Home About us Contact
|
Home About us Contact | ||
|
|
||
Subtilis Strains (subtili + strain)
Kinds of Subtilis Strains Selected AbstractsInduction of neutralizing antibodies in mice immunized with an amino-terminal polypeptide of Streptococcus mutans P1 protein produced by a recombinant Bacillus subtilis strainFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 2 2010Milene B. Tavares Abstract The oral pathogen Streptococcus mutans expresses a surface protein, P1, which interacts with the salivary pellicle on the tooth surface or with fluid-phase saliva, resulting in bacterial adhesion or aggregation, respectively. P1 is a target of protective immunity. Its N-terminal region has been associated with adhesion and aggregation functions and contains epitopes recognized by efficacious antibodies. In this study, we used Bacillus subtilis, a gram-positive expression host, to produce a recombinant N-terminal polypeptide of P1 (P139,512) derived from the S. mutans strain UA159. Purified P139,512 reacted with an anti-full-length P1 antiserum as well as one raised against intact S. mutans cells, indicating preserved antigenicity. Immunization of mice with soluble and heat-denatured P139,512 induced antibodies that reacted specifically with native P1 on the surface of S. mutans cells. The anti-P139,512 antiserum was as effective at blocking saliva-mediated aggregation of S. mutans cells and better at blocking bacterial adhesion to saliva-coated plastic surfaces compared with the anti-full-length P1 antiserum. In addition, adsorption of the anti-P1 antiserum with P139,512 eliminated its ability to block the adhesion of S. mutans cells to abiotic surfaces. The present results indicate that P139,512, expressed and purified from a recombinant B. subtilis strain, maintains important immunological features of the native protein and represents an additional tool for the development of anticaries vaccines. [source] Analysis of the germination of spores of Bacillus subtilis with temperature sensitive spo mutations in the spoVA operonFEMS MICROBIOLOGY LETTERS, Issue 1 2004Venkata Ramana Vepachedu Abstract A Bacillus subtilis strain with a base substitution in the ribosome-binding site of spoVAC was temperature sensitive (ts) in sporulation and spores prepared at the permissive temperature were ts in l -alanine-triggered germination, but not in germination with Ca2+ -dipicolinic acid (DPA) or dodecylamine. Spores of a ts spo mutant with a missense mutation in the spoVAC coding region were not ts for germination with l -alanine, dodecylamine or Ca2+ -DPA. These findings are discussed in light of the proposal that SpoVA proteins are involved not only in DPA uptake during sporulation, but also in DPA release during nutrient-mediated spore germination. [source] Role of lipopeptides produced by Bacillus subtilis GA1 in the reduction of grey mould disease caused by Botrytis cinerea on appleJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2004Y. Touré Abstract Aim:, Test of Bacillus subtilis strain GA1 for its potential to control grey mould disease of apple caused by Botrytis cinerea. Methods and Results:, GA1 was first tested for its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The potential of strain GA1 to reduce post-harvest infection caused by B. cinerea was tested on apples by treating artificially wounded fruits with endospore suspensions. Strain GA1 was very effective at reducing disease incidence during the first 5 days following pathogen inoculation and a 80% protection level was maintained over the next 10 days. Treatment of fruits with an extract of GA1 culture supernatant also exerted a strong preventive effect on the development of grey mould. Further analysis of this extract revealed that strain GA1 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families. A strong evidence for the involvement of such compounds in disease reduction arose from the recovery of fengycins from protected fruit sites colonized by bacterial cells. Conclusions:, The results presented here demonstrate that, despite unfavourable pH, B. subtilis endospores inoculated on apple pulp can readily germinate allowing significant cell populations to establish and efficient in vivo synthesis of lipopeptides which could be related to grey mould reduction. Significance and Impact of the Study:, This work enables for the first time to correlate the strong protective effect of a particular B. subtilis strain against grey mould with in situ production of fengycins in infected sites of apple fruits. [source] Secreted production of Renilla luciferase in Bacillus subtilisBIOTECHNOLOGY PROGRESS, Issue 2 2010Chung-Jen Chiang Abstract Luciferase (Rluc) from the soft coral Renilla reniformis has been widely used as a bioluminescent reporter, and its secreted production has been solely performed in mammalian cells thus far. To make the production more efficient, a series of approaches was attempted to overproduce Rluc extracellularly in Bacillussubtilis. First, Cys124 in the Rluc gene was substituted with Ala. The mutant gene was subsequently incorporated into a pUB110/R6K-based plasmid, consequently, fusing with the P43 promoter and the sacB signal peptide. With the nitrogen-rich medium, B. subtilis strain bearing the plasmid became able to secret a detectable amount of Rluc. Moreover, the secretion signal for the Rluc gene was replaced by the aprN leader peptide with or without the propeptide. The result led to a more than twofold increase in the secreted Rluc. Finally, by enhancing the transcription of the Rluc gene implementing the P43 and spac tandem promoter, it resulted in the secreted Rluc with a yield of 100 mg/L. Overall, this study illustrates a potential strategy for improving the secretion efficiency of heterologous proteins in B. subtilis. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source] Inhibition of the D -alanine:D -alanyl carrier protein ligase from Bacillus subtilis increases the bacterium's susceptibility to antibiotics that target the cell wallFEBS JOURNAL, Issue 12 2005Juergen J. May The surface charge as well as the electrochemical properties and ligand binding abilities of the Gram-positive cell wall is controlled by the d -alanylation of the lipoteichoic acid. The incorporation of d -Ala into lipoteichoic acid requires the d -alanine:d -alanyl carrier protein ligase (DltA) and the carrier protein (DltC). We have heterologously expressed, purified, and assayed the substrate selectivity of the recombinant proteins DltA with its substrate DltC. We found that apo-DltC is recognized by both endogenous 4,-phosphopantetheinyl transferases AcpS and Sfp. After the biochemical characterization of DltA and DltC, we designed an inhibitor (d -alanylacyl-sulfamoyl-adenosine), which is able to block the d -Ala adenylation by DltA at a Ki value of 232 nmin vitro. We also performed in vivo studies and determined a significant inhibition of growth for different Bacillus subtilis strains when the inhibitor is used in combination with vancomycin. [source] DETERMINATION OF HISTAMINE AND BACTERIAL ISOLATION IN MARLIN FILLETS (MAKAIRA NIGRICANS) IMPLICATED IN A FOODBORNE POISONINGJOURNAL OF FOOD SAFETY, Issue 3 2010H.C. CHEN ABSTRACT An incident of foodborne poisoning causing illness in seven victims due to ingestion of marlin fillets occurred in August, 2008, in Kaohsiung City, southern Taiwan. The two suspected marlin samples contained 47.8 and 43.5 mg/100 g of histamine, which is greater than the 5.0 mg/100 g allowable limit suggested by the U.S. Food and Drug Administration. Given the allergy-like symptoms of the victims and the high histamine content in the suspected marlin samples, this foodborne poisoning was strongly suspected to be due to histamine intoxication. Two histamine-producing bacterial strains capable of producing 3.10 ppm and 4.20 ppm of histamine in trypticase soy broth (TSB) supplemented with 1.0% l -histidine (TSBH) were identified as Bacillus subtilis by 16S rDNA sequencing with polymerase chain reaction amplification. However, major histamine-forming bacteria might have been killed during the preparation of fillets before serving and these two B. subtilis isolates might not be the main contributors to histamine accumulation in suspected fillets. PRACTICAL APPLICATIONS Based on the finding that high contents of histamine (>40 mg/100 g) were detected in the suspected marlin samples, we speculate the temperature abuse of the fillets before cooking contributed to the presence of high histamine levels in marlin fillets and resulted in foodborne poisoning. Although two histamine-producing Bacillus subtilis strains were isolated from suspected fish samples, both might not to be the main contributors to histamine accumulation because of low histamine production. These results re-emphasize proper handling temperature for seafoods and offer an important awareness which Makaira nigricans fillets could become a hazardous food item in causing histamine poisoning even though no quality deficiency was observed on the fillets. [source] CYTOTOXICITY ASSESSMENT OF BACILLUS STRAINS ISOLATED FROM STREET-VENDED FOODS IN JOHANNESBURG, SOUTH AFRICAJOURNAL OF FOOD SAFETY, Issue 2 2002F.M. MOSUPYE ABSTRACT Twenty-one isolates each of Bacillus (B.) cereus, B. licheniformis and B. subtilis from street foods, collected in central Johannesburg, were randomly selected to test for cytotoxicity against McCoy 5A Mouse cells using a 3-(4,5-dimethylthizol-2-yl)-2,5-diphenyltetrazolium bromide assay, and observation by confocal scanning laser microscopy (CSLM) and scanning electron microscopy (SEM). Forty-eight percent of B. cereus, 33% of B. licheniformis and 19% of B. subtilis strains produced cytotoxic compounds. For B. cereus strains, all supernatants exhibiting cytotoxic effects were inactivated by heat treatment at 121C for 15 min. By contrast, 24% of B. licheniformis and 10% of B. subtilis supernatants exhibited cytotoxic effects following heat treatment. CSLM and SEM showed that McCoy cells treated with cytotoxic supernatants exhibited leakage and necrosis. Presence of B. cereus, B. licheniformis and B. subtilis in street foods in high numbers may pose potetnial safety risks due to production of cytotoxic compounds. [source] Characterization of Bacillus subtilis strains in Thua nao, a traditional fermented soybean food in northern ThailandLETTERS IN APPLIED MICROBIOLOGY, Issue 3 2006Y. Inatsu Abstract Aims:, To clarify the diversity of Bacillus subtilis strains in Thua nao that produce high concentrations of products useful in food manufacturing and in health-promoting compounds. Method and Results:, Production of amylase, protease, subtilisin NAT (nattokinase), and gamma-polyglutamic acid (PGA) by the Bacillus subtilis strains in Thua nao was measured. Productivity of protease NAT by these strains tended to be higher than by Japanese commercial natto-producing strains. Molecular diversity of isolated strains was analysed via randomly amplified polymorphic DNA-PCR fingerprinting. The strains were divided into 19 types, including a type with the same pattern as a Japanese natto-producing strain. Conclusion:,B. subtilis strains that could be a resource for effective production of protease, amylase, subtilisin NAT, or PGA were evident in Thua nao produced in various regions in northern Thailand. Significance and Impact of the Study:, This study clearly demonstrated the value of Thua nao as a potential resource of food-processing enzymes and health-promoting compounds. [source] Maturation of the lantibiotic subtilin: matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to monitor precursors and their proteolytic processing in crude bacterial culturesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 2 2002Torsten Stein Bacillus subtilis synthesizes the lanthionine containing 32-amino-acid peptide antibiotic (lanti-biotic) subtilin from a ribosomally generated 56-amino-acid precursor pre-propeptide by extensive posttranslational modifications. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was used to monitor the production of matured subtilin within crude samples taken from B. subtilis culture media without prior fractionation. The processing reaction of subtilin was blocked with the serine protease inhibitor phenylmethylsulfonyl fluoride and different subtilin precursor peptides in the molecular mass range up to 6220 were observed. Two of these species were isolated by reversed-phase high-performance liquid chromatography (HPLC) and structurally analyzed by post-source decay MALDI-TOFMS. We provide evidence that the precursor species comprise the posttranslational modified C-terminal part of subtilin to which leader peptide moieties with different chain lengths are attached. These antimicrobial-inactive species could be processed to antibiotic-active subtilin by incubation with culture media of different subtilin-nonproducing B. subtilis strains as indicated by a combination of antimicrobial growth assays and MALDI-TOFMS analyses. These achievements are strong evidence for the sensitivity of MALDI-TOFMS methodology that allows straightforward investigations of analytes even in complex mixtures without time-consuming sample preparations. Copyright © 2001 John Wiley & Sons, Ltd. [source] Crystallization and preliminary crystallographic analysis of poly-,-glutamate hydrolase from bacteriophage ,NIT1ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2009Zui Fujimoto Particular Bacillus subtilis strains produce a capsular polypeptide poly-,-glutamate (,-PGA) that functions as a physical barrier against bacteriophage infection. Bacteriophage ,NIT1 can infect B. subtilis and produces a novel ,-PGA hydrolase PghP. PghP was overexpressed, purified and crystallized by the sitting-drop vapour-diffusion method. The crystals diffracted to a resolution of 2.4,Ĺ using a synchrotron X-ray source and were found to belong to space group P3121 or P3221. [source] Enhanced Hyaluronic Acid Production in Bacillussubtilis by Coexpressing Bacterial HemoglobinBIOTECHNOLOGY PROGRESS, Issue 5 2007Liang-Jung Chien Bacillus subtilis strains that can produce hyaluronic acid (HA) were constructed by integrating the HA synthase gene (hasA) and the UDP-glucose dehydrogenase gene of group C Streptococcus (hasB) or of B. subtilis itself (tauD) into the amyE locus of the B. subtilis chromosome. All of the inserted genes were under the control of a strong constitutive vegII promoter of B. subtilis. Although HA production could be achieved by expressing hasA alone, coexpressing hasB or tauD with hasA could enhance HA production at least 2-fold. To replenish the energy consumed for HA biosynthesis, Vitreoscilla hemoglobin (VHb) was coexpressed with the HA-expressing genes. With the expression of VHb, not only the cell concentration was enhanced 25%, but also HA production was further increased by 100%. About 1.8 g/L of HA was obtained by the recombinant strain B.subtilis carrying VHb, hasA, and tauD genes in the expression cassette after 30 h cultivation. [source] | |||||||||||||