sweden)

    Mohammad J Hossain

    2015-08-01

    Full Text Available To understand the growth-promoting and disease-inhibiting activities of plant growth-promoting rhizobacteria (PGPR strains, the genomes of 12 Bacillus subtilis group strains with PGPR activity were sequenced and analyzed. These B. subtilis strains exhibited high genomic diversity, whereas the genomes of B. amyloliquefaciens strains (a member of the B. subtilis group are highly conserved. A pairwise BLASTp matrix revealed that gene family similarity among Bacillus genomes ranges from 32- 90%, with 2,839 genes within the core genome of B. amyloliquefaciens subsp. plantarum. Comparative genomic analyses of B. amyloliquefaciens strains identified genes that are linked with biological control and colonization of roots and/or leaves, including 73 genes uniquely associated with subsp. plantarum strains that have predicted functions related to signaling, transportation, secondary metabolite production, and carbon source utilization. Although B. amyloliquefaciens subsp. plantarum strains contain gene clusters that encode many different secondary metabolites, only polyketide biosynthetic clusters that encode difficidin and macrolactin are conserved within this subspecies. To evaluate their role in plant pathogen biocontrol, genes involved in secondary metabolite biosynthesis were deleted in B. amyloliquefaciens subsp. plantarum strain, revealing that difficidin expression is critical in reducing the severity of disease, caused by Xanthomonas axonopodis pv. vesicatoria in tomato plants. This study defines genomic features of PGPR strains and links them with biocontrol activity and with host colonization.

  14. Antagonistic effects of Bacillus subtilis subsp. subtilis and B. amyloliquefaciens against Macrophomina phaseolina: SEM study of fungal changes and UV-MALDI-TOF MS analysis of their bioactive compounds.

    Science.gov (United States)

    Torres, M J; Brandan, C Pérez; Petroselli, G; Erra-Balsells, R; Audisio, M C

    2016-01-01

    The antifungal effect of Bacillus subtilis subsp. subtilis PGPMori7 and Bacillus amyloliquefaciens PGPBacCA1 was evaluated against Macrophomina phaseolina (Tassi) Goid. Cell suspension (CS), cell-free supernatant (CFS) and the lipopeptide fraction (LF) of PGPMori7 and PGPBacCA1 were screened against three different M. phaseolina strains. CS exhibited the highest inhibitory effect (around 50%) when compared to those of CFS and LF, regardless of the fungal strain studied. The synthesis of lipopeptides was studied by UV-MALDI TOF. Chemical analysis of Bacillus metabolite synthesis revealed that surfactin and iturin were mainly produced in liquid medium. Potential fengycin was also co-produced when both Bacillus were cultivated in solid medium. In co-culture assays, the bacterial colony-fungal mycelium interface at the inhibition zone was evaluated by both scanning electron microscopy (SEM) and UV-MALDI TOF, the former to determine the structural changes on M. phaseolina cells and the latter to identify the main bioactive molecules involved in the inhibitory effect. PGPBacCA1 produced surfactin, iturin and fengycin in the inhibition zone while PGPMori7 only produced these metabolites within its colony and not in the narrow inhibition zone. Interestingly, SEM revealed that PGPBacCA1 induced damage in M. phaseolina sclerotia, generating a fungicidal effect as no growth was observed when normal growth conditions were reestablished. In turn, PGPMori7 inhibited the growth of the Macrophomina mycelium without fungal injury, resulting only in a fungistatic activity. From these results, it was determined that the two bacilli significantly inhibited the growth of an important phytopathogenic fungus by at least two different mechanisms: lipopeptide synthesis and competition among microorganisms. PMID:26686611

  15. Isolation of Bacillus subtilis as indicator in the disinfection of residual water by means of gamma radiation; Aislamiento de Bacillus subtilis como indicador en la desinfeccion de aguas residuales mediante radiacion gamma

    Energy Technology Data Exchange (ETDEWEB)

    Mata J, M.; Colin C, A. [Facultad de Quimica, UAEM, Paseo Colon esq. Tollocan s/n, Toluca, 50000 Estado de Mexico (Mexico); Lopez V, H.; Brena V, M.; Carrasco A, H.; Pavon R, S. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)

    2002-07-01

    In the attempt to get more alternatives of disinfection of residual water, the Bacillus subtilis was isolated by means of gamma radiation as a bio indicator of disinfection since it turned out to be resistant to the 5 KGy dose, comparing this one with other usual microorganisms as biondicators like E. coli and S typhimurium which turn out more sensitive to such dose. (Author)

  16. Isolation and Identification of Lipopeptides Produced by Bacillus subtilis fmbJ%Bacillus subtilis fmbJ脂肽类抗菌物质的分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    别小妹; 吕凤霞; 陆兆新; 黄现青; 沈娟

    2006-01-01

    Bacillus subtilis fmbJ脂肽类抗菌物质的分离和鉴定进行了系统研究.通过HPLC层析确定Bacillus subtilis fmbJ抗菌物质由多种组分构成,其中含有保留时间与surfactin相似的成分.通过TLC层析和原位酸解确定Bacillus subtilis fmbJ抗菌物质含有两个具有闭合肽键类的物质,其中之一为迁移率Rf与标样surfactin非常相近的组分.通过ESI-MS分析检测到Bacillus subtilis fmbJ抗菌物质含有分子量与fengicin相同的m/z1449.9、m/z1463.8、m/z1477.8、m/z1491.9和m/z1505.9五种同系物,和分子量与surfactin相同的m/z1008.8、m/z1022.8和m/z1036.8三种同系物.

  17. Growth Inhibition of Colletotrichum gloeosporioides by Trichoderma harzianum, Trichoderma koningii, Bacillus subtilis and Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    Febrilia Nur ‘Aini

    2015-11-01

    Full Text Available Colletotrichum  gloeosporioides is  a  disease  which  can  cause  significant yield  loss  of  cocoa.  The  objective  of  this  research  is  to  investigate  the  abilityof  antagonist  microbes,  Trichoderma  harzianum,  Trichoderma  koningii,  Bacillus subtilis  and Pseudomonas  fluorescens  in  controlling  gloeosporioides  biologically  in  laboratorium  condition.  The  experiment  was  carried  out  in  Crop  Protection  Laboratory,  Indonesian  Coffee  and  Cocoa  Research  Institute.  Results of  this  research  showed  that  antagonist  fungi,  T.  harzianum,  T.  koningii,  had  a stronger  ability  in  inhibiting  growth  of  C.  gloeosporioides about  83%  compared  to  the  ability  of  antagonist  bacteria,  B.  subtilis  and P.  fluorescens,  only about  49%. Key words: Growth  inhibition,  Colletotrichum  gloeosporioides,  Trichoderma  harzianum, Trichoderma koningii,  Bacillus subtilis, Pseudomonas fluorescens.

  18. Effects of the electrolytic treatment on Bacillus subtilis Efeito do tratamento eletrolítico em Bacillus subtis

    Directory of Open Access Journals (Sweden)

    Rodolfo Tolentino-Bisneto

    2003-11-01

    Full Text Available Conventional processes of water disinfection can generate toxic composites. It is the case of the trihalomethanes (carcinogenic formed in the contact of chlorine with organic substances present in the water. The electrolytic treatment can be a substitute for the chlorination process without the need for addition of chemical substances to the process. The effect of the electrolytic treatment using carbon cathode on the viability of the microorganism Bacillus subtilis was tested to determine the death process. By means of electronic microscopy, it was observed that the main cause of the microorganism's death was the cellular lysis due to the electroporation in the cell membrane.Processos convencionais de desinfecção de águas podem gerar compostos tóxicos. Esse é o caso dos trialometanos formados na reação do cloro com compostos orgânicos presentes na água. O tratamento eletrolítico pode ser um substituto à cloração com vantagem de não requer a adição de nenhum composto na água. O efeito do tratamento eletrolítico, utilizando eletrodos de carbono, na viabilidade de Bacillus subtilis foi testado para se determinar o mecanismo de morte. Através de microscopia eletrônica, foi possível evidenciar que a morte do microrganismo se deu pela lise celular, provavelmente provocada pela eletroporação irreversível da membrana celular.

  19. In silico metabolic engineering of Bacillus subtilis for improved production of riboflavin, Egl-237, (R,R)-2,3-butanediol and isobutanol.

    Science.gov (United States)

    Hao, Tong; Han, Binbin; Ma, Hongwu; Fu, Jing; Wang, Hui; Wang, Zhiwen; Tang, Bincai; Chen, Tao; Zhao, Xueming

    2013-08-01

    Bacillus subtilis is a Gram-positive sporiferous bacterium widely used in a variety of industrial fields as a producer of high-quality vitamins, enzymes and proteins. Many genetic modifications and evolutionary engineering optimisations aiming at obtaining a better performing strain for its products have been studied. As genome-scale metabolic network models have gained significant popularity as effective tools in metabolic phenotype studies, we reconstructed a genome-scale metabolic network of B. subtilis-iBsu1147. The accuracy of iBsu1147 is validated by growth on various carbon sources, single gene knockout and large fragment non-essential gene knockout simulations. The model is used for the in silico metabolic engineering design of reactions over/underexpressed or knockout for increasing the production of four important products of B. subtilis: riboflavin, cellulase Egl-237, (R,R)-2,3-butanediol and isobutanol. The simulation predicted candidate reactions related to the improvement of strain performance on related products. The prediction is partly supported by previously published results. Due to the complexity of the biological system, it is difficult to manually find the factors that are not directly related to the production of the target compounds. The in silico predictions provide more choices for further strain improvement for these products. PMID:23666098

  20. Changes in the Acetylome and Succinylome of Bacillus subtilis in Response to Carbon Source.

    Directory of Open Access Journals (Sweden)

    Saori Kosono

    Full Text Available Lysine residues can be post-translationally modified by various acyl modifications in bacteria and eukarya. Here, we showed that two major acyl modifications, acetylation and succinylation, were changed in response to the carbon source in the Gram-positive model bacterium Bacillus subtilis. Acetylation was more common when the cells were grown on glucose, glycerol, or pyruvate, whereas succinylation was upregulated when the cells were grown on citrate, reflecting the metabolic states that preferentially produce acetyl-CoA and succinyl-CoA, respectively. To identify and quantify changes in acetylation and succinylation in response to the carbon source, we performed a stable isotope labeling by amino acids in cell culture (SILAC-based quantitative proteomic analysis of cells grown on glucose or citrate. We identified 629 acetylated proteins with 1355 unique acetylation sites and 204 succinylated proteins with 327 unique succinylation sites. Acetylation targeted different metabolic pathways under the two growth conditions: branched-chain amino acid biosynthesis and purine metabolism in glucose and the citrate cycle in citrate. Succinylation preferentially targeted the citrate cycle in citrate. Acetylation and succinylation mostly targeted different lysine residues and showed a preference for different residues surrounding the modification sites, suggesting that the two modifications may depend on different factors such as characteristics of acyl-group donors, molecular environment of the lysine substrate, and/or the modifying enzymes. Changes in acetylation and succinylation were observed in proteins involved in central carbon metabolism and in components of the transcription and translation machineries, such as RNA polymerase and the ribosome. Mutations that modulate protein acylation affected B. subtilis growth. A mutation in acetate kinase (ackA increased the global acetylation level, suggesting that acetyl phosphate-dependent acetylation is